EP2756063B1

EP2756063B1 - Detergent compositions comprising surfactant and enzyme

Info

Publication number: EP2756063B1
Application number: EP12756157.9A
Authority: EP
Inventors: Andrew Thomas COOK; Neil James Parry
Original assignee: Unilever PLC; Unilever NV
Current assignee: Unilever PLC; Unilever NV
Priority date: 2011-09-15
Filing date: 2012-08-30
Publication date: 2017-10-04
Anticipated expiration: 2032-08-30
Also published as: ZA201401260B; CN103946359A; WO2013037643A1; AR087847A1; BR112014005687A2; CL2014000636A1; EP2756063A1

Description

This invention relates to enzymatic detergent compositions.
Enzymes are used in detergent formulations to aid cleaning and stain removal. For example, US 2010/0075878 A1 , US 2003/0139319 A1 and US 6,271,190 B1 disclose detergent compositions comprising a surfactant system, one or more enzymes and one or more lignin compounds.
The objective of the invention is to improve enzyme performance in detergent formulations.
In a first aspect, the present invention provides an enzymatic detergent composition comprising the combination of:

(i) a surfactant system;
(ii) one or more enzymes
(iii) one or more lignin compounds, wherein the surfactant system comprises a biosurfactant. In a second aspect, the invention provides a process for cleaning a substrate, comprising the step of treating the substrate with the enzymatic detergent composition of the first aspect of the invention.
With the invention, cleaning performance is improved.
As used herein, the term "substrate" includes fabric, and clothing and laundry items. Accordingly, preferably the process is for cleaning a fabric, i.e. stain/soil removal from fabric.
Preferably the cleaning process takes place in a washing receptacle containing a wash liquor comprising water and the enzymatic detergent composition. The wash liquor may be applied to the substrate or the substrate may be immersed wholly or partially into the wash liquor

The cleaning process may alternatively comprise direct application of the enzymatic detergent composition (undissolved i.e. without addition of water) on to part or whole of the fabric, so as to directly treat a stain or stains on the fabric. Such a process is preferably a pre-treatment process, so may be followed by treatment with/in a wash liquor (e.g. as a `main' wash process). The main wash is preferably according to the second aspect of the invention.
Preferably the process duration is less than 60 minutes, more preferably less than 30 minutes. If it is a pre-treatment process, the pre-treatment step is preferably less than 5 mins, and more preferably less than 2 minutes (although cleaning by the enzyme so applied will continue during at least a part of any washing process which follows).
The synergistic combination of the invention is radically improved at low temperature where cleaning of oil and fat stain/soil is more problematic.
Preferably the wash liquor temperature of the process is less than 40°C and preferably less than 30°C and more preferably less than 25°C at all times. Low temperature wash liquor is advantageous environmentally and financially.
The enzymatic detergent composition is preferably a low temperature composition. Accordingly, the enzymatic detergent composition is preferably packaged with instructions to treat at low temperatures, the low temperatures being preferably less than 40°C, more preferably less than 30°C even more preferably less than 25°C.
The invention provides enzymatic performance of oily soil and/or stains in a low temperature cleaning processes (with low temperature wash liquor) without serious consideration to the temperature sensitivity of the enzyme. The enzyme can therefore be selected more freely, on the basis of other considerations.
The invention is especially advantageous for the particular situation where one requires enzymatic cleaning of oily soil and/or stains in a low temperature cleaning processes (with low temperature wash liquor) but where compositions are by necessity stored at higher temperatures. Psychrophilic enzymes are effective at low temperatures but are sensitive to raised temperatures due to their flexibility. Mesophilic (and thermophilic) enzymes are stable at raised temperatures, but have reduced performance. The invention affords low temperature enzymatic cleaning of a substrate using mesophilic enzymes without needing to expend effort in engineering psychrophilic enzymes which can withstand raised temperatures.
Accordingly, the enzyme system preferably comprises a mesophilic or thermophilic enzyme system. The enzyme system may even be a mesophilic and/or thermophilic enzyme system with the exclusion of pyschrophilic enzymes.
Enzymes may be from bacterial origin (derived from bacteria) or fungal origin (derived from fungus) however enzymes from bacterial origin are preferred.
The composition preferably comprises between 1 to 70 wt % of a surfactant, most preferably 10 to 30 wt %.
Preferably the surfactant system comprises at least 1 wt% (based on the cleaning composition) of a biosurfactant.
Preferably the biosurfactant is of bacterial origin.
Preferably the biosurfactant and the enzyme is of bacterial origin.
Chemically modified or protein engineered mutants are included.
The one or more enzymes may be provided as a system.
Preferably the one or more enzymes comprises a lipase. Preferred lipases include lipases from Humicola (synonym Thermomyces), e.g. from H. lanuginosa (T. lanuginosus) or from H. insolens, a Pseudomonas lipase, e.g. from P. alcaligenes or P. pseudoalcaligenes, P. cepacia, P. stutzeri, P. fluorescens, Pseudomonas sp. strain SD 705 ( WO 95/06720 and WO 96/27002 ), P. wisconsinensis, a Bacillus lipase, e.g. from B. subtilis (Dartois et al. (1993), Biochemica et Biophysica Acta, 1131, 253-360), B. stearothermophilus ( JP 64/744992 ) or B. pumilus ( WO 91/16422 ).
Commercially available lipase enzymes include Lipolase™ and Lipolase Ultra™, Lipex™ (Novozymes A/S) and the Bacterial enzyme, Lipomax® ex Genecor. This is a bacterially derived Lipase, of variant M21L of the lipase of Pseudomonas alcaligenes as described in WO 94/25578 to Gist-Brocades (M.M.M.J. Cox, H.B.M. Lenting, L.J.S.M. Mulleners and J.M. van der Laan ).
Preferred Phospholipases (EC 3.1.1.4 and/or EC 3.1.1.32) include enzymes which hydrolyse phospholipids. Phospholipases A₁ and A₂ which hydrolyze one fatty acyl group (in the sn-1 and sn-2 position, respectively) to form lysophospholipid; and lysophospholipase (or phospholipase B) which can hydrolyze the remaining fatty acyl group in lysophospholipid are included as are Phospholipase C and phospholipase D (phosphodiesterases)which release diacyl glycerol or phosphatidic acid respectively.
The term "phospholipase A" used herein in connection with an enzyme of the invention is intended to cover an enzyme with Phospholipase A₁ and/or Phospholipase A₂ activity. The phospholipase activity may be provided by enzymes having other activities as well, such as, e.g., a lipase with phospholipase activity.
The phospholipase may be of any origin, e.g., of animal origin (such as, e.g., mammalian), e.g. from pancreas (e.g., bovine or porcine pancreas), or snake venom or bee venom. Preferably the phospholipase may be of microbial origin, e.g., from filamentous fungi, yeast or bacteria, such as the genus or species Aspergillus, e.g., A. niger, Dictyostelium, e.g., D. discoideum; Mucor, e.g. M. javanicus, M. mucedo, M. subtilissimus; Neurospora, e.g. N. crassa; Rhizomucor, e.g., R. pusillus; Rhizopus, e.g. R. arrhizus, R. japonicus, R. stolonifer; Sclerotinia, e.g., S. libertiana; Trichophyton, e.g. T. rubrum; Whetzelinia, e.g., W. sclerotiorum; Bacillus, e.g., B. megaterium, B. subtilis; Citrobacter, e.g., C. freundii; Enterobacter, e.g., E. aerogenes, E. cloacae Edwardsiella, E. tarda; Erwinia, e.g., E. herbicola; Escherichia, e.g., E. coli; Klebsiella, e.g., K. pneumoniae; Proteus, e.g., P. vulgaris; Providencia, e.g., P. stuartii; Salmonella, e.g. S. typhimurium; Serratia, e.g., S. liquefasciens, S. marcescens; Shigella, e.g., S. flexneri; Streptomyces, e.g., S. violeceoruber, Yersinia, e.g., Y. enterocolitica. Thus, the phospholipase may be fungal, e.g., from the class Pyrenomycetes, such as the genus Fusarium, such as a strain of F. culmorum, F. heterosporum, F. solani, or a strain of F. oxysporum. The phospholipase may also be from a filamentous fungus strain within the genus Aspergillus, such as a strain of Aspergillus awamori, Aspergillus foetidus, Aspergillus japonicus, Aspergillus niger or Aspergillus oryzae.
Preferred phospholipases are derived from a strain of Humicola, especially Humicola lanuginosa or variant; and from strains of Fusarium, especially Fusarium oxysporum. The phospholipase may be derived from Fusarium oxysporum DSM 2672.
Preferably phospholipases comprise a phospholipase A₁ (EC. 3.1.1.32). or a phospholipase A₂ (EC.3.1.1.4.).
Examples of commercial phospholipases include LECITASE™ and LECITASE™ ULTRA, YIELSMAX, or LIPOPAN F (available from Novozymes A/S, Denmark).
Commercially available protease enzymes include Alcalase™, Savinase™, Primase™, Duralase™, Dyrazym™, Esperase™, Everlase™, Polarzyme™, and Kannase™, (Novozymes A/S), Maxatase™, Maxacal™, Maxapem™, Properase™, Purafect™, Purafect OxP™, FN2™, and FN3™ (Genencor International Inc.).
Other enzymes may be selected from the group comprising: cellulases, esterases, peroxidases/oxidases, oxidoreductases, pectases, lyases, mannanases and mixtures thereof.
Bacterial enzymes for use in the invention are cellulases, esterases, peroxidases/oxidases, pectases, lyases, and mannanases, or mixtures thereof. Bacterial genes encoding such enzymes can be transferred to preferred expression production hosts, which are not limited to bacterial and includes 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 bacterial cutinase is that from 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 an outer (sn-1) and the middle (sn-2) positions and esterified with phosphoric acid in the third position; the phosphoric acid, in turn, may 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 which hydrolyze one fatty acyl group (in the sn-1 and sn-2 position, respectively) to form lysophospholipid; and lysophospholipase (or phospholipase B) which can hydrolyze the remaining fatty acyl group in lysophospholipid. Phospholipase C and phospholipase D (phosphodiesterases) release diacyl glycerol or phosphatidic acid respectively.
The term phospholipase includes enzymes with phospholipase activity, e.g., phospholipase A (A₁ or A₂), phospholipase B activity, phospholipase C activity or phospholipase D activity. The term "phospholipase A" used herein in connection with an enzyme of the invention is intended to cover an enzyme with Phospholipase A₁ and/or Phospholipase A₂ activity. The phospholipase activity may be provided by enzymes having other activities as well, such as, e.g., a lipase with phospholipase activity. The phospholipase activity may, e.g., be from a lipase with phospholipase side 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 side activity.
Preferably, the phospholipase is of bacterial origin Bacillus, e.g., B. megaterium, B. subtilis; Citrobacter, e.g., C. freundii; Enterobacter, e.g., E. aerogenes, E. cloacae Edwardsiella, E. tarda; Erwinia, e.g., E. herbicola; Escherichia, e.g., E. coli; Klebsiella, e.g., K. pneumoniae; Proteus, e.g., P. vulgaris; Providencia, e.g., P. stuartii; Salmonella, e.g. S. typhimurium; Serratia, e.g., S. liquefasciens, S. marcescens; Shigella, e.g., S. flexneri;
Suitable cellulases are especially of bacterial origin. Chemically modified or protein engineered mutants are included. Suitable cellulases include cellulases from the genera Bacillus, Pseudomonas and Clostridia.
Suitable peroxidases/oxidases are especially of bacterial origin. Chemically modified or protein engineered mutants are included. An example of an oxidative bacterium is, but not limited to, are Aeromonas sp wherefrom oxidases can be sourced.
Examples of pectate lyases include pectate lyases that have been cloned from different bacterial genera such as Erwinia, Pseudomonas, Klebsiella and Xanthomonas, as well as from 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 several 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) describes a beta-mannanase derived from Bacillus subtilis. JP-A-03047076 discloses a beta-mannanase derived from Bacillus sp. JP-A-63056289 describes the production of an alkaline, thermostable beta-mannanase. JP-A-63036775 relates to the Bacillus microorganism FERM P-8856 which produces beta-mannanase and beta-mannosidase. JP-A-08051975 discloses alkaline beta-mannanases from alkalophilic Bacillus sp. AM-001. A purified mannanase from Bacillus amyloliquefaciens is disclosed in WO 97/11164 . WO 91/18974 describes a hemicellulase such as a glucanase, xylanase or mannanase active. Bacillus sp. mannanases concerned in the Examples in WO 99/64619 .
The composition may further comprise other enzymes of bacterial origin and/or enzymes that are not of bacterial origin.

Lignin compounds

Preferably the lignin compound comprises a lignin polymer and more preferably it is a modified lignin polymer. As used herein "a modified lignin polymer " is intended to mean lignin that has been subjected to a chemical reaction to covalently attach chemical moieties to the lignin. The attached chemical moieties are usually randomly substituted.
Preferred modified lignin polymers are lignins substituted with anionic, cationic or alkoxy groups, or mixtures thereof. Preferably the substitution occurs on the aliphatic portion of the lignin and is random. Preferably the modified lignin polymer is substituted with an anionic group, and preferably it is a sulfonate. A preferred cationic group is a quanternary amine. Preferred alkoxy groups are polyalkylene oxide chains having repeat units of alkoxy moieties in the range from 5 to 30, most preferably ethoxy. Preferably the modified lignin sulfonate is substituted with anionic or alkoxy groups. Modified lignin polymers are discussed in WO/2010/033743 . Most preferably the modified lignin polymer is lignin sulfonate (lignosulfonate). Lignin sulfonate may be obtained by the Howard process.
Exemplary lignin sulfonate may be obtained from a variety of sources including hardwoods, softwoods and recycling or effluent streams. The lignin sulfonate may be utilized in crude or pure forms, e.g., in an "as is" or whole liquor condition, or in a purified lignin sulfonate form from which or in which sugars and other saccharide constituents have been removed or destroyed, or from which or in which inorganic constituents have been partially or fully eliminated. The lignin sulfonate may be utilized in salt forms including calcium lignin sulfonate, sodium lignin sulfonate, ammonium lignin sulfonate, potassium lignin sulfonate, magnesium lignin sulfonate and mixtures or blends thereof.
The lignin sulfonate preferably has a weight average molecular weight of from 2000 to 100000. Their basic structural unit is phenylpropane. The degree of sulfonation is preferably from 0.3 and 1.0 sulfate groups per phenylpropane unit.
Commercially available Lignin sulfonates include Ultrazine from Borregaard LignoTech. Other suppliers include Georgia-Pacific Corporation, Lenzing AG and Tembec Inc. Lignin sulfonates are discussed in Lauten, R. A., Myrvold, B. O. and Gundersen, S. A. (2010) New Developments in the Commercial Utilization of Lignosulfonates, in Surfactants from Renewable Resources (eds M. Kjellin and I. Johansson), John Wiley & Sons, Ltd, Chichester, UK.

Surfactant

Surfactants

a) Bacterially derived Biosurfactants

Preferably the biosurfactant comprises a Rhamnolipid, which may be derived from Pseudomonas sp.
Other bacterially derived biosurfactants are available from "Mapping of Patents in Bioemulsifiers and biosurfactants - review, published in the Journal of Scientific and . Within the definition of bacterially produced biosurfactants, we include those where a bacterial gene is cloned and subsequently expressed from another organism as a manufacturing technique. For example, Rhamnolipids have been produced from E. coli in this way.

b) Biosurfactants from non-bacterial sources

Biosurfactants within the scope of this invention may also be derived from yeasts and fungi.
Biosurfactants from non-bacterial microbial sources include those derived from fungi and yeasts, e.g. sophorolipids 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, Towards 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.
Further information is given in Production, Characterisation and Applications of Biosurfactants Review - Biotechnology - Volume 7, 2008, page 370: Pattanathu, Rahman and Gakpe.
Surfactants not generally classified as "biological" may also be included in the invention.
Nonionic surfacants include, in particular, the reaction products of compounds having a hydrophobic group and a reactive hydrogen atom, for example, aliphatic alcohols, acids, amides or alkyl phenols with alkylene oxides, especially ethylene oxide either alone or with propylene oxide. Specific nonionic detergent compounds are C₆ to C₂₂ alkyl phenol-ethylene oxide condensates, generally 5 to 25 EO, i.e. 5 to 25 units of ethylene oxide per molecule, and the condensation products of aliphatic C₈ to C₁₈ primary or secondary linear or branched alcohols with ethylene oxide, generally 5 to 40 EO.
Nonionic detergent compounds which may be used are usually water-soluble alkali metal salts of organic sulphates and sulphonates having alkyl radicals containing from about 8 to about 22 carbon atoms, the term alkyl being used to include the alkyl portion of higher acyl radicals. Examples of suitable synthetic anionic detergent compounds are sodium and potassium alkyl sulphates, especially those obtained by sulphating higher C₈ to C₁₈ alcohols, produced for example from tallow or coconut oil, sodium and potassium alkyl C₉ to C₂₀ benzene sulphonates, particularly sodium linear secondary alkyl C₁₀ to C₁₅ benzene sulphonates; and sodium alkyl glyceryl ether sulphates, especially those ethers of the higher alcohols derived from tallow or coconut oil and synthetic alcohols derived from petroleum. The preferred anionic detergent compounds are sodium C₁₁ to C₁₅ alkyl benzene sulphonates and sodium C₁₂ to C₁₈ alkyl sulphates. Also applicable are surfactants such as those described in EP-A-328 177 (Unilever), which show resistance to salting-out, the alkyl polyglycoside surfactants described in EP-A-070 074 , and alkyl monoglycosides.
Preferred surfactant systems are mixtures of anionic with nonionic detergent active materials, in particular the groups and examples of anionic and nonionic surfactants pointed out in EP-A-346 995 (Unilever). Especially preferred is surfactant system that is a mixture of an alkali metal salt of a C₁₆ to C₁₈ primary alcohol sulphate together with a C₁₂ to C₁₅ primary alcohol 3 to 7 EO ethoxylate.
The nonionic detergent is preferably present in amounts greater than 10%, e.g. 25 to 90 wt % of the surfactant system. Anionic surfactants can be present for example in amounts in the range from about 5% to about 40 wt % of the surfactant system.
The detergent composition may comprise other ingredients commonly found in laundry liquids. Especially polyester substantive soil release polymers, hydrotropes, opacifiers, colorants, perfumes, other enzymes, other surfactants, microcapsules of ingredients such as perfume or care additives, softeners, polymers for anti redeposition of soil, bleach, bleach activators and bleach catalysts, antioxidants, pH control agents and buffers, thickeners, external structurants for rheology modification, visual cues, either with or without functional ingredients embedded therein and other ingredients known to those skilled in the art.
The invention will now be further described with reference to the following nonlimiting examples.

Examples

All values throughout are wt%.

Detergent formulation A

Ingredient	% by weight
Non-ionic surfactant Neodol 25-7	6.2
Anionic surfactant LAS acid	11.8
Anionic surfactant SLES 3EO	6.5
Lauric Fatty Acid P5908	5.2
Glycerol	5.0
Monopropylene Glycol	9.0
Citric acid	3.9
Minors	2.0
Water	balance to 100

Wherein:

Neodol 25-7 ex.Shell = C₁₂-C₁₅ alcohol 7-ethoxylate
LAS acid = C₁₀-C₁₄ alkyl benzene sulphonic acid;
SLES = C12-C13 alcohol 3-ethoxylate sulphate, Na salt: = sodium lauryl ether sulphate (with on average 3 ethylene oxide groups);

Lipolytic Enzyme (lipase)

Bacterial enzyme is Lipomax® ex Genecor. This is a bacterially derived Lipase, of variant M21L of the lipase of Pseudomonas alcaligenes as described in WO 94/25578 to Gist-Brocades (M.M.M.J. Cox, H.B.M. Lenting, L.J.S.M. Mulleners and J.M. van der Laan).

Rhamnolipid

The rhamnolipid is RBR425 (25% AM) ex Jeneil Biosurfactant Company.

Lignosulphonate

Lignosulphonate is Ultrazine NA ex Borregaard LignoTech.

Example 1

In this example, enzymatic detergent formulations according to the invention were tested to determine their ability to treat i.e. remove beef fat stains from cotton fabric.
CS61 (ex. CFT B.V. Vlaardingen, the Netherlands) which is coloured beef fat stain on cotton, was cut into round discs with a 96 well fabric punch and placed in the wells of a 96 micro titre well plate. Stains were washed in formulations in different combinations of:

(i) biosurfactant being rhamnolipid (RL) solution (solvent water) 0.9 g/L
(ii) lignin sulphonate (LS) solution (solvent water) - 3 concentrations: 10g/L; 5g/L; 2.5 g/L
(iii) bacterial lipase is 10mg/L when added : 172g of lipomax granules are added to 50mls water to make a 100mg/L concentration stock solution which is then diluted in the well to give final concentration of 10g/L.
(iv) detergent A formulation dissolved in water to give 6g/L stock solution.

The micro titre well layout was as follows 200µl total volume in well):

1) Detergent A 100%:- 100µl of detergent A (6g/L stock), 80µl water, 20µl enzyme (20µl water in no enzyme control wells)
2) Detergent A 70% & Rhamnolipid 0.9g/L 70µl A 6g/L stock, 30µl 24g/L Rhamnolipid (25% active), 80µl water, 20µl enzyme (20µl water in no enzyme control)
3) Detergent A 70% & Rhamnolipid 0.9g/L & 10g/L Sodium Lignosulphonate - 70µl Detergent A 6g/L stock, 30µl 24g/L Rhamnolipid (25% active), 80µl Sodium lignosulphonate 25g/L stock, 20µl enzyme (20µl water in no enzyme control)
4) Detergent A 70% & Rhamnolipid 0.9g/L & 5g/L Sodium Lignosulphonate - 70µl Detergent A 6g/L stock, 30µl 24g/L Rhamnolipid (25% active), 80µl Sodium lignosulphonate 12.5g/L stock, 20µl enzyme (20µl water in no enzyme control)
5) Detergent A 70% & Rhamnolipid 0.9g/L & 2.5g/L Sodium Lignosulphonate - 70µl Detergent A 6g/L stock, 30µl 24g/L Rhamnolipid (25% active), 80µl Sodium lignosulphonate 6.25g/L stock, 20µl enzyme (20µl water in no enzyme control)
6) Detergent A 100% & 10g/L Sodium Lignosulphonate - 100µl Detergent A 6g/L stock, 80µl Sodium lignosulphonate 25g/L stock, 20µl enzyme (20µl water in no enzyme control)
7) Detergent A 100% & 5g/L Sodium Lignosulphonate - 100µl Detergent A 6g/L stock, 80µl Sodium lignosulphonate 12.5g/L stock, 20µl enzyme (20µl water in no enzyme control)
8) Detergent A 100% & 2.5g/L Sodium Lignosulphonate - 100µl Detergent A 6g/L stock, 80µl Sodium lignosulphonate 6.25g/L stock, 20µl enzyme (20µl water in no enzyme control)

Washes were carried out at room temperature, at 1400 rpm agitation for an hour hour in an incubator shaker. Following the washing operation, the fabric discs were rinsed twice with 200µL demineralised water, prior to drying at room temperature in the dark overnight.
Stain removal from the fabric was measured at 410 nm using a flatbed remission spectrophotometer after the wash. The results are expressed as delta remission, which was generated using the CIEL*a*b (CIELAB) values generated using the Hunterlab Ultrascan VIS remission spectrophotometer.

Results are shown in Table 1:

Table 1 is graphically illustrated in Fig1.

		1	2	3	4	5	6	7	8

with Lipomax	average	37.64	44.85	58.04	53.58	49.09	39.66	37.60	37.77
	std dev	0.56	1.35	1.75	1.78	1.63	0.92	0.84	0.42

without Lipomax	average	37.15	36.10	35.86	35.42	35.97	35.67	36.27	36.30
	std dev	1.03	1.13	0.59	0.25	0.87	0.97	1.08	0.45

The results show that lignin sulphonate improves stain removal by a lipolytic enzyme (exemplified by Lipomax) at low temperature especially when a biosurfactant (exemplified by rhamnolipid) is incorporated.

Example 2

In this example, various enzyme/biosurfactant/lignin sulphonate compositions were examined to determine their ability to remove a range of fat and oil stains. The stains were (lard and violet dye and pigment vegetable fat stain -under medium scale wash conditions in tergotometers).

Stains used:

4x 1cm multi stain on woven cotton: lard and violet dye, ragu and 5% sunflower oil, green curry and instant gravy (only results for lard and violet dye are included), stains from Warwick Equest Limited.
7 x 7cm pigment vegetable fat stain on polyester from (ex. CFT B.V. Vlaardingen, the Netherlands)

Stains were washed together with woven cotton ballast (total cloth load 20g, 1:50 cloth to liquor ratio by weight) in duplicate in 1 Litre tergotometers at 20 °C (final temperature 23 °C. for 30 mins, 100 rpm agitation. Stains were washed in formulations in different combinations of:

(v) biosurfactant being rhamnolipid (RL) - 0.9 g/L when added
(vi) lignin sulphonate (LS)- 3 concentrations: 10, 5, 2.5 g/L when added
(vii) bacterial lipase 10mg/L when added
(viii) detergent A formulation dissolved in water to give varying concentrations of stock solutions.

The quantities of stock solutions added to the tergotometers were as follows:

1) Detergent A 100% : 20ml Detergent A (150g/L stock solution), 10ml Lipomax (1g/L) or 10ml water for the no enzyme control solutions
2) Detergent A 100% & 10g/L Sodium Lignosulphonate - 20ml Detergent A (150g/L), 10 grams sodium lignosulphonate and 10ml Lipomax (make up x100 stock at 1g/L) or 10ml water for the no enzyme control solutions
3) Detergent A & 70% Rhamnolipid 0.9g/L - 14ml Detergent A stock (150g/L), 50ml of Rhamnolipid stock (72g/L) and 10ml Lipomax stock (1g/L) or 10ml water for the no enzyme control solutions
4) Detergent A 70% Rhamnolipid 0.9g/L and 10g/L Sodium Lignosulphonate - 14ml Detergent A stock (150g/L), 50ml of Rhamnolipid stock (72g/L), 10 grams of Sodium Lignosulphonate and 10ml Lipomax stock (see above) or 10ml water for the no enzyme control solutions

Washes were carried out at room temperature, at 1400 rpm agitation for an hour. Following the washing operation, the fabric discs were rinsed twice with 200 µL demineralised water, prior to drying at room temperature in the dark overnight.

Colour remission of the stains was measured at 410nm using a Hunterlab Ultrascan VIS remission spectrophotometer before and after wash. The results are expressed as delta remission, which was generated using the CIEL*a*b (CIELAB) values generated.

Table 2 (illustrated in figure 2)

	Lard and violet dye
		MTS
100%	MTS	100% 10g/L NaL	MTS	70%/ Rham 0.9g/L	MTS	70%/ Rham 0.9g/L, 10g/L NaL
with Lipomax	Average	18.74	9.58	27.48	36.54
	Std Dev	1.11	2.76	3.01	3.88

Control	Average	12.73	9.56	13.32	12.78
	Std Dev	2.03	2.65	2.35	2.44

Table 3 (illustrated in figure 3)

		MTS 100%	MTS	100% 10g/L NaL	MTS	70%/ Rham 0.9g/L	MTS	70%/ Rham 0.9g/L, 10g/L NaL

with Lipomax	Average	72.64	70.59	70.87	74.44
	Std Dev	0.67	1.29	0.79	1.13

		MTS 100%	MTS	100% 10g/L NaL	MTS	70%/ Rham 0.9g/L	MTS	70%/ Rham 0.9g/L, 10g/L NaL

no Lipomax	Average	63.19	63.45	64.03	65.67
	Std Dev	0.49	0.69	0.77	0.91

The results show that lignin sulphonate improves lipase performance in a detergent composition especially when a rhamnolipid is incorporated.

Claims

An enzymatic detergent composition comprising the combination of :
(i) a surfactant system;

(ii) one or more enzymes; and

(iii) one or more lignin compounds, wherein the surfactant system comprises a biosurfactant.
An enzymatic detergent composition according to claim wherein the composition is packaged with instructions to treat at low temperatures, the low temperatures being less than 40°C and preferably less than 30°C and more preferably less than 25°C.
An enzymatic detergent composition according to claim 1 or claim 2 which comprises a mesophilic or thermophilic enzyme system.
An enzymatic detergent composition according to any preceding claim wherein the biosurfactant and the or each enzyme is bacterially derived.
A process for treating a substrate, comprising the steps of treating the substrate with the enzymatic detergent composition of claims 1 - 4.
A process according to claim 5 in which the enzymatic detergent composition is directly applied on to part or whole of the fabric, so as to treat a stain or stains on the fabric.
A process according to claim 5 or 6 wherein the substrate comprises a fabric.
A process according to any of claims 5 - 7 wherein the process duration is less than 60 minutes, preferably less than 30 minutes.
A process according to any of claims 5 - 8 wherein the wash liquor temperature of the process is less than 40°C and preferably less than 30°C and more preferably less than 25°C at all times.