BR112013000110B1 - cleaning compositions and cleaning process - Google Patents

Abstract

CLEANING COMPOSITIONS AND CLEANING PROCESS A cleaning composition comprising an effective amount of a surfactant system and an enzyme system characterized in that the surfactant system comprises at least 1% by weight (based on the cleaning composition) of a biosurfactant of bacterial origin and at least one enzyme of bacterial origin selected from the group comprising: cellulases, lipases, esterases, peroxidases/oxidases, oxidoreductases, pectases, lyases, mannanases and mixtures thereof.

Description

Field of Invention

This invention relates to detergent compositions comprising biosurfactant and enzyme.

Background of the Invention

A general description of biosurfactants is published by Rahman in Biotechnology 7(2): 360-370, 2008 ISSN 1682-296X “Production, Characterization and application of Biosurfactants - review”.

Enzymes have been used in detergent formulations as a cleaning aid for many years. They can be derived from bacterial sources or other sources. The most commonly used enzymes are proteases, amylases, mannanases, lipases and cellulases. They are often derived from fungal or yeast cultures.

Lipases are used in detergent formulations containing surfactant to aid in cleaning oily soils from fabrics. Despite their isolation and characterization a few decades ago, these enzymes have been difficult to formulate in conventional surfactant formulations because there is competition between enzyme and surfactant for the target substrate oil. Surfactants will win this competition for the surface and will outpace or displace enzymes from the oily surface and therefore reduce the enzyme's performance on those soils. Thus, the practical impact of lipases on detergent cleaning products is limited, especially when compared to the impact of other cleaning enzymes such as proteases and amylases.

The move to more sustainable chemicals reinforces a desire to reduce the level of surfactant in the wash. As a bioalternative, enzymes represent a weight efficient choice to maintain performance on oily dirt removal as surfactant levels are reduced. The use of biosurfactants has been proposed in many prior art documents. The following documents refer to combinations of biosurfactants and enzymes produced from bacteria.

DE102008038479A1 (Henkel) describes potential mixtures of alpha amylase enzyme that can be bacterially derived with surfactants that can be biosurfactants and can be produced from bacteria.

WO2006/031554A2 (Novozymes) describes as one of its examples a mixture of a bacterially derived protease with the biosurfactant surfactin. No importance seems to be attached to bacterially derived enzymes, most of the cited enzymes are derived from fungi.

“Lipase and biosurfactant production for utilization in bioremediation of vegetable oils and hydrocarbon”. Martins VG et al (2008) Quimica Nova No 31 15 vol 8, 1942-1947.

“Isolation and characterization of a lipid degrading bacterium and its application to lipid containing wastewater treatment”. Matsumiya Y. et al (2007) Journal of Bioscience and Bioengineering No 103, Vol 4, 325-330.

US2006106120 describes a mixture of microorganism, biosurfactant and a plastic degradation enzyme for the bioremediation of man-made materials. The biosurfactant can be derived from bacterial or other sources; the preferred enzyme used in the examples is a cutinase of bacterial origin. It can be co-expressed with amylase and hydrophobin. Compositions are not used for cleaning.

The following documents refer to combinations of biosurfactants and enzymes not specifically produced from bacteria for cleaning.

US2006080785A (Nero) describes carpet cleaning by applying a cleaning composition having biosurfactants and enzymes to the carpet; and surface cleaning of the material. Enzymes are derived from Kelp Forests and thus are not bacterially derived.

CN101126052 describes a biosurfactant containing cleaning composition which also contains a protease. The origin of the protease is a pineapple plant.

US5417879 (Unilever) describes synergistic dual surfactant laundry composition containing sophorolipid (from yeast), cellobiose lipid (from fungi) or rhamnolipid (from bacteria) glycolipid biosurfactant. Examples using these biosurfactants do not comprise any enzymes. In rows 24 to 25 of column 12, it is mentioned how possible to combine the biosurfactants with an undisclosed amount of enzyme of undisclosed origin.

US2004171512A (Igarashi Keisuke ; Hirata Yoshihiko ; Furuta Taro) describes low foam detergent compositions comprising a biosurfactant (yeast sophorolipid) which can replace a conventional low foam block polymer nonionic surfactant. According to the general description, the biosurfactant can be used with an undisclosed type of enzyme selected from amylase, protease, cellulose, lipase, pullulanase, isopululanase, isoamylase, catalase, peroxidase, or the like. The enzyme can be added by selecting appropriately in light of its substrate specificity. For example, protease can be selected for a protein dye, and amylase can be selected for a starch dye. Examples of the use of sophorolylpides for dishwashing (hard surface cleaning) in combination with Savinase 6.0T a protease from Novo Nordisk and Duramyl 60T a starch lytic enzyme (amylase) from Novo Nordisk. Duramyl is produced from Bacillus Licheniformis and Savinase is produced from Bacillus Clausii/lentus, both bacterial sources. These are not taught as being generically preferred sources in this document.

US2009188055A (Stepan Co) describes compositions comprising sulphonated stolides and other fatty acid derivatives. Table 20 provides prophetic examples of these surfactants in combination with other surfactants, including rhamnolipids. Enzymes are not included in these examples. Elsewhere in the document, it is said that cleaning performance on greasy soils is synergistically improved with stolides using lipases. Suitable lipase enzymes include those produced by microorganisms of the Pseudomonas group, such as Pseudomonas stutzeri ATCC 19154, as described in British Patent 1,372,034. Appropriate lipases include those that show an immunologically positive cross-reaction with the lipase antibody produced by the microorganisms Pseudomonas fluorescens IAM 1057. This lipase is available from Amano Pharmaceutical Co. Ltd., Nagoya, Japan, under the trade name Lipase P “Amano, ” hereinafter referred to as “Amano-P”. Further suitable lipases are lipases such as M1 Lipase.RTM and Lipomax.RTM (Gist-Brocades). Highly preferred lipases are the lipolytic enzyme D96L variant of native lipase derived from Humicola lanuginosa (a fungus) as described in U.S. 6,017,871 issued January 25, 2000 (P&G). Preferably, the strain of Humicola lanuginosa DSM 4106 is used. This enzyme is incorporated into the composition according to the present technology at a level of 50 LU to 8500 LU per liter of wash solution. Preferably, the D96L variant is present at a level of 100 LU to 7500 LU per liter of wash solution. More preferably at a level of 150 LU to 5000 LU per liter of wash solution.

One document suggests combinations of biosurfactants and bacterial-derived enzymes for washing. US2004072713A (Unilever) describes an article for use in an enzymatic fabric washing process, said article containing one or more types of harmless microorganisms capable of excreting enzymes useful in said fabric cleaning process. In one embodiment, the microorganism can be a bacterium, although fungal microorganisms are also exemplified. The examples all express bleaching enzymes. Although not used in the examples the document speculates that it is especially useful if, in addition to enzymes, microorganisms are also able to produce other chemical entities that contribute to the cleaning process, eg biosurfactants, eg lipopolysaccharides.

No liquid wash substance or concentrate comprising a mixture of bacterial-derived biosurfactants with “bacteria-derived enzymes” is currently described in this document. We are confident that the biosurfactant concentration would have been much less than 0.5 g/L.

Invention Summary

According to the present invention there is provided a cleaning composition comprising an effective amount of surfactant system and an enzyme system characterized in that the surfactant system comprises at least 1% by weight (based on the cleaning composition) of a biosurfactant of bacterial origin and at least one enzyme of bacterial origin selected from the group comprising: cellulases, lipases, esterases, peroxidases/oxidases, oxidoreductases, pectases, lyases, mannanases and mixtures thereof.

According to a second aspect of the invention there is provided a process for cleaning a substrate comprising the steps of immersing the substrate in water by adding a composition according to any one of the preceding claims to form a liquid washing substance and wash the substrate characterized by time. the wash cycle is less than 60 minutes, preferably less than 30 minutes and the water temperature is less than 35 °C at all times.

Amazing synergistic benefits in cleaning stains and dirt were discovered when certain bacterial-derived enzymes were combined with bacterial-derived biological surfactants (biosurfactants).

The combination can be used in any biological formulation. Lipases are a key enzyme for insertion into detergent compositions, especially laundry detergents, but also compositions designed to clean hard surfaces like dishwashing compositions, which clean everyday dirt and stains effectively at reduced surfactant levels to enable formulation concentration.

We tested three types of biosurfactants: (fungal, bacterial and yeast) in combination with two types of lipase enzyme (fungal and bacterial). The best result comes from a combination of bacterially derived enzyme with bacterially derived biosurfactant (Ramnolipid).

Detailed Description of the Invention

Enzymes Bacterial Enzymes Bacterial enzymes for use in the invention are cellulases, lipases, esterases, peroxidases/oxidases, pectases, lyases, and mannanases, or mixtures thereof. Bacterial genes encoding such enzymes can be transferred to preferred expressions in production hosts, which are not limited to bacterial and include, for example, other microbial hosts. The term bacterial enzyme as used herein includes enzymes originally from bacteria, however expressed.

The composition may comprise cutinase as classified in EC 3.1.1.74. An example of a bacterial cutinase is that of a strain of Pseudomonas, in particular Pseudomonas mendocina, or Pseudomonas putida.

The enzyme may be a phospholipase classified as EC 3.1.1.4 and/or EC 3.1.1.32. As used herein, the term phospholipase is an enzyme, which has activity towards phospholipids. Phospholipids, such as lecithin or phosphatidylcholine, consist of glycerol esterified with two fatty acids in the outer (sn-1) and middle (sn-2) positions and esterified with phosphoric acid in the third position; phosphoric acid, in turn, can be esterified to an amino alcohol. Phospholipases are enzymes that participate in the hydrolysis of phospholipids. Several types of phospholipase activity can be distinguished, including phospholipases A1 and A2 that hydrolyze a fatty acyl group (at the sn-1 and sn-2 positions, respectively) to form lysophospholipid; and lysophospholipase (or phospholipase B) which can hydrolyze the remaining fatty acyl group to lysophospholipid. Phospholipase C and phospholipase D (phosphodiesterases) release diacyl glycerol or phosphatidic acid respectively.

The term phospholipase includes enzymes with phospholipase activity, for example, phospholipase A (AT OR A2), phospholipase B activity, phospholipase C activity, or phospholipase D activity. invention is intended to cover an enzyme with Phospholipase AT and/or Phospholipase A2 activity. Phospholipase activity can be provided by enzymes having other activities as well, such as, for example, a lipase with phospholipase activity. The phospholipase activity can, for example, be a lipase with lateral phospholipase activity. In other embodiments of the invention, the phospholipase enzyme activity is provided by an enzyme having essentially only phospholipase activity and wherein the phospholipase enzyme activity is not a lateral activity.

Preferably, the phospholipase is of Bacillus bacterial origin, for example B. megaterium, B. subtilis; Citrobacter, for example C. freundii; Enterobacter, for example, E. aerogenes, E. cloacae Edwardsiella, for example, E. tarda; Erwinia, for example, E. herbicola; Escherichia, for example, E. coli; Klebsiella, for example K. pneumoniae; Proteus, for example, P. vulgaris; Providencia, for example, P. stuartii; Salmonella, for example S. typhimurium; Serratia, for example, S. liquefasciens, S. marcescens; Shigella, for example S. flexneri;

Appropriate cellulases are especially of bacterial origin. Engineered or chemically modified protein mutants are included. Suitable cellulases include cellulases from the Bacillus, Pseudomonas and Clostridia genera.

Appropriate peroxidases / oxidases are especially of bacterial origin. Engineered or chemically modified protein mutants are included. An example of an oxidative bacteria is, but not limited to, Aeromonas sp from which oxidases can originate.

Examples of lyase pectates include lyase pectates that have been cloned from different genera of bacteria such as Erwinia, Pseudomonas,

Klebsiella and Xanthomonas, as well as Bacillus subtilis (Nasser et al. (1993) FEBS Letts. 335:319-326) and Bacillus sp. YA-14 (Kim et al. (1994) Biosci. Biotech. Biochem. 58:947-949).

Examples of mannanases (EC 3.2.1.78) include those isolated from various bacteria, including Bacillus organisms. For example, Talbot et al., Appl. Environ. Microbiol., Vol.56, No. 11, pp. 3505-3510 (1990) describes a beta-mannanase derived from Bacillus stearothermophilus. Mendoza et al., World J. Microbiol. Biotech., Vol. 10, No. 5, pp. 551-555 (1994) deserves a beta-mannanase derived from Bacillus subtilis. JP-A-03047076 describes a beta-mannanase derived from Bacillus sp. JP-A-63056289 describes the production of an alkaline thermostable beta-mannanase. JP-A-63036775 refers to the microorganism Bacillus FERM P-8856 which produces beta-mannanase and beta-mannosidase. JP-A-08051975 describes alkaline beta-mannanases from Bacillus sp alkalophilic. AM-001. A purified mannanase from Bacillus amyloliquefaciens is described in WO 97/11164. WO 91/18974 describes a hemicellulase as an active glucanase, xylanase or mannanase. Mananases from Bacillus sp. discussed in the Examples in WO 99/64619.

The composition may further comprise other enzymes of bacterial origin and/or enzymes which are not of bacterial origin.

Biosurfactants

These are derived from bacteria. Other biosurfactants outside the scope of this invention can be derived from yeast and fungi. The term Biosurfactant in this patent specification does not include surfactants derived from plant material such as Alkyl Polyglucosides (APG).

a) Bacterial-derived biosurfactants

These are, for example, the Ramnolipids typically from Pseudomonas sp. Information on other bacterially derived biosurfactants is available from "Mapping of Patents in Bioemulsifiers and biosurfactants - journal, published in the Journal of Scientific and Industrial Research Vol 65, 2006, P91. Within the definition of bacterially produced biosurfactants, we include those in which a bacterial gene is is cloned and subsequently expressed from another organism as a manufacturing technique.For example, Rhamnolipids were produced from E. coli in this way.

b) Biosurfactants from non-bacterial sources

Biosurfactants from non-bacterial sources include those derived from fungi and yeasts, for example, sophorolylpides from Candida sp and Torulopsis sp. Candida apicola, Candida bombicola, Candida lipolytica, Candida bogoriensis. See: Environmental applications for biosurfactants - Environmental Pollution, Volume 133, 2005, Pages 183-198 Catherine N. Mulligan. See also, For Commercial Production of Microbial Surfactants - Trends in Biotechnology, Volume 24, 2006, Pages 509-515: Soumen Mukherjee, Palashpriya Das, Ramkrishna Sen.

Mannosylerythritol lipids are typically from Pseudozyma (formerly Candida) Antarctica. Cellobiose lipids are typically from Ustilago maydis. Trehalose lipids typically from Rhodococcus sp.

Additional information is given in Journal Production, Characterization and Applications of Biosurfactants - Biotechnology - Volume 7, 2008, page 370: Pattanathu, Rahman and Gakpe.

The detergent composition may comprise other ingredients commonly found in laundry liquids. Especially dirt-releasing polymers of polyester substances, hydrotropes, opacifiers, dyes, perfumes, other enzymes, other surfactants, microcapsules of ingredients such as perfume or treatment additives, softeners, polymers for anti-dirt redeposition, bleach, bleach activators and bleach catalysts, antioxidants, pH control agents and buffers, thickening agents, external builders for rheology modification, visual stimuli, with or without functional ingredients embedded in that place, and other ingredients known to those skilled in the art. The composition is preferably a liquid and is advantageously packaged in a multi-dose bottle or in a soluble unit dose pouch.

The invention will now be further described with reference to the following non-limiting examples.

Example 1

In this example, various enzyme/biosurfactant compositions were tested to determine their ability to remove a colored beef stain from the cotton cloth.

Wash solutions were prepared by dispersing the lipase at a concentration of 4mg protein per liter with “the detergent surfactant at a required concentration in phosphate buffered saline (PBS) adjusted to pH 8 and 12° FH water hardness. 10 ml of the wash solution was mixed in 25 ml plastic flasks at 37 °C with shaking at 200 rpm in an orbital incubator for 30 minutes. Samples (approximately 1 cm2 ) of cotton cloth stained with Sudan Red colored beef fat were then added to the vials and returned to the shaking incubator. Samples were removed at timed intervals, rinsed in cold water and dried at 37°C. Residual color was monitored using a Macbeth Color Eye, and compared to untreated stained cloths. Results are shown in Table 1 for 30 minutes and Table 2 for 4 hours.

The bacterial enzyme is "Lipomax", a bacterially derived lipase variant M21L of the lipase from Pseudomonas alcaligenes as described in WO 94/25578 for Gist-Brocades (MMMJ Cox, HBM Lenting, LJSM Mulleners and JM van der Laan). The fungal enzyme is "Lipolase", derived from Humicola languginosa as described in EP 0 258 068 and available from NovoZymes A/S. Details of the surfactants were as follows: SL = Sophorolipid: a biosurfactant of fungal origin AC = Accell: a biosurfactant derived from a yeast RL = Ramnolipid: a biosurfactant of bacterial origin.

The bacterial enzyme consistently outperforms the fungal enzyme in all stain types. For sophorolipids the presence of the fungal enzyme provides no benefit over the surfactant used without any enzyme.

Enzymes were all dosed at the same level by determining the amount of active enzyme protein in each of the samples using a standard BCA protein assay kit (ex Pierce) following the manufacturer's protocol.

Example 2

In this example, various enzyme/biosurfactant compositions were examined to determine their ability to remove a colored beef stain.

The same experiment was performed as in example 1 except that 5 the rhamnolipid material was separated into its mono-rhamnolipid and di-rhamnolipid components. Di-rhamnolipid having two rhamnose sugars on the acyl group. The notation R1 is used for the mono-ramnolipid material and R2 for the di-ramnolipid. The cleaning results for 1 hour and 4 hours are given in tables 3 and 4.

Claims (5)

1. Cleaning composition comprising a surfactant system and an enzyme system, characterized in that the surfactant system comprises at least 1% by weight (based on the cleaning composition) of a biosurfactant of bacterial origin and at least one enzyme of bacterial origin selected from group comprising: cellulases, lipases, esterases, peroxidases/oxidases, oxidoreductases, pectases, lyases, mannanases and mixtures thereof, in which the biosurfactant is a rhamnolipid. 2. Composition according to claim 1, characterized in that the enzyme is selected from the group consisting of: lipase, cellulase, mannanase, esterase, oxidoreductase and mixtures thereof. Composition according to claim 2, characterized in that the enzyme comprises lipase and/or esterase, preferably lipase. Composition according to any one of claims 1 to 3, characterized in that the rhamnolipid comprises two or more rhamnose units in the acyl chain, preferably it is at least 60% d-ramnolipid. A process for cleaning a substrate comprising the steps of immersing the substrate in water by adding a composition as defined in any one of claims 1 to 4 to water to form a washing liquid and washing the substrate, characterized by the time of wash cycle is less than 60 minutes, preferably less than 30 minutes and the water temperature is less than 35 °C at all times.