DETERGENT POUCH COMPRISING METALLOPROTEASES
Reference to sequence listing
This application contains a Sequence Listing in computer readable form. The computer readable form is incorporated herein by reference.
Field of the invention
The invention relates to a multi-compartment pouch comprising a selected combination of ingredients in different compartments as well as a washing process and the use of the multi compartment pouch for laundry applications, such as for washing and cleaning of textiles, or for dishwashing applications.
Background of the invention
The detergent formulators are constantly looking for new detergent forms with improved cleaning profile. Lately, products in unit dose form have become one of the preferred forms for the user due to the ease of use, particularly water-soluble pouches which present the added advantage of no need to unwrap.
Products having size and geometry constrictions -as in the case of dishwasher products that need to be dosed through the dispenser and therefore are restricted not only by the size of the dispenser but also by the shape- and ingredients in different physical forms are always challenging from the formulation view point. It is also more challenging when it is desired that the product provides controlled and/or differential release.
Enzymatic detergent compositions normally contain a protease. One group of proteases, the metalloproteases, have not yet found widespread use in detergent industry, however, mainly due to low stability in detergent compositions as well as under the conditions during the wash process. For example, an M4 family metal loprotease from Bacillus amyloliquefaciens known as Neutrase® has been used for many years as an additive in various food and feed products. In spite of the fact, that this and other metalloproteases have been described for use in detergent and cleaning compositions and processes, they have not been used in detergent compositions in practice. Thus, although a number of metalloproteases are known to have activity that in theory would make them suitable for detergent use, their instability in detergent compositions and wash conditions has in practice prevented such use.
The object of the present invention is to provide an improved way of delivering an enzyme such as a metalloprotease in a multi-compartment pouch.
Summary of the invention
The invention relates to an enzyme containing water-soluble films, and their use in detergents.
The present invention relates to a multi-compartment pouch comprising at least two compartments, wherein said first compartment comprises an enzyme-containing composition and a second compartment comprises a detergent composition.
The present invention relates to a method of releasing an enzyme in a cleaning process comprising steps of: a. providing a multicompartment pouch with at least two compartments, the first compartment comprising enzyme-containing composition and the second compartment comprising a detergent composition; b. releasing the detergent composition from the second compartment and subsequently releasing the enzyme-containing composition from the first compartment into the wash liquor.
The present invention also relates to a method of treating a substrate, where the method includes the step of contacting the substrate with the pouch in the presence of water, wherein the substrate is a fabric or a hard surface.
The present invention also relates to a method of cleaning comprising contacting a surface and/or a fabric with the pouch.
The present invention also relates to a method for removing and/or reducing soil and/or for reducing redeposition on a surface and/or textile comprising contacting the surface and/or textile with the pouch.
The present invention also relates to a method of laundering or dishwashing in a washing machine comprising the steps of placing the pouch into the product dispenser and releasing it during the wash cycle.
The present invention also relates to use of the pouch for cleaning of an item, wherein the item is a textile or a surface.
The present invention also relates to use of the pouch in a cleaning process, such as laundry, hard surface cleaning, dish wash or automated dish wash.
The present invention also relates to use of the pouch for removing or releasing a stain from a textile having a stain.
Definitions
In accordance with the detailed description, the following abbreviations and definitions apply. Note that the singular forms“a”,“an”, and“the” include plural referents unless the context clearly indicates otherwise. Thus, for example, reference to“an enzyme” includes a plurality of such enzymes, and reference to“the dosage” includes reference to one or more dosages and equivalents thereof known to those skilled in the art, and so forth.
Certain ranges are presented herein with numerical values being preceded by the term “about”. The term“about” as used herein, is to provide literal support for the exact number that it precedes, as well as a number that is near to or approximately the number that the term precedes. In determining whether a number is near to or approximately a specifically recited number, the near or approximating unrecited number may be a number which, in the context in which it is presented, provides the substantial equivalent of the specifically recited number. For example, for a specific numerical value, the term“about” refers to a range of -10% to +10% of the numerical value, unless term is otherwise specifically defined in context. In another example, the phrase a “pH value of about 9” refers to pH values of from 8.1 to 9.9, unless the pH value is specifically defined otherwise.
The term“protease activity” or“peptidase activity” is defined herein as the ability to break down the amide bond of a protein by hydrolysis of the peptide bonds that link amino acids together in a polypeptide chain.
“Protease activity” can be measured using any assay in which a substrate is employed that includes peptide bonds relevant for the specificity of the protease in question. Assay-pH and assay-temperature are likewise to be adapted to the protease in question. Examples of assay- pH-values are pH 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , or 12. Examples of assay-temperatures are 15, 20, 25, 30, 35, 37, 40, 45, 50, 55, 60, 65, 70, 80, 90, or 95°C. Examples of general protease substrates are casein, bovine serum albumin and haemoglobin. In the classical Anson and Mirsky method, denatured haemoglobin is used as substrate and after the assay incubation with the protease in question, the amount of trichloroacetic acid soluble haemoglobin is determined as a measurement of protease activity (Anson, M.L. and Mirsky, A.E., 1932, J. Gen. Physiol. 16: 59 and Anson, M.L., 1938, J. Gen. Physiol. 22: 79).
For the purpose of the present invention, protease activity was determined using assays which are described in“Materials and Methods”, such as the Protazyme OL assay.
The term“metalloprotease” as used herein refers to a protease having one or more metal ions in the binding/active site.
The terms“cleaning compositions” and "cleaning formulations," refer to compositions that find use in the removal of undesired compounds from items to be cleaned, such as fabric, carpets, dishware including glassware, contact lenses, hard surfaces such as tiles, zincs, floors, and table surfaces, hair (shampoos), skin (soaps and creams), teeth (mouthwashes, toothpastes), etc. The terms encompass any materials/compounds selected for the particular type of cleaning composition desired and the form of the product (e.g., liquid, gel, granule, or spray compositions). The specific selection of cleaning composition materials is readily made by considering the surface, item or fabric to be cleaned, and the desired form of the composition for the cleaning conditions during use. These terms further refer to any composition that is suited for cleaning,
bleaching, disinfecting, and/or sterilizing any object and/or surface. It is intended that the terms include, but are not limited to detergent composition (e.g., liquid and/or solid laundry detergents and fine fabric detergents; hard surface cleaning formulations, such as for glass, wood, ceramic and metal counter tops and windows; carpet cleaners; oven cleaners; fabric fresheners; fabric softeners; and textile and laundry pre-spotters, as well as dishwash detergents).
The term“DNase” means a polypeptide with DNase activity that catalyses the hydrolytic cleavage of phosphodiester linkages in the DNA backbone, thus degrading DNA. DNase activity may be determined according to the procedure described in the Assay I described in“Materials and Methods”.
The term“detergent composition” refers to compositions that find use in the removal of undesired compounds from items to be cleaned, such as hard surfaces or dishware. The detergent composition may be used to e.g. clean dishware for both household cleaning and industrial cleaning. The terms encompass any materials/compounds selected for the particular type of cleaning composition desired and the form of the product (e.g., liquid, gel, powder, granulate, paste, or spray compositions) and includes, but is not limited to, detergent compositions (e.g., liquid and/or solid laundry detergents and fine fabric detergents; fabric fresheners; fabric softeners; and textile and laundry pre-spotters/pretreatment). In addition to containing the enzyme of the invention, the detergent formulation may contain one or more additional enzymes (such as other proteases, amylases, lipases, cutinases, cellulases, endoglucanases, xyloglucanases, pectinases, pectin lyases, xanthanases, peroxidaes, haloperoxygenases, DNase, catalases and mannanases, or any mixture thereof), and/or detergent components such as surfactants, builders, chelators or chelating agents, bleach system or bleach components, polymers, fabric conditioners, foam boosters, suds suppressors, dyes, perfume, tannish inhibitors, optical brighteners, bactericides, fungicides, soil suspending agents, anti-corrosion agents, enzyme inhibitors or stabilizers, enzyme activators, transferase(s), hydrolytic enzymes, oxido reductases, bluing agents and fluorescent dyes, antioxidants, and solubilizers.
The term "fabric" encompasses any textile material. Thus, it is intended that the term encompass garments, as well as fabrics, yarns, fibers, non-woven materials, natural materials, synthetic materials, and any other textile material.
The term“textile” means any textile material including yarns, yarn intermediates, fibers, non-woven materials, natural materials, synthetic materials, and any other textile material, fabrics made of these materials and products made from fabrics (e.g., garments and other articles). The textile or fabric may be in the form of knits, wovens, denims, non-wovens, felts, yarns, and toweling. The textile may be cellulose based such as natural cellulosics, including cotton, flax/linen, jute, ramie, sisal or coir or manmade cellulosics (e.g. originating from wood pulp)
including viscose/rayon, cellulose acetate fibers (tricell), lyocell or blends thereof. The textile or fabric may also be non-cellulose based such as natural polyamides including wool, camel, cashmere, mohair, rabbit and silk or synthetic polymers such as nylon, aramid, polyester, acrylic, polypropylene and spandex/elastane, or blends thereof as well as blends of cellulose based and non-cellulose based fibers. Examples of blends are blends of cotton and/or rayon/viscose with one or more companion material such as wool, synthetic fiber (e.g. polyamide fiber, acrylic fiber, polyester fiber, polyvinyl chloride fiber, polyurethane fiber, polyurea fiber, aramid fiber), and/or cellulose-containing fiber (e.g. rayon/viscose, ramie, flax/linen, jute, cellulose acetate fiber, lyocell). Fabric may be conventional washable laundry, for example stained household laundry. When the term fabric or garment is used it is intended to include the broader term textiles as well. In the context of the present invention, the term“textile” also covers fabrics.
The term "effective amount of enzyme" refers to the quantity of enzyme necessary to achieve the enzymatic activity required in the specific application, e.g., in a defined detergent composition. Such effective amounts are readily ascertained by one of ordinary skill in the art and are based on many factors, such as the particular enzyme used, the cleaning application, the specific composition of the detergent composition, and whether a liquid or dry (e.g., granular, bar) composition is required, and the like.
The term "relevant washing conditions" is used herein to indicate the conditions, particularly washing temperature, time, washing mechanics, detergent concentration, type of detergent and water hardness, actually used in households in a detergent market segment.
The term "wash performance" of an enzyme refers to the contribution of an enzyme to washing that provides additional cleaning performance to the detergent without the addition of the enzyme to the composition. Wash performance is compared under relevant washing conditions. Wash performance of enzymes is conveniently measured by their ability to remove certain representative stains under appropriate test conditions. In these test systems, other relevant factors, such as detergent composition, detergent concentration, water hardness, washing mechanics, time, pH, and/or temperature, can be controlled in such a way that conditions typical for household application in a certain market segment are imitated.
The term“water hardness” or“degree of hardness” or“dH” or“°dH” as used herein refers to German degrees of hardness. One degree is defined as 10 milligrams of calcium oxide per litre of water.
The term "improved wash performance" is used to indicate that a better end result is obtained in stain removal from items washed (e.g., fabrics or dishware and/or cutlery) under relevant washing conditions as compared to no enzyme or to a reference enzyme, or that less enzyme, on weight basis, is needed to obtain the same end result relative to no enzyme or to a reference enzyme. Improved wash performance could in this context also be that the same effect,
e.g., stain removal effect is obtained in shorter wash time, e.g., the enzymes provide their effect more quickly under the tested conditions.
The term“wash cycle” is defined herein as a washing operation wherein fabric is exposed to the wash liquor for a period of time by circulating the wash liquor and spraying the wash liquor onto the fabric in order to clean the fabric and finally the superfluous wash liquor is removed. A wash cycle may be repeated one, two, three, four, five or even six times at the same or at different temperatures. Hereafter the fabric is generally rinsed and dried.
The term“wash liquor” is defined herein as the solution or mixture of water and detergent components optionally including enzymes used for laundrering textiles, for hard surface cleaning or for dishwashing.
The term“wash time” is defined herein as the time it takes for the entire washing process; i.e. the time for the wash cycle(s) and rinse cycle(s) together.
The term“enzyme detergency” or“detergency” or“detergency effect” is defined herein as the advantageous effect an enzyme may add to a detergent compared to the same detergent without the enzyme. Important detergency benefits which can be provided by enzymes are stain removal with no or very little visible soils after washing and/or cleaning, prevention or reduction of redeposition of soils released in the washing process an effect that also is termed anti redeposition, restoring fully or partly the whiteness of textiles, which originally were white but after repeated use and wash have obtained a greyish or yellowish appearance an effect that also is termed whitening. Textile care benefits, which are not directly related to catalytic stain removal or prevention of redeposition of soils, are also important for enzyme detergency benefits. Examples of such textile care benefits are prevention or reduction of dye transfer from one fabric to another fabric or another part of the same fabric an effect that is also termed dye transfer inhibition or anti- back staining, removal of protruding or broken fibers from a fabric surface to decrease pilling tendencies or remove already existing pills or fuzz an effect that also is termed anti-pilling, improvement of the fabric-softness, color clarification of the fabric and removal of particulate soils which are trapped in the fibers of the fabric or garment. Enzymatic bleaching is a further enzyme detergency benefit where the catalytic activity generally is used to catalyze the formation of bleaching component such as hydrogen peroxide or other peroxides.
The term“hard surface cleaning” is defined herein as cleaning of hard surfaces, such as reducing or removing biofilm from a hard surface, wherein hard surfaces may include floors, tables, walls, roofs etc. as well as surfaces of hard objects such as cars (car wash) and dishes (dish wash). Hard surface cleaning also includes cleaning the interior of washing machines, such as the interior of laundry washing machines or dishwashing machines, this includes cleaning soap intake box, walls, windows, baskets, racks, nozzles, pumps, sump, filters, pipelines, tubes, joints, seals, gaskets, fittings, impellers, drums, drains, traps, coin traps inlet and outlets. Dish washing
includes but are not limited to cleaning of plates, cups, glasses, bowls, pots, cutlery, spoons, knives, forks, serving utensils, ceramics, plastics, cutting boards, china and glass ware.
The term“laundering” relates to both household laundering and industrial laundering and means the process of treating textiles with a solution containing a cleaning or detergent composition of the present invention. The laundering process can for example be carried out using e.g. a household or an industrial washing machine or can be carried out by hand.
DETAILED DESCRIPTION OF THE INVENTION
The present invention relates to a multi-compartment pouch comprising at least two compartments, wherein said first compartment comprises an enzyme-containing composition and a second compartment comprises a detergent composition.
The present invention also relates to a method of releasing an enzyme in a cleaning process comprising steps of: a. providing a multi-compartment pouch with at least two compartments, the first compartment comprising enzyme-containing composition and the second compartment comprising a detergent composition; b. releasing the detergent composition from the second compartment and subsequently releasing the enzyme-containing composition from the first compartment into the wash liquor.
In one embodiment, the present invention envisages a multi-compartment pouch. The water-soluble multi-compartment pouch of the invention (herein referred to as pouch) comprises at least two compartments and may for example be in the form of two side-by-side compartments superposed onto another compartment. In one embodiment of the present invention, the pouch is typically a closed structure, made of materials described herein, enclosing a volume space. The volume space is separated into at least two compartments. In one embodiment of the present invention, the pouch can be of any form, shape and material which is suitable to hold the compositions, e.g without allowing the release of the composition from the pouch prior to contact of the pouch with water. The exact execution will depend, for example on the type and amount of the composition.
In one embodiment of the present invention, the multi-compartment pouch comprises at least two compartments, wherein said first compartment comprises an enzyme-containing composition and a second compartment comprises a detergent composition.
In another embodiment of the present invention, the multi-compartment pouch can have more than two compartments, which can be in any disposition, side-by-side, superposed or compartment-inside-compartment. Especially preferred are: i) pouches having two compartments on side-by-side disposition superposed onto a single compartment; and ii) pouches having two side-by-side compartments superposed onto two other side-by-side compartments.
In one embodiment of the present invention, each compartment of the multi-compartment
pouch may contain a detergent composition or part thereof in any physical form, including solid (loose or densified powder, tablet, pre-formed discrete particles, etc.), liquids (gels, aqueous liquids, non-aqueous liquids, etc.), liquids with solid suspended on them, etc.
In one embodiment of the present invention, the multi-compartment pouch is very effective in terms of separation of incompatible ingredients. The invention also envisages a method of automatic dishwashing using the pouch of the invention. The pouch of the invention can contain any kind of detergent composition, preferably the composition is a laundry or dishwashing composition.
In one embodiment of the present invention, the multi-compartment pouch may be formed by water-soluble films that form the different compartments can be the same but preferably the films have different solubility and are suitable for delivering the content of different compartments at different points in time of the wash cycle or during the wash and during the rinse cycle.
The pouch of the invention is very effective in terms of separation of incompatible ingredients. The pouches of the invention are robust, compact and have a great flexibility in terms of ingredients separation and controlled release. The inventors of the present invention have seen a surprising effect when using the multi-compartment pouch. The multi-compartment pouch of the invention an advantageous wash and cleaning capacity is obtained, especially when an enzyme such as metalloprotease is sensitive in presence of detergent components. The wash and cleaning capacity of a preferred multi-compartment pouch of the invention with sequential release of detergent composition followed by the release of enzyme-containing composition is significantly enhanced in comparison with multi-compartment pouches of the invention in which detergent components and enzyme such as metalloprotease on the other hand are released simultaneously.
The term “water-soluble film” or “water-soluble polymer” encompasses films which disintegrates when in contact with water by forming a solution and/or dispersion of polymer in water. The pouch is preferably made of a film material wherein the film material is soluble in water, and has water solubility of at least 50%, at least 60%, at least 70%, preferably at least 75%, at least 80%, at least 90% or even at least 95%.
Preferred pouch materials are polymeric materials, preferably polymers which are formed into a film or sheet. The pouch material can, for example, be obtained by casting, blow moulding, extrusion or blown extrusion of the polymeric material, as known in the art.
The pouch of the present invention may be made using any suitable equipment and method.
In one embodiment of the present invention, the multi-compartment pouch may be of such a size that it conveniently contains either a unit dose amount of the composition herein,
suitable for the required operation, for example one wash, or only a partial dose, to allow the consumer greater flexibility to vary the amount used, for example depending on the size and/or degree of soiling of the wash load. In another embodiment of the present invention, the pouch is made from a water-soluble film which encloses an inner volume; wherein the inner volume is divided into the compartments of the pouch.
In one embodiment of the present invention, the multi-compartment pouch herein defined are closed structures, made from a water-soluble film, which encloses a volume space which comprises the solid component or the liquid component of the composition. Said volume space is preferably enclosed by a water-soluble film in such a manner that the volume space is separated from the outside environment. In a preferred embodiment of the invention, the pouches are made by a process involving stretching the material used for the pouch.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch.
In one embodiment of the present invention, the enzyme-containing composition present in the first compartment of the multi-compartment pouch comprises a protease.
In one embodiment of the present invention, the enzyme-containing composition present in the first compartment of the multi-compartment pouch comprises a metalloprotease.
In one embodiment of the present invention, the enzyme-containing composition present in the first compartment of the multi-compartment pouch comprises a metalloprotease selected from a group consisitng of EC classes EC 3.4.22 or EC3.4.24.
In one embodiment of the present invention, the enzyme-containing composition present in the first compartment of the multi-compartment pouch comprising the metalloprotease is selected from a group consisting of a. a thermolysin and/or variant thereof; b. a metalloprotease from M4 family, that is not a thermolysin and variant thereof; c. a metalloprotease from the M7 family; and d. a metalloprotease from the M35 metalloprotease.
In one embodiment of the present invention, the enzyme-containing composition present in the first compartment of the multi-compartment pouch comprising the metalloprotease is an enzyme (E.C 3.4.24) that hydrolyse bonds within peptides and proteins, in the laundry context this leads to enhanced removal of protein or peptide containing stains. Metalloproteases (metallopeptidases or metalloproteinases) is a class of hydrolases which cleave peptide bonds by the action of a water molecule which is activated by complexing to at least one bivalent metal ions belonging to the group of zinc, manganese, cobalt, nickel or copper ions, preferably zinc. According to the classification of proteases based on protein structure and homology implemented in the MEROPS database (http://merops.sanger.ac.uk/cgi- bin/family_index?type=P), metalloproteases are found in 14 different families. Preferably the protease is selected from the M4, M7 or M35 family, more preferably an M4 metalloprotease,
most preferably a neutral metalloprotease. The metalloproteases that may be used in this invention includes any of those which may be used in a homecare application. These metalloproteases are, for example, derived from bacterium selected from the group consisting of Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus stearothermophilus, and Bacillus thermoproteolyticus, and fungi selected from the group consisting of Aspergillus oryzae and Aspergillus niger.
Especially preferred metalloproteases for use herein belong to EC classes EC 3.4.22 or EC3.4.24, more preferably they belong to EC classes EC3.4.22.2, EC3.4.24.28 or EC3.4.24.27.
The term“M4 Metalloprotease Family” or“M4 Metalloprotease” or“M4” as used herein means a polypeptide falling into the M4 metalloprotease family according to Rawlings et al., Biochem. J., 290, 205-218 (1993) and as further described in MEROPS - (Rawlings et al., MEROPS: the peptidase database, Nucl Acids Res, 34 Database issue, D270-272, 2006). The M4 metalloproteases are neutral metalloproteases containing mainly endopeptidases. All peptidases in the family bind a single, catalytic zinc ion. M4 metalloprotease family members include the common HEXXH motif, where the histidine residues serve as zinc ligands and glutamate is an active site residue. M4 metalloproteases have a pH optimum mainly at neutral pH. The M4 metalloprotease family includes, e.g., Neutrase® (classified as MEROPS subclass M04.014), Thermolysin, Bacillolysin, vibriolysin, pseudolysin, Msp peptidase, coccolysin, aureolysin, vimelysin, lambda toxin neutral peptidase B, PA peptidase (Aeromonas-type), griselysin, stearolysin, Mprlll ( Alteromonas sp. strain 0-7), pap6 peptidase, neutral peptidase ( Thermoactinomyces-type ), ZmpA peptidase ( Burkholderia sp.), zpx peptidase, PrtS peptidase (Photorhabdus luminescens), protealysin, ZmpB peptidase ( Burkholderia sp.). The M4 metalloprotease family of polypeptides have been further characterized and presently includes, according to MEROPS, at least twenty-two subclasses for which a distinct MEROPS ID (i.e., an identifier of the formula M04.xxx) has been assigned, as well as non-peptidase homologues and unassigned peptidases.
The term “Thermolysin-Like Metalloprotease” as used herein means (a) an M4 metalloprotease of the MEROPS subclass M04.001 ; (b) an M4 metalloprotease of the MEROPS subclass M04.018; (c) an M4 metalloprotease of the MEROPS subclass M04.021 ; (d) an M4 metalloprotease having an active cleft motif TG[TS][QS]DNGGVH[TI] as shown in SEQ ID NO: 65; (e) an M4 metalloprotease having an active cleft motif
DPDHSKRYTG[TS][QS]DNGGVH[TI]NSGI as shown in SEQ ID NO: 66 and (f) an M4 metalloprotease having an active cleft motif NT[TS][QS]DNGGVH[TI]NSGI as shown in SEQ ID NO: 67. In these motifs, the accepted lUPAC single letter amino acid abbreviation is employed. Also in the above motifs, the use of brackets indicates alternative amino acid choices at a particular position. The Thermolysin-Like Metalloproteases are described, e.g., in Barret et al.
(1998) Handbook of proteolytic enzymes. Academic Press, pp. 350-369. The amino acid sequences of several TLPs have been determined, and the three-dimensional structures of several TLPs have been solved. Focus has been on increasing the thermostability of the TLP and a number of publications describe the thermostability of Bacillus TLPs, e.g., Veltman et al., (1998) Biochemistry 37(15):5312-9. The TLPs consist of an alpha-helical C-terminal domain and an N- terminal domain mainly consisting of beta-strands. The domains are connected by a central alpha- helix. This helix is located at the bottom of the active site cleft and contains several of the catalytically important residues such as four substrate binding pockets S2, S1 , ST and S2' have been identified (Hangauer et al. (1984) Biochemistry 23:5730-5741).
The term“M7 Metalloprotease Family” or“M7 Metalloprotease” or“M7” or“snapalysin family” as used herein means a polypeptide falling into the M7 metalloprotease family according to Rawlings et al., Biochem. J., 290, 205-218 (1993) and as further described in MEROPS - (Rawlings et al., MEROPS: the peptidase database, Nucl Acids Res, 34 Database issue, D270- 272, 2006). The protease family M7 contains a metalloendopeptidase, snapalysin. Snapalysin is active at neutral pH. The only known activity is cleavage of proteins of skimmed milk to form clear plaques around the growing bacterial colonies. The Zinc is bound by two histidines and an aspartate in an HEXXHXXGXXD sequence motif as shown in SEQ ID NO: 68; the glutamate is a catalytic residue. The M7 proteases have clear signal peptides recognized by the SignalP prediction program. They also all have a propeptide that is cleaved off.
The term “M35 Metalloprotease Family” or “M35 Metalloprotease” or “M35” or “deuterolysin family” as used herein means a polypeptide falling into the M35 metalloprotease family according to Proteolysis in Cell Function, pp13-21 , IOS Press, Amsterdam (1997), Rawlings et al., Biochem. J., 290, 205-218 (1993) and as further described in MEROPS - (Rawlings et al., MEROPS: the peptidase database, Nucl Acids Res, 34 Database issue, D270- 272, 2006). Family M35 members contain two zinc binding histidine’s and a catalytic glutamate in an HEXXH motif as shown in SEQ ID NO: 69. There is a third zinc ligand, an Asp, found in a GTXDXXYG motif as shown in SEQ ID NO: 70 C-terminal to the His zinc ligands (see the Alignment). For this reason, the peptidases in this family are sometimes termed“aspzincins”, although peptidases in which the third ligand of zinc is Asp also occur in families M6, M7 and M64.
Metalloproteases can be derived from animals, plants, bacteria or fungi. Suitable metalloprotease can be selected from the group of neutral metalloproteases and Myxobacter metalloproteases. Suitable metalloproteases can include collagenases, hemorrhagic toxins from snake venoms and thermolysin from bacteria. Preferred thermolysin enzyme variants include an M4 peptidase, more preferably the thermolysin enzyme variant is a member of the PepSY~Peptidase_M4~Peptidase_M4_C family.
In some embodiments, the thermolysin enzyme variant is from a genus selected from the group consisting of Bacillus, Geobacillus, Alicyclobacillus, Lactobacillus, Exiguobacterium, Brevibacillus, Paenibacillus, Herpetosiphon, Oceanobacillus, Shewanella, Clostridium, Staphylococcus, Flavobacterium, Stigmatella, Myxococcus, Vibrio, Methanosarcina, Chryseobacterium, Streptomyces, Kribbella, Janibacter, Nocardioides, Xanthamonas, Micromonospora, Burkholderia, Dehalococcoides, Croceibacter, Kordia, Microscilla, Thermoactinomyces, Chloroflexus, Listeria, Plesiocystis, Haliscomenobacter, Cytophaga, Hahella, Arthrobacter, Brachybacterium, Clavibacter, Micro bacterium ntrasporangium, Frankia, Meiothermus, Pseudomonas, Ricinus, Catenulispora, Anabaena, Nostoc, Halomonas, Chromohalobacter, Bordetella, Variovorax, Dickeya, Pectobacterium, Citrobacter, Enter obacter, Salmonella, Erwinia, Pantoea, Rahnella, Serratia, Geodermatophilus, Gemmata, Xenorhabdus, Photorhabdus, Aspergillus, Neosartorya, Pyrenophora, Saccharopolyspora, Nectria, Gibberella, Metarhizium, Waddlia, Cyanothece, Cellulphaga, Providencia, Bradyrhizobium, Agrobacterium, Mucilaginibacter, Serratia, Sorangium, Streptosporangium, Renibacterium, Aeromonas, Reinekea, Chromobacterium, Moritella, Haliangium, Kangiella, Marinomonas, Vibrionales, Listonella, Salinivibrio, Photobacterium, Alteromonadales, Legionella, Teredinibacter, Reinekea, Hydrogenivirga and Pseudoalteromonas. In some embodiments, the thermolysin enzyme variant is from a genus selected from the group consisting of Bacillus, Geobacillus, Alicyclobacillus, Lactobacillus, Exiguobacterium, Brevibacillus, Paenibacillus, Herpetosiphon, Oceanobacillus, Shewanella, Clostridium, Staphylococcus, Flavobacterium, Stigmatella, Myxococcus, Vibrio, Methanosarcina, Chryseobacterium, and Pseudoalteromonas.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises at least one metalloprotease having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NOs: 1-36.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease wherein the metalloprotease comprises of amino acid sequence having at least 60% at least 65%, e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, but less than 100% sequence identity to a polypeptide selected from the group consisting of SEQ ID Nos: 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35 and 36.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at
least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 1. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 1.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, or 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 2. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 2.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 3. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 3.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least or 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 4. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 4.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent, but less than 100 percent sequence identity to SEQ ID NO: 5. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 5.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 6. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 6.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 7. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 7.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 8. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 8.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 9. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 9.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent,
at least 97 percent, at least 98 percent, at least 99 percent, but less than 100 percent sequence identity to SEQ ID NO: 10. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 10.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 11. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 1 1.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent, but less than 100 percent sequence identity to SEQ ID NO: 12. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 12.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 13. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 13.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent, but less than 100 percent sequence identity to SEQ ID NO: 14. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 14.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 15. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 15.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 16. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 16.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 17. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 17.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 18. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 18.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent,
at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 19. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 19.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 20. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 20.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 21. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 21.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 22. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 22.
In an embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 23. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 23.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 24. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 24.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 25. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 25.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 26. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 26.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 27. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 27.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent,
at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 28. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 28.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 29. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 29.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 30. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 30.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 31. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 31.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 32. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 32.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 33. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 33.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 34. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 34
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent at least 97 percent at least 98 percent at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 35. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 35.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 36. In one embodiment, the present invention relates to a metalloprotease wherein the metalloprotease comprises or consists of amino acid sequence as set forth in SEQ ID NO: 36.
Suitable commercially available metalloproteases enzymes include those sold under the trade names Neutrase® by Novozymes A/S (Denmark), the Corolase® range including Corolase® 2TS, Corolase® N, Corolase® L10, Corolase® LAP and Corolase® 7089 from AB Enzymes, Protex
14L and Protex 15L from DuPont (Palo Alto, California), those sold as thermolysin from Sigma and the Thermoase range (PC10F and CIOO) and thermolysin enzyme from Amano enzymes.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease and further comprises one or more additional enzymes.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease wherein the metalloprotease comprises of amino acid sequence having at least 60% at least 65%, e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, but less than 100% sequence identity to a polypeptide selected from the group consisting of SEQ ID Nos: 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35 and 36 and further comprising one or more additional enzymes.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease wherein the metalloprotease comprises of amino acid sequence having at least 60% at least 65%, e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, but less than 100% sequence identity to a polypeptide selected from the group consisting of SEQ ID Nos: 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35 and 36 and further comprising one or more additional enzymes, wherein the additional enzyme is selected from a group consisitng of is selected from the group consisting of an alpha-amylase, a beta-amylase, a pullulanase, a lipase, a cellulase, an oxidase, another protease, a phospholipase, a perhydrolase, a xylanase, a pectate lyase, a pectinase, a galacturanase, a hemicellulase, a xyloglucanase, a nuclease, a mannanase and mixtures thereof.
In one embodiment, the enzyme-containing composition in the first compartment of the pouch is present in a liquid, solid or gel form.
In one embodiment of the present invention, the enzyme of the present invention, in particular a metalloprotease, may be added in an amount corresponding to 0.001-100 mg of protein, such as 0.01-100 mg of protein, preferably 0.005-50 mg of protein, more preferably 0.01- 25 mg of protein, even more preferably 0.05-10 mg of protein, most preferably 0.05-5 mg of protein, and even most preferably 0.01-1 mg of protein per liter of wash liquor.
In another embodiment of the present invention, the detergent composition is present in the second compartment of the multi-compartment pouch.
In an embodiment of the present invention, the detergent composition present in the second compartment of the multi-compartment pouch comprises at least one enzyme.
In an embodiment of the present invention, the detergent composition present in the second compartment of the multi-compartment pouch comprising the enzyme is preferably a DNase.
In an embodiment of the present invention, the detergent composition is present in the second compartment of the multi-compartment pouch comprises at least one DNase.
DNase
The term “DNase” means a polypeptide with DNase (deoxyribonuclease) activity that catalyzes the hydrolytic cleavage of phosphodiester linkages in a DNA backbone, thus degrading DNA. Exodeoxyribonuclease cut or cleaves residues at the end of the DNA back bone where endo-deoxyribonucleases cleaves or cut within the DNA backbone. A DNase may cleave only double-stranded DNA or may cleave double stranded and single stranded DNA. The term “DNases” and the expression “a polypeptide with DNase activity” are used interchangeably throughout the application. For purposes of the present invention, DNase activity is determined according to the procedure described in the Assay I.
Preferably the DNase is selected from any of the enzyme classes E.C.3.1 , preferably E.C.3.1.21 , e.g. such as E.C.3.1.21.X, where X = 1 , 2, 3, 4, 5, 6, 7, 8 or 9, or e.g. Deoxyribonuclease I, Deoxyribonuclease IV, Type I site-specific deoxyribonuclease, Type II site- specific deoxyribonuclease, Type III site-specific deoxyribonuclease, CC-preferring endo- deoxyribonuclease, Deoxyribonuclease V, T(4) deoxyribonuclease II, T(4) deoxyribonuclease IV or E.C. 3.1.22.Y where Y = 1 , 2, 4 or 5, e.g. Deoxyribonuclease II, Aspergillus deoxyribonuclease K(1), Crossover junction endo-deoxyribonuclease, Deoxyribonuclease X.
Preferably, the polypeptide having DNase activity is obtained from a microorganism and the DNase is a microbial enzyme. The DNase is preferably of fungal or bacterial origin.
The DNase may be obtainable from Bacillus e.g. such as a Bacillus licheniformis, Bacillus subtilis, Bacillus horikoshii, Bacillus horneckiae, Bacillus cibi, Bacillus idriensis, Bacillus algicola, Bacillus vietnamensis, Bacillus hwajinpoensis, Bacillus indicus, Bacillus marisflavi or Bacillus luciferensis.
In one embodiment of the invention, the detergent composition present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
Other suitable DNases obtained from Bacillus includes a polypeptide having DNase activity, wherein the polypeptide comprises an amino acid sequence having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to the polypeptide shown in SEQ ID No: 61 and 62.
In one embodiment of the invention, the detergent composition present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the invention, the detergent composition present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
The DNase may also be fungal, preferably obtained from Aspergillus e.g. Aspergillus oryzae. One embodiment of the invention the pouch comprises a polypeptide having DNase activity, wherein the polypeptide comprises an amino acid sequence having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 63.
In one embodiment of the invention, the detergent composition present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
The fungal DNase may also be obtained from Trichoderma e.g. Trichoderma Harzianum e.g. One embodiment of the invention the pouch comprises a polypeptide having DNase activity, wherein the polypeptide comprises an amino acid sequence having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 64.
In one embodiment of the invention, the detergent composition present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition present in a first compartment of a multi-compartment pouch comprises at least one metalloprotease and the detergent composition is present in a second compartment of a multi-compartment pouch comprises at least one DNase.
In one embodiment of the present invention, the enzyme-containing composition present in a first compartment of a multi-compartment pouch comprises at least one metalloprotease and the detergent composition present in a second compartment of a multi-compartment pouch comprises at least one DNase, wherein the metalloprotease is classified in EC 3.4.22 or EC3.4.24.
In one embodiment of the present invention, the enzyme-containing composition present in a first compartment of a multi-compartment pouch comprises at least one metalloprotease and the detergent composition present in a second compartment of a multi-compartment pouch comprises at least one DNase and wherein the metalloprotease is an M4, M7 or M35.
In one embodiment of the present invention, the enzyme-containing composition is present in a first compartment of a multi-compartment pouch comprises at least one metalloprotease and the detergent composition present in a second compartment of a multi compartment pouch comprises at least one DNase, wherein the DNase is obtained from Bacillus.
In one embodiment of the present invention, the enzyme-containing composition is present in a first compartment of a multi-compartment pouch comprises at least one metalloprotease and the detergent composition present in a second compartment of a multi compartment pouch comprises at least one DNase, wherein the DNase is obtained from Aspergillus.
In one embodiment of the present invention, the enzyme-containing composition is present in a first compartment of a multi-compartment pouch comprises at least one metalloprotease and the detergent composition present in a second compartment of a multi compartment pouch comprises at least one DNase, wherein the DNase is obtained from Trichoderma.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 sequence percent identity to SEQ ID NOs: 1-36 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NOs: 60-64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 1 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 2 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 3 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 4 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 5 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 6 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 7 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 8 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 9 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 10 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 1 1 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 12 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 13 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 14 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 15 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 16 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 17 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 18 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 19 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 20 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 21 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 22 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 23 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 24 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 25 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 26 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 27 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 28 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 29 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 30 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 31 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 32 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 33 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 34 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 35 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 36 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 60.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 1 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 2 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 3 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 4 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 5 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 6 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 7 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 8 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 9 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 10 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 1 1 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 12 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 13 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 14 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 15 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 16 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 17 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 18 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 19 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 20 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 21 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 22 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 23 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 24 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 25 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 26 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 27 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 28 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 29 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 30 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 31 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 32 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 33 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 34 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 35 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 36 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 61.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent sequence identity to SEQ ID NO: 1 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 2 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 3 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 4 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 5 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 6 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 7 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 8 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 9 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 10 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 1 1 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 12 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 13 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 14 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 15 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 16 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 17 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 18 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 19 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 20 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 21 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 22 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 23 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 24 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 25 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 26 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 27 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 28 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 29 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 30 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 31 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 32 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 33 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 34 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 35 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 36 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 62.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 1 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 2 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 3 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 4 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 5 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 6 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 7 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 8 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 9 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 10 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 1 1 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 12 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 13 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 14 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 15 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 16 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 17 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 18 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 19 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 20 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 21 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 22 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 23 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 24 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 25 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 26 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 27 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 28 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 29 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 30 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 31 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 32 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 33 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 34 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 35 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 36 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 63.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 1 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 2 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 3 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 4 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 5 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 6 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 7 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 8 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 9 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 10 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 1 1 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 12 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 13 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 14 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 15 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 16 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 17 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 18 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 19 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 20 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 21 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 22 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 23 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 24 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 25 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 26 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 27 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 28 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 29 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 30 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 31 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 32 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 33 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 34 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 35 and the detergent composition is present in the second compartment of the multi compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 percent identity to SEQ ID NO: 36 and the detergent composition present in the second compartment of the multi compartment pouch comprises a DNase and having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 64.
In an embodiment of the present invention, the detergent composition is present in the second compartment of the multi-compartment pouch further comprises one or more detergent components selected from the group consisting of surfactants, builders, flocculating aid, chelating agents, dye transfer inhibitors, enzyme stabilizers, enzyme inhibitors, catalytic materials, bleach activators, hydrogen peroxide, sources of hydrogen peroxide, preformed peracids, polymeric dispersing agents, clay soil removal/anti-redeposition agents, brighteners, suds suppressors, dyes, perfumes, structure elasticizing agents, fabric softeners, carriers, hydrotropes, builders and co-builders, fabric hueing agents, anti-foaming agents, dispersants, processing aids, and/or pigments.
Surfactants
In one embodiment of the present invention, the detergent composition present in the second compartment of the multi-compartment pouch comprises a surfactant.
In one embodiment of the present invention, the detergent composition present in the second compartment of the multi-compartment pouch comprises a surfactant and wherein the surfactant is an anionic and/or a nonionic surfactant.
In one embodiment of the present invention, the detergent composition is present in the second compartment of the multi-compartment pouch comprises an anionic surfactant selected from linear alkylbenzenesulfonates (LAS) isomers of LAS, alcohol ether sulfate (AEO, AEOS) and sodium lauryl ether sulfate and sodium laureth sulfate (SLES).
In one embodiment of the present invention, the detergent composition present in the second compartment of the multi-compartment pouch comprises a nonionic surfactant is selected from alcohol ethoxylates (AE or AEO), alcohol propoxylates, alcohol propoxylates, propoxylated fatty alcohols (PFA), alkoxylated fatty acid alkyl esters, such as ethoxylated and/or propoxylated fatty acid alkyl esters, alkylphenol ethoxylates (APE), nonylphenol ethoxylates (NPE), alkylpolyglycosides (APG), alkoxylated amines, fatty acid monoethanolamides (FAM), fatty acid diethanolamides (FADA), ethoxylated fatty acid monoethanolamides (EFAM), propoxylated fatty acid monoethanolamides (PFAM), polyhydroxyalkyl fatty acid amides, /V-acyl N- alkyl derivatives of glucosamine (glucamides, GA, or fatty acid glucamides, FAGA) and combinations thereof.
The detergent composition present in the second compartment of the multi-compartment pouch may comprise one or more surfactants, which may be anionic and/or cationic and/or non-ionic and/or semi-polar and/or zwitterionic, or a mixture thereof. In a particular embodiment, the detergent composition includes a mixture of one or more nonionic surfactants and one or more anionic surfactants. The surfactant(s) is typically present at a level of from about 0.1 % to 60% by weight, such as about 1 % to about 40%, or about 3% to about 20%, or about 3% to about 10%. The surfactant(s) is chosen based on the desired cleaning application, and may include any conventional surfactant(s) known in the art.
When included therein the detergent will usually contain from about 1 % to about 40% by weight of an anionic surfactant, such as from about 5% to about 30%, including from about 5% to about 15%, or from about 15% to about 20%, or from about 20% to about 25% of an anionic surfactant. Non-limiting examples of anionic surfactants include sulfates and sulfonates, in particular, linear alkylbenzenesulfonates (LAS), isomers of LAS, branched alkylbenzenesulfonates (BABS), phenylalkanesulfonates, alpha-olefinsulfonates (AOS), olefin sulfonates, alkene sulfonates, alkane- 2, 3-diylbis(sulfates), hydroxyalkanesulfonates and disulfonates, alkyl sulfates (AS) such as sodium dodecyl sulfate (SDS), fatty alcohol sulfates (FAS), primary alcohol sulfates (PAS), alcohol
ethersulfates (AES or AEOS or FES, also known as alcohol ethoxysulfates or fatty alcohol ether sulfates), secondary alkanesulfonates (SAS), paraffin sulfonates (PS), ester sulfonates, sulfonated fatty acid glycerol esters, alpha-sulfo fatty acid methyl esters (alpha-SFMe or SES) including methyl ester sulfonate (MES), alkyl- or alkenylsuccinic acid, dodecenyl/tetradecenyl succinic acid (DTSA), fatty acid derivatives of amino acids, diesters and monoesters of sulfo-succinic acid or salt of fatty acids (soap), and combinations thereof.
When included therein the detergent will usually contain from about 1 % to about 40% by weigh of a cationic surfactant, for example from about 0.5% to about 30%, in particular from about 1 % to about 20%, from about 3% to about 10%, such as from about 3% to about 5%, from about 8% to about 12% or from about 10% to about 12%. Non-limiting examples of cationic surfactants include alkyldimethylethanolamine quat (ADMEAQ), cetyltrimethylammonium bromide (CTAB), dimethyldistearylammonium chloride (DSDMAC), and alkylbenzyldimethylammonium, alkyl quaternary ammonium compounds, alkoxylated quaternary ammonium (AQA) compounds, ester quats, and combinations thereof.
When included therein the detergent will usually contain from about 0.2% to about 40% by weight of a nonionic surfactant, for example from about 0.5% to about 30%, in particular from about 1 % to about 20%, from about 3% to about 10%, such as from about 3% to about 5%, from about 8% to about 12%, or from about 10% to about 12%. Non-limiting examples of nonionic surfactants include alcohol ethoxylates (AE or AEO), alcohol propoxylates, propoxylated fatty alcohols (PFA), alkoxylated fatty acid alkyl esters, such as ethoxylated and/or propoxylated fatty acid alkyl esters, alkylphenol ethoxylates (APE), nonylphenol ethoxylates (NPE), alkylpolyglycosides (APG), alkoxylated amines, fatty acid monoethanolamides (FAM), fatty acid diethanolamides (FADA), ethoxylated fatty acid monoethanolamides (EFAM), propoxylated fatty acid monoethanolamides (PFAM), polyhydroxyalkyl fatty acid amides, or N-acyl N-alkyl derivatives of glucosamine (glucamides, GA, or fatty acid glucamides, FAGA), as well as products available under the trade names SPAN and TWEEN, and combinations thereof.
When included therein the detergent will usually contain from about 0.01 % to about 10 % by weight of a semipolar surfactant. Non-limiting examples of semipolar surfactants include amine oxides (AO) such as alkyldimethylamineoxide, N-(coco alkyl)-N,N-dimethylamine oxide and N- (tallow-alkyl)-N,N-bis(2-hydroxyethyl)amine oxide and combinations thereof.
When included therein the detergent will usually contain from about 0.01 % to about 10 % by weight of a zwitterionic surfactant. Non-limiting examples of zwitterionic surfactants include betaines such as alkyldimethylbetaines, sulfobetaines, and combinations thereof.
The detergent composition present in the second compartment of the multi-compartment pouch preferably comprises surfactant in an amount to provide from 0.001 ppm to 5,000 ppm surfactant in the wash liquor during the laundering process.
Builders and Co-Builders
In one embodiment of the present invention, the detergent composition is present in the second compartment of the multi-compartment pouch comprises a builder.
The etergent composition present in the second compartment of the multi-compartment pouch comprises a builder, wherein the builder is selected from a group consisting of phosphates, sodium citrate builders, sodium carbonate, sodium silicate, sodium and zeolites.
The detergent composition present in the second compartment of the multi-compartment pouch may contain about 0-65% by weight, such as about 5% to about 50% of a detergent builder or co-builder, or a mixture thereof. In a dish wash detergent, the level of builder is typically 40-65%, particularly 50-65%. The builder and/or co-builder may particularly be a chelating agent that forms water-soluble complexes with Ca and Mg. Any builder and/or co-builder known in the art for use in laundry/ADW/hard surface cleaning detergents may be utilized. Non-limiting examples of builders include zeolites, diphosphates (pyrophosphates), triphosphates such as sodium triphosphate (STP or STPP), carbonates such as sodium carbonate, soluble silicates such as sodium metasilicate, layered silicates (e.g., SKS-6 from Hoechst), ethanolamines such as 2-aminoethan-1-ol (MEA), diethanolamine (DEA, also known as 2,2’-iminodiethan-1-ol), triethanolamine (TEA, also known as 2,2’,2”-nitrilotriethan-1-ol), and (carboxymethyl)inulin (CMI), and combinations thereof.
The detergent composition present in the second compartment of the multi-compartment pouch may also contain 0-50% by weight, such as about 5% to about 30%, of a detergent co-builder. The detergent composition may comprise a co-builder alone, or in combination with a builder, for example a zeolite builder. Non-limiting examples of co-builders include homopolymers of polyacrylates or copolymers thereof, such as poly(acrylic acid) (PAA) or copoly(acrylic acid/maleic acid) (PAA/PMA). Further non-limiting examples include citrate, chelators such as aminocarboxylates, aminopolycarboxylates and phosphonates, and alkyl- or alkenylsuccinic acid. Additional specific examples include 2,2’,2”-nitrilotriacetic acid (NTA), ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), iminodisuccinic acid (IDS), ethylenediamine-N,N’-disuccinic acid (EDDS), methylglycinediacetic acid (MGDA), glutamic acid- N,N-diacetic acid (GLDA), 1-hydroxyethane-1 ,1-diphosphonic acid (HEDP), ethylenediaminetetra(methylenephosphonic acid) (EDTMPA), diethylenetriaminepentakis(methylenephosphonic acid) (DTMPA or DTPMPA), N-(2- hydroxyethyl)iminodiacetic acid (EDG), aspartic acid-N-monoacetic acid (ASMA), aspartic acid-N,N- diacetic acid (ASDA), aspartic acid-N-monopropionic acid (ASMP), iminodisuccinic acid (IDA), N-(2- sulfomethyl)-aspartic acid (SMAS), N-(2-sulfoethyl)-aspartic acid (SEAS), N-(2-sulfomethyl)- glutamic acid (SMGL), N-(2-sulfoethyl)-glutamic acid (SEGL), N-methyliminodiacetic acid (MIDA), a- alanine-N,N-diacetic acid (a-ALDA), serine-N,N-diacetic acid (SEDA), isoserine-N,N-diacetic acid
(ISDA), phenylalanine-N,N-diacetic acid (PHDA), anthranilic acid-N,N-diacetic acid (ANDA), sulfanilic acid-N,N-diacetic acid (SLDA) , taurine-N,N-diacetic acid (TUDA) and sulfomethyl-N,N- diacetic acid (SMDA), N-(2-hydroxyethyl)ethylenediamine-N,N’,N”-triacetic acid (HEDTA), diethanolglycine (DEG), diethylenetriamine penta(methylenephosphonic acid) (DTPMP), aminotris(methylenephosphonic acid) (ATMP), and combinations and salts thereof. Further exemplary builders and/or co-builders are described in, e.g., WO 09/102854, US 5977053.
Hydrotropes
In one embodiment, the detergent composition present in the second compartment of the multi-compartment pouch comprises one or more hydrotrope.
A hydrotrope is a compound that solubilises hydrophobic compounds in aqueous solutions (or oppositely, polar substances in a non-polar environment). Typically, hydrotropes have both hydrophilic and a hydrophobic character (so-called amphiphilic properties as known from surfactants); however the molecular structure of hydrotropes generally do not favor spontaneous self-aggregation, see e.g., review by Hodgdon and Kaler (2007), Current Opinion in Colloid & Interface Science 12: 121-128. Hydrotropes do not display a critical concentration above which self-aggregation occurs as found for surfactants and lipids forming miceller, lamellar or other well defined meso-phases. Instead, many hydrotropes show a continuous-type aggregation process where the sizes of aggregates grow as concentration increases. However, many hydrotropes alter the phase behavior, stability, and colloidal properties of systems containing substances of polar and non-polar character, including mixtures of water, oil, surfactants, and polymers. Hydrotropes are classically used across industries from pharma, personal care, food, to technical applications. Use of hydrotropes in detergent compositions allow for example more concentrated formulations of surfactants (as in the process of compacting liquid detergents by removing water) without inducing undesired phenomena such as phase separation or high viscosity.
The detergent present in the second compartment of the multi-compartment pouch may contain 0-10% by weight, such as about 0.5 to about 5%, or about 3% to about 5%, of a hydrotrope. Any hydrotrope known in the art for use in detergents may be utilized. Non-limiting examples of hydrotropes include sodium benzene sulfonate, sodium p-toluene sulfonate (STS), sodium xylene sulfonate (SXS), sodium cumene sulfonate (SCS), sodium cymene sulfonate, amine oxides, alcohols and polyglycolethers, sodium hydroxynaphthoate, sodium hydroxynaphthalene sulfonate, sodium ethylhexyl sulfate, and combinations thereof.
Bleaching Systems
In one embodiment of the present invention, the detergent composition present in the
second compartment of the multi-compartment pouch comprises one or more bleaching system.
The detergent composition present in the second compartment of the multi-compartment pouch may contain 0-30% by weight, such as about 1 % to about 20%, of a bleaching system. Any bleaching system known in the art for use in laundry/ADW/hard surface cleaning detergents may be utilized. Suitable bleaching system components include sources of hydrogen peroxide; sources of peracids; and bleach catalysts or boosters.
Sources of hydrogen peroxide:
Suitable sources of hydrogen peroxide are inorganic persalts, including alkali metal salts such as sodium percarbonate and sodium perborates (usually mono- or tetra hydrate), and hydrogen peroxide— urea (1/1).
Sources of peracids:
Peracids may be (a) incorporated directly as preformed peracids or (b) formed in situ in the wash liquor from hydrogen peroxide and a bleach activator (perhydrolysis) or (c) formed in situ in the wash liquor from hydrogen peroxide and a perhydrolase and a suitable substrate for the latter, e.g., an ester.
a) Suitable preformed peracids include, but are not limited to, peroxycarboxylic acids such as peroxybenzoic acid and its ring-substituted derivatives, peroxy-a-naphthoic acid, peroxyphthalic acid, peroxylauric acid, peroxystearic acid, e-phthalimidoperoxycaproic acid [phthalimidoperoxyhexanoic acid (PAP)], and o-carboxybenzamidoperoxycaproic acid; aliphatic and aromatic diperoxydicarboxylic acids such as diperoxydodecanedioic acid, diperoxyazelaic acid, diperoxysebacic acid, diperoxybrassylic acid, 2-decyldiperoxybutanedioic acid, and diperoxyphthalic, -isophthalic and -terephthalic acids; perimidic acids; peroxymonosulfuric acid; peroxydisulfuric acid; peroxyphosphoric acid; peroxysilicic acid; and mixtures of said compounds. It is understood that the peracids mentioned may in some cases be best added as suitable salts, such as alkali metal salts (e.g., Oxone®) or alkaline earth-metal salts.
b) Suitable bleach activators include those belonging to the class of esters, amides, imides, nitriles or anhydrides and, where applicable, salts thereof. Suitable examples are tetraacetylethylenediamine (TAED), sodium 4-[(3, 5, 5-trimethylhexanoyl)oxy]benzene-1 -sulfonate (ISONOBS), sodium 4-(dodecanoyloxy)benzene-1 -sulfonate (LOBS), sodium 4- (decanoyloxy) benzene- 1 -sulfonate, 4-(decanoyloxy)benzoic acid (DOBA), sodium 4- (nonanoyloxy)benzene-l-sulfonate (NOBS), and/or those disclosed in W098/17767. A particular family of bleach activators of interest was disclosed in EP624154 and particularly preferred in that family is acetyl triethyl citrate (ATC). ATC or a short chain triglyceride like triacetin has the advantage that they are environmentally friendly. Furthermore, acetyl triethyl citrate and triacetin have good hydrolytical stability in the product upon storage and are efficient bleach activators. Finally, ATC is multifunctional, as the citrate released in the perhydrolysis reaction may function
as a builder.
Bleach catalysts and boosters
In one embodiment of the present invention, the detergent composition present in the second compartment of the multi-compartment pouch comprises one or more bleach catalyst and/or booster.
The bleaching system may also include a bleach catalyst or booster.
Some non-limiting examples of bleach catalysts that may be used in the compositions of the present invention include manganese oxalate, manganese acetate, manganese-collagen, cobalt-amine catalysts and manganese triazacyclononane (MnTACN) catalysts; particularly preferred are complexes of manganese with 1 ,4,7-trimethyl-1 ,4,7-triazacyclononane (Me3- TACN) or 1 ,2,4,7-tetramethyl-1 ,4,7-triazacyclononane (Me4-TACN), in particular Me3-TACN, such as the dinuclear manganese complex [(Me3-TACN)Mn(0)3Mn(Me3-TACN)](PF6)2, and [2,2',2"-nitrilotris(ethane-1 ,2-diylazanylylidene-KN-methanylylidene)triphenolato- K30]manganese(lll). The bleach catalysts may also be other metal compounds; such as iron or cobalt complexes.
In some embodiments, where a source of a peracid is included, an organic bleach catalyst or bleach booster may be used having one of the following formulae:
(iii) and mixtures thereof; wherein each R1 is independently a branched alkyl group containing from 9 to 24 carbons or linear alkyl group containing from 11 to 24 carbons, preferably each R1 is independently a branched alkyl group containing from 9 to 18 carbons or linear alkyl group containing from 11 to 18 carbons, more preferably each R1 is independently selected from the group consisting of 2-propylheptyl, 2-butyloctyl, 2-pentylnonyl, 2-hexyldecyl, dodecyl, tetradecyl, hexadecyl, octadecyl, isononyl, isodecyl, isotridecyl and isopentadecyl.
Other exemplary bleaching systems are described, e.g. in W02007/087258, W02007/087244, W02007/087259, EP1867708 (Vitamin K) and W02007/087242. Suitable photobleaches may for example be sulfonated zinc or aluminium phthalocyanines.
Metal care agents
In one embodiment of the present invention, the detergent composition present in the
second compartment of the multi-compartment pouch comprises one or more metal care agents. Metal care agents may prevent or reduce the tarnishing, corrosion or oxidation of metals, including aluminium, stainless steel and non-ferrous metals, such as silver and copper. Suitable examples include one or more of the following:
(a) benzatriazoles, including benzotriazole or bis-benzotriazole and substituted derivatives thereof. Benzotriazole derivatives are those compounds in which the available substitution sites on the aromatic ring are partially or completely substituted. Suitable substituents include linear or branch-chain Ci-C20- alkyl groups (e.g., C1-C20- alkyl groups) and hydroxyl, thio, phenyl or halogen such as fluorine, chlorine, bromine and iodine.
(b) metal salts and complexes chosen from the group consisting of zinc, manganese, titanium, zirconium, hafnium, vanadium, cobalt, gallium and cerium salts and/or complexes, the metals being in one of the oxidation states II , III, IV, V or VI. In one aspect, suitable metal salts and/or metal complexes may be chosen from the group consisting of Mn(ll) sulphate, Mn(ll) citrate, Mn(ll) stearate, Mn(ll) acetylacetonate, KATiF6 (e.g., K2T 6), KAZrF6 (e.g., K2ZrF6), CoS04, Co(NOs)2 and Ce(NOs)3, zinc salts, for example zinc sulphate, hydrozincite or zinc acetate.;
(c) silicates, including sodium or potassium silicate, sodium disilicate, sodium metasilicate, crystalline phyllosilicate and mixtures thereof.
Further suitable organic and inorganic redox-active substances that act as silver/copper corrosion inhibitors are disclosed in WO 94/26860 and WO 94/26859. Preferably the composition of the invention comprises from 0.1 to 5% by weight of the composition of a metal care agent, preferably the metal care agent is a zinc salt.
Polymers
In one embodiment of the present invention, the detergent composition present in the second compartment of the multi-compartment pouch comprises one or more polymers.
The detergent composition may contain 0-10% by weight, such as 0.5-5%, 2-5%, 0.5-2% or 0.2-1 % of a polymer. Any polymer known in the art for use in detergents may be utilized. The polymer may function as a co-builder as mentioned above, or may provide antiredeposition, fiber protection, soil release, dye transfer inhibition, grease cleaning and/or anti-foaming properties. Some polymers may have more than one of the above-mentioned properties and/or more than one of the below- mentioned motifs. Exemplary polymers include (carboxymethyl)cellulose (CMC), poly(vinyl alcohol) (PVA), poly(vinylpyrrolidone) (PVP), poly(ethyleneglycol) or poly(ethylene oxide) (PEG), ethoxylated poly(ethyleneimine), carboxymethyl inulin (CMI), and polycarboxylates such as PAA, PAA/PMA, poly-aspartic acid, and lauryl methacrylate/acrylic acid copolymers , hydrophobically modified CMC (HM-CMC) and silicones, copolymers of terephthalic acid and oligomeric glycols, copolymers of poly(ethylene terephthalate) and poly(oxyethene terephthalate) (PET-POET), PVP,
poly(vinylimidazole) (PVI), poly(vinylpyridine-N-oxide) (PVPO or PVPNO) and polyvinylpyrrolidone- vinylimidazole (PVPVI). Further exemplary polymers include sulfonated polycarboxylates, polyethylene oxide and polypropylene oxide (PEO-PPO) and diquaternium ethoxy sulfate. Other exemplary polymers are disclosed in, e.g., WO 2006/130575. Salts of the above-mentioned polymers are also contemplated.
Fabric hueing agents
In one embodiment of the present invention, the detergent composition present in the second compartment of the multi-compartment pouch comprises one or more fabric hueing agent.
The detergent composition may also comprise fabric hueing agents such as dyes or pigments, which when formulated in detergent compositions can deposit onto a fabric when said fabric is contacted with a wash liquor comprising said detergent compositions and thus altering the tint of said fabric through absorption/reflection of visible light. Fluorescent whitening agents emit at least some visible light. In contrast, fabric hueing agents alter the tint of a surface as they absorb at least a portion of the visible light spectrum. Suitable fabric hueing agents comprise dyes and dye- clay conjugates, and may also comprise pigments. Suitable dyes comprise small molecule dyes and polymeric dyes. Suitable small molecule dyes include small molecule dyes selected from the group consisting of dyes falling into the Colour Index (C.l.) classifications of Direct Blue, Direct Red, Direct Violet, Acid Blue, Acid Red, Acid Violet, Basic Blue, Basic Violet and Basic Red, or mixtures thereof, for example as described in W02005/03274, W02005/03275, W02005/03276 and EP1876226 (hereby incorporated by reference). The detergent composition present in the second compartment of the multi-compartment pouch preferably comprises from about 0.00003 wt% to about 0.2 wt%, from about 0.00008 wt% to about 0.05 wt%, or even from about 0.0001 wt% to about 0.04 wt% fabric hueing agent. The detergent composition present in the second compartment of the multi compartment pouch may comprise from 0.0001 wt% to 0.2 wt% fabric hueing agent, this may be especially preferred when the composition is in the form of a unit dose pouch. Suitable hueing agents are also disclosed in, e.g. WO 2007/087257 and W02007/087243.
Adjunct materials
In one embodiment of the present invention, the detergent composition present in the second compartment of the multi-compartment pouch comprises one or more adjunct material.
Any detergent components known in the art for use in laundry/ADW/hard surface cleaning detergents may also be utilized. Other optional detergent components include anti-corrosion agents, anti-shrink agents, anti-soil redeposition agents, anti-wrinkling agents, bactericides, binders, corrosion inhibitors, disintegrants/disintegration agents, dyes, enzyme stabilizers (including boric acid, borates, CMC, and/or polyols such as propylene glycol), fabric conditioners including clays,
fillers/processing aids, fluorescent whitening agents/optical brighteners, foam boosters, foam (suds) regulators, perfumes, soil-suspending agents, softeners, suds suppressors, tarnish inhibitors, and wicking agents, either alone or in combination. Any ingredient known in the art for use in laundry/ADW/hard surface cleaning detergents may be utilized. The choice of such ingredients is well within the skill of the artisan.
Dispersants
In one embodiment of the present invention, the detergent composition present in the second compartment of the multi-compartment pouch comprises one or more dispersants.
The detergent composition can also contain dispersants. In particular powdered detergents may comprise dispersants. Suitable water-soluble organic materials include the homo- or co-polymeric acids or their salts, in which the polycarboxylic acid comprises at least two carboxyl radicals separated from each other by not more than two carbon atoms. Suitable dispersants are for example described in Powdered Detergents, Surfactant science series volume 71 , Marcel Dekker, Inc.
Dye Transfer Inhibiting Agents
In one embodiment of the present invention, the detergent composition present in the second compartment of the multi-compartment pouch comprises one or more dye transfer inhibiting agents.
Suitable polymeric dye transfer inhibiting agents include, but are not limited to, polyvinylpyrrolidone polymers, polyamine N-oxide polymers, copolymers of N-vinylpyrrolidone and N-vinylimidazole, polyvinyloxazolidones and polyvinylimidazoles or mixtures thereof. When present in a subject composition, the dye transfer inhibiting agents may be present at levels from about 0.0001 % to about 10%, from about 0.01 % to about 5% or even from about 0.1 % to about 3% by weight of the composition.
Fluorescent whitening agent
In one embodiment of the present invention, the detergent composition present in the second compartment of the multi-compartment pouch comprises one or more fluorescent whitening agent.
The detergent composition present in the second compartment of the multi-compartment pouch will preferably also contain additional components that may tint articles being cleaned, such as fluorescent whitening agent or optical brighterners. Any fluorescent whitening agent suitable for use in a laundry detergent composition may be used in the composition of the present invention. The most commonly used fluorescent whitening agents are those belonging to the
classes of diaminostilbene-sulphonic acid derivatives, diarylpyrazoline derivatives and bisphenyl- distyryl derivatives. Examples of the diaminostilbene-sulphonic acid derivative type of fluorescent whitening agents include the sodium salts of:
4,4'-bis-(2-diethanolamino-4-anilino-s-triazin-6-ylamino) stilbene-2,2'-disulphonate,
4,4'-bis-(2,4-dianilino-s-triazin-6-ylamino) stilbene-2.2'-disulphonate,
4,4'-bis-(2-anilino-4(N-methyl-N-2-hydroxy-ethylamino)-s-triazin-6-ylamino) stilbene-2,2'- disulphonate, 4,4'-bis-(4-phenyl-2,1 ,3-triazol-2-yl)stilbene-2,2'-disulphonate,
4,4'-bis-(2-anilino-4(1-methyl-2-hydroxy-ethylamino)-s-triazin-6-ylamino) stilbene-2,2'- disulphonate and, 2-(stilbyl-4"-naptho-1 ,2':4,5)-1 , 2, 3-trizole-2"-sulphonate. Preferred fluorescent whitening agents are Tinopal DMS and Tinopal CBS available from Ciba-Geigy AG, Basel, Switzerland. Tinopal DMS is the disodium salt of 4,4'-bis-(2-morpholino-4 anilino-s-triazin-6- ylamino) stilbene disulphonate. Tinopal CBS is the disodium salt of 2,2'-bis-(phenyl-styryl) disulphonate. Also preferred are fluorescent whitening agents is the commercially available Parawhite KX, supplied by Paramount Minerals and Chemicals, Mumbai, India. Other fluorescers suitable for use in the invention include the 1-3-diaryl pyrazolines and the 7- alkylaminocoumarins.
Suitable fluorescent brightener levels include lower levels of from about 0.01 , from 0.05, from about 0.1 or even from about 0.2 wt% to upper levels of 0.5 or even 0.75 wt%.
Soil release polymers
In one embodiment of the present invention, the detergent composition present in the second compartment of the multi-compartment pouch comprises one or more soil release polymer.
The detergent composition present in the second compartment of the multi-compartment pouch may also include one or more soil release polymers which aid the removal of soils from fabrics such as cotton and polyester based fabrics, in particular the removal of hydrophobic soils from polyester based fabrics. The soil release polymers may for example be nonionic or anionic terephthalte based polymers, polyvinyl caprolactam and related copolymers, vinyl graft copolymers, polyester polyamides see for example Chapter 7 in Powdered Detergents, Surfactant science series, volume 71 , Marcel Dekker, Inc. Another type of soil release polymers is amphiphilic alkoxylated grease cleaning polymers comprising a core structure and a plurality of alkoxylate groups attached to that core structure. The core structure may comprise a polyalkylenimine structure or a polyalkanolamine structure as described in detail in WO 2009/087523. Furthermore, random graft co-polymers are suitable soil release polymers Suitable graft co-polymers are described in more detail in WO 2007/138054, WO 2006/108856 and WO 2006/113314. Other soil release polymers are substituted polysaccharide structures especially
substituted cellulosic structures such as modified cellulose deriviatives such as those described in EP 1 867 808 or WO 2003/040279. Suitable cellulosic polymers include cellulose, cellulose ethers, cellulose esters, cellulose amides and mixtures thereof. Suitable cellulosic polymers include anionically modified cellulose, nonionically modified cellulose, cationically modified cellulose, zwitterionically modified cellulose, and mixtures thereof. Suitable cellulosic polymers include methyl cellulose, carboxy methyl cellulose, ethyl cellulose, hydroxyl ethyl cellulose, hydroxyl propyl methyl cellulose, ester carboxy methyl cellulose, and mixtures thereof.
Anti-redeposition agents
In one embodiment of the present invention, the detergent composition present in the second compartment of the multi-compartment pouch comprises one or more anti-redeposition agents.
The detergent composition present in the second compartment of the multi-compartment pouch may also include one or more anti-redeposition agents such as carboxymethylcellulose (CMC), polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), polyoxyethylene and/or polyethyleneglycol (PEG), homopolymers of acrylic acid, copolymers of acrylic acid and maleic acid, and ethoxylated polyethyleneimines. The cellulose based polymers described under soil release polymers above may also function as anti-redeposition agents.
Other suitable adjunct materials include, but are not limited to, anti-shrink agents, anti wrinkling agents, bactericides, binders, carriers, dyes, enzyme stabilizers, fabric softeners, fillers, foam regulators, hydrotropes, perfumes, pigments, sod suppressors, solvents, structurants for liquid detergents and/or structure elasticizing agents.
Rheology Modifiers
In one embodiment of the present invention, the detergent composition present in the second compartment of the multi-compartment pouch comprises one or more rheology modifiers.
The detergent composition present in the second compartment of the multi-compartment pouch may also comprise one or more rheology modifiers, structurants or thickeners, as distinct from viscosity reducing agents. The rheology modifiers are selected from the group consisting of non polymeric crystalline, hydroxy-functional materials, polymeric rheology modifiers which impart shear thinning characteristics to the aqueous liquid matrix of a liquid detergent composition. The rheology and viscosity of the detergent can be modified and adjusted by methods known in the art, for example as shown in EP 2169040.
Other suitable adjunct materials
In one embodiment of the present invention, the detergent composition present in the
second compartment of the multi-compartment pouch comprises one or more other suitable adjunct material. Other suitable adjunct materials include, but are not limited to, anti-shrink agents, anti-wrinkling agents, bactericides, binders, carriers, dyes, enzyme stabilizers, fabric softeners, fillers, foam regulators, hydrotropes, perfumes, pigments, sod suppressors, solvents, and structurants for liquid detergents and/or structure elasticizing agents.
Enzymes
In one embodiment, the enzyme-containing present in the first compartment of the multi compartment pouch further comprises one or more additional enzymes.
In one embodiment, the detergent composition present in the second compartment of the multi-compartment pouch further comprises one or more additional enzymes.
In one embodiment, the enzyme-containing present in the first compartment of the multi compartment pouch further comprises one or more additional enzymes and/or the detergent composition present in the second compartment of the multi-compartment pouch further comprises one or more additional enzymes.
In one embodiment, the enzyme-containing present in the first compartment of the multi compartment pouch and/or the detergent composition present in the second compartment of the multi-compartment pouch comprises an additional enzyme selected from the group consisting of an alpha-amylase, a beta-amylase, a protease, a pullulanase, a lipase, a cellulase, an oxidase, a phospholipase, a perhydrolase, a xylanase, a pectate lyase, a pectinase, a galacturanase, a hemicellulase, a xyloglucanase, a nuclease, a mannanase and mixtures thereof.
In general, the properties of the selected enzyme(s) should be compatible with the selected detergent, (i.e., pH-optimum, compatibility with other enzymatic and non-enzymatic ingredients, etc.), and the enzyme(s) should be present in effective amounts.
Cellulases:
Suitable cellulases comprise those of bacterial or fungal origin. Chemically modified or protein engineered mutants are included. Suitable cellulases include cellulases from the genera Bacillus, Pseudomonas, Humicola, Fusarium, Thielavia, Acremonium, e.g., the fungal cellulases produced from Humicola insolens, Myceliophthora thermophila and Fusarium oxysporum disclosed in US 4,435,307, US 5,648,263, US 5,691 , 178, US 5,776,757 and WO 89/09259.
Especially suitable cellulases are the alkaline or neutral cellulases having colour care benefits. Examples of such cellulases are cellulases described in EP 0 495 257, EP 0 531 372, WO 96/1 1262, WO 96/29397, WO 98/08940. Other examples are cellulase variants such as those described in WO 94/07998, EP 0 531 315, US 5,457,046, US 5,686,593, US 5,763,254, WO 95/24471 , WO 98/12307 and W099/001544.
Other cellulases are endo-beta-1 ,4-glucanase enzyme having a sequence of at least 97% identity to the amino acid sequence of position 1 to position 773 of SEQ ID NO:2 of WO 2002/099091 or a family 44 xyloglucanase, which a xyloglucanase enzyme having a sequence of at least 60% identity to positions 40-559 of SEQ ID NO: 2 of WO 2001/062903.
Commercially available cellulases include Celluzyme™, and Carezyme™ (Novozymes A/S) Carezyme Premium™ (Novozymes A/S), Celluclean ™ (Novozymes A/S), Celluclean Classic™ (Novozymes A/S), Cellusoft™ (Novozymes A/S), Whitezyme™ (Novozymes A/S), Clazinase™, and Puradax HA™ (Genencor International Inc.), and KAC-500(B)™ (Kao Corporation).
Cellulase in particular an enzyme exhibiting endo-beta-1 ,4-glucanase activity. Cellulose is a polymer of glucose linked by beta-1 ,4-glucosidic bonds. Cellulose chains form numerous intra- and intermolecular hydrogen bonds, which results in the formation of insoluble cellulose micro-fibrils. Microbial hydrolysis of cellulose to glucose involve three major classes of cellulases: (i) endo-glucanases (EC 3.2.1.4) which cleave beta-1 , 4-glucosidic links randomly throughout cellulose molecules, also called endo-beta-1 ,4-glucanases; (ii) cellobiohydrolases (EC 3.2.1.91) which digest cellulose from the non-reducing end, releasing cellobiose; and (iii) beta-glucosidases (EC 3.2.1.21) which hydrolyse cellobiose and low molecular-weight cellodextrins to release glucose. The cellulases useful in the cleaning composition of the invention are preferably endo- glucanases (EC 3.2.1.4). Beta-1 , 4-glucosidic bonds are also present beta-glucans from plants such as barley and oats. In some cases, endo-glucanases also provide hydrolysis of such non cellulose polymers. The cellulases are placed into different families of glycosyl hydrolases; fungal and bacterial glycosyl hydrolases have been grouped into 35 families (Henrissat, B.: A classification of glycosyl hydrolases based on amino acid sequence similarities. Biochem. J. 280 (1991), 309-316. Henrissat, B., and Bairoch, A.: New families in the classification of glycosyl hydrolases based on amino acid sequence similarities. Biochem. J. 293 (1993), 781 -788.). Most cellulases consist of a cellulose-binding domain (CBD) and a catalytic domain (CAD) separated by a linker which may be rich in proline and hydroxy amino acid residues. Another classification of cellulases has been established on the basis of the similarity of their CBDs (Gilkes et al. (1991)) giving five families of glycosyl hydrolases (l-V). Cellulases are synthesized by a large number of microorganisms which include fungi, actinomycetes, myxobacteria and true bacteria but also by plants. Especially endo-beta-1 ,4-glucanases of a wide variety of specificities have been identified. Many bacterial endo-glucanases have been described (Gilbert, H.J. and Hazlewood, G.P. (1993) J. Gen. Microbiol. 139:187-194. Henrissat, B., and Bairoch, A.: New families in the classification of glycosyl hydrolases based on amino acid sequence similarities. Biochem. J. 293 (1993), 781 - 788.). One preferred cellulase includes endo-beta-1 ,4-glucanase activity (EC 3.2.1.4), preferably obtained from Bacillus sp. AA349 (DSM 12648), as described in W02002/099091. Other
preferred cellulases include those described in W01996/029397, which discloses family 45 endoglucanases e.g. cellulases from Thielavia in particular a strain of Thielavia terrestris and the cellulases described in WO1991/017243, which discloses endoglucanases from e.g. of Humicola such as Humicola insolens.
One embodiment of the invention, the detergent composition in the second compartment of the pouch comprises a cellulase selected from the group consisting of:
a) a cellulase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 45, preferably obtained from Bacillus e.g. Bacillus subtilis; b) a cellulase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 46, preferably obtained from Bacillus e.g. Bacillus subtilis ; c) a cellulase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 47, preferably obtained from Humicola insolens ; and d) a cellulase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 48, preferably obtained from Thielavia terrestris.
Mannanases
Suitable mannanases include those of bacterial or fungal origin. Chemically or genetically modified mutants are included. The mannanase may be an alkaline mannanase of Family 5 or 26. It may be a wild-type from Bacillus or Humicola, particularly B. agaradhaerens, B. licheniformis, B. halodurans, B. clausii, or H. insolens. Suitable mannanases are described in WO 1999/064619.
One embodiment of the invention, the detergent composition in the second compartment of the pouch comprises a mannanase selected from the group consisting of:
a) a mannanase, wherein the mannanase belongs to the Glycoside Hydrolase Family 5 mannanases;
i. a mannanase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 50, preferably from Bacillus e.g. Bacillus bogoriensis ;
b) a mannanase wherein the mannanase belongs to the Glycoside Hydrolase Family 26 mannanases;
I. a mannanase having at least 60%, at least 65%, at least 70%, at least 75%,
at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 51 , preferably obtained from Paenibacillus woosongensis;
II. a mannanase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 52, preferably obtained from Paenibacillus illinoisensis·,
III. a mannanase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 53, preferably obtained from Neobulgaria sp.;
IV. a mannanase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 54, preferably obtained from Preussia aemulans ;
V. a mannanase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 54, preferably obtained from Yunnania penicillata·,
VI. a mannanase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 56, preferably obtained from Myrothecium roridunr,
VII. a mannanase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 57, preferably obtained from Chaetomium brasiliense ;
VIII. a mannanase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 58, preferably obtained from Ascobolus stictoideus ; and
IX. a mannanase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 59, preferably obtained from Chaetomium virescens.
A commercially available mannanase is Mannaway (Novozymes A/S).
Lipases and Cutinases:
Suitable lipases and cutinases include those of bacterial or fungal origin. Chemically modified or protein engineered mutant enzymes are included. Examples include lipase from Thermomyces, e.g. from T. lanuginosus (previously named Humicola lanuginosa) as described in EP258068 and EP305216, cutinase from Humicola, e.g. H. insolens (WO96/13580), lipase from strains of Pseudomonas (some of these now renamed to Burkholderia), e.g. P. alcaligenes or P. pseudoalcaligenes (EP218272), P. cepacia (EP331376), P. sp. strain SD705 (W095/06720 & W096/27002), P. wisconsinensis (WO96/12012), GDSL-type Streptomyces lipases (W010/065455), cutinase from Magnaporthe grisea (W010/107560), cutinase from Pseudomonas mendocina (US5,389,536), lipase from Thermobifida fusca (W011/084412), Geobacillus stearothermophilus lipase (W011/084417), lipase from Bacillus subtilis (W011/084599), and lipase from Streptomyces griseus (WO11/150157) and S. pristinaespiralis (W012/137147).
Other examples are lipase variants such as those described in EP407225, WO92/05249, WO94/01541 , WO 94/25578, W095/14783, WO95/30744, W095/35381 , W095/22615,
W096/00292, W097/04079, W097/07202, WO00/34450, WO00/60063, W001/92502,
W007/87508 and WO09/109500.
One embodiment of the invention, the detergent composition in the second compartment of the pouch comprises a lipase selected from the group consisting of:
a) a lipase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 49, b) a lipase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 49 comprising one or more of the substitutions selected from the group consisting of D27R, G38A, G91A/Q, D96E, G163K, T231 R, N233R, D254S and P256T, compared to SEQ ID NO 49; and
c) a lipase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 49, wherein the lipase comprises one or both the substitutions T231 R and/or N233R, wherein the position corresponds to the positions of SEQ ID NO 49.
Preferred commercial lipase products include Lipolase™, Lipex™; Lipolex™ and Lipoclean™ (Novozymes A/S), Lumafast (originally from Genencor) and Lipomax (originally from Gist-Brocades).
Still other examples are lipases sometimes referred to as acyltransferases or perhydrolases, e.g. acyltransferases with homology to Candida antarctica lipase A
(WO10/1 11 143), acyltransferase from Mycobacterium smegmatis (WO05/56782), perhydrolases from the CE 7 family (WO09/67279), and variants of the M. smegmatis perhydrolase in particular the S54V variant used in the commercial product Gentle Power Bleach from Huntsman Textile Effects Pte Ltd (W010/100028).
Proteases:
Suitable proteases include those of bacterial, fungal, plant, viral or animal origin e.g. vegetable or microbial origin. Microbial origin is preferred. Chemically modified or protein engineered mutants are included. It may be an alkaline protease, such as a serine protease. A serine protease may for example be of the S1 family, such as trypsin, or the S8 family such as subtilisin.
The term "subtilases" refers to a sub-group of serine protease according to Siezen et al., Protein Engng. 4 (1991) 719-737 and Siezen et al. Protein Science 6 (1997) 501-523. Serine proteases are a subgroup of proteases characterized by having a serine in the active site, which forms a covalent adduct with the substrate. The subtilases may be divided into 6 sub-divisions, i.e. the Subtilisin family, the Thermitase family, the Proteinase K family, the Lantibiotic peptidase family, the Kexin family and the Pyrolysin family.
Examples of subtilases are those derived from Bacillus such as Bacillus lentus, B. alkalophilus, B. subtilis, B. amyloliquefaciens, Bacillus pumilus and Bacillus gibsonii described in; US7262042 and W009/021867, and subtilisin lentus, subtilisin Novo, subtilisin Carlsberg, Bacillus licheniformis, subtilisin BPN’, subtilisin 309, subtilisin 147 and subtilisin 168 described in WO89/06279 and protease PD138 described in (WO93/18140). Other useful proteases may be those described in WO 92/175177, WO 01/016285, WO 02/026024 and WO 02/016547. Examples of trypsin-like proteases are trypsin (e.g. of porcine or bovine origin) and the Fusarium protease described in WO 89/06270, WO 94/25583 and WO 05/040372, and the chymotrypsin proteases derived from Cellumonas described in WO 05/052161 and WO 05/052146.
A further preferred protease is the alkaline protease from Bacillus lentus DSM 5483, as described for example in WO 95/23221 , and variants thereof which are described in WO 92/21760, WO 95/23221 , EP 1921147 and EP 1921148.
Examples of useful proteases are the variants described in: WO 92/19729, WO 96/034946, WO 98/201 15, WO 98/201 16, WO 99/011768, WO 01/44452, WO 03/006602, WO 04/03186, WO 04/041979, WO 07/006305, WO 1 1/036263, WO 11/036264, especially the variants with substitutions in one or more of the following positions: 3, 4, 9, 15, 24, 27, 42, 55, 59, 60, 66, 74, 85, 96, 97, 98, 99, 100, 101 , 102, 104, 1 16, 1 18, 121 , 126, 127, 128, 154, 156, 157, 158, 161 , 164, 176, 179, 182, 185, 188, 189, 193, 198, 199, 200, 203, 206, 211 , 212, 216, 218,
226, 229, 230, 239, 246, 255, 256, 268 and 269 wherein the positions correspond to the positions of the Bacillus lentus protease shown in SEQ ID NO 1 of WO 2016/001449. More preferred the subtilase variants may comprise the mutations: S3T, V4I, S9R, S9E, A15T, S24G, S24R, K27R, N42R, S55P, G59E, G59D, N60D, N60E, V66A, N74D, N85S, N85R, , G96S, G96A, S97G, S97D, S97A, S97SD, S99E, S99D, S99G, S99M, S99N, S99R, S99H, S101A, V102I, V102Y, V102N, S104A, G1 16V, G1 16R, H118D, H118N, N120S, S126L, P127Q, S128A, S154D, A156E, G157D, G157P, S158E, Y161A, R164S, Q176E, N179E, S182E, Q185N, A188P, G189E, V193M, N198D, V199I, Y203W, S206G, L211Q, L211 D, N212D, N212S, M216S, A226V, K229L, Q230H, Q239R, N246K, N255W, N255D, N255E, L256E, L256D T268A, R269H. The protease variants are preferably variants of the Bacillus lentus protease (Savinase®) shown in SEQ ID NO 1 of WO 2016/001449, the Bacillus amyloliquefaciens protease (BPN’) shown in SEQ ID NO 2 of W02016/001449. The protease variants preferably have at least 80 % sequence identity to SEQ ID NO 1 or SEQ ID NO 2 of WO 2016/001449.
A protease variant comprising a substitution at one or more positions corresponding to positions 171 , 173, 175, 179, or 180 of SEQ ID NO: 1 of W02004/067737, wherein said protease variant has a sequence identity of at least 75% but less than 100% to SEQ ID NO: 1 of WO 2004/067737.
Suitable commercially available protease enzymes include those sold under the trade names Alcalase®, DuralaseTm, DurazymTm, Relase®, Relase® Ultra, Savinase®, Savinase® Ultra, Primase®, Polarzyme®, Kannase®, Liquanase®, Liquanase® Ultra, Ovozyme®, Coronase®, Coronase® Ultra, Blaze®, Blaze Evity® 100T, Blaze Evity® 125T, Blaze Evity® 150T, Everlase® and Esperase® (Novozymes A/S), those sold under the tradename Maxatase®, Maxacal®, Maxapem®, Purafect Ox®, Purafect OxP®, Puramax®, FN2®, FN3®, FN4®, Excellase®, Excellenz P1000™, Excellenz P1250™, Eraser®, Preferenz P100™, Purafect Prime®, Preferenz P1 10™, Effectenz P1000™, Purafect®™, Effectenz P1050™, Purafect Ox®™, Effectenz P2000™, Purafast®, Properase®, Opticlean® and Optimase® (Danisco/DuPont), Axapem™ (Gist-Brocases N.V.), BLAP (sequence shown in Figure 29 of US5352604) and variants hereof (Henkel AG) and KAP ( Bacillus alkalophilus subtilisin) from Kao.
Amylases:
Suitable amylases include alpha-amylases and/or glucoamylases and may be of bacterial or fungal origin. Chemically modified or protein engineered mutants are included. Amylases include, for example, alpha-amylases obtained from Bacillus, e.g., a special strain of Bacillus licheniformis, described in more detail in GB 1 ,296,839.
Suitable amylases include amylases having SEQ ID NO: 2 in WO 95/10603 or variants having 90% sequence identity to SEQ ID NO: 3 thereof. Preferred variants are described in WO
94/02597, WO 94/18314, WO 97/43424 and SEQ ID NO: 4 of WO 99/019467, such as variants with substitutions in one or more of the following positions: 15, 23, 105, 106, 124, 128, 133, 154, 156, 178, 179, 181 , 188, 190, 197, 201 , 202, 207, 208, 209, 211 , 243, 264, 304, 305, 391 , 408, and 444.
Different suitable amylases include amylases having SEQ ID NO: 6 in WO 02/010355 or variants thereof having 90% sequence identity to SEQ ID NO: 6. Preferred variants of SEQ ID NO: 6 are those having a deletion in positions 181 and 182 and a substitution in position 193.
Other amylases which are suitable are hybrid alpha-amylase comprising residues 1-33 of the alpha-amylase derived from B. amyloliquefaciens shown in SEQ ID NO: 6 of WO 2006/066594 and residues 36-483 of the B. licheniformis alpha-amylase shown in SEQ ID NO: 4 of WO 2006/066594 or variants having 90% sequence identity thereof. Preferred variants of this hybrid alpha-amylase are those having a substitution, a deletion or an insertion in one of more of the following positions: G48, T49, G107, H156, A181 , N 190, M197, 1201 , A209 and Q264. Most preferred variants of the hybrid alpha-amylase comprising residues 1-33 of the alpha-amylase derived from B. amyloliquefaciens shown in SEQ ID NO: 6 of WO 2006/066594 and residues 36- 483 of SEQ ID NO: 4 are those having the substitutions:
M197T;
H 156Y+A 181 T+ N 190F+A209V+Q264S; or
G48A+T49I+G107A+ H 156Y+A 181 T+ N 190F+ 1201 F+A209V+Q264S.
Further amylases which are suitable are amylases having SEQ ID NO: 6 in WO 99/019467 or variants thereof having 90% sequence identity to SEQ ID NO: 6. Preferred variants of SEQ ID NO: 6 are those having a substitution, a deletion or an insertion in one or more of the following positions: R181 , G182, H183, G184, N195, I206, E212, E216 and K269. Particularly preferred amylases are those having deletion in positions R181 and G182, or positions H183 and G184.
Additional amylases which can be used are those having SEQ ID NO: 1 , SEQ ID NO: 3, SEQ ID NO: 2 or SEQ ID NO: 7 of WO 96/023873 or variants thereof having 90% sequence identity to SEQ ID NO: 1 , SEQ ID NO: 2, SEQ ID NO: 3 or SEQ ID NO: 7. Preferred variants of SEQ ID NO: 1 , SEQ ID NO: 2, SEQ ID NO: 3 or SEQ ID NO: 7 are those having a substitution, a deletion or an insertion in one or more of the following positions: 140, 181 , 182, 183, 184, 195, 206, 212, 243, 260, 269, 304 and 476, using SEQ ID 2 of WO 96/023873 for numbering. More preferred variants are those having a deletion in two positions selected from 181 , 182, 183 and 184, such as 181 and 182, 182 and 183, or positions 183 and 184. Most preferred amylase variants of SEQ ID NO: 1 , SEQ ID NO: 2 or SEQ ID NO: 7 are those having a deletion in positions 183 and 184 and a substitution in one or more of positions 140, 195, 206, 243, 260, 304 and 476.
Other amylases which can be used are amylases having SEQ ID NO: 2 of WO 08/153815, SEQ ID NO: 10 in WO 01/66712 or variants thereof having 90% sequence identity to SEQ ID NO: 2 of WO 08/153815 or 90% sequence identity to SEQ ID NO: 10 in WO 01/66712. Preferred variants of SEQ ID NO: 10 in WO 01/66712 are those having a substitution, a deletion or an insertion in one of more of the following positions: 176, 177, 178, 179, 190, 201 , 207, 21 1 and 264.
Further suitable amylases are amylases having SEQ ID NO: 2 of WO 09/061380 or variants having 90% sequence identity to SEQ ID NO: 2 thereof. Preferred variants of SEQ ID NO: 2 are those having a truncation of the C-terminus and/or a substitution, a deletion or an insertion in one of more of the following positions: Q87, Q98, S125, N128, T131 , T165, K178, R180, S181 , T182, G183, M201 , F202, N225, S243, N272, N282, Y305, R309, D319, Q320, Q359, K444 and G475. More preferred variants of SEQ ID NO: 2 are those having the substitution in one of more of the following positions: Q87E,R, Q98R, S125A, N128C, T131 I, T165I, K178L, T182G, M201 L, F202Y, N225E.R, N272E.R, S243Q,A,E,D, Y305R, R309A, Q320R, Q359E, K444E and G475K and/or deletion in position R180 and/or S181 or of T182 and/or G183. Most preferred amylase variants of SEQ ID NO: 2 are those having the substitutions:
N 128C+K178L+T182G+Y305R+G475K;
N128C+K178L+T182G+F202Y+Y305R+D319T+G475K;
S125A+N 128C+K178L+T182G+Y305R+G475K; or
S125A+N 128C+T131 I+T165I+K178L+T182G+Y305R+G475K wherein the variants are C-terminally truncated and optionally further comprises a substitution at position 243 and/or a deletion at position 180 and/or position 181.
Further suitable amylases are amylases having SEQ ID NO: 1 of W013184577 or variants having 90% sequence identity to SEQ ID NO: 1 thereof. Preferred variants of SEQ ID NO: 1 are those having a substitution, a deletion or an insertion in one of more of the following positions: K176, R178, G179, T180, G181 , E187, N 192, M199, I203, S241 , R458, T459, D460, G476 and G477. More preferred variants of SEQ ID NO: 1 are those having the substitution in one of more of the following positions: K176L, E187P, N192FYH, M199L, I203YF, S241 QADN, R458N, T459S, D460T, G476K and G477K and/or deletion in position R178 and/or S179 or of T180 and/or G181. Most preferred amylase variants of SEQ ID NO: 1 are those having the substitutions:
E187P+I203Y+G476K
E187P+I203Y+R458N+T459S+D460T+G476K
wherein the variants optionally further comprise a substitution at position 241 and/or a deletion at position 178 and/or position 179.
Further suitable amylases are amylases having SEQ ID NO: 1 of W010104675 or variants having 90% sequence identity to SEQ ID NO: 1 thereof. Preferred variants of SEQ ID NO: 1 are
those having a substitution, a deletion or an insertion in one of more of the following positions: N21 , D97, V128 K177, R179, S180, 1181 , G182, M200, L204, E242, G477 and G478. More preferred variants of SEQ ID NO: 1 are those having the substitution in one of more of the following positions: N21 D, D97N, V128I K177L, M200L, L204YF, E242QA, G477K and G478K and/or deletion in position R179 and/or S180 or of 1181 and/or G182. Most preferred amylase variants of SEQ ID NO: 1 are those having the substitutions:
N21 D+D97N+V128I
wherein the variants optionally further comprise a substitution at position 200 and/or a deletion at position 180 and/or position 181.
Other suitable amylases are the alpha-amylase having SEQ ID NO: 12 in WO01/66712 or a variant having at least 90% sequence identity to SEQ ID NO: 12. Preferred amylase variants are those having a substitution, a deletion or an insertion in one of more of the following positions of SEQ ID NO: 12 in WO01/66712: R28, R1 18, N174; R181 , G182, D183, G184, G186, W189, N195, M202, Y298, N299, K302, S303, N306, R310, N314; R320, H324, E345, Y396, R400, W439, R444, N445, K446, Q449, R458, N471 , N484. Particular preferred amylases include variants having a deletion of D183 and G184 and having the substitutions R1 18K, N195F, R320K and R458K, and a variant additionally having substitutions in one or more position selected from the group: M9, G149, G182, G186, M202, T257, Y295, N299, M323, E345 and A339, most preferred a variant that additionally has substitutions in all these positions.
Other examples are amylase variants such as those described in WO2011/098531 , WO2013/001078 and WO2013/001087.
One embodiment of the invention, the detergent composition in the second compartment of the pouch comprises an amylase selected from the group consisting of:
a) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO: 37, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO: 37 comprising a two amino acid deletion in the sequence region R180, S181 , T182, G183, compared to SEQ ID NO: 37, wherein each position corresponds to the position in SEQ ID NO: 37; b) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO: 37, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO: 37 comprising one or preferably all the alterations sets selected from the group consisting of:
a. R180*, S181 *, S243Q, G475K;
b. R180*, T182*, S243Q, G475K;
c. R180*, T182*, G183S, S243Q, G475K; and
d. R180*, S181*, Y242F, S243Q, F266Y, G475K compared to SEQ ID NO: 37, wherein each position corresponds to the position in SEQ ID NO: 37; c) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO: 38, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO: 38, comprising a two amino acid deletion in the sequence region R178, G179, T180, G181 compared to SEQ ID NO:
38, wherein each position corresponds to the position in SEQ ID NO: 38;
d) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO: 38, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO: 38, comprising one or more, preferably all of the alterations sets selected from the group consisting of:
I. R178*, G179*, E187P, I203Y, G476K;
II. R178*, G179*, E187P, M199L, I203Y, G476K;
III. R178*, G179*, E187P, I203Y R458N, T459S, D460T, G476K;
IV. N126Y, F153W, R178*, G179*, T180H, I203Y, S241 Q;
V. N126Y, F153W, R178*, G179*, T180H, I203Y, S241 Q, S362A, R375Y;
VI. T38N, N126Y, T129I, F153W, R178*, G179*, T180D, E187P, I203Y, G476K, G477E; and
VII. N126Y, F153W, R178*, G179*, T180H, E187P, I203Y, S241Q, G476K, G477E, compared to SEQ ID NO: 34, wherein each position corresponds to the position in SEQ ID NO: 38;
e) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO: 39, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO: 39, comprising a two amino acid deletion in the sequence region R181 , G182, D183, G184 compared to SEQ ID NO:
39, wherein each position corresponds to the position in SEQ ID NO: 39;
f) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO: 39, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO: 39 comprising an alteration at one or more, preferably at all of the position(s) selected from 3, 4, 5, 1 18, 167, 170, 177, 195, 202, 204, 271 , 320, 330, 377, 385, 445, 458, 475, 476, 314, 315 or 316, compared to SEQ ID NO: 39, wherein each position corresponds to the position in SEQ ID NO: 39;
g) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO: 40, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO: 40 preferably comprising a two amino acid deletion in the sequence region R181 , G182, D183, G184, compared to SEQ ID NO: 40, wherein each position corresponds to the position in SEQ ID NO: 40; h) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO: 40, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO: 40 comprising one or more, preferably all of the alterations selected from the group consisting of
a. D183*, G184*, N195F, Y243F;
b. D183*, G184*, N 195F, V206Y, Y243F;
c. W140Y, D183*, G184*, N195F, V206Y, Y243F, E260G, G304R, G476K; d. W140Y, D183*, G184*, N195F, V206Y, Y243F, E260G, G477E;
e. W140Y, D183*, G184*, N195F, V206Y, Y243F, W284D;
f. W140Y + N195F + V206Y + Y243F + E260G + G477E;
g. G109A + W140Y + N195F + V206Y + Y243F + E260G;
h. T51 I S52Q N54K G109A W140Y N195F V206Y Y243F E260G G476E;
i. W140Y N195F V206Y Y243F E260G W284R G477K;
j. W140Y N195F V206Y Y243F E260G W284F G477R; and
k. H 1 *, G7A, G109A, W140Y, D183*, G184*, N195F, V206Y, Y243F, E260G, N280S, G304R, E391A, G476K, compared to SEQ ID NO: 40, wherein each position corresponds to the position in SEQ ID NO: 40;
i) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity
to SEQ ID NO: 41 , or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO: 41 , comprising a two amino acid deletion in the sequence region R181 , G182, D183, G184, compared to SEQ ID NO:
41 , wherein each position corresponds to the position in SEQ ID NO: 41 ;
j) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO: 41 , or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO: 41 , comprising one or more, preferably all of the alterations selected from the group consisting of
I. R1 18K, D183*, G184*, N195F, R320K, R458K;
II. M9I, D183*, G184*, R118K, N195F, M202L, R320K, M323T, R458K;
III. M9L, G149A, R118K, G182T, D183*, G184*, G186A, N195F, M202L, T257I, Y295F, N299Y, M323T, A339S, E345R, R458K;
IV. M9L, G149A, R1 18K, G182T, D183*, G184*, G186A, N195F, T246V, T257I, Y295F, N299Y, M323T, A339S, E345R, R458K; and
V. M9L, G149A, G182T, D183*, G184*, G186A, M202L, T257I, Y295F, N299Y, M323T, A339S, E345R, N471 E, compared to SEQ ID NO: 41 , wherein each position corresponds to the position in SEQ ID NO: 41 ; k) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO: 42, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO: 42, comprising a two amino acid deletion in the sequence region R181 , G182, G182, D183, compared to SEQ ID NO:
42, wherein each position corresponds to the position in SEQ ID NO: 42;
L) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO: 42, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO: 42, comprising one or more, preferably all of the alterations selected from the group consisting of
a. D183*, G184*, N195F, V206Y, R320K, R458K;
b. D183*, G184*, N 195F, M202L, V206L, R320K, R458K;
c. G149A, G182T, D183*, G184*, N195F, M202L, V206L, T257I, Y295F, Q299Y,
A339S, Q345R, R458K;
d. G149A, G182T, D183*, G184*, N195F, V206L, M246V, T257I, Y295F, Q299Y, A339S, Q345R, R458K;
e. G149A, G182T, D183*, G184*, M202L, V206L, T257I, Y295F, Q299Y, A339S, Q345R, H471 E; and
f. H1A, N54S, V56T, K72R, G109A, F113Q, R116Q, W167F, Q172G, A174S, G182*, D183*, G184T, N 195F, V206L, K391A, P473R and G476K, compared to SEQ ID NO: 42, wherein each position corresponds to the position in SEQ ID NO: 42;
m) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO: 43, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO: 43, comprising a two amino acid deletion in the sequence region R181 , G182, H183, G184, compared to SEQ ID NO: 43, wherein each position corresponds to the position in SEQ ID NO: 43;
n) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO: 43, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO: 43, comprising one or more, preferably all of the alterations selected from the group consisting of
a. H183*, G184*, I405L, A421 H, A422P, A428T;
b. R1 18K, H183*, G184*, N195F, R320K, R458K;
c. M9I, H183*, G184*, R118K, N195F, M202L, R320K, S323T, R458K;
d. M9L, G149A, R118K, G182T, H183*, G184*, N195F, M202L, T257I, Y295F, N299Y, A339S, E345R, R458K;
e. M9L, G149A, R1 18K, G182T, H183*, G184*, N195F, T246V, T257I, Y295F, N299Y, A339S, E345R, R458K; and
f. M9L, G149A, G182T, H183*, G184*, M202L, T257I, Y295F, N299Y, S323T, A339S, E345R, N471 E, compared to SEQ ID NO: 43, wherein each position corresponds to the position in SEQ ID NO: 43;
o) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO: 44, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but
less than 100% sequence identity to SEQ ID NO: 44, comprising a two amino acid deletion in the sequence region R181 , G182, G182, D183, compared to SEQ ID NO: 44, wherein each position corresponds to the position in SEQ ID NO: 44;
p) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO: 44, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO: 44, comprising one or more, preferably all of the alterations selected from the group consisting of
a. H 1 *, D183*, G184*, N195F, V206Y;
b. H 1 *, D183*, G184*, N195F, M202L, V206L, R320K, R458K;
c. G149A, G182T, D183*, G184*, N195F, M202L, V206L, T257I, Y295F, Q299Y, A339S, Q345R, R458K;
d. G149A, G182T, D183*, G184*, N195F, V206L, M246V, T257I, Y295F, Q299Y, A339S, Q345R, R458K;
e. G149A, G182T, D183*, G184*, M202L, V206L, T257I, Y295F, Q299Y, A339S, Q345R, H471 E;
f. H 1 *, N54S, V56T, G109A, Q169E, Q172K, A174*, G182*, D183*, N195F, V206L, K391A, G476K;
g. G182*, D183*, N195F, W140Y, N260G, G304R, R320A, G476K, V410I, V429I, F451W, C474V;
h. H 1 *, N54S, V56T, K72R, G109A, F1 13Q, R116Q, W167F, Q172G, A174S, G182*, D183*, G184T, N195F, V206L, K391A, P473R, G476K;
i. H 1 *, N54S, V56T, G109A, Q169E, Q172K, A174*, G182*, D183*, N195F, V206L, K391A, G476K;
j. H 1 *, N54S, V56T, G109A, R116H, A174S, G182*, D183*, N195F, V206L, K391A, G476K;
k. H 1 *, N54S, V56T, K72R, G109A, F1 13Q, R116Q, W167F, Q172G, A174S, G182*, D183*, G184T, N195F, V206L, K391A, P473R, G476K;
L. H 1 *, N54S, V56T, G109A, F1 13Q, R1 16Q, Q172N, A174S, G182*, D183*, N 195F, V206L, A265G, K391A, P473R, G476K;
m. H 1 *, N54S, V56T, K72R, G109A, F1 13Q, W167F, Q172R, A174S, G182*, D183*, N195F, V206L, K391A, G476K;
n. H 1 *, N54S, V56T, K72R, G109A, R116H, T134E, W167F, Q172G, L173V, A174S, G182*, D183*, N 195F, V206L, G255A, K391A, G476K;
o. H 1 *, N54S, V56T, K72R, G109A, R116H, T134E, W167F, Q172G, L173V,
A174S, G182*, D183*, N195F, V206L, G255A, K391A, Q395P, T444Q, P473R, G476K;
p. H 1 *, N54S, V56T, G109A, T134E, A174S, G182*, D183*, N195F, V206L, K391A, G476K;
q. H 1 *, N54S, V56T, K72R, G109A, A174S, G182*, D183*, N195F, V206L, G255A, K391A, G476K;
r. H 1 *, N54S, V56T, G109A, W167F, Q172E, L173P, A174K, G182*, D183*, N195F, V206L, K391A, G476K;
s. H 1 *, N54S, V56T, G109A, R116Q, V120L, Q172G, 1173V, A174S, G182*, D183*, G184T, N195F, V206L, I405L, A421 H, A422P, A428T; and
t. H 1 *, N54S, V56T, G109A, F113Q, R1 16Q, W167F, Q172G, 1173V, A174S, G182*, D183*, G184T, N195F, V206L, I405L, A421 H, A422P, A428T, compared to SEQ ID NO: 44, wherein each position corresponds to the position in SEQ ID NO: 44.
Commercially available amylases are Duramyl™, Termamyl™, Fungamyl™, Stainzyme ™, Stainzyme Plus™, Natalase™, Liquozyme X and BAN™ (from Novozymes A/S), and Rapidase™, Purastar™/Effectenz™, Powerase, Preferenz S1000, Preferenz S100 and Preferenz S110 (from Genencor International Inc./DuPont).
Peroxidases/Oxidases
Suitable peroxidases/oxidases include those of plant, bacterial or fungal origin. Chemically modified or protein engineered mutants are included. Examples of useful peroxidases include peroxidases from Coprinopsis, e.g., from C. cinerea (EP 179,486), and variants thereof as those described in WO 93/24618, WO 95/10602, and WO 98/15257. A suitable peroxidase includes a haloperoxidase enzyme, such as chloroperoxidase, bromoperoxidase and compounds exhibiting chloroperoxidase or bromoperoxidase activity. Haloperoxidases are classified according to their specificity for halide ions. Chloroperoxidases (E.C. 1.1 1.1.10) catalyze formation of hypochlorite from chloride ions. Preferably, the haloperoxidase is a vanadium haloperoxidase, i.e. , a vanadate-containing haloperoxidase. Haloperoxidases have been isolated from many different fungi, in particular from the fungus group dematiaceous hyphomycetes, such as Caldariomyces, e.g., C. fumago, Alternaria, Curvularia, e.g., C. verruculosa and C. inaequalis, Drechslera, Ulocladium and Botrytis.
Haloperoxidases have also been isolated from bacteria such as Pseudomonas, e.g., P. pyrrocinia and Streptomyces, e.g., S. aureofaciens.
A suitable oxidase includes in particular, any laccase enzyme comprised by the enzyme classification EC 1.10.3.2, or any fragment derived therefrom exhibiting laccase activity, or a
compound exhibiting a similar activity, such as a catechol oxidase (EC 1.10.3.1), an o- aminophenol oxidase (EC 1.10.3.4), or a bilirubin oxidase (EC 1.3.3.5). Preferred laccase enzymes are enzymes of microbial origin. The enzymes may be derived from plants, bacteria or fungi (including filamentous fungi and yeasts). Suitable examples from fungi include a laccase derivable from a strain of Aspergillus, Neurospora, e.g., N. crassa, Podospora, Botrytis, Collybia, Pomes, Lentinus, Pleurotus, Trametes, e.g., T. villosa and T. versicolor, Rhizoctonia, e.g., R. solani, Coprinopsis, e.g., C. cinerea, C. comatus, C. friesii, and C. plicatilis, Psathyrella, e.g., P. condelleana, Panaeolus, e.g., P. papilionaceus, Myceliophthora, e.g., M. thermophila, Schytalidium, e.g., S. thermophilum, Polyporus, e.g., P. pinsitus, Phlebia, e.g., P. radiata (WO 92/01046), or Coriolus, e.g., C. hirsutus (JP 2238885). Suitable examples from bacteria include a laccase derivable from a strain of Bacillus. A laccase derived from Coprinopsis or Myceliophthora is preferred; in particular, a laccase derived from Coprinopsis cinerea, as disclosed in WO 97/08325; or from Myceliophthora thermophila, as disclosed in WO 95/33836.
Nucleases
Suitable nucleases include deoxyribonucleases (DNases) and ribonucleases (RNases) which are any enzyme that catalyzes the hydrolytic cleavage of phosphodiester linkages in the DNA or RNA backbone respectively, thus degrading DNA and RNA. There are two primary classifications based on the locus of activity. Exonucleases digest nucleic acids from the ends. Endonucleases act on regions in the middle of target molecules.
In an embodiment, the detergent composition present in the second compartment of the multi-compartment pouch is a solid or liquid detergent composition.
In an embodiment, the detergent composition present in the second compartment of the multi-compartment pouch is selected from a group comprising: liquid detergents, gel detergents, powder detergents, granule detergents.
In one embodiment, the detergent composition present in the second compartment of the multi-compartment pouch is an anionic detergent composition, cationic detergent composition or non-ionic composition and zwitterionic detergent composition.
In some preferred embodiments, the detergent composition present in the second compartment of the multi-compartment pouch provided herein are typically formulated such that, during use in aqueous cleaning operations, the wash water has a pH of from about 5.0 to about 1 1.5, or in alternative embodiments, even from about 6.0 to about 10.5, such as from about 5 to about 11 , from about 5 to about 10, from about 5 to about 9, from about 5 to about 8, from about 5 to about 7, from about 6 to about 1 1 , from about 6 to about 10, from about 6 to about 9, from about 6 to about 8, from about 6 to about 7, from about 7 to about 11 , from about 7 to about 10, from about 7 to about 9, or from about 7 to about 8. In some preferred embodiments, granular or liquid laundry products are formulated such that the wash water has a pH from about 5.5 to about
1 1. Techniques for controlling pH at recommended usage levels include the use of buffers, alkalis, acids, etc., and are well known to those skilled in the art.
In one embodiment, the enzyme-containing composition present in the first compartment of the multi-compartment pouch and detergent composition present in the second compartment of the multi-compartment pouch may comprise low amount of water.
In another embodiment, composition present in the first compartment of the multi compartment pouch contains no water or only traces of water.
Enzyme components weights are based on total active protein. All percentages and ratios are calculated by weight unless otherwise indicated. All percentages and ratios are calculated based on the total composition unless otherwise indicated. In the exemplified detergent composition, the enzymes levels are expressed by pure enzyme by weight of the total composition and unless otherwise specified, the detergent ingredients are expressed by weight of the total composition.
The enzyme(s) of the invention may be stabilized using conventional stabilizing agents, e.g., a polyol such as propylene glycol or glycerol, a sugar or sugar alcohol, lactic acid, boric acid, or a boric acid derivative, e.g., an aromatic borate ester, or a phenyl boronic acid derivative such as 4-formylphenyl boronic acid, and the composition may be formulated as described in, for example, WO92/19709 and WO92/19708.
In certain markets different wash conditions and, as such, different types of detergents are used. This is disclosed in e.g. EP 1 025 240. For example, In Asia (Japan) a low detergent concentration system is used, while the United States uses a medium detergent concentration system, and Europe uses a high detergent concentration system.
Non-limiting examples of useful compositions in the multi-compartment pouch include light duty and heavy-duty liquid detergent compositions, hard surface cleaning compositions, fabric enhancers, detergent gels commonly used for laundry, and bleach and laundry additives, shampoos, body washes, and other personal care compositions. Compositions of use in the present pouches may take the form of a liquid, solid or a powder. Liquid compositions may comprise a solid. Solids may include powder or agglomerates, such as micro-capsules, beads, noodles or one or more pearlized balls or mixtures thereof. Such a solid element may provide a technical benefit, through the wash or as a pre-treat, delayed or sequential release component.
The choice of additional components is within the skill of the artisan and includes conventional ingredients, including the exemplary non-limiting components set forth below. The choice of components may include, for fabric care, the consideration of the type of fabric to be cleaned, the type and/or degree of soiling, the temperature at which cleaning is to take place, and the formulation of the detergent product.
Although components mentioned below are categorized by general header according to a
particular functionality, this is not to be construed as a limitation, as a component may comprise additional functionalities as will be appreciated by the skilled artisan.
Method
The present invention also relates to a method of releasing enzyme in a cleaning process.
In one embodiment, the present invention relates to a method of releasing enzymes in a cleaning process comprising steps of: a. providing a multicompartment pouch with at least two compartments the first comprising at least one enzyme containing composition and the second at least one detergent composition, b. releasing of the detergent composition from the second compartment during the main-wash cycle and subsequently releasing the enzyme from the first compartment into the wash liquor.
In one embodiment of the present invention, the multicompartment pouch with at least two compartments the first comprising an enzyme-containing composition and the second compartment comprising a detergent composition.
In one embodiment of the present invention, the enzyme-containing composition present in the first compartment of the multi-compartment pouch comprises a protease.
In one embodiment of the present invention, the enzyme-containing composition present in the first compartment of the multi-compartment pouch comprises a metal loprotease.
In one embodiment of the present invention, the enzyme-containing composition present in the first compartment of the multi-compartment pouch comprises a metalloprotease selected from a group consisitng of EC classes EC 3.4.22 or EC3.4.24.
In one embodiment of the present invention, the enzyme-containing composition present in the first compartment of the multi-compartment pouch comprising the metalloprotease is selected from a group consisting of a. a thermolysin and/or variant thereof; b. a metalloprotease from M4 family, that is not a thermolysin and variant thereof; c. a metalloprotease from the M7 family; and d. a metalloprotease from the M35 metalloprotease.
In an embodiment of the present invention, the enzyme-containing composition present in the first compartment of the multi-compartment pouch comprises of metalloprotease having amino acid sequence having at least 60% at least 65%, e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, but less than 100% sequence identity to a polypeptide selected from the group consisting of SEQ ID
Nos: 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35 and 36.
In an embodiment of the present invention, the enzyme-containing composition present in the first compartment of the multi-compartment pouch comprises of at least one metalloprotease having amino acid sequence having at least 60% at least 65%, e.g., at least 70%, at least 75%,
at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, but less than 100% sequence identity to a polypeptide selected from the group consisting of SEQ
ID Nos: 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35 and 36.
In an embodiment of the present invention, the enzyme-containing composition present in the first compartment of the multi-compartment pouch comprises or consists of metalloprotease having amino acid sequence having at least 60% at least 65%, e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, but less than 100% sequence identity to a polypeptide selected from the group consisting of SEQ
ID Nos: 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26,
27, 28, 29, 30, 31 , 32, 33, 34, 35 and 36.
In one embodiment of the present invention, the enzyme-containing composition present in the first compartment of the multi-compartment pouch further comprises one or more additional enzymes.
In another embodiment of the present invention, the detergent composition is present in the second compartment of the multi-compartment pouch.
In an embodiment of the present invention, the detergent composition present in the second compartment of the multi-compartment pouch comprises at least one enzyme.
In an embodiment of the present invention, the detergent composition present in the second compartment of the multi-compartment pouch comprising the enzyme is preferably a DNase.
In an embodiment of the present invention, the detergent composition present in the second compartment of the multi-compartment pouch comprises at least one DNase.
In one embodiment of the present invention, the enzyme-containing composition present in a first compartment of a multi-compartment pouch comprises at least one metalloprotease and the detergent composition is present in a second compartment of a multi-compartment pouch comprises at least one DNase.
In one embodiment of the present invention, the enzyme-containing composition present in a first compartment of a multi-compartment pouch comprises at least one metalloprotease and further one or more additional enzymes and the detergent composition is present in a second compartment of a multi-compartment pouch comprises at least one DNase.
In one embodiment of the present invention, the enzyme-containing composition present in a first compartment of a multi-compartment pouch comprises at least one metalloprotease and the detergent composition is present in a second compartment of a multi-compartment pouch comprises at least one DNase and further one or more additional enzymes.
In one embodiment of the present invention, the enzyme-containing composition present in a first compartment of a multi-compartment pouch comprises at least one metalloprotease and further one or more additional enzymes and the detergent composition is present in a second compartment of a multi-compartment pouch comprises at least one DNase and further one or more additional enzymes.
In one embodiment of the present invention, the enzyme-containing composition present in a first compartment of a multi-compartment pouch comprises at least one metalloprotease and the detergent composition present in a second compartment of a multi-compartment pouch comprises at least one DNase, wherein the metalloprotease is classified in EC 3.4.22 or EC3.4.24.
In one embodiment of the present invention, the enzyme-containing composition present in a first compartment of a multi-compartment pouch comprises at least one metalloprotease and the detergent composition present in a second compartment of a multi-compartment pouch comprises at least one DNase and wherein the metalloprotease is an M4, M7 or M35.
In one embodiment of the present invention, the enzyme-containing composition is present in a first compartment of a multi-compartment pouch comprises at least one metalloprotease and the detergent composition present in a second compartment of a multi compartment pouch comprises at least one DNase, wherein the DNase is obtained from Bacillus.
In one embodiment of the present invention, the enzyme-containing composition is present in a first compartment of a multi-compartment pouch comprises at least one metalloprotease and the detergent composition present in a second compartment of a multi compartment pouch comprises at least one DNase, wherein the DNase is obtained from Aspergillus.
In one embodiment of the present invention, the enzyme-containing composition is present in a first compartment of a multi-compartment pouch comprises at least one metalloprotease and the detergent composition present in a second compartment of a multi compartment pouch comprises at least one DNase, wherein the DNase is obtained from Trichoderma.
In one embodiment of the present invention, the enzyme-containing composition is present in the first compartment of the multi-compartment pouch comprises a metalloprotease having at least 60 percent, at least 65 percent, at least 70 percent, at least 75 percent, at least 80 percent, at least 85 percent, at least 90 percent, at least 95 percent, at least 96 percent, at least 97 percent, at least 98 percent, at least 99 percent but less than 100 sequence percent identity to SEQ ID NOs: 1-36 and the detergent composition is present in the second compartment of the multi-compartment pouch comprises a DNase having at least 60%, at least 65%, at least 70%, at
least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NOs: 60-64.
In one embodiment of the present invention, the multi-compartment pouch is suitable for delivering different compositions at different points in time of the wash-cycle of a laundry or automatic dishwashing machine. Difference in solubility can be achieved by means of films of different thickness or films which solubility is temperature dependent.
In one embodiment of the present invention, the detergent composition present in the second compartment of the multi-compartment pouch is first released into the water containing in the washing machine or in the dishwashing machine to form a washing liquor/a dishwashing liquor comprising water and the laundry to be washed and cleaned or the dishes to be cleaned to said washing liquor or dishwashing liquor.
In another embodiment of the present invention, the enzyme-containing composition in the first compartment of the multi-compartment pouch is subsequently released i.e after the release of the detergent composition in the second compartment of the multi-compartment pouch into washing liquor contained in the washing machine together with the laundry to be washed and cleaned or into the dishwashing liquor contained in the dishwashing machine together with the dishes to be cleaned.
In one embodiment of the present invention, the films enclosing the enzyme-containing composition dissolves after to the films enclosing the detergent composition in the multi compartment pouch during the main-wash cycle of a washing machine, thereby releasing the enzyme-containing composition into the wash liquor after the delivery of the detergent composition. This gives the enzymes the possibility to operate under optimum condition, avoiding interactions with highly concentrated detergent activities. The pouch provides excellent cleaning.
In one embodiment of the present invention, the detergent composition in the multi compartment pouch will react in water to release its contents before the enzyme-containing composition in the multi-compartment pouch. To further enhance this simultaneous or sequential release, the detergent composition may be more water-soluble than the enzyme-containing composition. This can for example be achieved by using different type of material for the detergent compartment and enzyme compartment of the pouch, for example, the detergent compartment is made of a material having a different type of polymer, different plasticizer, different levels components in the material, different coating of the film material, different thickness of the film material.
The multi-compartment pouch according to the invention can be used advantageously for washing and cleaning of textiles or for automatic dishwashing and preferably for removing enzyme-sensitive and bleachable soiling’s on textiles.
In one embodiment of the present invention, the enzyme-containing composition present in the first compartment of the multi-compartment pouch is at a concentration of 0.01 to 100 ppm, preferably 0.05 to 80 ppm, most preferably 0.05 to 20 ppm.
The present invention relates to a method of cleaning a fabric, a dishware or hard surface with a multi-compartment pouch comprising enzyme and detergent composition.
A preferred embodiment concerns a method of cleaning, said method comprising the steps of: contacting a fabric or object with a composition comprising a metalloprotease under conditions suitable for cleaning said fabric or object.
In a preferred embodiment relates to a method for removing stains from fabric which comprises contacting said a fabric with a composition comprising a metalloprotease under conditions suitable for cleaning said object.
The detergent pouch of the present invention is ideally suited for use in laundry applications. Accordingly, the present invention includes a method for laundering a fabric. The method comprises the steps of contacting a fabric to be laundered with the detergent pouch according to the invention.
The method of the present invention comprises a step of contacting the fabric with an aqueous wash liquor. The aqueous wash liquor is formed by the addition of a detergent composition to water. The detergent composition added to water to form the aqueous wash liquor. The fabric may be contacted with the composition in a hand washing step or even a wash liquor in a machine wash cycle.
In the method of the present invention, in step (b) the enzyme is delivered during the main wash cycle wherein the fabric may be washed in a conventional wash step in which an aqueous wash liquor is formed by the addition of a detergent composition to water.
The wash cycle settings for automatic washing machines generally have somewhat similar parameters of wash time and wash temperature. For instance, "Regular" wash cycle settings in the United States are generally between about 12 minutes to about 18 minutes at about 33 C to about 43 C ("Warm" setting), while those in European washing machines generally are between about 100 minutes to about 140 minutes ("Normal" setting) at about 40°C to about 60 C. Wash settings in Asia generally are about 15 minutes at about 25 C.
In one embodiment of the present invention, the enzyme-containing composition in first compartment of the multi-compartment pouch is released at least 30 seconds later than the detergent composition present in the second compartment releases of the multi-compartment pouch.
In one embodiment of the present invention, the method of cleaning comprising contacting a surface and/or a fabric with the multi-compartment pouch comprising at least two compartments, wherein said first compartment comprises an enzyme-containing composition and a second
compartment comprises a detergent composition.
In one embodiment of the present invention, the method for removing and/or reducing soil and/or for reducing redeposition on a surface and/or textile comprising contacting the surface and/or textile with multi-compartment pouch comprising at least two compartments, wherein said first compartment comprises an enzyme-containing composition and a second compartment comprises a detergent composition.
In one embodiment of the present invention, the method of dishwashing in an automatic dishwashing machine comprising the steps of placing the multi-compartment pouch comprising at least two compartments, wherein said first compartment comprises an enzyme-containing composition and a second compartment comprises a detergent composition into the product dispenser and releasing it during the wash cycle.
In one embodiment, of the present invention the method of treating a fabric and/or textile comprising contacting the fabric and/or textile with the multi-compartment pouch comprising at least two compartments, wherein said first compartment comprises an enzyme-containing composition and a second compartment comprises a detergent composition.
In one embodiment of the present invention relates to a method of treating a substrate, where the method includes the step of contacting the substrate with the multi-compartment pouch comprising at least two compartments, wherein said first compartment comprises an enzyme- containing composition and a second compartment comprises a detergent composition in the presence of water, wherein the substrate is a fabric or a hard surface.
In one embodiment of the present invention relates to a method of cleaning comprising contacting a surface and/or a fabric with the multi-compartment pouch comprising at least two compartments, wherein said first compartment comprises an enzyme-containing composition and a second compartment comprises a detergent composition.
In one embodiment of the present invention relates to a method for removing and/or reducing soil and/or for reducing redeposition on a surface and/or textile comprising contacting the surface and/or textile with the pouch comprising at least two compartments, wherein said first compartment comprises an enzyme-containing composition and a second compartment comprises a detergent composition.
In one embodiment of the present invention relates to a method of laundering or dishwashing in a washing machine comprising the steps of placing the multi-compartment pouch comprising at least two compartments, wherein said first compartment comprises an enzyme- containing composition and a second compartment comprises a detergent composition into the product dispenser and releasing it during the wash cycle.
Use
The present invention relates to a use of a multi-compartment pouch in a cleaning process.
The soils and stains that are important for detergent formulators are composed of many different substances, and a range of different enzymes, all with different substrate specificities have been developed for use in detergents both in relation to laundry and hard surface cleaning. These enzymes are considered to provide an enzyme detergency benefit, since they specifically improve stain removal in the cleaning process they are applied in as compared to the same process without enzymes.
In one embodiment of the present invention, use of a multi-compartment pouch in a cleaning process, such as laundry, hard surface cleaning, dish wash or automated dish wash.
In one embodiment of the present invention, use of a multi-compartment pouch comprising at least two compartments, wherein said first compartment comprises an enzyme-containing composition and a second compartment comprises a detergent composition for cleaning of an item, wherein the item is a textile or a surface.
In one embodiment of the present invention, use of a multi-compartment pouch comprising at least two compartments, wherein said first compartment comprises an enzyme-containing composition and a second compartment comprises a detergent composition for removing or releasing a stain from a textile having a stain.
In one aspect, the present invention concerns the use of metalloproteases in detergent compositions and cleaning processes, such as laundry and hard surface cleaning. Thus, in one aspect, the present invention demonstrates the detergency effect of a variety of exemplary metalloproteases on various stains and under various conditions.
The cleaning process or the textile care process may for example be a laundry process, a dishwashing process or cleaning of hard surfaces such as bathroom tiles, floors, table tops, drains, sinks and washbasins. Laundry processes can for example be household laundering, but it may also be industrial laundering. Furthermore, the invention relates to a process for laundering of fabrics and/or garments where the process comprises treating fabrics with a washing solution containing a detergent composition, and at least one metalloprotease. The cleaning process or a textile care process can for example be carried out in a machine-washing process or in a manual washing process. The washing solution can for example be an aqueous washing solution containing a detergent composition.
The fabrics and/or garments subjected to a washing, cleaning or textile care process of the present invention may be conventional washable laundry, for example household laundry. Preferably, the major part of the laundry is garments and fabrics, including knits, woven, denims, non-woven, felts, yarns, and towelling. The fabrics may be cellulose based such as natural cellulosics, including cotton, flax, linen, jute, ramie, sisal or coir or manmade cellulosics (e.g., originating from wood pulp) including viscose/rayon, ramie, cellulose acetate fibers (tricell), lyocell or blends thereof. The fabrics may also be non-cellulose based such as natural polyamides
including wool, camel, cashmere, mohair, rabbit and silk or synthetic polymer such as nylon, aramid, polyester, acrylic, polypropylene and spandex/elastane, or blends thereof as well as blend of cellulose based and non-cellulose based fibers. Examples of blends are blends of cotton and/or rayon/viscose with one or more companion material such as wool, synthetic fibers (e.g., polyamide fibers, acrylic fibers, polyester fibers, polyvinyl alcohol fibers, polyvinyl chloride fibers, polyurethane fibers, polyurea fibers, aramid fibers), and cellulose-containing fibers (e.g., rayon/viscose, ramie, flax, linen, jute, cellulose acetate fibers, lyocell).
The invention further concerns the use of multicompartment pouch in a proteinaceous stain removing processes. The proteinaceous stains may be stains such as food stains, e.g., baby food, sebum, cocoa, egg, blood, milk, ink, grass, or a combination thereof.
The invention is further described in the following paragraphs:
1. A multi-compartment pouch comprising at least two compartments, wherein said first compartment comprises an enzyme-containing composition and a second compartment comprises a detergent composition.
2. The pouch according to paragraph 1 , wherein the enzyme-containing composition comprises at least one enzyme.
3. The pouch according to paragraphs 1-2, wherein the enzyme is a protease.
4. The pouch according to any of the preceding paragraphs, wherein the protease is a metalloprotease, preferably selected from a group consisting of:
a. a thermolysin variant;
b. a metalloprotease from M4 family, that is not a thermolysin variant;
c. a metalloprotease from the M7 family; and
d. a metalloprotease from the M35.
5. The pouch according to any of the preceding paragraphs wherein the metalloprotease comprises of amino acid sequence having at least 60%, at least 65%, e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, but less than 100% sequence identity to a polypeptide selected from the group consisting of SEQ ID Nos: 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35 and 36.
6. The pouch according to any of the preceding paragraphs wherein the metalloprotease comprises or consists of amino acid sequence as shown in SEQ ID Nos: 1 , 2, 3, 4, 5, 6,
7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 ,
32, 33, 34, 35 and 36.
7. The pouch according to any of the preceding paragraphs, wherein the enzyme-containing composition further comprises one or more additional enzymes.
8. The pouch according to any of the preceding paragraphs, wherein the enzyme is present in liquid, solid, powder, or gel form.
9. The pouch according to any of the preceding paragraphs, wherein the second compartment comprising the detergent composition comprises at least one enzyme.
10. The pouch according to any of the preceding paragraphs wherein the enzyme is preferably a DNase having at least 60 %, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the amino acid sequence shown in SEQ ID NO: 60 or SEQ ID NO:61 or SEQ ID NO:62 or SEQ ID NO:63 or SEQ ID NO:64.
1 1. The pouch according to any of the preceding paragraphs, wherein the detergent composition further comprises one or more detergent components which is selected from the group consisting of surfactants, builders, flocculating aid, chelating agents, dye transfer inhibitors, enzyme stabilizers, enzyme inhibitors, catalytic materials, bleach activators, hydrogen peroxide, sources of hydrogen peroxide, preformed peracids, polymeric dispersing agents, clay soil removal/anti-redeposition agents, brighteners, suds suppressors, dyes, perfumes, structure elasticizing agents, fabric softeners, carriers, hydrotropes, builders and co-builders, fabric hueing agents, anti-foaming agents, dispersants, processing aids, pigments and mixtures thereof.
12. The pouch according to any of the preceding paragraphs, wherein the builder is selected from a group consisting of phosphates, sodium citrate builders, sodium carbonate, sodium silicate, sodium and zeolites.
13. The pouch according to any of the preceding paragraphs, wherein the surfactant is anionic and/or nonionic.
14. The pouch according to any of the preceding paragraphs, wherein the anionic surfactant is selected from linear alkylbenzenesulfonates (LAS) isomers of LAS, alcohol ether sulfate (AEO, AEOS) and sodium lauryl ether sulfate and sodium laureth sulfate (SLES).
15. The pouch according to any of the preceding paragraphs, wherein the nonionic surfactant is selected from alcohol ethoxylates (AE or AEO), alcohol propoxylates, alcohol propoxylates, propoxylated fatty alcohols (PFA), alkoxylated fatty acid alkyl esters, such as ethoxylated and/or propoxylated fatty acid alkyl esters, alkylphenol ethoxylates (APE), nonylphenol ethoxylates (NPE), alkylpolyglycosides (APG), alkoxylated amines, fatty acid monoethanolamides (FAM), fatty acid diethanolamides (FADA), ethoxylated fatty acid monoethanolamides (EFAM), propoxylated fatty acid monoethanolamides (PFAM), polyhydroxyalkyl fatty acid amides, /V-acyl /V-alkyl derivatives of glucosamine (glucamides, GA, or fatty acid glucamides, FAGA) and combinations thereof.
16. The pouch according to any of the preceding paragraphs, wherein the detergent composition further comprises one or more additional enzymes.
17. The pouch according to any of the preceding paragraphs, wherein the enzyme is selected from the group consisting of an alpha-amylase, a beta-amylase, a pullulanase, a lipase, a cellulase, an oxidase, another protease, a phospholipase, a perhydrolase, a xylanase, a pectate lyase, a pectinase, a galacturanase, a hemicellulase, a xyloglucanase, a nuclease, a mannanase and mixtures thereof.
18. The pouch according to any of the preceding paragraphs wherein the enzyme is an amylase selected from
a. an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO: 37, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO: 37 comprising a two amino acid deletion in the sequence region R180, S181 , T182, G183, compared to SEQ ID NO: 37, wherein each position corresponds to the position in SEQ ID NO: 37;
b. an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO: 37, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100%
sequence identity to SEQ ID NO: 37 comprising one or preferably all the alterations sets selected from the group consisting of:
1) R180*, S181 *, S243Q, G475K;
2) R180*, T182*, S243Q, G475K;
3) R180*, T182*, G183S, S243Q, G475K; and
4) R180*, S181*, Y242F, S243Q, F266Y, G475K compared to SEQ ID NO: 37, wherein each position corresponds to the position in SEQ ID NO: 37;
c. an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO: 38, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 38, comprising a two amino acid deletion in the sequence region R178, G179, T180, G181 compared to SEQ ID NO 38, wherein each position corresponds to the position in SEQ ID NO 38;
d. an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 38, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 38, comprising one or more, preferably all of the alterations sets selected from the group consisting of:
I. R178*, G179*, E187P, I203Y, G476K;
II. R178*, G179*, E187P, M199L, I203Y, G476K;
III. R178*, G179*, E187P, I203Y R458N, T459S, D460T, G476K;
IV. N126Y, F153W, R178*, G179*, T180H, I203Y, S241 Q;
V. N126Y, F153W, R178*, G179*, T180H, I203Y, S241 Q, S362A, R375Y;
VI. T38N, N126Y, T129I, F153W, R178*, G179*, T180D, E187P, I203Y, G476K, G477E; and
VII. N126Y, F153W, R178*, G179*, T180H, E187P, I203Y, S241Q, G476K, G477E, compared to SEQ ID NO 38, wherein each position corresponds to the position in SEQ ID NO 38;
e. an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 39, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 39, comprising a two amino acid deletion in the sequence region R181 , G182, D183, G184 compared to SEQ ID NO 39, wherein each
position corresponds to the position in SEQ ID NO 39;
f. an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 39, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 39 comprising an alteration at one or more, preferably at all of the position(s) selected from 3, 4, 5, 118, 167, 170, 177, 195, 202, 204, 271 , 320, 330, 377, 385, 445, 458, 475, 476, 314, 315 or 316, compared to SEQ ID NO 35, wherein each position corresponds to the position in SEQ ID NO 39;
g. an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 40, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 40 preferably comprising a two amino acid deletion in the sequence region R181 , G182, D183, G184, compared to SEQ ID NO 40, wherein each position corresponds to the position in SEQ ID NO 40;
h. an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 40, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 40 comprising one or more, preferably all of the alterations selected from the group consisting of
a. D183*, G184*, N195F, Y243F;
b. D183*, G184*, N 195F, V206Y, Y243F;
c. W140Y, D183*, G184*, N195F, V206Y, Y243F, E260G, G304R, G476K; d. W140Y, D183*, G184*, N195F, V206Y, Y243F, E260G, G477E; e. W140Y, D183*, G184*, N195F, V206Y, Y243F, W284D;
f. W140Y + N195F + V206Y + Y243F + E260G + G477E;
g. G109A + W140Y + N195F + V206Y + Y243F + E260G;
h. T51 I S52Q N54K G109A W140Y N195F V206Y Y243F E260G G476E; i. W140Y N195F V206Y Y243F E260G W284R G477K;
j. W140Y N195F V206Y Y243F E260G W284F G477R; and
k. H 1 *, G7A, G109A, W140Y, D183*, G184*, N195F, V206Y, Y243F, E260G, N280S, G304R, E391A, G476K, compared to SEQ ID NO 40, wherein each position corresponds to the position in SEQ ID NO 40; i. an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%,
at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 41 , or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 41 , comprising a two amino acid deletion in the sequence region R181 , G182, D183, G184, compared to SEQ ID NO 41 , wherein each position corresponds to the position in SEQ ID NO 41 ;
j. an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 41 , or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 41 , comprising one or more, preferably all of the alterations selected from the group consisting of
I. R1 18K, D183*, G184*, N195F, R320K, R458K;
II. M9I, D183*, G184*, R118K, N195F, M202L, R320K, M323T, R458K;
III. M9L, G149A, R1 18K, G182T, D183*, G184*, G186A, N195F, M202L, T257I, Y295F, N299Y, M323T, A339S, E345R, R458K;
IV. M9L, G149A, R1 18K, G182T, D183*, G184*, G186A, N195F, T246V, T257I, Y295F, N299Y, M323T, A339S, E345R, R458K; and
V. M9L, G149A, G182T, D183*, G184*, G186A, M202L, T257I, Y295F, N299Y, M323T, A339S, E345R, N471 E, compared to SEQ ID NO 41 , wherein each position corresponds to the position in SEQ ID NO 41 ; k. an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 42, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 42, comprising a two amino acid deletion in the sequence region R181 , G182, G182, D183, compared to SEQ ID NO 42, wherein each position corresponds to the position in SEQ ID NO 42;
L. an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 42, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 42, comprising one or more, preferably all of the alterations selected from the group consisting of
L. D183*, G184*, N 195F, V206Y, R320K, R458K;
m. D183*, G184*, N 195F, M202L, V206L, R320K, R458K;
n. G149A, G182T, D183*, G184*, N195F, M202L, V206L, T257I, Y295F, Q299Y, A339S, Q345R, R458K;
o. G149A, G182T, D183*, G184*, N195F, V206L, M246V, T257I, Y295F, Q299Y, A339S, Q345R, R458K;
p. G149A, G182T, D183*, G184*, M202L, V206L, T257I, Y295F, Q299Y, A339S, Q345R, H471 E; and
q. H1A, N54S, V56T, K72R, G109A, F113Q, R1 16Q, W167F, Q172G, A174S, G182*, D183*, G184T, N195F, V206L, K391A, P473R and G476K, compared to SEQ ID NO 42, wherein each position corresponds to the position in SEQ ID NO 42;
m. an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 43, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 43, comprising a two amino acid deletion in the sequence region R181 , G182, H183, G184, compared to SEQ ID NO 43, wherein each position corresponds to the position in SEQ ID NO 43;
n. an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 43, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 43, comprising one or more, preferably all of the alterations selected from the group consisting of
r. H183*, G184*, I405L, A421 H, A422P, A428T;
s. R1 18K, H183*, G184*, N195F, R320K, R458K;
t. M9I, H183*, G184*, R118K, N195F, M202L, R320K, S323T, R458K; u. M9L, G149A, R1 18K, G182T, H183*, G184*, N195F, M202L, T257I, Y295F, N299Y, A339S, E345R, R458K;
v. M9L, G149A, R118K, G182T, H183*, G184*, N195F, T246V, T257I, Y295F, N299Y, A339S, E345R, R458K; and
w. M9L, G149A, G182T, H183*, G184*, M202L, T257I, Y295F, N299Y, S323T, A339S, E345R, N471 E, compared to SEQ ID NO 43, wherein each position corresponds to the position in SEQ ID NO 43; o. an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 44, or an amylase having at least 60%, at least 65%, at least 70%, at least
75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 44, comprising a two amino acid deletion in the sequence region R181 , G182, G182, D183, compared to SEQ ID NO 44, wherein each position corresponds to the position in SEQ ID NO 44;
p. an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 44, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 44, comprising one or more, preferably all of the alterations selected from the group consisting of
x. H 1 *, D183*, G184*, N195F, V206Y;
y. H 1 *, D183*, G184*, N195F, M202L, V206L, R320K, R458K; z. G149A, G182T, D183*, G184*, N195F, M202L, V206L, T257I, Y295F, Q299Y, A339S, Q345R, R458K;
aa. G149A, G182T, D183*, G184*, N195F, V206L, M246V, T257I, Y295F, Q299Y, A339S, Q345R, R458K;
bb. G149A, G182T, D183*, G184*, M202L, V206L, T257I, Y295F, Q299Y, A339S, Q345R, H471 E;
cc. H 1 *, N54S, V56T, G109A, Q169E, Q172K, A174*, G182*, D183*, N195F, V206L, K391A, G476K;
dd. G182*, D183*, N195F, W140Y, N260G, G304R, R320A, G476K, V410I, V429I, F451W, C474V;
ee. H 1 *, N54S, V56T, K72R, G109A, F113Q, R116Q, W167F, Q172G, A174S, G182*, D183*, G184T, N195F, V206L, K391A, P473R, G476K; ff. H 1 *, N54S, V56T, G109A, Q169E, Q172K, A174*, G182*, D183*, N195F, V206L, K391A, G476K;
gg. H 1 *, N54S, V56T, G109A, R116H, A174S, G182*, D183*, N195F, V206L, K391A, G476K;
hh. H 1 *, N54S, V56T, K72R, G109A, F113Q, R116Q, W167F, Q172G, A174S, G182*, D183*, G184T, N195F, V206L, K391A, P473R, G476K; ii. H 1 *, N54S, V56T, G109A, F1 13Q, R116Q, Q172N, A174S, G182*, D183*, N195F, V206L, A265G, K391A, P473R, G476K; jj. H 1 *, N54S, V56T, K72R, G109A, F113Q, W167F, Q172R, A174S, G182*, D183*, N 195F, V206L, K391A, G476K;
kk. H 1 *, N54S, V56T, K72R, G109A, R1 16H, T134E, W167F, Q172G, L173V, A174S, G182*, D183*, N 195F, V206L, G255A, K391A, G476K;
II. H 1 *, N54S, V56T, K72R, G109A, R116H, T134E, W167F, Q172G,
L173V, A174S, G182*, D183*, N195F, V206L, G255A, K391A, Q395P, T444Q, P473R, G476K;
mm. H 1 *, N54S, V56T, G109A, T134E, A174S, G182*, D183*, N195F, V206L, K391A, G476K;
nn. H 1 *, N54S, V56T, K72R, G109A, A174S, G182*, D183*, N195F, V206L, G255A, K391A, G476K;
oo. H 1 *, N54S, V56T, G109A, W167F, Q172E, L173P, A174K, G182*, D183*, N195F, V206L, K391A, G476K;
pp. H 1 *, N54S, V56T, G109A, R116Q, V120L, Q172G, 1173V, A174S, G182*, D183*, G184T, N195F, V206L, I405L, A421 H, A422P, A428T; and qq. H 1 *, N54S, V56T, G109A, F113Q, R116Q, W167F, Q172G, 1173V, A174S, G182*, D183*, G184T, N195F, V206L, I405L, A421 H, A422P, A428T, compared to SEQ ID NO 44, wherein each position corresponds to the position in SEQ ID NO 44.
19. The pouch according to any of the preceding paragraphs, wherein the detergent composition is selected from a group comprising: liquid detergents, solid detergents, gel detergents, powder detergents and granule detergents.
20. The pouch according to any of the preceding paragraphs, wherein the multicompartment pouch is a water-soluble pouch.
21. The pouch according to any of the preceding paragraphs, wherein upon dissolution in an aqueous solution the second compartment and first compartment of the pouch dissolve sequentially.
22. The pouch according to any of the preceding paragraphs, wherein the metalloprotease present in the first compartment is at a concentration of 0.01 to 100 ppm, preferably 0.05 to 80 ppm, most preferably 0.05 to 20 ppm.
23. The pouch according to any of the preceding paragraphs, wherein the pouch comprises two side-by-side compartments superposed onto another compartment wherein at least two different compartments contain two different compositions.
24. The pouch according to any of the preceding paragraphs, wherein the weight of the pouch is
about 5 to about 40g.
25. The pouch according to any of the preceding paragraphs, wherein the volume of enzyme- containing composition is in the range of volume from 5ml to 25ml.
26. The pouch according to any of the preceding paragraphs, wherein the volume of detergent composition is in the range of volume from 5ml to 25ml.
27. The pouch according to any of the preceding paragraphs, wherein the total volume in the pouch ranges from 10ml to 80ml.
28. A method of releasing an enzyme in a cleaning process comprising steps of: a. providing a multicompartment pouch with at least two compartments, the first compartment comprising enzyme-containing composition and the second compartment comprising a detergent composition; b. releasing the detergent composition from the second compartment and subsequently releasing the enzyme-containing composition from the first compartment into the wash liquor.
29. The method according to paragraph 28, wherein the detergent composition disperses and/or dissolves at the beginning of or during the main wash step of the wash cycle.
30. The method according to paragraphs 28-29, wherein the enzyme-containing composition disperses and/or dissolves during the main wash and/or at the last rinse step of the wash cycle.
31. The method according to paragraphs 28-30, wherein the wash cycle is performed at temperatures of between 5 and 95 degrees centigrade.
32. The method according to paragraphs 28-31 , wherein the length of the main wash is between 5 and 20 minutes.
33. The method according to paragraphs 28-32, wherein the wash cycle has pH between 4 and 12, preferably 5 and 11.
34. The method according to paragraphs 28-33, wherein the detergent composition in the second compartment and the enzyme-containing composition in the first compartment of the pouch dissolve sequentially.
35. The method according to paragraph 28-34, wherein the enzyme-containing composition comprises at least one enzyme.
36. The method according to paragraphs 28-35, wherein the enzyme is a protease.
37. The method according to paragraph 28-36, wherein the protease is a metalloprotease.
38. The method according to any of paragraphs 28-37, wherein the metalloprotease is selected from a group consisting of
a. a thermolysin variant;
b. a metalloprotease from M4 family, that is not a thermolysin variant;
c. a metalloprotease from the M7 family; and
d. a metalloprotease from the M35 metalloprotease.
39. The method according to any of paragraphs 28-38, wherein the metalloprotease comprises of amino acid sequence having at least 60%, at least 65%, e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, but less than 100% sequence identity to a polypeptide selected from the group consisting of SEQ ID
Nos: 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35 and 36.
40. The method according to any of paragraphs 28-39, wherein the metalloprotease comprises or consists of amino acid sequence as shown in SEQ ID Nos: 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35 and 36.
41. The method according to any of paragraphs 28-40, wherein the enzyme is present in liquid, solid, powder or gel form.
42. The method according to any of paragraphs 28-41 , wherein the detergent composition comprises at least one enzyme, preferably a DNase.
43. The method according to any of paragraphs 28-42, wherein the detergent composition further comprises one or more detergent components selected from the group consisting of surfactants, builders, flocculating aid, chelating agents, dye transfer inhibitors, enzyme stabilizers, enzyme inhibitors, catalytic materials, bleach activators, hydrogen peroxide, sources of hydrogen
peroxide, preformed peracids, polymeric dispersing agents, clay soil removal/anti-redeposition agents, brighteners, suds suppressors, dyes, perfumes, structure elasticizing agents, fabric softeners, carriers, hydrotropes, builders and co-builders, fabric hueing agents, anti-foaming agents, dispersants, processing aids, and/or pigments.
44. The method according to any of paragraphs 28-43, wherein the builder is selected from a group consisting of phosphates, sodium citrate builders, sodium carbonate, sodium silicate, sodium and zeolites.
45. The method according to any of paragraphs 28-44, wherein the surfactant is anionic and/or nonionic.
46. The method according to any of paragraphs 28-45, wherein the anionic surfactant is selected from linear alkylbenzenesulfonates (LAS) isomers of LAS, alcohol ether sulfate (AEO, AEOS) and sodium lauryl ether sulfate and sodium laureth sulfate (SLES).
47. The method according to any of paragraphs 28-46, wherein the nonionic surfactant is selected from alcohol ethoxylates (AE or AEO), alcohol propoxylates, alcohol propoxylates, propoxylated fatty alcohols (PFA), alkoxylated fatty acid alkyl esters, such as ethoxylated and/or propoxylated fatty acid alkyl esters, alkylphenol ethoxylates (APE), nonylphenol ethoxylates (NPE), alkylpolyglycosides (APG), alkoxylated amines, fatty acid monoethanolamides (FAM), fatty acid diethanolamides (FADA), ethoxylated fatty acid monoethanolamides (EFAM), propoxylated fatty acid monoethanolamides (PFAM), polyhydroxyalkyl fatty acid amides, N-acyl N-alkyl derivatives of glucosamine (glucamides, GA, or fatty acid glucamides, FAGA) and combinations thereof.
48. The method according to any of paragraphs 28-47, wherein the detergent composition further comprises one or more additional enzymes.
49. The method according to any of paragraphs 28-48, wherein the additional enzyme is selected from the group consisting of an alpha-amylase, a beta-amylase, a pullulanase, a lipase, a cellulase, an oxidase, another protease, a phospholipase, a perhydrolase, a xylanase, a pectate lyase, a pectinase, a galacturanase, a hemicellulase, a xyloglucanase, a nuclease, a mannanase and mixtures thereof.
50. The method according to any of paragraphs 28-49, wherein the enzyme is an amylase selected from the group consisting of:
a) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO: 37, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO: 37 comprising a two amino acid deletion in the sequence region R180, S181 , T182, G183, compared to SEQ ID NO:
37, wherein each position corresponds to the position in SEQ ID NO: 37;
b) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO: 37, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO: 37 comprising one or preferably all the alterations sets selected from the group consisting of:
a. R180*, S181 *, S243Q, G475K;
b. R180*, T182*, S243Q, G475K;
c. R180*, T182*, G183S, S243Q, G475K; and
d. R180*, S181*, Y242F, S243Q, F266Y, G475K compared to SEQ ID NO: 37, wherein each position corresponds to the position in SEQ ID NO: 37;
c) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO: 38, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 38, comprising a two amino acid deletion in the sequence region R178, G179, T180, G181 compared to SEQ ID NO
38, wherein each position corresponds to the position in SEQ ID NO 38;
d) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 38, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 38, comprising one or more, preferably all of the alterations sets selected from the group consisting of:
I. R178*, G179*, E187P, I203Y, G476K;
II. R178*, G179*, E187P, M199L, I203Y, G476K;
III. R178*, G179*, E187P, I203Y R458N, T459S, D460T, G476K;
IV. N126Y, F153W, R178*, G179*, T180H, I203Y, S241 Q;
V. N126Y, F153W, R178*, G179*, T180H, I203Y, S241 Q, S362A, R375Y;
VI. T38N, N126Y, T129I, F153W, R178*, G179*, T180D, E187P, I203Y, G476K, G477E; and
VII. N126Y, F153W, R178*, G179*, T180H, E187P, I203Y, S241Q, G476K, G477E, compared to SEQ ID NO 38, wherein each position corresponds to the position in SEQ ID NO 38;
e) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 39, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 39, comprising a two amino acid deletion in the sequence region R181 , G182, D183, G184 compared to SEQ ID NO 39, wherein each position corresponds to the position in SEQ ID NO 39;
f) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 39, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 39 comprising an alteration at one or more, preferably at all of the position(s) selected from 3, 4, 5, 118, 167, 170, 177, 195, 202, 204, 271 , 320, 330, 377, 385, 445, 458, 475, 476, 314, 315 or 316, compared to SEQ ID NO 35, wherein each position corresponds to the position in SEQ ID NO 39; g) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 40, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 40 preferably comprising a two amino acid deletion in the sequence region R181 , G182, D183, G184, compared to SEQ ID NO 40, wherein each position corresponds to the position in SEQ ID NO 40;
h) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 40, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 40 comprising one or more, preferably all of the alterations selected from the group consisting of
a. D183*, G184*, N195F, Y243F;
b. D183*, G184*, N 195F, V206Y, Y243F;
c. W140Y, D183*, G184*, N195F, V206Y, Y243F, E260G, G304R, G476K;
d. W140Y, D183*, G184*, N195F, V206Y, Y243F, E260G, G477E; e. W140Y, D183*, G184*, N195F, V206Y, Y243F, W284D;
f. W140Y + N195F + V206Y + Y243F + E260G + G477E;
g. G109A + W140Y + N195F + V206Y + Y243F + E260G;
h. T51 I S52Q N54K G109A W140Y N195F V206Y Y243F E260G G476E;
i. W140Y N195F V206Y Y243F E260G W284R G477K;
j. W140Y N195F V206Y Y243F E260G W284F G477R; and
k. H 1 *, G7A, G109A, W140Y, D183*, G184*, N195F, V206Y, Y243F, E260G, N280S, G304R, E391A, G476K, compared to SEQ ID NO 40, wherein each position corresponds to the position in SEQ ID NO 40;
i) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 41 , or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 41 , comprising a two amino acid deletion in the sequence region R181 , G182, D183, G184, compared to SEQ ID NO 41 , wherein each position corresponds to the position in SEQ ID NO 41 ;
j) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 41 , or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 41 , comprising one or more, preferably all of the alterations selected from the group consisting of
VI. R1 18K, D183*, G184*, N195F, R320K, R458K;
VII. M9I, D183*, G184*, R118K, N195F, M202L, R320K, M323T, R458K;
VIII. M9L, G149A, R118K, G182T, D183*, G184*, G186A, N195F, M202L, T257I, Y295F, N299Y, M323T, A339S, E345R, R458K;
IX. M9L, G149A, R1 18K, G182T, D183*, G184*, G186A, N195F, T246V, T257I, Y295F, N299Y, M323T, A339S, E345R, R458K; and
X. M9L, G149A, G182T, D183*, G184*, G186A, M202L, T257I, Y295F, N299Y, M323T, A339S, E345R, N471 E, compared to SEQ ID NO 41 , wherein each position corresponds to the position in SEQ ID NO 41 ; k) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 42, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but
less than 100% sequence identity to SEQ ID NO 42, comprising a two amino acid deletion in the sequence region R181 , G182, G182, D183, compared to SEQ ID NO
42, wherein each position corresponds to the position in SEQ ID NO 42;
L) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 42, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 42, comprising one or more, preferably all of the alterations selected from the group consisting of
a. D183*, G184*, N195F, V206Y, R320K, R458K;
b. D183*, G184*, N 195F, M202L, V206L, R320K, R458K;
c. G149A, G182T, D183*, G184*, N195F, M202L, V206L, T257I, Y295F, Q299Y, A339S, Q345R, R458K;
d. G149A, G182T, D183*, G184*, N195F, V206L, M246V, T257I, Y295F, Q299Y, A339S, Q345R, R458K;
e. G149A, G182T, D183*, G184*, M202L, V206L, T257I, Y295F, Q299Y, A339S, Q345R, H471 E; and
f. H1A, N54S, V56T, K72R, G109A, F1 13Q, R116Q, W167F, Q172G, A174S, G182*, D183*, G184T, N195F, V206L, K391A, P473R and G476K, compared to SEQ ID NO 42, wherein each position corresponds to the position in SEQ ID NO 42;
m) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 43, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 43, comprising a two amino acid deletion in the sequence region R181 , G182, H 183, G184, compared to SEQ ID NO
43, wherein each position corresponds to the position in SEQ ID NO 43;
n) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 43, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 43, comprising one or more, preferably all of the alterations selected from the group consisting of
a. H183*, G184*, I405L, A421 H, A422P, A428T;
b. R1 18K, H183*, G184*, N195F, R320K, R458K;
c. M9I, H183*, G184*, R118K, N195F, M202L, R320K, S323T, R458K;
d. M9L, G149A, R118K, G182T, H183*, G184*, N195F, M202L, T257I, Y295F, N299Y, A339S, E345R, R458K;
e. M9L, G149A, R1 18K, G182T, H183*, G184*, N195F, T246V, T257I, Y295F, N299Y, A339S, E345R, R458K; and
f. M9L, G149A, G182T, H183*, G184*, M202L, T257I, Y295F, N299Y, S323T, A339S, E345R, N471 E, compared to SEQ ID NO 43, wherein each position corresponds to the position in SEQ ID NO 43;
o) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 44, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 44, comprising a two amino acid deletion in the sequence region R181 , G182, G182, D183, compared to SEQ ID NO 44, wherein each position corresponds to the position in SEQ ID NO 44;
p) an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to SEQ ID NO 44, or an amylase having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% but less than 100% sequence identity to SEQ ID NO 44, comprising one or more, preferably all of the alterations selected from the group consisting of
a. H 1 *, D183*, G184*, N195F, V206Y;
b. H 1 *, D183*, G184*, N195F, M202L, V206L, R320K, R458K;
c. G149A, G182T, D183*, G184*, N195F, M202L, V206L, T257I, Y295F, Q299Y, A339S, Q345R, R458K;
d. G149A, G182T, D183*, G184*, N195F, V206L, M246V, T257I, Y295F, Q299Y, A339S, Q345R, R458K;
e. G149A, G182T, D183*, G184*, M202L, V206L, T257I, Y295F, Q299Y, A339S, Q345R, H471 E;
f. H 1 *, N54S, V56T, G109A, Q169E, Q172K, A174*, G182*, D183*, N 195F, V206L, K391A, G476K;
g. G182*, D183*, N195F, W140Y, N260G, G304R, R320A, G476K, V410I, V429I, F451W, C474V;
h. H 1 *, N54S, V56T, K72R, G109A, F1 13Q, R1 16Q, W167F, Q172G, A174S, G182*, D183*, G184T, N195F, V206L, K391A, P473R, G476K;
i. H 1 *, N54S, V56T, G109A, Q169E, Q172K, A174*, G182*, D183*, N 195F, V206L,
K391A, G476K;
j. H 1 *, N54S, V56T, G109A, R116H, A174S, G182*, D183*, N195F, V206L, K391A, G476K;
k. H 1 *, N54S, V56T, K72R, G109A, F1 13Q, R1 16Q, W167F, Q172G, A174S, G182*, D183*, G184T, N195F, V206L, K391A, P473R, G476K;
L. H 1 *, N54S, V56T, G109A, F1 13Q, R1 16Q, Q172N, A174S, G182*, D183*, N 195F, V206L, A265G, K391A, P473R, G476K;
m. H 1 *, N54S, V56T, K72R, G109A, F113Q, W167F, Q172R, A174S, G182*, D183*, N195F, V206L, K391A, G476K;
n. H 1 *, N54S, V56T, K72R, G109A, R1 16H, T134E, W167F, Q172G, L173V, A174S, G182*, D183*, N195F, V206L, G255A, K391A, G476K;
o. H 1 *, N54S, V56T, K72R, G109A, R1 16H, T134E, W167F, Q172G, L173V, A174S, G182*, D183*, N195F, V206L, G255A, K391A, Q395P, T444Q, P473R, G476K; p. H 1 *, N54S, V56T, G109A, T134E, A174S, G182*, D183*, N195F, V206L, K391A, G476K;
q. H 1 *, N54S, V56T, K72R, G109A, A174S, G182*, D183*, N195F, V206L, G255A, K391A, G476K;
r. H 1 *, N54S, V56T, G109A, W167F, Q172E, L173P, A174K, G182*, D183*, N195F, V206L, K391A, G476K;
s. H 1 *, N54S, V56T, G109A, R1 16Q, V120L, Q172G, 1173V, A174S, G182*, D183*, G184T, N195F, V206L, I405L, A421 H, A422P, A428T; and
t. H 1 *, N54S, V56T, G109A, F1 13Q, R1 16Q, W167F, Q172G, 1173V, A174S, G182*, D183*, G184T, N195F, V206L, I405L, A421 H, A422P, A428T, compared to SEQ ID NO 44, wherein each position corresponds to the position in SEQ ID NO 44.
51. The method according to any of the preceding paragraphs 28-50, wherein the DNase is microbial, preferably obtained from bacteria or fungi.
52. The method according to any of the preceding paragraphs 28-51 , wherein the DNase comprises a polypeptide having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to the amino acid sequence shown in SEQ ID NO 60.
53. The method according to any of the preceding paragraphs 28-52, wherein the DNase comprises a polypeptide having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to the amino acid sequence shown in SEQ ID NO: 61.
54. The method according to any of the preceding paragraphs 28-53, wherein the DNase comprises a polypeptide having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to the amino acid sequence shown in SEQ ID NO: 62.
55. The method according to any of the preceding paragraphs 28-54, wherein the DNase is fungal, preferably obtained from Aspergillus and even more preferably from Aspergillus oryzae and wherein the DNase comprises a polypeptide having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to the amino acid sequence shown in SEQ ID NO: 63.
56. The method according to any of the preceding paragraphs 28-55, wherein the DNase is fungal, preferably obtained from Trichoderma and even more preferably from Trichoderma harzianum and wherein the DNase comprises a polypeptide having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% sequence identity to the amino acid sequence shown in SEQ ID NO: 64.
57. The method according to any of paragraphs 28-56, wherein the first compartment releases the enzyme composition at least 30 seconds later than the second compartment releases the detergent composition.
58. A method of treating a substrate, where the method includes the step of contacting the substrate with the multi-compartment pouch of paragraphs 1-27 in the presence of water, wherein the substrate is a fabric or a hard surface.
59. A method of cleaning comprising contacting a surface and/or a fabric with the multi compartment pouch of any of the paragraphs 1-27.
60. A method for removing and/or reducing soil and/or for reducing redeposition on a surface and/or textile comprising contacting the surface and/or textile with the pouch of any of paragraphs 1-27.
61. A method of laundering or dishwashing in a washing machine comprising the steps of placing the multi-compartment pouch of any of the paragraphs 1-27 into the product dispenser and releasing it during the wash cycle.
62. Use of a multi-compartment pouch according to paragraphs 1-27 for cleaning of an item, wherein the item is a textile or a surface.
63. Use of a multi-compartment pouch according to paragraphs 1-27 in a cleaning process, such as laundry, hard surface cleaning, dish wash or automated dish wash.
64. Use of a multi-compartment pouch according to paragraphs 1-27 for removing or releasing a stain from a textile having a stain.
The present invention is further described by the following examples that should not be construed as limiting the scope of the invention.
Examples
Materials and methods
Protazyme OL characterization assay:
Substrate: Protazyme OL tablet (AZCL-collagen, Megazyme T-PROL 1000).
Temperature: Controlled (assay temperature).
Assay buffers: 100mM succinic acid, 100mM HEPES, 100mM CHES, 100mM CABS,
1 mM CaCI2, 150mM KCI, 0.01 % Triton X-100 adjusted to pH-values 2.0,
3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, 10.0 and 11.0 with HCI or NaOH.
A Protazyme OL tablet is suspended in 2.0 ml 0.01 % Triton X-100 by gentle stirring. 500 mI of this suspension and 500 mI assay buffer are dispensed in an Eppendorf tube and placed on ice. 20 mI protease sample (diluted in 0.01 % Triton X-100) is added to the ice cold tube. The assay is initiated by transferring the Eppendorf tube to an Eppendorf thermomixer, which is set to the assay temperature. The tube is incubated for 15 minutes on the Eppendorf thermomixer at its highest shaking rate (1400 rpm). The incubation is stopped by transferring the tube back to the ice bath. Then the tube is centrifuged in an ice cold centrifuge for a few minutes and 200 mI supernatant is transferred to a microtiter plate. OD650 is read as a measure of protease activity. A buffer blind is included in the assay (instead of enzyme).
Automatic Mechanical Stress Assay (AMSA) for laundry
In order to assess the wash performance in laundry washing experiments are performed, using the Automatic Mechanical Stress Assay (AMSA). With the AMSA, the wash performance of a large quantity of small volume enzyme-detergent solutions can be examined. The AMSA plate
has a number of slots for test solutions and a lid firmly squeezing the laundry sample, the textile to be washed against all the slot openings. During the washing time, the plate, test solutions, textile and lid are vigorously shaken to bring the test solution in contact with the textile and apply mechanical stress in a regular, periodic oscillating manner. For further description see W002/42740 especially the paragraph "Special method embodiments" at page 23-24.
A typical laundry experiments are conducted under the experimental conditions specified below:
Model detergents and test materials are as follows:
All test materials are obtained from EMPA Testmaterials AG Movenstrasse 12, CH-9015 St. Gallen, Switzerland or from Center For Testmaterials BV, P.O. Box 120, 3133 KT Vlaardingen, the Netherlands.
Water hardness is adjusted to 15°dH by addition of CaCL, MgCL, and NaHCOs (Ca2+:Mg2+:NaHC03= 4:1 :7.5) to the test system. After washing the textiles were flushed in tap water and dried.
The wash performance was measured as the brightness of the colour of the textile washed. Brightness can also be expressed as the intensity of the light reflected from the sample when illuminated with white light. When the sample is stained the intensity of the reflected light is lower than that of a clean sample. Expressed another way, a cleaner sample will reflect more light and will have a higher intensity. Therefore, the intensity of the reflected light can be used to measure wash performance.
Color measurements are made with a professional flatbed scanner (Kodak iQsmart, Kodak, Midtager 29, DK-2605 Brondby, Denmark), which is used to capture an image of the washed textile.
To extract a value for the light intensity from the scanned images, 24-bit pixel values from the image are converted into values for red, green and blue (RGB). The intensity value (Int) is calculated by adding the RGB values together as vectors and then taking the length of the resulting vector:
Example 1 : Relative wash performance of different metalloproteases
In order to demonstrate that separating the metalloproteases from the detergent ingredients until the ingredients are dissolved in water enables the use of metalloproteases in detergents, different metalloproteases were added to a liquid model detergent, incubated for 30 minutes at room temperature and then diluted with water hardness according to the detergent dosage. Afterwards wash performance was investigated in AMSA. This experimental setup reflects the addition of a metalloprotease to a nowadays commonly used liquid detergent. The results from this setup were compared with the results from a second setup, where the liquid model detergent was diluted with water hardness according to the detergent dosage before the metalloproteases were added. This setup reflects the usage of a pouch, where the detergent is dissolved in water prior metalloprotease release. Wash performance of the detergent solution with metalloprotease was investigated in AMSA after 30min incubation of the wash liquor at room temperature.
The metalloprotease’s as listed in the table 1 , is added to both, the undiluted/undissolved detergent and a diluted detergent solution (= wash liquor). The metalloprotease shows significant wash performance when added to the diluted detergent solution (= wash liquor). On the other hand, when added to the undiluted/undissolved detergent, then a drastic performance drop is observed as only a small metalloprotease performance is detectable. It may be observed that the metalloprotease must be kept separated from the undiluted/undissolved detergent. Further, the metalloprotease should get into the wash liquor when the detergent is already diluted. Table 1 shows the wash performance of the different metalloproteases from setup 1 (metalloproteases
added to detergent and then after 30 min diluted with water hardness according to detergent dosage) relative to setup 2 (detergent first diluted with water hardness according to detergent dosage and then metalloproteases added) in percent. Table 1 : Relative wash performance of different metalloproteases
From the data, it shows that metalloproteases have a significantly reduced wash performance when getting in contact with undiluted detergent for a short time (relative to having contact with diluted detergent for the same time period), meaning that metalloproteases suffer from a significant performance loss when getting in contact with undiluted detergent. Therefore, it is suggested that in pouches where the metalloprotease is released after the detergent is diluted with water, is a good solution.
The invention described and claimed herein is not to be limited in scope by the specific aspects herein disclosed, since these aspects are intended as illustrations of several aspects of the invention. Any equivalent aspects are intended to be within the scope of this invention. Indeed, various modifications of the invention in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims. In the case of conflict, the present disclosure including definitions will control.