EP4189051B1

EP4189051B1 - Use of an enzyme and surfactant for inhibiting microorganisms

Info

Publication number: EP4189051B1
Application number: EP21751536.0A
Authority: EP
Inventors: Gurpreet Singh Kohli; Srilaxmi Venkata Medepalli; Yugandhar B.S. REDDY; Asha Telkar
Original assignee: Unilever Global IP Ltd; Unilever IP Holdings BV
Current assignee: Unilever Global IP Ltd; Unilever IP Holdings BV
Priority date: 2020-07-27
Filing date: 2021-07-26
Publication date: 2024-02-28
Anticipated expiration: 2041-07-26
Also published as: WO2022023250A1; EP4189051A1; CN116057158A

Description

Field of the invention

The present invention relates to the use of an enzyme and a surfactant in a detergent composition for inhibiting microorganisms. The present invention particularly relates to the use of an enzyme and a surfactant in detergent composition for antiviral activity.

Background of the invention

Human health is impacted by a variety of microorganisms encountered on a daily basis. Viruses and bacteria cause a wide variety of sicknesses and ailments.
Sanitizing or disinfecting skin and inanimate surfaces is a very important aspect for ensuring healthy condition. Areas of focus for ensuring disinfection include personal uses like hand and body hygiene and hygiene of hard surfaces like doorknobs and soft surfaces such as clothing.
Bacteria, virus and protozoa are the three common microorganisms known to cause diseases in human and other mammals. Cleaning skin and other animate and inanimate surfaces to reduce microbial populations is a first defense in removing such pathogens and minimizing the risk of infection. There are compositions available for disinfection against each of these types of organisms. While many antibacterial actives and compositions are available and widely used, killing of virus is more difficult and often requires harsher chemicals like chlorine or alcohol.
Viruses are a category of pathogens of primary concern and viral infections are among the greatest causes of human morbidity. Virus infections of respiratory tract are generally spread from person to person by direct contact with virus-contaminated respiratory secretions. Typically, this contact is in the form of physical contact with a contaminated surface or via inhalation of airborne viral particles. Viruses can survive on environmental surfaces for hours after initial contamination, and infection is readily transmitted by finger-to-finger contact, and by contaminated environmental surface-to-finger contact, if the newly contaminated finger is then used to rub an eye or touch the nasal mucosa. Therefore, minimizing virus contamination of skin and environmental surfaces which includes inanimate and animate surfaces may prove to be effective in reducing the risk of transmitting the infection to the general population.
Viruses propagate only within living cells. The principal obstacle encountered by a virus is gaining entry into the cell, which is protected by a cell membrane of thickness comparable to the size of the virus. In order to penetrate a cell, a virus first must become attached to the cell surface. Therefore, in order to control a viral infection, it is important to rapidly kill a virus that contacts the skin, and ideally to provide a persistent antiviral activity on the skin, or inanimate surface, in order to control viral infections.
For example, rhinoviruses, influenza viruses, coronaviruses and adenoviruses are known to cause respiratory infections. Coronaviruses primarily infect the respiratory tract and gastrointestinal tract of mammals and birds. Coronaviruses are enveloped viruses with a positive-sense single-stranded RNA genome and with a nucleocapsid of helical symmetry. Coronavirus infection begins with attachment of the spike protein with its cognate cell receptor. It is desirable find new ways capable of deactivating viruses specifically coronaviruses.
Virus control poses a more difficult problem than bacterial control. By sufficiently reducing bacterial populations, the risk of bacterial infection is reduced to acceptable levels. Therefore, a rapid antibacterial kill is desired. With respect to viruses, however, not only is a rapid kill desired, but a persistent antiviral activity also is required. This difference is because merely reducing a virus population is insufficient to reduce infection. In theory, a single virus can cause infection. Therefore, an essentially total, and persistent, antiviral activity is required, or at least desired, for an effective antiviral cleansing composition.
EP1065265 B1 (Kao Corporation ) discloses a germicidal detergent composition having a pH of 6.5 to 7.5 and which includes protease.
WO 2010/069812 A2 (Henkel ) discloses a process for disinfecting textiles and or hard surfaces by contacting with a virucidal treatment solution having at least one hydrolytic enzyme.
WO98/50512 A1 (Procter & Gamble ) discloses a laundry or cleaning product having a hexosaminidase enzyme which shows antimicrobial activity.
WO 2017/207770 A1 (Novozymes ) discloses a cleaning detergent composition having an enzyme with deoxyribonuclease activity and an enzyme having hexosaminidase activity for deep cleaning e.g., reduction/removing of biofilm components from a textile surface.
US 2019/024024 A1 (Sava Alex ) discloses a liquid composition having high level of disinfection to treat laundries and includes an enzyme, a biocidal quaternary ammonium biocide and an anionic hydrotrope.
While detergent compositions which generally have pH in the range from 8 to 13 are known to impart stain removal benefits, they were not known to impart antimicrobial, particularly antibacterial and viral inactivation benefits.
Accordingly, it is an object of the present invention to achieve inactivation of microorganisms particularly viruses using a detergent composition having a pH from 10 to 13 from an article during a laundering process.
A further object is that the composition does not contain harsh chemicals which lead to undesirable affects that can be considered to be harmful to the consumer.

Summary of the invention

We have now found that the use of a combination of alkyl benzene sulphonate surfactant, hydrolytic enzyme and an alkaline source in a detergent composition having a pH from 8 to 13 provides for inactivation of microorganisms on a textile article. Particularly for inactivation of bacteria and viruses.
The invention in a first aspect relates to the use of a combination of alkyl benzene sulphonate surfactant, hydrolytic enzyme and an alkaline source in a solid detergent composition having a pH from 10 to 13 as measured at 25°C and 10% aqueous concentration in deionised water for inactivating microorganisms on a textile article during a laundering process wherein the hydrolytic enzyme comprises a serine protease and wherein the solid detergent composition does not contain a peroxygen bleach and wherein the alkaline source comprises from 10 wt.% to 40 wt.% sodium carbonate and wherein the wash liquor prepared by adding the solid detergent composition in water has an alkaline source concentration ranging from 200 ppm to 2500 ppm.
In a second aspect, the invention provides a method for inactivation of microorganisms from a textile article, the method comprising the steps of:
i) applying a solid detergent composition to the textile article, wherein the solid detergent composition comprises a combination of alkyl benzene sulphonate, hydrolytic enzyme comprising a serine protease and an alkaline source and the detergent composition has a pH from 10 to 13; ii) allowing the textile article to be in contact with the solid detergent composition for a period of at least 30 minutes, still preferably at least one hour; wherein the solid detergent composition does not contain a peroxygen bleach and wherein the alkaline source comprises from 10 wt.% to 40 wt.% sodium carbonate and wherein the wash liquor prepared by adding the solid detergent composition in water has an alkaline source concentration ranging from 200 ppm to 2500 ppm.
As used herein, the term "solid detergent composition" includes granular, powder, tablets, or bar composition. The composition is a solid laundry detergent composition.

Detailed description of the invention

All percentages mentioned herein are by weight calculated on the total composition, unless specified otherwise. The abbreviation 'wt%' is to be understood as % by weight of the total composition.

Alkyl benzene sulphonate surfactant

According to the first aspect of the present invention, provided is the use of alkyl benzene sulphonate surfactant in a detergent composition.
Linear alkylbenzene sulfonate or LAS are linear alkylbenzenes that has been sulfonated to include an acidic sulfonate group attached to the benzene ring to form a parent acid, linear alkylbenzene sulfonic acid. The linear alkylbenzene sulfonic acid by neutralization using any of alkali metal hydroxides, alkaline earth hydroxides, ammonium hydroxides, alkylammonium hydroxides, alkanolamine or any chemical agent known by those skilled in the art forms water-soluble linear alkylbenzene sulfonates.
The composition comprises alkyl benzene sulphonate, preferably a linear or branched, substituted or unsubstituted, C₈ to C₂₄ alkyl benzene sulphonate. The C₈ to C₂₄ alkyl benzene sulphonate can be a modified alkylbenzene sulphonate (MLAS) as described in more detail in WO 99/05243 , WO 99/05242 , WO 99/05244 , WO 99/05082 , WO 99/05084 , WO 99/05241 , WO 99/07656 , WO 00/23549 , and WO 00/23548 . Highly preferred C₈ to C₂₄ alkyl benzene sulphonates are linear C₁₀ to C₁₆ alkylbenzene sulphonates. Especially preferred are linear C₁₀ to C₁₃ alkylbenzene sulphonates that are obtainable, preferably obtained, by sulphonating commercially available linear alkyl benzenes (LAB); suitable LAB include low 2-phenyl LAB, such as those supplied by Sasol under the tradename Isochem^® or those supplied by Petresa under the tradename Petrelab^®, other suitable LAB include high 2-phenyl LAB, such as those supplied by Sasol under the tradename Hyblene^®. Preferably, the composition comprises alkyl benzene sulphonate, wherein the alkyl benzene sulphonate comprises at least 25 wt.% of the 2-phenyl isomer. A suitable alkyl benzene sulphonate having this feature is obtained by DETAL synthesis.
Examples of suitable synthetic anionic detergent compounds are sodium and potassium salts, especially those obtained by sulphating alcohols, produced for example from tallow or coconut oil, sodium and potassium alkyl C₁₀ to C₂₀ benzene sulphonates, particularly sodium linear secondary alkyl C₁₀ to C₁₅ benzene sulphonates; are sodium C₁₁ to C₁₅ alkyl benzene sulphonates.
Preferably the detergent composition according to the present invention comprises from 2 wt.% to 40 wt.% alkyl benzene sulphonate surfactant. Preferably the detergent composition comprises at least 6 wt.% alkyl benzene sulphonate surfactant based on the weight of the detergent composition, still preferably at least 7 wt.%, still preferably at least 8 wt.%, most preferably at least 10 wt.%, but typically not more than 35 wt.%, still preferably not more than 30 wt.%, more preferably not more than 25 wt.% and most preferably not more than 15 wt.% alkyl benzene sulphonate surfactant in the detergent composition.
Preferably the use according to the first aspect of the present invention involves inactivating the microorganisms in a wash liquor prepared by addition of the solid detergent composition in water, wherein the alkyl benzene sulphonate surfactant is present at a concentration from 40 ppm to 2000 ppm. Preferably concentration is at least 50 ppm, still preferably at least 100 ppm, still preferably at least 200 ppm, most preferably at least 250 ppm, but typically not more than 1600ppm, still preferably not more than 1000 ppm, more preferably not more than 800 ppm and most preferably not more than 600 ppm in the wash liquor.

Hydrolytic enzyme

According to the first aspect present invention discloses the use of a hydrolytic enzyme, wherein the hydrolytic enzyme comprises a serine protease, in a detergent composition for treating textile article for inactivation of microorganisms.
A hydrolytic enzyme is a hydrolase enzyme which hydrolytically cleaves esters, ethers, peptides, glycosides, acid anhydrides or C-C bonds in a reversible reaction. The hydrolytic enzyme catalyzes the hydrolytic cleavage of substances.
The hydrolytic enzyme can further comprise an enzyme selected from the group consisting of protease, lipase, cellulase, amylase, mannanase or combinations thereof. Preferred further enzyme for use in the present invention includes but is not limited to the group consisting of glycosidase, hemicellulases, xylanase, pectinase, glucosidase, carrageenase, or combination thereof. Preferably the further enzyme is a protease enzyme.
The hydrolytic enzyme is present in the solid detergent composition of the invention in a preferred level of from 0.0001 to about 1%, more preferably from about 0.001 to about 0.5% and especially from about 0.005 to about 0.6% of active hydrolytic enzyme.
Preferably the wash liquor prepared by diluting the solid detergent composition according to the present invention in water comprises from 0.0001 ppm to 30 ppm of pure hydrolytic enzyme, still preferably from 0.0001 ppm to 20 ppm of pure hydrolytic enzyme. Preferably the detergent composition comprises at least 0.0005 ppm, still preferably at least 0.001 ppm, still preferably at least 0.002 ppm, most preferably at least 0.005 ppm, still more preferably 0.4 ppm, but typically not more than 22 ppm, preferably not more than 20 ppm, still preferably not more than 15 ppm, most preferably not more than 10 ppm.

Protease:

Preferably the hydrolytic enzyme is a serine protease. Suitable proteases include those of bacterial, fungal, plant, viral or animal origin, preferably of vegetable or microbial origin. Microbial origin is preferred. Chemically modified or protein engineered mutants are included. Serine protease may for example be of the S1 family, such as trypsin, or the S8 family such as subtilisin. A metalloproteases protease may for example be a thermolysin from e.g., family M4 or other metalloprotease such as those from M5, M7 or M8 families.
Serine proteases are a subgroup of carbonyl hydrolases comprising a diverse class of enzymes having a wide range of specificities and biological functions. The term "subtilases" refers to a sub-group of serine protease according to Siezen et al., 1991, Protein Engng. 4: 719-737 and Siezen et al., 1997, Protein Science 6: 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; US 7,262,042 and WO 2009/021867 , and subtilisin lentus, subtilisin Novo, subtilisin Carlsberg, Bacillus licheniformis, subtilisin BPN', subtilisin 309, subtilisin 147 and subtilisin 168 described in WO 89/06279 and protease PD138 described in ( WO 93/18140 ). Other useful proteases may be those described in WO 92/175177 , WO 01/16285 , WO 02/026024 , US8753861B2 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 2005/040372 , and the chymotrypsin proteases derived from Cellumonas described in WO 2005/052161 and WO 2005/052146 .
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 metalloproteases are the neutral metalloprotease as described in WO 2007/044993 (Genencor Int. ) such as those derived from Bacillus amyloliquefaciens.
Examples of useful proteases are the variants described in: WO92/19729 , WO96/034946 , WO98/20115 , WO98/20116 , WO99/011768 , WO01/44452 , WO03/006602 , WO2004/03186 , WO2004/041979 , WO2007/006305 , WO201 1/036263 , WO2011/036264 , especially the variants with substitutions in one or more of the following positions: 3, 4, 9, 15, 27, 36, 57, 68, 76, 87, 95, 96, 97, 98, 99, 100, 101 , 102, 103, 104, 106, 1 18, 120, 123, 128, 129, 130, 160, 167, 170, 194, 195, 199, 205, 206, 217, 218, 222, 224, 232, 235, 236, 245, 248, 252 and 274 using the BPN' numbering. More preferred the subtilase variants may comprise the mutations: S3T, V41, S9R, A15T, K27R, *36D, V68A, N76D, N87S,R, *97E, A98S, S99G,D,A, S99AD, S101 G,M,R S103A, V104I,Y,N, S106A, G1 18V,R, H120D,N, N123S, S128L, P129Q, S130A, G160D, Y167A, R170S, A194P, G195E, V199M, V205I, L217D, N218D, M222S, A232V, K235L, Q236H, Q245R, N252K, T274A (using BPN' numbering).
Suitable commercially available protease enzymes include those sold under the trade names Alcalase^™, Duralase^™, Durazym^™, Relase^™, Relase^™ Ultra, Savinase^™, Savinase^™ Ultra, Primase^™, Polarzyme^™, Kannase^™, Liquanase^™, Liquanase^™ Ultra, Ovozyme^™, Coronase^™, Coronase^™ Ultra, Neutrase^™, Everlase^™ and Esperase^™ (Novozymes A/S), those sold under the tradename Maxatase^™, Maxaca^™, Maxapem^™, Purafect^™, Purafect Prime^™, Preferenz^™, Purafect MA^™, Purafect Ox^™, Purafect OxP^™, Puramax^™, Properase^™, Effectenz^™, FN2^™, FN3^™ , FN4^™, Excellase^™,, 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.

Lipase:

Lipase is an enzyme which catalyses hydrolysis of ester bonds of edible fats and oils, i.e. triglycerides, into free fatty acids, mono- and diglycerides and glycerol.
Cleaning lipases are discussed in Enzymes in Detergency edited by Jan H. Van Ee, Onno Misset and Erik J. Baas (1997 Marcel Dekker, New York). The lipase may be selected from lipase enzymes in E.C. class 3.1, 3.2 or a combination thereof.

Preferably the cleaning lipases selected is a Triacylglycerol lipases (E.C. 3.1.1.3). Suitable triacylglycerol lipases can be selected from variants of the Humicola lanuginosa (Thermomyces lanuginosus) lipase. Other suitable triacylglycerol lipases can be selected from variants of Pseudomonas lipases, e.g., from P. alcaligenes or P. pseudoalcaligenes (EP 218 272), P. cepacia (EP 331 376), P. stutzeri (GB 1,372,034), P. fluorescens, Pseudomonas sp. strain SD 705 (WO 95/06720 and WO 96/27002), P. wisconsinensis 25 (WO 96/12012), Bacillus lipases, 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).

Further examples of EC 3.1.1.3 lipases include those described in WIPO publications WO 00/60063 , WO 99/42566 , WO 02/062973 , WO 97/04078 , WO 97/04079 and US 5,869,438 . Preferred lipases are produced by Absidia reflexa, Absidia corymbefera, Rhizmucor miehei, Rhizopus deleman Aspergillus niger, Aspergillus tubigensis, Fusajum oxysporum, Fusarium heterosporum, Aspergillus oryzea, Penicilium camembertii, Aspergillus foetidus, Aspergillus niger, Thermomyces lanoginosus (synonym: Humicola lanuginosa) and Landerina penisapora, particularly Thermomyces lanoginosus.
Certain preferred lipases are supplied by Novozymes and includes those under the tradenames of Lipolase^®, Lipolase Ultra^®, Lipoprime^®, Lipoclean^® and Lipex^® (registered tradenames of Novozymes) and LIPASE P "AMANO^®" available from Areario Pharmaceutical Co. Ltd., Nagoya, Japan, AMANO-CES^®, commercially available from Toyo Jozo Co., Tagata, Japan; and further Chromobacter viscosum lipases from Amersham Pharmacia Biotech., Piscataway, New Jersey, U.S.A. and Diosynth Co., Netherlands, and other lipases such as Pseudomonas gladioli.
Other examples are lipase variants such as those described in EP407225 , WO92/05249 , WO94/01541 , WO94/25578 , WO95/14783 , WO95/30744 , WO95/35381 , WO95/22615 , WO96/00292 , WO97/04079 , WO97/07202 , WO00/34450 , WO00/60063 , WO01/92502 , WO07/87508 and WO09/109500 .
Preferred commercial lipase products include Lipolase^™, Lipex^™; Lipolex^™ and Lipoclean^™ (Novozymes A/S), Lumafast^™ (originally from Genencor) and Lipomax^™ (originally from Gist-Brocades). LIPEX^® is particularly preferred, and LIPEX^® 100 TB is further particularly preferred.
Additional useful lipases are described in WIPO publications WO 02062973 , WO 2004/101759 , WO 2004/101760 , and WO 2004/101763 . In one embodiment, suitable lipases include the "first cycle lipases" described in WO 00/60063 and U.S. Patent 6,939,702 BI, preferably a variant of SEQ ID No. 2, more preferably a variant of SEQ ID No. 2 having at least 90% homology to SEQ ID No. 2 comprising a substitution of an electrically neutral or negatively charged amino acid with R or K at any of positions 3, 224, 229, 231 and 233, with a most preferred variant comprising T23 IR and N233R mutations, such most preferred variant being sold under the tradename Lipex^® (Novozymes).
The aforementioned lipases can be used in combination (any mixture of lipases can be used). Suitable lipases can be purchased from Novozymes, Bagsvaerd, Denmark; Areario Pharmaceutical Co. Ltd., Nagoya, Japan; Toyo Jozo Co., Tagata, Japan; Amersham Pharmacia Biotech., Piscataway, New Jersey, U.S.A; Diosynth Co., Oss, Netherlands and/or made in accordance with the examples contained herein.
Lipase with reduced potential for odour generation and a good relative performance, are particularly preferred, as described in WO2007/087243 . These include lipoclean^® (Novozyme).

Amylase:

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, 21 1, 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, N190, 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; H156Y+A181T+N190F+A209V+Q264S; or G48A+T49I+G107A+H156Y+ A181T+N190F+I201 F+A209V+Q264S.
Further amylases which are suitable are amylases having SEQ ID NO: 6 in WO99/019467 or variants thereof having 90% sequence identity to SEQ ID NO: 6. Preferred variants of SEQ I D 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 I D 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 WO08/153815 , 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, 211 and 264.
Other examples are amylase variants such as those described in WO2011/098531 , WO2013/001078 and WO2013/001087 .
Commercially available amylases are Duramyl^™, Termamyl^™, Fungamyl^™, Stainzyme^™, Stainzyme Plus^™, Natalase^™, Liquozyme X^™ and BAN^™ (from Novozymes AS), and Rapidase^™, Purastar^™/Effectenz^™, Powerase^™, Preferenz S1000^™, Preferenz S100^™ and Preferenz S110^™ (from Genencor International Inc./DuPont).

Lyases:

The lyase may be a pectate lyase derived from Bacillus, particularly B. licheniformis or B. agaradhaerens, or a variant derived of any of these, e.g. as described in US 6124127 , WO 99/27083 , WO 99/27084 , WO 02/006442 , WO 02/092741 , WO 03/095638 , Commercially available pectate lyases are XPect^™; Pectawash^™ and Pectaway^™ (Novozymes A/S).

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 . A commercially available mannanase is Mannaway^™ (Novozymes A/S).

Cellulase:

Suitable cellulases include 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, Thielavia terrestris, Myceliophthora thermophila, and Fusarium oxysporum disclosed in US 4,435,307 , US 5,648,263 , US 5,691 ,178 , US 5,776,757 , WO 89/09259 , WO 96/029397 , and WO 98/012307 .
Commercially available cellulases include Celluzyme^™, Carezyme^™, Celluclean^™, Endolase^™, Renozyme^™ (Novozymes A/S), Clazinase^™ and Puradax HA^™ (Genencor International Inc.), and KAC-500(B)^™ (Kao Corporation).
Preferably the use according to the first aspect of the present invention is substantially free of hydrolytic enzyme selected from the group consisting of deoxyribonuclease, hexosaminidase or combination thereof. The term substantially free means that there is no intentionally added deoxyribonuclease and/or hexosaminidase hydrolytic enzyme in the composition, preferably their amount is 0 wt.%. It is highly preferred that all the hydrolytic enzyme is a detersive hydrolytic enzyme.

Alkaline source

According to the first aspect, present invention discloses the use of an alkaline source in a detergent composition for treating textile article for inactivation of microorganisms.
The alkaline source comprises sodium carbonate. Examples of alkaline source include alkali metal or alkaline earth metal salts of carbonate, bicarbonate, silicate, metasilicates or combination thereof.
In a preferred embodiment the alkaline source is a mixture of sodium carbonate with bicarbonate and/or sesqui-carbonate.
The carbonate and bicarbonate preferably have an amorphous structure. Preferably the carbonate and bicarbonates are coated with coating materials. The particles of carbonate and bicarbonate can have a mean particle size of 250 micrometers or greater, preferably 500 micrometers or greater.
Preferably the carbonate salt of alkali metal and/or alkaline earth metal are present in the detergent composition of the present invention in an amount ranging from 10 wt.% to 35 wt.%. The term carbonates include bicarbonates and sesquicarbonates.
The detergent composition according to the present invention comprises from 10 wt.% to 40 wt.% sodium carbonate alkaline source. Preferably the detergent composition comprises at least 12 wt.% carbonate alkaline source based on the weight of the detergent composition, still preferably at least 15 wt.%, still preferably at least 18 wt.%, most preferably at least 20 wt.%, but typically not more than 35 wt.%, still preferably not more than 30 wt.%, more preferably not more than 25 wt.% carbonate alkaline source in the solid detergent composition.
The alkalinity system may include other components, such as a silicate. Preferably the silicate is present in the detergent composition in an amount ranging from 1 wt.% to 10wt.%. Suitable silicates include the water-soluble sodium silicates with an SiO₂: Na₂O ratio of from 1.0 to 2.8, with ratios of from 1.6 to 2.0 being preferred, and 2.0 ratio being most preferred. The silicates may be in the form of either the anhydrous salt or a hydrated salt. Sodium silicate with an SiO₂: Na₂O ratio of 2.0 is the most preferred silicate. Preferably the silicates have an amorphous structure. Alkali metal persilicates or metasilicates are also suitable sources of silicate herein.
Preferred crystalline layered silicates for use herein have the general formula NaMSixO2_X+l.yH2O wherein M is sodium or hydrogen, x is a number from 1.9 to 4 and y is a number from 0 to 20. Crystalline layered sodium silicates of this type are disclosed in EP-A-0164514 and methods for their preparation are disclosed in DE-A-3417649 and DE-A-3742043 . Herein, x in the general formula above preferably has a value of 2, 3 or 4 and is preferably 2. The most preferred material is δ-Na₂Si₂O₅, available from Hoechst AG as NaSKS-6.
Preferably the detergent composition according to the present invention comprises from 10 wt.% to 40 wt.% alkaline source. Preferably the detergent composition comprises at least 12 wt.% alkaline source based on the weight of the detergent composition, still preferably at least 15 wt.%, still preferably at least 18 wt.%, most preferably at least 20 wt.%, but typically not more than 35 wt.%, still preferably not more than 30 wt.%, more preferably not more than 25 wt.% alkaline source in the detergent composition.
The use according to the first aspect of the present invention involves inactivating the microorganisms in a wash liquor prepared by addition of the solid detergent composition in water, wherein the alkaline source is present at a concentration from 200 ppm to 2500 ppm. Preferably concentration is at least 500 ppm, most preferably at least 600 ppm, but typically not more than 1800ppm, still preferably not more than 1600 ppm, more preferably not more than 1500 ppm and most preferably not more than 1000 ppm in the wash liquor. Preferably the alkaline source is a combination of sodium carbonate and sodium silicate.

Detergent composition

The combination of alkyl benzene sulphonate surfactant, hydrolytic enzyme wherein the hydrolytic enzyme comprises a serine protease and alkaline source according to the present invention can be employed in any suitable detergent composition having a pH from 10 to 13.
Preferably the pH of the composition is from 10.2 to 13, still preferably from 10.5 to 13, still preferably from 10.2 to 12, more preferably from 10.2 to 11, still more preferably 10.2 to 11 and most preferably from 10.5 to about 11, as measured at 25°C and 10% aqueous concentration in deionized water. The pH of the composition can be adjusted using pH modifying ingredients known in the art.
The detergent composition must be suitable for use with a soft surface, preferably textile article.
The detergent composition is solid and may be granular, powder, or in the form of a unit dose product where the solid detergent composition is at least partially enclosed in a water-soluble film.
The solid detergent composition in particulate form may include agglomerate, a spray-dried powder, an extrudate, a flake, a needle, a noodle, a bead, or any combination thereof. The composition may be in compacted-particulate form, such as in the form of a tablet or bar. Then composition may be some other unit dose form, such as a pouch; typically being at least partially, preferably essentially completely, enclosed by a water-soluble film, such as polyvinyl alcohol. Preferably, the composition is in free-flowing particulate form; by free-flowing particulate form, it is typically meant that the composition is in the form of separate discrete particles. The solid composition may be made by any suitable method including agglomeration, spray-drying, extrusion, mixing, dry-mixing, liquid spray-on, roller compaction, spheronisation, tabletting or any combination thereof.
The solid detergent composition typically has a bulk density of from 450 g/l to 1,000 g/l, preferred low bulk density detergent compositions have a bulk density of from 550 g/l to 650 g/l and preferred high bulk density detergent compositions have a bulk density of from 750 g/l to 900 g/l. During the laundering process, the composition is typically contacted with water to give a wash liquor having a pH of from 8 to 13, preferably from 8.5 to less than 11.
The compositions may be used only to deliver alkyl benzene sulphonate surfactant, hydrolytic enzyme and an alkaline source during the laundering process, or they may have additional functions such as cleaning.

Inactivation of microorganisms

The first aspect of the present invention discloses a use of a combination of alkyl benzene sulphonate surfactant, hydrolytic enzyme and an alkaline source in a solid detergent composition having a pH from 10 to 13 as measured at 25°C and 10% aqueous concentration in deionised water for inactivating microorganisms on a textile article during a laundering process wherein the hydrolytic enzyme comprises a serine protease and wherein the solid detergent composition does not contain a peroxygen bleach and wherein the alkaline source comprises from 10 wt.% to 40 wt.% sodium carbonate and wherein the wash liquor prepared by adding the solid detergent composition in water has an alkaline source concentration ranging from 200 ppm to 2500 ppm.
The term inactivation is understood in the context of the present invention to be an activity against at least one virus (antiviral efficacy) or bacteria species.
Antiviral efficacy is understood to mean any reduction in the virus titer and the associated infectivity of a virus, the infectivity being the ability of a virus to infect a host. Antiviral efficacy is therefore advantageously achieved by damaging the virus or viruses, in particular with regard to the ability to adhere to a host cell and / or to introduce the genetic material into a host cell and / or to replicate the genetic material in a host cell.
Antimicrobial activity is assessed as the log reduction, or alternatively the percent reduction, in microbial populations provided by the antimicrobial composition. A 1 to 3 log reduction is preferred, a log reduction of 3 to 5 is most preferred, whereas a log reduction of less than 1 is least preferred, for a particular contact time, generally ranging from 15 seconds to 5 minutes, more preferably the contact time ranges from 15 seconds to at least one hour. Thus, a highly preferred antimicrobial composition exhibits a 3 to 5 log reduction against a broad spectrum of microorganisms in a short contact time.
Preferably the use according to the first aspect of the invention provides for log 1 to log 5 reduction, still preferably log 2 to log 5 reduction in viruses. Preferably the use according to the present invention provides at least 50%, still preferably at least 60%, 70%, 80%, 90%, 95%, 96%, 97 %, 98%, 99% and particularly preferably around 99.999% (corresponds to at least 5 log10 levels) reduction in the virus titer. Preferably the contact time with the aqueous liquor of the detergent composition according to the first aspect for at least 30 minutes to at least 2 hours, preferably for at least 1 hour.
Viruses (singular: virus) mean intracellular, but non-cellular parasites that can infect cells of living beings. Viruses contain the genetic program (genetic material) in the form of at least one nucleic acid (deoxyribonucleic acid (DNA) or ribonucleic acid (RNA)) and optionally also other auxiliary components for their multiplication and spread. The viruses may be enveloped or non-enveloped.
The use according to the first aspect provides for inactivation of viruses which may be selected from the group consisting of Adenoviridae, Alphaherpesvirinae, Astroviridae, Betaherpesvirinae, Birnaviridae, Bornaviridae, Bunyaviridae, Caliciviridae, Chordopoxoviridae, Gordopoxoviridae, Chordopoxviridae, Chordopoxoviridae, Hepadnaviridae, Herpesviridae, Iridoviridae, Orthomyxoviridae, Orthoretroviridae, Papillomaviridae, Paramyxovirinae, Parvovirinae, Picornaviridae, Pneumovirinae, Polyomaviridae, Reoviridae, Rhabdoviridae, Togiviririridae, Coronavirinae, and Roniviririridae.
This includes, in particular, a virus that belongs to one of the following genera:
Alphapapillomavirus, Alpharetrovirus, Alphavirus, Aphthovirus, Aquabirnavirus, Aquareovirus, adenovirus, Avibirnavirus, Avulavirus, Betapapillomavirus, Betaretrovirus, Bocavirus, Bornavirus, Cardovirusomovirus, Colivirus Delta Virus, Delta Virus, Dependovirus, Ebola Virus, Enterovirus, Ephemerovirus, Epsilonretrovirus, Erbovirus, Erythrovirus, Fijivirus, Flavivirus, Fungal Prions, Gammapapillomavirus, Gammaretrovirus, Hantavirus, Henipavirus, Hepacivalirus Virus, Influenza Virus C, Iridovirus, Kobuvirus, Lentivirus, Lymphocryptovirus, Lyssavirus, Mamastrovirus, Marburgvirus, Mastadenovirus, Megalocytivirus, Morbillivirus, Mupapillomavirus, Muromegalovirus, Mycoreovirus, Nairovirus, Norovirus, coronavirus, murine norovirus, bovine coronavirus, Novirhabapunovirus, Novirhabapvirus, orthoreovirus, oryzavirus, parechovirus, parvovirus, pestivirus, phlebovirus, phytoreovirus, pneumovirus, polyomavirus, respirovirus, rhadinovirus, rhinovirus an roseolovirus.
The use according to the present invention includes applying the solid detergent composition to the textile article to be laundered in a liquid diluted form, preferably diluted with water to form a wash liquor. The use according to the first aspect of the present invention provides for inactivation of microorganisms on an article which may be either a hard surface or a textile surface. The textile includes all types of fabrics, including different compositions, for example made of cotton, wool, silk, other natural fibers, polyester and mixed fabrics of all types. Preferred textiles are laundry. This includes all washable textiles. The textile may be woven or non-woven.

Additional surfactants

The detergent composition may preferably include other surfactants in addition to the alkyl benzene sulphonate surfactant.
Detersive surfactants utilized can be of the anionic, nonionic, zwitterionic, ampholytic or cationic type or can comprise compatible mixtures of these types. More preferably surfactants are selected from the group consisting of anionic, nonionic, cationic surfactants and mixtures thereof. Detergent surfactants useful herein are described in U.S. Patent 3,664,961, Norris, issued May 23, 1972 , U.S. Patent 3,919,678, Laughlin et al., issued December 30, 1975 , U.S. Patent 4,222,905, Cockrell, issued September 16, 1980 , and in U.S. Patent 4,239,659, Murphy, issued December 16, 1980 . Anionic and nonionic surfactants are preferred.
Preferably the compositions are substantially free of salts of alkylsulfosuccinic acid which are also referred to as sulfosuccinates or as sulfosuccinic acid esters, and which are monoesters and / or diesters of sulfosuccinic acid with alcohols, preferably fatty alcohols and in particular ethoxylated fatty alcohols.
Useful anionic surfactants can themselves be of several different types. For example, water-soluble salts of the higher fatty acids, i.e., "soaps". This includes alkali metal soaps such as the sodium, potassium, ammonium, soluble salts of organic bases, such as mono-, di- or triethanolamine and alkyl ammonium salts of higher fatty acids containing from about 8 to about 24 carbon atoms, and preferably from about 12 to about 18 carbon atoms. Soaps can be made by direct saponification of fats and oils or by the neutralization of free fatty acids. Soap includes the sodium and potassium salts of the mixtures of fatty acids derived from coconut oil and tallow, i.e., sodium or potassium tallow and coconut soap. Soaps includes saturated fatty acid soaps are particularly suitable, such as the salts of lauric acid, myristic acid, palmitic acid, stearic acid, hydrogenated erucic acid and behenic acid, and in particular soap mixtures derived from natural fatty acids, for example coconut, palm kernel or tallow fatty acids.
Preferably the detergent composition of the present invention are substantially free of soap.
Additional non-soap anionic surfactants which are suitable for use herein include the water-soluble salts, preferably the alkali metal, and ammonium salts, of organic sulfuric reaction products having in their molecular structure an alkyl group containing from about 10 to about 20 carbon atoms and a sulfonic acid or sulfuric acid ester group. (Included in the term "alkyl" is the alkyl portion of acyl groups.) Examples of this group of synthetic surfactants are a) the sodium, potassium and ammonium alkyl sulfates, especially those obtained by sulfating the higher alcohols (Csto C₁₈ carbon atoms) such as those produced by reducing the glycerides of tallow or coconut oil; b) the sodium, potassium and ammonium alkyl polyethoxylate sulfates, particularly those in which the alkyl group contains from 10 to 22, preferably from 12 to 18 carbon atoms, and wherein the polyethoxylate chain contains from 1 to 15, preferably 1 to 6 ethoxylate moieties; and c) the sodium and potassium alkylbenzene sulfonates in which the alkyl group contains from about 9 to about 15 carbon atoms, in straight chain or branched chain configuration, e.g., those of the type described in U.S. Patents 2,220,099 and 2,477,383 .
Preferred nonionic surfactants are those of the formula R¹(OC₂H₄)nOH, wherein R¹ is a C₁₀-C₁₆ alkyl group or a C₈ to C₁₂ alkyl phenyl group, and n is from 3 to about 80. Particularly preferred are condensation products of C₁₂ to C₁₅ alcohols with from about 5 to about 20 moles of ethylene oxide per mole of alcohol, e.g., C₁₂ to C₁₃ condensed with about 6.5 moles of ethylene oxide per mole of alcohol. Preferred nonionic surfactant which may be used include the reaction products of compounds having a hydrophobic group and a reactive hydrogen atom, for example, aliphatic alcohols, acids, amides with alkylene oxides, especially ethylene oxide either alone or with propylene oxide. Specific nonionic detergent compounds are the condensation products of aliphatic primary or secondary linear or branched alcohols with ethylene oxide, generally 5 to 40 EO, preferably 7EO to 9EO.
In all aspects of the invention, preferably an additional anionic surfactant is present which may be selected from C₁₀ to C₂₀ alkyl sulphates, C₁₀ to C₂₀ alkyl ether sulphates, and mixtures thereof. More preferably the additional anionic surfactant is different from C₁₀ to C₂₀ linear alkylbenzene sulphonates and includes a mixture of anionic surfactants as previously specified. Preferred additional anionic surfactant 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 alcohols, 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 additional anionic detergent compounds sodium C₁₂ to C₁₄ 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.
Further preferably the additional surfactant may be selected from cationic surfactant, nonionic surfactants, amphoteric surfactants, zwitterionic surfactants or combinations thereof.
In general, the nonionic and anionic surfactants of the surfactant system may be chosen from the surfactants described "Surface Active Agents" Vol. 1, by Schwartz & Perry, Interscience 1949, Vol. 2 by Schwartz, Perry & Berch, Interscience 1958, in the current edition of "McCutcheon's Emulsifiers and Detergents" published by Manufacturing Confectioners Company or in "Tenside-Taschenbuch", H. Stache, 2nd Edn, Carl Hauser Verlag, 1981. Preferably the surfactants used are saturated.
Preferred zwitterionic surfactants include cocamidopropyl betaine. Preferred levels of zwitterionc surfactants are from 0.1 to 5 wt.%, preferably from 0.5 to 4 wt.%.

Further preferred ingredients

The detergent composition, may comprises further preferred ingredients which may include complexing agents, fluorescers, dyes, enzyme stabilizers, further builders, perfume, polymers, enzyme stabilizers, fillers selected from sulphate salts, bleaching agent and combinations thereof.
The solid detergent composition includes preferably from 1 to 3 wt.% moisture, still preferably from 1 to 2 wt.% moisture content.

Further Builders:

Builder materials may be selected from 1) calcium sequestrant materials, 2) precipitating materials, 3) calcium ion-exchange materials and 4) mixtures thereof.
Examples of calcium sequestrant builder materials include alkali metal polyphosphates, such as sodium tripolyphosphate and organic sequestrants, such as ethylene diamine tetra-acetic acid.
Examples of precipitating builder materials include sodium orthophosphate.
Examples of calcium ion-exchange builder materials include the various types of water-insoluble crystalline or amorphous aluminosilicates, of which zeolites are the best known representatives, e.g. zeolite A, zeolite B (also known as zeolite P), zeolite C, zeolite X, zeolite Y and also the zeolite P-type as described in EP-A-0,384,070 .
The composition may also contain 0 wt.% to 65 wt.% of an organic builder or complexing agent such as ethylenediaminetetraacetic acid, diethylenetriamine-pentaacetic acid, alkyl- or alkenylsuccinic acid, nitrilotriacetic acid or the other builders mentioned below. Many builders are also bleach-stabilising agents by virtue of their ability to complex metal ions.
Zeolite, bicarbonate and sesquicarbonate are examples of preferred builders.
The composition may contain as builder a crystalline aluminosilicate, preferably an alkali metal aluminosilicate, more preferably a sodium aluminosilicate. This is typically present at a level of less than 15 wt.%. Aluminosilicates are materials having the general formula: 0.8-1.5 M₂₀. Al₂O₃. 0.8-6 Si0₂
where M is a monovalent cation, preferably sodium. These materials contain some bound water and are required to have a calcium ion exchange capacity of at least 50 mg CaO/g. The preferred sodium aluminosilicates contain 1.5-3.5 SiO₂ units in the formula above. They can be prepared readily by reaction between sodium silicate and sodium aluminate, as amply described in the literature. The ratio of surfactants to alumuminosilicate (where present) is preferably greater than 5:2, more preferably greater than 3:1.
Alternatively, or additionally to the aluminosilicate builders, phosphate builders may be used. In this art the term 'phosphate' embraces diphosphate, triphosphate, and phosphonate species. Other forms of builder include silicates, such as soluble silicates, metasilicates, layered silicates (e.g., SKS-6 from Hoechst).
When a laundry composition, preferably the laundry detergent formulation is a non-phosphate-built laundry detergent formulation, i.e., contains less than 1 wt.% of phosphate.

Fluorescent Agent:

These materials may be particularly useful in liquid laundry detergent compositions for hand wash. The composition preferably comprises a fluorescent agent (optical brightener).
Fluorescent agents are well known and many such fluorescent agents are available commercially. Usually, these fluorescent agents are supplied and used in the form of their alkali metal salts, for example, the sodium salts. The total amount of the fluorescent agent or agents used in the composition is generally from 0.005 to 2 wt.%, more preferably 0.01 to 0.1 wt.%. Preferred classes of fluorescer are: Di-styryl biphenyl compounds, e.g. Tinopal (Trade Mark) CBS-X, Di-amine stilbene di-sulphonic acid compounds, e.g. Tinopal DMS pure Xtra and Blankophor (Trade Mark) HRH, and Pyrazoline compounds, e.g. Blankophor SN. Preferred fluorescers are: sodium 2 (4-styryl-3-sulfophenyl)-2H-napthol[1 ,2-d]triazole, disodium 4,4'-bis{[(4-anilino-6-(N methyl-N-2 hydroxyethyl) amino 1 ,3,5-triazin-2-yl)]amino}stilbene-2-2' disulfonate, disodium 4,4'-bis{[(4-anilino-6-morpholino-1 ,3,5-triazin-2-yl)]amino} stilbene-2-2' disulfonate, and disodium 4,4'-bis(2-sulfostyryl)biphenyl.
It is preferred that the aqueous solution used in the method has a fluorescer present. When a fluorescer is present in the aqueous solution used in the method it is preferably in the range from 0.0001 g/l to 0.1 g/l, preferably 0.001 to 0.02 g/l.

Dye:

The composition preferably comprises a dye. Dyes are discussed in K.Hunger (ed). Industrial Dyes: Chemistry, Properties, Applications (Weinheim: Wiley-VCH 2003). Organic dyes are listed in the colour index (Society of Dyers and Colourists and the American Association of Textile Chemists and Colorists)
Preferred dye chromophores are azo, azine, anthraquinone, phthalocyanine and triphenylmethane.
Azo, anthraquinone, phthalocyanine and triphenylmethane dyes preferably carry a net anionic charged or are uncharged. Azine dyes preferably carry a net anionic or cationic charge.
Preferred non-shading dyes are selected are selected from blue dyes, most preferably anthraquinone dyes bearing sulphonate groups and triphenylmethane dye bearing sulphonate groups. Preferred compounds are acid blue 80, acid blue 1, acid blue 3; acid blue 5, acid blue 7, acid blue 9, acid blue 1 1, acid blue 13, acid blue 15, acid blue 17, acid blue 24, acid blue 34, acid blue 38, acid blue 75, acid blue 83, acid blue 91, acid blue 97, acid blue 93, acid blue 93:1, acid blue 97, acid blue 100, acid blue 103, acid blue 104, acid blue 108, acid blue 109, acid blue 110, and acid blue 213. On dissolution granules with non-shading dyes provide an attractive colour to the wash liquor.
Blue or violet Shading dyes are most preferred. Shading dyes deposit to fabric during the wash or rinse step of the washing process providing a visible hue to the fabric. In this regard the dye gives a blue or violet colour to a white cloth with a hue angle of 240 to 345, more preferably 260 to 320, most preferably 270 to 300. The white cloth used in this test is bleached non-mercerised woven cotton sheeting.
Shading dyes are discussed in WO 2005/003274 , WO 2006/032327(Unilever ), WO 2006/032397(Unilever ), WO 2006/045275(Unilever ), WO 2006/027086(Unilever ), WO 2008/017570(Unilever ), WO 2008/141880(Unilever ), WO 2009/132870(Unilever ), WO 2009/141 173 (Unilever ), WO 2010/099997(Unilever ), WO 2010/102861 (Unilever ), WO 2010/148624(Unilever ), WO 2008/087497 (P&G) , WO 2011/011799 (P&G ), WO 2012/054820 (P&G ), WO 2013/142495 (P&G ) and WO 2013/151970 (P&G ).
A mixture of shading dyes may be used.
The shading dye chromophore is most preferably selected from mono-azo, bis-azo, anthraquinone, and azine.
Mono-azo dyes preferably contain a heterocyclic ring and are most preferably thiophene dyes. The mono-azo dyes are preferably alkoxylated and are preferably uncharged or anionically charged at pH=7. Alkoxylated thiophene dyes are discussed in WO 2013/142495 and WO 2008/087497 .
Most preferred shading dyes are selected from Direct Violet 9, Direct Violet 99, Direct Violet 35, Solvent Violet 13, Disperse Violet 28, dyes of the structure

Perfume:

Preferably the composition comprises a perfume. The perfume is preferably in the range from 0.001 to 3 wt.%, most preferably 0.1 to 1 wt.%. Many suitable examples of perfumes are provided in the CTFA (Cosmetic, Toiletry and Fragrance Association) 1992 International Buyers Guide, published by CFTA Publications, and OPD 1993 Chemicals Buyers Directory 80th Annual Edition, published by Schnell Publishing Co.
It is commonplace for a plurality of perfume components to be present in a formulation. In the compositions of the present invention it is envisaged that there will be four or more, preferably five or more, more preferably six or more or even seven or more different perfume components.
In perfume mixtures preferably 15 to 25 wt.% are top notes. Top notes are defined by Poucher (Journal of the Society of Cosmetic Chemists 6(2):80 [1955]). Preferred top-notes are selected from citrus oils, linalool, linalyl acetate, lavender, dihydromyrcenol, rose oxide and cis-3-hexanol.
The laundry treatment composition does not contain a peroxygen bleach, e.g., sodium percarbonate, sodium perborate, and peracid.

Polymers:

The composition may comprise one or more further polymers. Examples are carboxymethylcellulose, poly(ethylene glycol), polyvinyl alcohol), polycarboxylates such as polyacrylates, maleic/acrylic acid copolymers and lauryl methacrylate/acrylic acid copolymers. Polymers present to prevent dye deposition, for example poly(vinylpyrrolidone), poly(vinylpyridine-N-oxide), and poly(vinylimidazole), may be present in the formulation.
Thickening polymers such as anionic acrylic polymers may be included, examples include Acusol 820.

Enzyme Stabilizers:

Any enzyme present in the composition 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 e.g. WO 92/19709 and WO 92/19708 .
Where alkyl groups are sufficiently long to form branched or cyclic chains, the alkyl groups encompass branched, cyclic and linear alkyl chains. The alkyl groups are preferably linear or branched, most preferably linear.
The indefinite article "a" or "an" and its corresponding definite article "the" as used herein means at least one, or one or more, unless specified otherwise.
The invention will be further described with the following non-limiting examples.

Sulphate salt:

In the present invention the ratio of sulphate salt to the acidic dispersing aid is preferably from 12:1 to 1:1, most preferably from 11:1 to 2:1.
The sulphate salt can be present in the detergent composition in any form, preferably it is an inorganic sulphate salt, such as sodium sulphate, magnesium sulphate, ammonium sulphate or mixtures of various forms of sulphate. The sulphate preferably is substantially anhydrous, (i.e. generally no greater than 50% by weight of the sulphate salt containing water, preferably no greater than 25%, more preferably no greater than 15%, most preferably no greater than 10%), preferably it is anhydrous sodium sulphate. This is preferably combined with a small amount of magnesium sulphate, preferably of from 0.2% to 5% by weight of the composition.

Bleaching agent:

Use according to the first aspect of the present invention may preferably include a bleaching agent, provided that the solid detergent composition does not contain a peroxygen bleach.
Bleaching agents not according to the invention include hydrogen peroxide, or substances that can generate perhydroxyl radical, such as inorganic or organic peroxides. Generally, a peroxygen bleach compound or hydrogen peroxide must be activated. The solid detergent composition preferably includes a bleach, provided that it does not contain a peroxygen bleach. Still preferably the bleach is present along with a bleach activator.
Examples of bleach activators are tetraacetylethylenediamine (TAED) and sodium nonanoyloxybenzenesulphonate(NOBS). The bleach activators react with the perhydroxide anion (OOH-) of the hydrogen peroxide released by the peroxygen bleach compound in the aqueous solution to form a peroxyacid which is more reactive as a bleaching agent than the peroxide bleach alone.
Hydrogen peroxide sources are well known in the art. Hydrogen peroxide sources are described in details in Kirk Othmer's Encyclopedia of Chemical Technology, 4th Ed (1992, John Wiley & Sons), Vol. 4, pp. 271 -300 "Bleaching Agents (Survey)", and include the alkali metal salts of sodium perborates and sodium percarbonates, including various coated and modified forms. Peroxygen bleach compounds include hydrogen peroxide or any of its solid adducts such as organic peroxides example; urea peroxide and inorganic persalts such as the alkali metal perborates, percarbonates, perphosphates, persilicates and persulphates. inorganic persalts are sodium perborate monohydrate and tetrahydrate, and sodium percarbonate. Hydrogen peroxide is especially preferred in liquid cleaning compositions, which are not according to the invention.
Percarbonate is stable during storage and dissolves quickly in the cleaning liquor and is especially preferred. It is believed that such rapid dissolution results in the formation of higher levels of percarboxylic acid and, thus, enhances substrate bleaching performance. Percarbonate is in uncoated or coated form. Preferably the average particle size of uncoated and coated percarbonate ranges from about 400 to about 1200 micrometres, most preferably from about 400 to about 600 micrometres. If coated percarbonate is used, the preferred coating materials include mixtures of carbonate and sulphate, zeolite, precipitated silica, waxes, borates, polymers, citrates, silicate, borosilicate or fatty acids.
Preferred bleach activators includes compound from the class of polyacylated alkylenediamines preferably tetraacetylethylenediamine (TAED). Yet another preferred bleach activator is hydroxybenzoic acid derivative of the formula (I)
in which R is C₈ to C₁₂ alkyl group. Preferably the bleach activator is decanoyloxybenzoic acid (DOBA) and derivatives thereof.
The following non-limiting examples further illustrate the preferred embodiments of the invention. All percentages referred to in the examples and throughout this specification are by weight based on total weight unless otherwise indicated.
According to the second aspect, disclosed is a method of inactivating microorganism from a textile article, the method comprising the steps of:

i) applying a solid detergent composition to the textile article, wherein the solid detergent composition comprises a combination of alkyl benzene sulphonate, hydrolytic enzyme comprising a serine protease and an alkaline source and the detergent composition has a pH from 10 to 13;
ii) allowing the textile article to be in contact with the solid detergent composition for a period of at least 30 minutes, still preferably at least one hour; wherein the solid detergent composition does not contain a peroxygen bleach and wherein the alkaline source comprises from 10 wt.% to 40 wt.% sodium carbonate and wherein the wash liquor prepared by adding the solid detergent composition in water has an alkaline source concentration ranging from 200 ppm to 2500 ppm.

Preferably the solid detergent composition is applied in a liquid diluted form. The liquid is preferably water. The solid detergent composition is diluted to form a wash liquor. The wash liquor preferably includes alkyl benzene sulphonate surfactant in a concentration from 40 ppm to 2000 ppm, hydrolytic enzyme in a concentration from 0.0001 ppm to 30 ppm and the alkaline source in a concentration from 200 ppm to 2500 ppm.

Examples

Example 1: Evaluation of the virus inactivation using different detergent compositions.

Solid powder laundry detergent compositions as shown in Table 1 was prepared and used for the evaluation of the efficacy for inhibiting the growth of the virus. 2 different solid detergent composition were taken for the evaluation with different levels of the LAS surfactant as shown in the table 1 below.

Table 1

Ingredients	Ex 1 (wt.%)	Ex 2 (wt.%)
LAS (Linear alkyl benzene sulphonate surfactant)	13.2	7.7
SLES/MES/PAS	0	1.644
Total AD	13.2	9.34
sodium carbonate	30.58	11.6
Sulphate	42.098	68
zeolite	2.351	3.11
Savinase 12T (protease)	0.267	0.14
Miscellaneous (includes ingredients such as perfume, dyes, colorants, NDOM, moisture and other non-microorganism inactivating ingredients)	Upto 100	Upto 100

Antiviral study:

Each of the detergent composition as shown in Table 1 was individually diluted using sterilized hard water according to the EN144476 standards described below to prepare aqueous liquor with concentrations of 4 grams/Litre, 2 grams/Litre and 0.4 grams/Litre.

Aqueous liquor with 3 different concentrations as described above were prepared from the detergent composition of Ex 1 and was tested for virucidal efficacy in accordance to the European Standard method EN14476: 2013+A2:2019 (chemical disinfectants and antiseptics - Quantitative suspension test for the evaluation of virucidal activity in the medical area- Test method and requirements (Phase 2/Step 1). The same procedure was also followed for the detergent composition of Ex 2.

Experimental conditions:

The virucidal efficacy test was set up with the three concentrations of test product solution as described above and a 60 minute contact time was given. The test temperature was maintained at 27°C and the interfering condition was Clean with low levels of organic soil.
The virucidal efficacy of the test composition was tested for Adeno virus, Murine Norovirus, Poliovirus & bovine coronavirus and the final virucidal efficacy is expressed as log reduction.

Similarly, aqueous liquor was prepared and tested for Ex 2 and the test results are provided in Table 2.

Table 2

Detergent composition @4GPL dose levels	LAS surfactant (ppm)	Protease (ppm)	Alkaline* source (ppm)	Poliovirus	Adenovirus	Murine norovirus	Bovine Coronavirus
	Concentration in 4gpl liquor			Log reduction
Ex 1 (pH 10.71)	516	10.8	1233	2.00 ± 0.000	> 5.00 ± 0.000	2.17 ± 0.17	≥4.00±0.000
Ex 2 (pH 10.2)	308	4	464	0.84 ± 0.27	1.57 ± 0.27	1.00 ± 0.28	≥4.67±0.200
*alkaline source is sodium carbonate

Detergent composition @2GPL dose levels	LAS surfactant (ppm)	Protease (ppm)	Alkaline* source (ppm)	Poliovirus	Adenovirus	Murine norovirus	Bovine Coronavirus
	Concentration in 2 gpl liquor			Log reduction
Ex 1 (pH 10.25)	226	5.4	622	2.00 ± 0.000	1.00 ± 0.000	1 ± 0.25	≥4.00±0.000
Ex 2 (pH 9.63)	154	2	234	0.67 ± 0.28	0.33 ± 0.2	0.84 ± 0.270	≥4.67±0.200

*alkaline source is sodium carbonate
Detergent composition @0.4 GPL dose levels	LAS surfactant (ppm)	Protease (ppm)	Alkaline source (ppm)	Poliovirus	Adenovirus	Murine norovirus	Bovine Coronavirus
	Concentration in 0.4 gpl liquor			Log reduction
Ex 1 (pH 9.24)	45	1.08	124	2.00 ± 0.000	0 ± 0.00	0.5 ± 0.27	≥4.00±0.000
Ex 2 (pH 8.15)	30	0.4	46	0.5 ± 0.27	0.33 ± 0.2	0.67 ± 0.200	0.67±0.200

* alkaline source is sodium carbonate.

The example using detergent composition @0.4 GPL dose levels is a reference example.
The results on Table 2 show that the use of a solid detergent composition with a pH ranging from 10 to 13 and having a combination of LAS surfactant, alkaline source and Protease ( a hydrolytic enzyme selected from the group consisting of protease, lipase, cellulase, amylase, mannanase or combinations thereof) provides improved virucidal efficacy (log kill of at least 1) on both enveloped and non-enveloped viruses.

Example 2: Evaluation of the bacterial inactivation using different detergent compositions

A detergent composition according to Table 3 was diluted using sterilized hard water according to the EN1276 standards described below to prepare an aqueous liquor with concentrations of 7.5 grams/litre. Table 3 Comparative example

Ingredients Ex 3 (wt.%)

LAS (Linear alkyl benzene sulphonate surfactant) 11.27

sodium carbonate 31.09

Sulphate 55

zeolite 2.35

Hydrolytic enzyme 0.3
The aqueous liquor as described above was prepared from the detergent composition of Ex 3 and was tested for antibacterial efficacy in accordance to the European Standard method EN1276 test.

Experimental conditions:

The antibacterial efficacy test was set up with a 7.5 grams/Litre concentration test product solution as described above and a 60 minutes contact time was provided. The test temperature was maintained at 27°C and the interfering condition was clean with low levels of organic soil. The antibacterial efficacy of the test composition was tested using Staphylococcus aureus (S. aureus), Enterococcus hirae (E. hirae) and the final antibacterial efficacy was expressed as log reduction. The test results are provided in Table 4.

Table 4

Detergent composition @7.5GPL dose levels	LAS surfactant (PPm)	Hydrolytic enzyme^& (ppm)	Alkaline* source (ppm)	S. aureus	E. hirae
	Concentration in 7.5 gpl liquor			Log reduction
Ex 3 (10.8 pH)	845	22.5	2331	4.8	3.51
* alkaline source is sodium carbonate
^& hydrolytic enzyme is protease

The results on Table 4 show that the use of a solid detergent composition with a pH ranging from 10 to 13 and having a combination of LAS surfactant, alkaline source, and a hydrolytic enzyme ( a hydrolytic enzyme selected from the group consisting of protease, lipase, cellulase, amylase, mannanase or combinations thereof) provides improved bactericidal efficacy (log kill of at least 1) on bacteria.

Claims

Use of a combination of alkyl benzene sulphonate surfactant, hydrolytic enzyme and an alkaline source in a solid detergent composition having a pH from 10 to 13 as measured at 25°C and 10% aqueous concentration in deionised water for inactivating microorganisms on a textile article during a laundering process wherein the hydrolytic enzyme comprises a serine protease and wherein the solid detergent composition does not contain a peroxygen bleach and wherein the alkaline source comprises from 10 wt.% to 40 wt.% sodium carbonate and wherein the wash liquor prepared by adding the solid detergent composition in water has an alkaline source concentration ranging from 200 ppm to 2500 ppm.
Use according to claim 1 wherein the hydrolytic enzyme further comprises an enzyme selected from the group consisting of protease, lipase, cellulase, amylase, mannanase or combinations thereof.
Use according to claim 1, wherein the alkaline source is further selected from a group consisting of alkali metal or alkaline earth metal salts of carbonate (other than sodium carbonate), bicarbonate, silicate, metasilicates or combination thereof.
Use according to claim 1, wherein the detergent composition having combination of alkyl benzene sulphonate surfactant, hydrolytic enzyme and an alkaline source is applied to the textile article to be laundered in a liquid diluted form, preferably diluted with water to form a wash liquor.
Use according to any one of the preceding claims wherein the microorganism is a virus or a bacteria.
Use according to any one of the preceding claims wherein the microorganism is in intimate contact with the detergent composition for a period of at least 30 minutes, still preferably at least 1 hour.
Use according to any one of the preceding claims, wherein the combination of alkyl benzene sulphonate surfactant, hydrolytic enzyme and an alkaline source has virus inactivating efficacy against an enveloped and/or non-enveloped virus.
Use according to any one of the preceding claims, wherein the combination of alkyl benzene sulphonate surfactant, hydrolytic enzyme and an alkaline source has virus inactivating efficacy against a virus selected from the group consisting of Adenoviridae, Alphaherpesvirinae, Astroviridae, Betaherpesvirinae, Birnaviridae, Bornaviridae, Bunyaviridae, Caliciviroviridae, Chordopoxiridae, Chordopoxiridae , Flaviviridae, Gammaherpesvirinae, Hepadnaviridae, Herpesviridae, Iridoviridae, Orthomyxoviridae, Orthoretroviridae, Papillomaviridae, Paramyxovirinae, Parvovirinae, Picornaviridae, Pneumovirinae,Romovaviridairidae, Polyomaviriridae, Coronaviridae, Arteriviridae, Mesoniviridae, Torovirinae, and Roniviridae.
Use according to any one of the preceding claims wherein the detergent composition achieves log 1 to log 5 reduction of the virus when the contact time is at least 30 minutes, still preferably at least 1 hour.
Use according to any one of the preceding claims wherein the alkyl benzene sulphonate surfactant is present at a concentration from 2 wt.% to 40 wt.% in the detergent composition.
Use according to any one of the preceding claims, wherein the hydrolytic enzyme is present at a concentration of from 0.0001 ppm to 30 ppm in the wash liquor during the laundering process.
Use according to any preceding claims, wherein the solid detergent composition is in the form of a unit dose product where the solid detergent composition is at least partially enclosed by a water-soluble film.
A method of inactivating microorganism from a textile article, the method comprising the steps of:
i) applying a solid detergent composition to the textile article, wherein the solid detergent composition comprises a combination of alkyl benzene sulphonate, hydrolytic enzyme comprising a serine protease and an alkaline source and the detergent composition has a pH from 10 to 13;

ii) allowing the textile article to be in contact with the solid detergent composition for a period of at least 30 minutes, still preferably at least one hour.
wherein the solid detergent composition does not contain a peroxygen bleach and wherein the alkaline source comprises from 10 wt.% to 40 wt.% sodium carbonate and wherein the wash liquor prepared by adding the solid detergent composition in water has an alkaline source concentration ranging from 200 ppm to 2500 ppm.