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(11) | EP 4 428 218 A1 |
| (12) | EUROPEAN PATENT APPLICATION |
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| (54) | BIODEGRADABLE TRANSGLUTAMINASE-BASED DETERGENT COMPOSITION FOR REGENERATION OF MICRODAMAGE AND REGULATION OF ELASTICITY OF DELICATE FIBER |
| (57) The invention relates to a composition comprising transglutaminase and α-amylase.
Said composition can be used as a laundry washing composition, especially as a hand
cleaning washing composition, to effectively remove starch and/or polysaccharide residue-based
stains, even at a pH of 7.5 to 10.5. In addition, the composition repairs animal protein-based
fabric fibres, and can be used in a method for cleaning fabrics and/or for fabric
care.
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FIELD
[0001] The invention relates to a biodegradable transglutaminase-based detergent composition for regeneration of microdamage of surface and deep layers of composite delicate fiber caused by preceding chemical treatments and, which is more important, proteases in detergents and for regulation of elasticity by changing the tensile strength of protein fibers in delicate fabrics. The composition is to be included in fabric care detergents use of which makes it possible to increase effectiveness of protection of delicate fibers against microdamage because of inappropriate use of synthetic detergents in home washing and to regulate color fastness and maintain smoothness of fabrics. The composition is biodegradable, safe for skin and can be used for making detergents for sensitive skin such as dry, liquid, concentrated laundry detergents, while maintaining continuous cleanness of fabrics. Use of the composition can contribute to reducing of damaging effect on skin of the consumer with allergic diseases, while preserving skin hydration and elasticity during hand washing.
BACKGROUND
[0002] Daily, a large quantity of laundry detergents for various fabrics is used, such as washing powders for white and colored clothes, liquid detergents and capsules, conditioners, stain removers and bleaching agents. Developing of household chemicals with improved consumer properties is one of promising directions in the category of home care products. According to analysts' evaluation, nowadays consumers pay more attention to ingredients of laundry detergents and thoroughly examine them in terms of safety for people and environment. Besides, consumers look for more natural products without harmful chemical additives [Kantar Profiles/Mintel; A year of innovation in fabric and dish care, 2021, Mintel; KuRunData/Mintel; A year of innovation in fabric and dish care, 2021, Mintel]. This creates demand for more ecological products and changing of ingredients of household chemicals towards natural components. Thus, by 2025 there is going to be a trend for sustainable products with a high content of natural components.
[0003] A modern detergent shall combine a great number of functions within itself: removing general soils and specific stains, preventing formation of pills, maintaining fabric color brightness, preventing redeposition of soils in the washing process as well as having impact on aesthetic perception of clothes. The demand for new laundry solutions is determined by frequent use of short wash cycles and low-temperature cycles, the increasing demand for liquid laundry detergents with natural and ecological ingredients. Consumers want their clothes to look new longer and to be worn longer [https://www.savers.com/sites/default/files/reuse_report _2017_sav.pdf]. 55% of consumers are ready to pay more for products that improve quality of clothes and prolong their wear, which is directly connected with restoration of fabrics in the process of washing, wearing and drying. After the first wash delicate clothes start to look faded, lose softness, no longer look new, get damaged, while numerous washes lead to formation of pills and other undesirable defects such as microdamage invisible to the eye. In the research among European consumers carried out by Efficience, 54% of respondents said that they consider their clothes to be "worn out" as early as after 20 washes. Consumers might have a wish to replace "defective" clothes with new ones, but their frequent replacement is harmful for the environment and budget. In connection therewith, a detergent with new properties can significantly prolong wear of clothes. The most attractive advantage for consumers is care of fabrics, specifically, the care of fabrics that is obvious and visible to the eye [Survery Novozymes/Conjointly consumer survey Europe 2019, N=1902]. Thus, there is a need in developing of high-quality laundry detergents that would fulfill the specified tasks in an efficient way and would not harm the environment. One of such solutions is use of biotechnological components, specifically, enzymes; however, they have a number of restrictions of use in laundry detergents.
[0004] There are various types of fibers included in textiles. Wool and silk are most widely spread. Wool and silk are materials based on proteins of animal origin. These fibers have moderate strength, flexibility, elasticity, acid fastness and moisture absorption. Like any other proteins, proteins of wool and silk consist of polypeptides. Polypeptide chains in wool and silk make up a fibrous structure. Fibrous proteins are mainly characterised by their protein primary (amino acid sequence and composition) and secondary structure (interaction of hydrogen bonds between polypeptides). The secondary structure is an alpha-helix for keratin-based fibers [Hopkins GE. The Structure of the Wool Fiber. Textile Research Journal. 1949;19(12):816-821.] such as wool or hair. In case of silk, polypeptide chains have a beta-sheet in the secondary structure [Peng, Zhangchuan; Yang, Xi; Liu, Chun; Dong, Zhaoming; Wang, Feng; Wang, Xin; Hu, Wenbo; Zhang, Xia; Zhao, Ping; Xia, Qingyou. Structural and mechanical properties of silk from different instars of Bombyx mori. Biomacromolecules, 2019, 20, 3, 1203-1216.]. This structure explains exposure of fibers to impact of various enzymes. While some enzymes can cause useful modifications leading to changes in properties of fibers and increasing of their strength, others - such as proteases - can cause cleavage of protein molecules and, as a result, damage textiles on the microlevel. Preservation and restoration of properties of wool fibers require special care and gentle cleaning that are not provided by common laundry detergents.
[0005] Thus, one of the priority tasks of companies engaged in development of laundry detergents is introduction of new components that preserve and restore initial properties of various fibers, effectively remove soils and are ecological. One of such solutions is use of biotechnological enzymes. According to requirements of the international ecological certification Ecolabel EU, enzymes are welcome in the composition of detergents both from the point of view of soil removal performance and sustainable development. They are biodegradable and have numerous positive qualities. Thus, according to the European Commission, use of enzymes has positive economic and ecological consequences - reduction of wash temperature and water consumption, reduction of wash duration and quantity of toxic substances that get into sewage. Among enzymes, of special interest is transglutaminase, which has not been used in household products before.
[0006] Transglutaminase, or acyltransferase (enzyme classification code EC 2.3.2.13., CAS 80146-85-6), is a class of enzymes that catalyse formation of a covalent bond between the γ-carboxamide group of glutamine (acyl donors) and the free amino group of lysine (acyl acceptors) of proteins and peptides. If there is no amino group-bearing substrate, water molecules can act as acyl acceptors.
[0007] Transglutaminases can encapsulate lipids and/or fat-soluble materials, form heat- and water-resistant films, improve elasticity and water-retaining capacity of modified proteins, which explains their use in food industry. As regards textile, transglutaminase is used only for protein fixation on wool fibers, which contributes to restoration of wool damage, color fixation during dyeing as well as development of hydrophilic properties of wool fibers. Silk proteins grafted by means of transglutaminase restore textile fibers from wool, silk, cashmere, cotton or polyester, however there are no data about use of transglutaminase as a monosubstance for regeneration of microdamage and regulation of elasticity of delicate fiber. The activity of transglutaminase is known to greatly depend on their origin as most enzymes of this class are calcium-dependent.
[0008] Transglutaminases have optimum pH 5-8 [L. Zhang; L. Zhang; H. Yi; M. Du; C. Ma; X. Han; Z. Feng; Y. Jiao; Y. Zhang. Enzymatic characterization of transglutaminase from Streptomyces mobaraensis DSM 40587 in high salt and effect of enzymatic cross-linking of yak milk proteins on functional properties of stirred yogurt. 2012, 95(7), 3559-3568], are activated by bivalent calcium cations, inhibited by iron ions in hard water and strong complexing (chelating) agents, for example, EDTA, MGDA, GLDA, sodium citrate capable of effective bonding of polyvalent metal cations. The activity of transglutaminase is maintained within the range of temperatures of 20-60°C, the optimal temperature is 40-55°C, which restricts use of transglutaminase in low-temperature wash conditions. Transglutaminase is stable at pH from 5.0 to 9.0, 37°C during 30 min. The activity of TGase gradually decreases at an acidic pH and quickly decreases at a basic pH by 80% in 60 minutes, which negatively affects general performance in the system and demonstrates low stability of the component in products [L. Zhang; L. Zhang; H. Yi; M. Du; C. Ma; X. Han; Z. Feng; Y. Jiao; Y. Zhang. Enzymatic characterization of transglutaminase from Streptomyces mobaraensis DSM 40587 in high salt and effect of enzymatic cross-linking of yak milk proteins on functional properties of stirred yogurt. 2012, 95(7), 3559-3568]. Transglutaminase-containing systems do not require a great amount of water, which enables producing a concentrated detergent and reducing water consumption during its use.
[0009] The activity of transglutaminase is known to influence the activity of alpha-amylase in case of co-presence and reduce effect of its use [Wee, May & Henry, Jeya. (2019). Effects of Transglutaminase on the Protein Network and In Vitro Starch Digestibility of Asian Wheat Noodles. Foods. 8. 607. doi:10.3390/foods8120607]. The cause thereof might be that starchy soils are strongly held on the surface of different fabrics, including delicate fabrics, and it is difficult to remove them in low-temperature wash cycles. Residues of starch soils are held in delicate fabrics as transglutaminase forms a dense protein mesh for restoration of the delicate fabric structure, which restricts enzymatic access of alpha-amylase and destruction of these soils. In addition, starch soils make textiles stiff, which is felt by skin surface during daily wear. Also, the activity of alpha-amylase can decrease during product storage [Wang, HW., Kim, I.H., Park, CS. et al. Immobilization of α-amylase from Bacillus licheniformis on developed support using microbial transglutaminase. Korean J. Chem. Eng.. 25, 801-803 (2008)]https://doi.org/10.1007/s11814-008-0131-1, as the optimal pH range for alpha-amylase is 6-9.
[0010] Use of transglutaminases from microorganisms, specifically, bacteria, in laundry detergents is innovative and insufficiently studied. The authors suggested that products based on transglutaminase biotechnologically obtained from bacteria and alpha-amylase containing no additional proteins (silk proteins) would improve durability of delicate fabrics and restore microdamage of fibers. This microdamage arises during wear and as a result of impact of other protease-containing detergents. The authors unexpectedly discovered that transglutaminase is compatible with alpha-amylase in the non-trivial pH range 7.5-10.5. It was shown that presence of transglutaminase does not reduce effectiveness of alpha-amylase with respect to soils based on strongly held starch-based polysaccharides but, on the opposite, increases and demonstrates synergism for significant improvement of stain-removal performance. Often, performance of laundry detergents is ensured by basic pH 10-12 or a high content of soap, however, this is not desirable for delicate fabrics because of their damage and does not contribute to effective stain removal. Presence of transglutaminase enables maintaining the activity of alpha-amylase and increasing its performance in liquid laundry detergents.
[0011] The composition of the liquid delicate laundry detergent with transglutaminase and alpha-amylase of the invention makes it possible to effectively remove stains and soils from delicate fabrics and hard-to-reach areas, thus reducing the total quantity of surfactants in household chemicals, ensuring dermatological safety for people and achieving almost complete removal of biological stains based on simple and complex starch compounds. Removal of starch compounds has a hygienic function, prevents excessive sorption of other soils on surfaces of delicate fabrics (antiresorbing effect) and increases wear of textiles in the long run. Addition of transglutaminase and alpha-amylase enables increasing performance of removal of food stains (chocolate drink, cocoa, grass, starch) and blood stains without impairing the look of delicate fabrics. The technical result of the innovative complex based on transglutaminase and alpha-amylase consists in effective regeneration of microdamage, regulation of elasticity of delicate fiber, improvement of light-resistance of delicate materials during the cycle and maintaining of enzymatic activity of enzymes upon co-presence in laundry detergents. The complex is active in the basic pH range, specifically, 7.5-10.5 units, and the temperature range, specifically, from +15 to +60°C, which expands the sphere of use in ecological products for preservation of planet resources and improvement of energy efficiency of wash and demonstrates stability in extreme conditions for both enzymes. Transglutaminase and alpha-amylase are materials containing corresponding pure active components as well as additional ingredients and technical impurities that could have been formed in the process of production of target raw materials.
[0012] The combination of components is known to provide for synergetic effect with respect to stubborn soils and preserve effectiveness at a lower percentage of introduction of separate components. Transglutaminase obtained by modern biotechnology methods without use of genetically modified organisms is an active enzyme cross-linking amines or proteins of delicate fabrics without additional use of other agents. Alpha-amylase acts as an additive enzyme for destruction of glucoside bonds in most widely spread soils of starch origin as well as in polysaccharide residues. An additional property of the innovative complex is regeneration of microdamage of delicate fabrics and improvement of elasticity of delicate fibers.
[0013] The innovative complex including transglutaminase and α-amylase is directed at effective regeneration of microdamage and improvement of elasticity of delicate fabric fibers, removal of biological soils by cleavage of glucoside bonds in stains, improvement of washing performance of detergents. The complex based on natural and biodegradable components is effective within the basic pH range 7.5-10.5 upon presence of different synthetic and natural components, which makes it possible to use it in ecological delicate laundry detergents. The components have a targeted effect on the structure of different fabrics, specifically, delicate ones, and have a softening effect on fabrics making them more pleasant to the touch. Thus, combined use of the components in one product results in greater cleavage of complex biological soils by increasing activity of α-amylase and regeneration of microdamage of delicate fabrics thanks to transglutaminase, which makes it possible to enable fast removal of complex stubborn soils even in conditions of cold water use and to preserve consumer properties of delicate fabric garments for a long period of time. A distinctive feature is that the components in the claimed concentrations are effective only against complex soils and do not spoil the look of most fabrics, specifically, delicate ones. The composition has no aggressive inorganic surfactants and organic solvents, occlusive filming agents and alcohols, quaternary ammonium bases, silicones, optical bleaching agents, chlorine and phosphorus derivatives, therefore it is possible to use detergents with this composition on a regular basis without any harm for human skin. The combined use of these components has a synergetic effect ensuring complex care of delicate fabrics within one detergent for daily use.
[0014] Thus, the invention, generally, relates to the composition and its use enabling achieving of such technical results as regeneration of microdamage of delicate fibers with simultaneous improvement of elasticity and removal of stubborn polysaccharide-based soils, while preserving the look of materials for a longer period of time, maintaining continuous cleanness and brightness of color fabrics in ΔL∗a∗b coordinates, which are not achieved or are insufficiently achieved by modern commercially available products in this pertinent art.
SUMMARY OF THE INVENTION
[0015] In the first aspect, the invention relates to a composition that is intended for use in laundry detergents and is active at pH 7.5-10.5 and water hardness 0-15° dH consisting of:
- (A) Biotransglutaminase obtained biotechnologically from bacteria or yeast-like fungi resistant to presence of protease, wherein the said transglutaminase has biological activity of not less than 100 U/g at 37°C, viscosity of 0-1000 sPas at 37°C;
- (B) Alpha-amylase obtained biotechnologically from microorganisms, with activity of not less than 100 U/g, at 37°C, viscosity of 0-1000 sPas at 37°C;
[0016] In the second aspect, the invention relates to use of the composition of the invention in a laundry detergent. The laundry detergent of the invention can contain 0.10-1.0 % wt. composition of the invention.
[0017] The composition is characterized in that the quantity of transglutaminase in the said mass ratio of transglutaminase and alpha-amylase (0.001-0.1):(0.0025-0.05) is 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09 or 0.10.
[0018] The composition is characterized in that the quantity of alpha-amylase in the said mass ratio of transglutaminase and alpha-amylase (0.001-0.1):(0.0025-0.05) is 0.0025, 0.0030, 0.0035, 0.0040, 0.0045, 0.0050, 0.0055, 0.0060, 0.0065, 0.0070, 0.0075, 0.0080, 0.0085, 0.0090, 0.0095, 0.01, 0.02, 0.03, 0.04 or 0.05.
[0019] The composition is characterized in that the said transglutaminase is in a hydroglyceric or hydrosorbitol or hydroglycerol-sorbitol or hydropropyleneglycol or hydroglycerol-maltitol solution.
[0020] The composition is characterized in that the said hydroglycerol-maltitol solution of transglutaminase is the commercially available product Sternzym PT 100L.
[0022] The composition is characterized in that the said alpha-amylase is in a hydroglyceric or hydrosorbitol or hydroglycerol-sorbitol or hydropropyleneglycol or hydroglycerol-maltitol solution.
[0023] The composition is characterized in that the said hydroglycerol-maltitol solution of transglutaminase is the commercially available product Amplify® Prime 100L. Alpha-amylase is available, specifically, from Novozymes and can be identified, for example, by < https://biosolutions.novozymes.com/en/laundry/products/amplifyr-prime-100-l>.
[0024] The composition is characterized in that the activity of the said transglutaminase is at least 100 U/g at pH 8.0-10.0.
[0025] The composition is characterized in that the activity of the said alpha-amylase is at least 100 U/g at pH 8.0-10.0.
[0026] The composition is characterized in that the said detergent is selected from a laundry detergent, including a delicate laundry detergent or a baby laundry detergent, a laundry conditioner, a stain remover for laundry pre-treatment and wash, a liquid detergent and a laundry conditioner.
[0027] In another aspect the invention relates to a laundry detergent containing 0.10-1.00% wt. composition of the invention.
[0028] The laundry detergent is characterized in that the said detergent is selected from a delicate laundry detergent and a baby laundry detergent.
[0029] The laundry detergent is characterized in that the said detergent is selected from a membrane laundry detergent and a sports laundry detergent.
[0031] In another aspect the invention relates to use of the composition of the invention for regeneration of microdamage of delicate fabrics and regulation of elasticity of delicate fiber.
[0032] The use is characterized in that the said fabric is selected from delicate fabrics (wool, silk, cashmere, merino wool, down, feather and their mixtures).
[0033] The invention will be disclosed in detail in specific cases of its embodiment and illustrated by examples of implementation.
DESCRIPTION OF THE INVENTION
[0034] In the first aspect the invention relates to biotransglutaminase intended for use in detergents for regeneration of microdamage and improvement of elasticity of delicate fiber, wherein the said transglutaminase has a biological activity of at least 100 U/g at 37°C.
[0035] The mass content of transglutaminase in the said detergents can be 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09 or 0.10 or any values between the said ones.
[0036] Transglutaminase is characterized in that biotransglutaminase is transglutaminase obtained biotechnologically from microorganisms, specifically, fungi or bacteria. Transglutaminase can be a substance or a commercially available product with the registration number CAS 80146-85-6.
[0037] The present invention also relates to use of transglutaminase of the invention in detergents.
[0038] The detergent can contain 0.001-0.1 wt.% transglutaminase of the invention. For example, the detergent can contain 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09 or 0.10% wt. or any values between the said ones of transglutaminase of the invention.
[0039] The present invention also relates to a delicate laundry detergent containing 0.001-0.1% wt. transglutaminase of the invention.
[0040] The present invention also relates to a laundry detergent, a sports laundry detergent or a baby laundry detergent containing 0.001-0.1% wt. transglutaminase of the invention.
[0041] The present invention also relates to a white and color laundry powder containing 0.001-0.1% wt. transglutaminase of the invention.
[0042] The detergent of the invention can contain, %wt.:
| Biotransglutaminase Transglutaminase | 0.001-0.1 wt.%. |
| Alpha-amylase Amylase | 0.0025-0.05 wt.% |
[0043] The present invention also relates to use of transglutaminase and alpha-amylase of the invention for regeneration of microdamage and improvement of elasticity of delicate fiber caused by preceding chemical treatments or proteases in the composition of detergents.
[0044] The detergents, preferably, do not contain other active and/or auxiliary substances such as washing active agents and/or acceptable auxiliary substances, however, the detergents can still contain them. Such substances represent or can represent agents conventionally used in this field, that are well known to a person skilled in the art. Addition of the said agents in the composition of the complex of the invention does not invalidate achievement of the declared technical results, but can improve them.
[0045] The invention also encompasses such detergents as a delicate laundry detergent, a sports laundry detergent, a baby laundry detergent, a laundry powder.
[0046] All mass fractions, mass parts, mass percentages, as well as volume fractions, volume parts and volume percentages in the present disclosure are given in relation to the detergent, agent, composition or product they relate to in the given context.
[0047] In the detergent of the invention auxiliary acceptable substances can be selected from the following categories of components.
[0048] Anionic surfactants:
Salts of higher carboxylic acids of the general formula: R1-CO2X1, wherein R1 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 5 to 21 carbon atoms, and X1 is a cation of an alkali and/or alkali-earth metal, ammonium, alkylammonium, alkanolammonium, glucoammonium, basic amino acid;
Alkyl polyethyleneglycol sulfate of the general formula: R2-O(-CH2-CH2-O) n1SO3X2, wherein n1 is from 1 to 10 and denotes the quantity of polyethyleneglycol groups, R2 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 6 to 22 carbon atoms, and X2 is a cation of an alkali and/or alkali-earth metal, ammonium, alkylammonium, alkanolammonium, glucoammonium;
Alkyl sulfate of the general formula R3-OSO3X3, wherein R3 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 6 to 22 carbon atoms, and X3 is a cation of an alkali and/or alkali-earth metal, ammonium, alkylammonium, alkanolammonium, glucoammonium;
Salt of higher fatty acid amide and methylglycine of the general formula R4-C(O)-N(-CH3)-CH2-CO2X4, wherein R4 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 5 to 21 carbon atoms, and X4 is a cation of an alkali and/or alkali-earth metal, ammonium, alkylammonium, alkanolammonium, glucoammonium;
Alkyl polyethyleneglycol carboxylate of the general formula: R5-O(-CH2-CH2-O-)n2CH2-CO2X5, wherein n2 is from 1 to 15 and denotes the quantity of polyethylene glycol groups, R5 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 6 to 22 carbon atoms, and X5 is a cation of an alkali and/or alkali-earth metal, ammonium, alkylammonium, alkanolammonium, glucoammonium;
Disubstituted salt of 2-sulfocarboxylic acid of the general formula: R6-CH(-SO3X6)-CO2X6, wherein R6 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 4 to 20 carbon atoms, and X6 is a cation of an alkali and/or alkali-earth metal, ammonium, alkylammonium, alkanolammonium, glucoammonium;
Mono or disubstituted salt of higher carboxylic acid amide and glutamic acid of the general formula: R7-C(O)-NH-CH(-CH2-CH2-CO2X7)-CO2X7, wherein R7 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 5 to 21 carbon atoms, and X7 is a cation of an alkali and/or alkali-earth metal, ammonium, alkylammonium, alkanolammonium, glucoammonium or hydrogen;
Salt of higher fatty acid amide and glycine of the general formula R8-C(O)-NH-CH2-CO2X8, wherein R8 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 5 to 21 carbon atoms, and X8 is a cation of an alkali and/or alkali-earth metal, ammonium, alkylammonium, alkanolammonium, glucoammonium;
Salt of higher fatty acid amide and alanine of the general formula R9-C(O)-NH-CH(-CH3)-CO2X9,, wherein R9 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 5 to 21 carbon atoms, and X9 is a cation of an alkali and/or alkali-earth metal, ammonium, alkylammonium, alkanolammonium, glucoammonium;
Salt of higher fatty acid amide and 2-aminomethylethanesulfonic acid of the general formula R10-C(O)-N(-CH3)-CH2-CH2-SO3X10, wherein R10 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 5 to 21 carbon atoms, and X10 is a cation of an alkali and/or alkali-earth metal, ammonium, alkylammonium, alkanolammonium, glucoammonium;
Alkylpolyglucoside hydroxypropylsulfonate of the general formula: R11-O-[G]p1-O-CH2-CH(-OH)-CH2-SO3X11, wherein R11 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 6 to 22 carbon atoms, G is a saccharide fragment containing 5 or 6 carbon atoms, p1 is from 1 to 4, and X11 is a cation of an alkali and/or alkali-earth metal, ammonium, alkylammonium, alkanolammonium, glucoammonium;
Alkylpolyglucoside carboxylate of the general formula: R12-O-[G]p2-O-CH2-CO2X12, wherein R12 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 6 to 22 carbon atoms, G is a saccharide fragment containing 5 or 6 carbon atoms, p2 is from 1 to 4, and X12 is a cation of an alkali and/or alkali-earth metal, ammonium, alkylammonium, alkanolammonium, glucoammonium;
Salt of higher fatty acid amide and threonine of the general formula: R13-C(O)-NH-CH(-CH(-OH)-CH3)-CO2X13, wherein R13 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 5 to 21 carbon atoms, and X13 is a cation of an alkali and/or alkali-earth metal, ammonium, alkylammonium, alkanolammonium, glucoammonium;
Salt of higher fatty acid amide and amino acid obtained by hydrolysis of proteins of vegetable raw materials of the general formula: R14-C(O)-AAX14, wherein R14 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 5 to 21 carbon atoms, AA is an amino acid or peptide obtained during hydrolysis of vegetable protein (possible protein sources: apple, soybeans, wheat, cotton, etc.), and X14 is a cation of an alkali and/or alkali-earth metal, ammonium, alkylammonium, alkanolammonium, glucoammonium.
[0049] Amphoteric surfactants:
Disubstituted salt of acylamphodiacetate of the general formula: R15-C(O)-NH-CH2-CH2-N(-CH2-CO2X15)-CH2-CH2-O-CH2-CO2X15, wherein R15 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 5 to 21 carbon atoms, and X15 is a cation of an alkali and/or alkali-earth metal, ammonium, alkylammonium, alkanolammonium, glucoammonium;
Salt of acylamphoacetate of the general formula: R16-C(O)-NH-CH2-CH2-N(-CH2-CO2X16)-CH2-CH2-OH, wherein R16 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 5 to 21 carbon atoms, and X16 is a cation of an alkali and/or alkali-earth metal, ammonium, alkylammonium, alkanolammonium, glucoammonium;
Salt of alkylamphoacetate of the general formula: R17-C(=N-CH2-CH2-N((-CH2-CH2-OH)-CH2-CO2X17)-), wherein R17 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 5 to 21 carbon atoms, and X17 is a cation of an alkali and/or alkali-earth metal, ammonium, alkylammonium, alkanolammonium, glucoammonium;
Acylamidoalkylbetaine of the general formula: R18-C(O)-NH-R19-N(-CH3)2)-CH2-CO2 , wherein R18 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 5 to 21 carbon atoms, R19 is an alkyl group with the hydrocarbon chain length from 1 to 4 carbon atoms;
Acylamidoalkylhydroxysultaine of the general formula: R20-C(O)-NH-R21-N(-CH3)2-CH2-CH(-OH)-CH2-SO3, wherein R20 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 5 to 21 carbon atoms, R21 is an alkyl group with the hydrocarbon chain length from 1 to 4 carbon atoms;
Acylamidoalkylamine oxide of the general formula: R22-C(O)-NH-R23-N(-CH3)2-O, wherein R22 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 5 to 21 carbon atoms, R23 is an alkyl group with the hydrocarbon chain length from 1 to 4 carbon atoms;
Alkylbetaine of the general formula: R24-N(-CH3)2)-CH2-CO2, wherein R24 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 5 to 21 carbon atoms;
Alkylhydroxysultaine of the general formula: R25-N(-CH3)2-CH2-CH(-OH)-CH2-SO3, wherein R25 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 6 to 22 carbon atoms;
Alkylsultaine of the general formula: R26-N(-CH3)2-CH2-CH2-CH2-SO3, wherein R26 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 6 to 22 carbon atoms;
Alkylamine oxide of the general formula: R27-N(-CH3)2-O, wherein R26 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 6 to 22 carbon atoms.
[0050] Non-ionic surfactants:
Alkylglucoside of the general formula: R28-O-[G]p3, wherein R28 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 4 to 22 carbon atoms, G is a saccharide fragment containing 5 or 6 carbon atoms, p3 is from 1 to 4;
Alkylpolyethyleneglycol of the general formula: R29-O(-CH2-CH2-O-)n3H, wherein n3 is fom 2 to 20 and denotes the quantity of polyethyleneglycol groups, R29 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 6 to 22 carbon atoms;
Alkylpolyethylene/propyleneglycol of the general formula: R30-O(-CH2-CH2-O-)n4(-CH(-CH3)-CH2-O-)n5H, wherein n4 is from 2 to 20 and denotes the quantity of polyethyleneglycol groups, n5 is from 2 to 20 and denotes the quantity of polypropyleneglycol groups, R30 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 6 to 22 carbon atoms;
Dialkylpolyethyleneglycol of the general formula: R31-O(-CH2-CH2-O-)n6R32, wherein n6 is from 2 to 20 and denotes the quantity of polyethyleneglycol groups, R31 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 6 to 22 carbon atoms, R32 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 1 to 12 carbon atoms;
Dialkylpolyethylene/propyleneglycol of the general formula: R33-O(-CH2-CH2-O-)n7(-CH(-CH3)-CH2-O-)n8-R34, wherein n7 is from 2 to 20 and denotes the quantity of polyethyleneglycol groups, n8 is from 2 to 20 and denotes the quantity of polyepropyleneglycol groups, R33 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 6 to 22 carbon atoms, R34 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 1 to 12 carbon atoms.
[0051] Dispersed medium for the polysaccharide/solvent:
Organic alcohol of the general formula: R35(-OH)s1, wherein R35 is an alkyl group with the hydrocarbon chain length from 3 to 12 carbon atoms, S1 is from 1 to 12 and denotes the quantity of hydroxyl groups located in the hydrocarbon radical in a random order in relation to each other;
Alkylpolypropyleneglycol of the general formula: H(-CH(-CH3)-CH2-O-)n9R36, wherein n9 is from 2 to 10 and denotes the quantity of polypropyleneglycol groups, R36 is an alkyl group with the hydrocarbon chain length from 1 to 10 carbon atoms.
[0052] pH adjustors:
Organic acids of the general formula: R37(-OH)s2(-COOH)m1, wherein R37 is an alkyl group with the hydrocarbon chain length from 1 to 12 carbon atoms, S2 is from 1 to 12 and denotes the quantity of hydroxyl groups located in the hydrocarbon radical in a random order in relation to each other, M1 is from 1 to 4 and denotes the quantity of carboxyl groups located in the hydrocarbon radical in a random order in relation to each other;
Solutions of hydroxides of alkali or alkali-earth metals, ammonia, primary and tertiary alkylamines, primary and tertiary alkanolamines, primary and tertiary glucamines, basic amino acids, disodium salt of citric acid, trisodium salt of citric acid.
[0053] Chelating agent:
Trisodium salt of methylglycinediacetic acid, tetrasodium salt of glutaminediacetic acid, trisodium salt of ethylenediamine-(N,N)-disuccinate;
Esters of phosphonic acids of the general formula RP(O)(OR'1)n(OH)2-n, wherein R, R' are organic radicals, specifically alkyl, alkenyl or aryl radical, wherein, depending on hydroxyl groups, they can be primary (n=1, acidic phosphonates) and secondary (n=2, complete phosphonates);
Organic acids as well as salts of alkali metals, ammonium, alkylammonium, alkanolammonium, glucoammonium corresponding to the following acids: citric acid, malic acid, tartaric acid, glutaric acid, adipic acid, glucuronic acid, galacturonic acid, galactaric acid, gluconic acid, phytic acid, polytaconic acid, polyacrylic acid, polymethacrylic acid, a copolymer from acrylic and maleic acids as well as oganic acids of the general formula R38(-OH)s3(-COOH)m2, wherein R38 is an alkyl group with the hydrocarbon chain length from 1 to 12 carbon atoms, S3 is from 1 to 12 and denotes the quantity of hydroxyl groups located in the hydrocarbon radical in a random order in relation to each other, M2 is from 1 to 4 and denotes the quantity of carboxyl groups located in the hydrocarbon radical in a random order in relation to each other.
[0054] Soil redeposition inhibitors:
Derivatives of polysaccharides: sodium salt of carboxymethylpolysaccharide, hydroxyalkylpolysaccharide, alkylpolysaccharide;
Polyvinylpyrrolidone and its derivatives, copolymers of polyvinylpyrrolidone and vinylimidazole;
Water-soluble salts of polyacrylic acid, polymethacrylic acid, copolymer of acrylic/methacrylic and maleic acid.
[0055] Defoaming agents:
Higher carboxylic acids of the general formula: R39-CO2H, wherein R39 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 5 to 21 carbon atoms;
Higher carboxylic alcohols of the general formula: R40-COH, wherein R40 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 5 to 21 carbon atoms;
Ethers of higher carboxylic alcohols of the general formula: R41-O-R42, wherein R41, R42 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 4 to 22 carbon atoms;
Bisamides of alkyldiamines and higher carboxylic acids of the general formula: R43-C(O)-NH-R44-NH-C(O)-R45, wherein R43, R45 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 5 to 21 carbon atoms, and R44 is alkyl radical with the hydrocarbon chain length from 1 to 12 carbon atoms.
[0056] Preservatives:
Organic acids and salts of alkali and alkali-earth metals, ammonium, alkylammonium, alkanolammonium, glucoammonium corresponding to the following acids: benzoic acid, sorbic acid, 4-methoxybenzoic acid, salicylic acid, undecylenic acid;
Organic alcohols and phenols: phenoxyethanol, benzyl alcohol, caprylyl glycol, ethylhexylglycerin, phenethyl alcohol, 3-methyl-4-isopropylphenol, 2,4-dichlorobenzyl alcohol;
Broad-spectrum biocydes: benzisothiazolinone, dodecyldipropylene triamine, methylisothiazolinone.
Fungicides: sodium pyrithione, climbazole.
Enzymes: pectate lyase, mannanase, mannosidase, cellulase, aminooxidase, feruloyl esterase, beta-glucanase, tannase, alpha-glycosidase, beta-glycosidase, alpha-galactosidase, beta-galactosidase, manganese peroxidase, licheninase, xylanase and other commercially available enzymes used in laundry detergents.
Perfumes with essential oils or essential oils in pure form or as mixtures in different ratios of: orange, bergamot, lemon, lime, tangerine, grapefruit, neroli, rosewood, yuja, lemongrass, lavender, sage, rosemary, thyme, melissa, mint of different kinds, manuka, eucalyptus, cypress, pine wood, cedar, sandal wood, vetiver, black pepper, rose pepper, cinnamon, cardamom, coriander, jasmine, rose, peony, blue chamomile, ylang-ylang, monoi, palmarose and other commercially available essential oils.
[0057] In another aspect the invention relates to use of biotransglutaminase and alpha-amylase of the invention for regeneration of microdamage and improvement of elasticity of delicate fiber caused by preceding chemical treatments or proteases in the composition of detergents. The use is characterized in that the said delicate fabric is selected from wool, silk, cashmere, merino wool, down, feather and their mixtures or other protein-based material.
EXPERIMENTAL PART
[0058] The examples included in the present description are non-limiting for the invention and are given only with the purpose of illustration and proof of achievement of the expected technical results. These examples are ones of many experimental data obtained by the inventors that prove high performance of the detergents within the scope of the invention.
Example 1.
[0059] Components to be included in the composition of the invention were studied as part of laundry detergents. A liquid detergent, specifically, a liquid concentrated delicate laundry detergent was prepared within the scope of the invention (Table 1).
Table 1. Ingredients of the liquid delicate laundry detergent with the claimed composition
| No. | Component | Content, % wt. |
| 1 | Purified water | up to 100.00 |
| 2 | Alkyl polyethyleneglycol sulfate of the general formula R1-O(-CH2-CH2-O)n1 (SO3) n2 X1, wherein n1 is from 0 to 10 and denotes the quantity of polyethylene groups, R1 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 5 to 22 carbon atoms, n2 is from 0 to 1 and denotes the quantity of sulfate groups, X1 is a cation of an alkali and/or alkali-earth metal, ammonium, alkylammonium, alkanolammonium, glucoammonium, basic amino acid; | 2.5-10.00 |
| 3 | Alkylglucoside of the general formula: R28-O-[G]p3, wherein R28is an alkyl and/or alkenyl group with the hydrocarbon chain length from 4 to 22 carbon atoms, G is a saccharide fragment containing 5 or 6 carbon atoms, p3 is from 1 to 4, specifically, C10-16 alkylpolyglucoside and C8-10 alkylpolyglucoside | 1.5-10.00 |
| 4 | Salts of higher carboxylic acids of the general formula: R1-CO2X1, wherein R1 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 5 to 21 carbon atoms, and X1 is a cation of an alkali and/or alkali-earth metal, ammonium, alkylammonium, alkanolammonium, glucoammonium, basic amino acid; | 1.0-5.00 |
| 5 | Biotransglutaminase - expressed as active substance | 0.001-0.1 |
| 6 | Alpha-amylase - expressed as active substance | 0.0025-0.05 |
| 7 | Complexing agent MGDA | 0.1-1.50 |
| 8 | Natural-origin glycerol | 1.0-5.00 |
| 9 | Sodium chloride | 0.05-5.00 |
| 10 | Cotton seed extract | 0.005-0.50 |
| 11 | Preservative | 0.005-0.75 |
| 12 | Sodium hydroxide or potassium hydroxide for pH adjustment | 0.005-1.00 |
| 13 | pH adjustor, for example, citric acid or another acid | 0.005-1.00 |
| 14 | Auxiliary substances if needed | 0.10-10.00 |
[0060] The prepared liquid laundry and stain-removing detergent enables high regeneration of microdamage, improves elasticity of delicate fabrics and effectively removes stubborn starch-based stains at any running water hardness 0-15° dH at at any wash temperature from +15°C to +60°C. The average pH range of the product is 7.5-9.5. It does not change fabric color or wash out dyes, preserves the look of the garment, leaves no streaks, is fully washed away from the fabric surface, suits for baby laundry and for people with sensitive skin surface, remains stable during storage of 24 months (observation period).
Example 2.
[0061] Components to be included in the composition of the invention were studied as part of laundry detergents. A liquid detergent, specifically, a universal concentrated delicate laundry stain remover was prepared within the scope of the invention (Table 2).
Table 2. Ingredients of the liquid delicate laundry stain remover with the claimed
composition
| No. | Component | Content, % wt. |
| 1 | Purified water | up to 100.00 |
| 2 | Alkyl polyethyleneglycol sulfate of the general formula: R2-O(-CH2-CH2-O) n1SO3X2, wherein n1 is from 1 to 10 and denotes the quantity of polyethyleneglycol groups, R2 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 6 to 22 carbon atoms, and X2 is a cation of an alkali and/or alkali-earth metal, ammonium, alkylammonium, alkanolammonium, glucoammonium | 2.5-10.00 |
| 3 | Alkylglucoside of the general formula: R28-O-[G]p3, wherein R28is an alkyl and/or alkenyl group with the hydrocarbon chain length from 4 to 22 carbon atoms, G is a saccharide fragment containing 5 or 6 carbon atoms, p3 is from 1 to 4, specifically, C10-16 alkylpolyglucoside and C8-10 alkylpolyglucoside | 1.5-10.00 |
| 5 | Acylamidoalkylbetaine of the general formula: R18-C(O)-NH-R19-N(-CH3)2)-CH2-CO2, where R18 is an alkyl and/or alkenyl group with the hydrocarbon chain length from 5 to 21 carbon atoms, R19 is an alkyl group with the hydrocarbon chain length from 1 to 4 carbon atoms | 0.5-7.00 |
| 6 | Alpha-amylase - expressed as active substance | 0.0025-0.05 |
| 7 | Biotransglutaminase - expressed as active substance | 0.001-0.10 |
| 8 | Additional enzymes, for example, lipase, pectinase, mannanase and others if required | 0.0-1.0 |
| 9 | Natural-origin glycerol | 0.0-5.00 |
| 10 | Sodium chloride | 0.05-5.00 |
| 10 | Cotton seed extract | 0.005-0.50 |
| 11 | Preservative | 0.005-0.75 |
| 12 | Sodium hydroxide or potassium hydroxide for pH adjustment | 0.005-1.00 |
| 13 | Citric acid or another acid for pH adjustment | 0.005-1.00 |
| 14 | Auxiliary substances if needed | 0.10-10.00 |
[0062] The prepared liquid laundry and stain-removing detergent provides for highly effective regeneration of microdamage and improves elasticity of delicate fiber, at any running water hardness 0-15° dH and at any wash temperature from +15°C to +60°C. The average pH range of the product is 7.0-9.0. It does not change fabric color or wash out dyes, preserves the look of the garment, leaves no streaks, is fully washed away from the fabric surface, suits for baby laundry and for people with sensitive skin surface, remains stable during storage of 24 months (observation period).
Example 3.
[0063] There was a study of visualization of fibers of 2 types of fabrics (wool+ cashmere 80%:20%, silk 100%) under electronic microscope to evaluate damage/restoration of fabrics after wash with the liquid concentrated delicate laundry detergent using the composition of the invention. The liquid concentrated delicate laundry detergent given in Table 1 was used as the base for introduction of components.
[0064] The test method was based on the examination of samples of wool and silk fabrics before and after washes (1, 5, 10) to determine effectiveness of regeneration of microdamage of fabric fibers. The examination was held by high-resolution visualization of different areas of samples with search for damaged fabric fibers with topograpical and/or compositional contrast. Effects of combinations of enzymes on fiber structures of 26 fabric samples (wool+ cashmere 80%:20% and silk 100%) were studied.
[0065] At the first stage of the experiment samples of the selected delicate fabrics (wool+ cashmere 80%:20% and silk 100%) were treated by protease (subtilisin), transglutaminase and proteins of silk in different sequences and combinations to evaluate mutual impact on the fabric structure. Thereafter the fabric samples were sent to the laboratory to examine the surface and structure under an electronic microscope with the aim of visualization of the damage/restoration after wash with enzymes. 3 pieces sized 1×1 cm2 were cut out from the initial analysed fabric, placed on a carbon tape attached to an aluminum stage and fixed with an adhesive tape along the sample edges. Then, a thin layer of gold was sprayed on the surface of the samples for electrostatic discharge and afterwards the sample was placed into the electronic microscope chamber. A preliminary search for most damaged/restored areas was carried out for further high-resolution imaging by the scanning electron microscope FEI Teneo. Fabric surface visualization was carried out by means of the electron-ion microscope FEI Helios NanoLab 660 (double-beam scanning electron microscope) with localization of the most damaged/restored areas. The microscope chamber was pumped to the pressure of ≤10-3 Pa in high-vacuum mode. The navigational camera was used to take an overall optical image of samples placed in the chamber for convenience of their positioning during the tests. Accelerating voltage and current of the electron beam, detector and its operation mode were selected to take images that enable best visualization of relevant objects of the sample. To estimate uniformity of distribution of relevant objects on the sample surface and determine the area of interest the sample was preliminarily viewed at a low magnification (x100-x200). The area of interest was selected based on the need in displaying areas localized at the previous experiment stage.
[0066] The test parameters are given in Table 3.
Table 3. Test parameters
| Accelerating voltage of primary electron beam | 1-10 kV |
| Primary electron beam current | 0.40 nA |
| Type of detector in use | T1, CBS |
| Two-dimensional image pixel size | 22.5 nm - 1.70 µm |
| Two-dimensional projection horizontal field of view | 34.5 µm - 2.76 mm |
| Sample inclination angle | 0° |
[0067] The study results were used to compare and analyse the obtained images, evaluate defects of fabric samples: soils/presence of particles; damage of fibers/loss of integrity of fibers; frizz after impact of selective active substances: transglutaminase (abbreviated as TG), protease (subtilisin), silk proteins.
[0068] The samples were washed in a Miele washing machine once. The following conditions were selected as the test conditions: temperature 30°C, water hardness 10.9 °dH and hand wash mode (wool+cashmere and silk). The concentration of the liquid concentrated delicate laundry detergent was 5.0 g/l per standard washing machine load of 2-4 kg. Upon completion of the experiment the best combination of effects of enzymes on the fiber structure was selected. The wash results were evaluated by visualization on a scale from one to five comparing to the initial one:
5 - fiber structure was fully restored;
4 -minimum quantity of fiber defects;
3 -significant fiber structure cross-linking is visible;
2 -fabric delamination is seen;
1 -a large quantity of fabric defects
Results.
[0069] Based on the results of evaluation of effectiveness of regeneration of delicate fiber microdamage it was established that the composition of the invention included in the liquid delicate laundry detergent (Table 1) has a pronounced effect and as early as after first wash the garment preserves its original form, no microdamage perceived as small stretches or texture damage is seen on it comparing to fabrics washed with references, with protease (subtilisin) or silk proteins.
[0070] Visually there were fewer microdefects in the form of tears, microruptures of fibers invisible to the eye, using visualization by means of a focused beam of ions (22.5 nm) (see the microscopic examination in Figures 1 and 2).
[0071] One-time wash of silk fabric samples with addition of transglutaminase after fabric treatment with protease during 40 minutes made it possible to eliminate fabric damage caused by protease by more than 85%, which speaks of well-expressed regeneration of fiber microdamage caused by preceding chemical treatments and, more importantly, proteases (Figure 1). One-time wash of fabrics first with transglutaminase, then protease did not enable achieving such effectiveness and could not cross-link fiber structure, which speaks of a negative effect of the protease enzyme on the fabric structure. Addition of silk proteins has no impact on improvement of fabric structure as the effect is mediated by activity of transglutaminase in the composition of the delicate laundry detergent.
[0072] One-time wash of wool fabric samples with addition of transglutaminase before fabric treatment with protease (subtilisin) during 40 minutes made it possible to eliminate fabric damage by protease by more than 85%, which speaks of well-expressed regeneration of wool: cashmere fiber microdamage (Figure 2). One-time wash of fabrics first with protease, then transglutaminase did not enable achieving the same effectiveness and strengthening and restoring of fiber structure fully, which speaks of a negative effect of protease enzyme on the structure of delicate fabrics. Addition of silk proteins has no impact on improvement of fabric structure as the effect is mediated by activity of transglutaminase in the composition of the delicate laundry detergent.
Table 4. Evaluation of effectiveness of restoration of the structure of silk 100%
and wool:cashmere 80%:20% after treatment with transglutaminase per 1 wash
| Tested sample | Structure restoration | |||||||
| 0.025% protease, then 0.0025% TG | 0.0025% TG then 0.025% protease | 0.025% protease, then 0.25% silk proteins with 0.0025% TG | 0.25% silk proteins with 0.0025% TG, then 0.025% protease | |||||
| before | after | before | after | before | after | before | after | |
| Sample 1 Silk | 1 | 4.5∗ | 4.5∗ | 3 | 3 | 4 | 3.5 | 2.5 |
| Sample 2 Wool | 2 | 4.5∗ | 4.5∗ | 3 | 3 | 2.5 | 3 | 2.5 |
| *statistically significant result (p<0.05) |
[0073] Upon completion of the laboratory study there were pronounced changes of the evaluated parameter of regeneration of microdamage of delicate fabric fibers. The scale layer of wool and silk fibers after pre-treatment with protease is partially damaged, revealing more active groups: amino group, primary hydroxyl group and phenol hydroxyl group existing in serine and tyrosine residues. According to the results of electronic microscopy, addition of high-performance biotransglutaminase reduced the quantity of microdamage of fibers of selected fabrics (wool:cashmere 80%:20%, silk), by restoring damaged active groups, self-regeneration of fiber and strengthening of its structure. It was established for woolen fabric that pre-treatment with transglutaminase enables reducing further destruction by protease. The composition with 0.0025% biotransglutaminase enabled reducing the area of fabric fiber damage after a single wash. Use of protease demonstrated a greater quantity of scale frizz, lamination and microtears of fibers, which speaks of the need in using transglutaminase for levelling of this effect and regeneration of microdamage of delicate fabric fibers.
Example 4.
[0074] Visualization of fibers of 2 types of fabrics using an electronic microscope was carried out to evaluate damage or restoration of fabrics after multiple washes with the liquid concentrated delicate laundry detergent using different concentrations of transglutaminase and without the enzyme. The aim of the experiment was to prove regeneration of fiber microdamage after multiple washes of delicate garments and determine the range of effective concentrations of the selected transglutaminase of the invention to provide for targeted action in terms of cross-linking active groups existing in serine and tyrosine residues of delicate fibers. The liquid concentrated delicate laundry detergent given in Table 1 was used as the base for introduction of components. The experiment was carried out using the method described in Example 3, in the same conditions. Transglutaminase was added to the composition of the liquid detergent (pH 8.0-8.5) in concentrations of 0.001%, 0.0025% and 0.005%. The exposure cycle time was 40 minutes, which corresponds to the hand wash cycle and recommended wash time in an automatic washing machine. The cycle temperature was 30 degrees as wool and silk are among the most temperature-sensitive fibers. During the experiment performance of transglutaminase at selected concentrations after multiple washes (up to 10 washes) was tested.
[0075] The wash results were evaluated via visualization on a scale from 1 to 5 comparing with the initial value:
5 - fiber structure was fully restored;
4 -minimum quantity of fiber defects;
3 -significant fiber structure cross-linking is visible;
2 -fabric delamination is seen;
1 -a large quantity of fabric defects
Results.
[0076] Based on the results of evaluation of fiber microdamage regeneration it was established that the analysed transglutaminase-based composition included in the liquid delicate laundry detergent has a pronounced effect of self-restoration. After multiple use a delicate garment does not lose its original shape, microdamage and tears looking like small stretches or structure damage are eliminated. Based on the results of evaluation of fiber microdamage regeneration it was established that the optimal concentration of transglutaminase is within the range from 0.0025% to 0.005% as these enzyme contents result in self-cross-linking of damaged protein fibers without use of additional agents. At the minimum transglutaminase concentration of 0.001% in the composition minimum visible restoration of keratine and fibroin fibers in the composition of delicate fibers takes place. When a focused ion beam (22.5 nm) is used during visual examination, the quantity of defects in the form of tears, ruptures of fibers reduces proportionally to transglutaminase concentration, with the optimal transglutaminase concentration from 0.001% to 0.005%, and, more precisely, within the optimal content range from 0.0025% to 0.005% (see Figures 3a and 3b).
[0077] The ability of transglutaminase to catalyse formation of an isopeptide bond between γ-carboxamide groups ( -(C=O)NH2) of side chains of the glutamine residue and ε-aminogroups ( -NH2) of side chains of the lysine residue significantly expands the area of potential use of the enzyme in detergents for self-regeneration of delicate fiber microdamage. After wash the scale layer of wool fibers is partially damaged, revealing an amino group, primary hydroxyl and phenol hydroxyl group. Use of transglutaminase as a cross-linking monocomponent enables formation of numerous covalent bonds between fabric fibers in home conditions, preserving the structure, look, color and increasing light-resistance of garments. 5 washes of samples of delicate fabrics (wool: cashmere and silk) with addition of transglutaminase of the invention at the concentration of 0.0025% during 40 minutes enabled elimination of fabric damage by more than 80% comparing to the original look, which speaks of evident regeneration of microdamage and increase of strength by cross-linking of fiber structure (Table 5).
Table 5. Evaluation of effectiveness of restoration of the structure of fibers based
on protein substances after wash with transglutaminase
| Tested sample | Restoration of fabric structure, evaluation in scores | |||
| Effect after 5 washes | ||||
| Detergent base w/o TG | Composition with 0.001% transglutaminase | Composition with 0.0025% transglutaminase | Composition with 0.005% transglutaminase | |
| Sample 1 Silk | 2.5 | 2.5 | 4.0 | 4.5∗ |
| Sample 2 Wool | 2.0 | 2.5 | 4.5∗ | 4.5∗ |
| ∗statistically significant result (p<0.05) | ||||
[0078] Upon completion of the laboratory study there were obvious changes of the evaluated parameters, namely, elimination of microdamage in the form of tears, ruptures of fibers. Based on the results of electronic microscopy, addition of highly-effective biotransglutaminase at the concentration of 0.0025% (expressed as active substance) enabled self-regeneration of fibers of selected fabrics (wool: cashmere, silk) after 10 washes. The composition with 0.0025% biotransglutaminase effectively restored damaged fibers of delicate fabrics even after numerous washes and did not spoil the look of the garments. The samples of the liquid concentrated laundry detergent that did not contain transglutaminase demonstrated a greater quantity of scale frizz, laminations and fiber tears, which speaks of no effect of restoration of fabrics and their damage during washing of delicate fabrics. Multiple use of transglutaminase at the concentration of 0.0025% demonstrates a positive accumulative effect, enabling restoration up to 90% of wool and silk fibers during 10 washes. Multiple use of the liquid concentrated delicate laundry detergent has an aggressive impact on the structure of fibers on the microscopic and macroscopic levels, impairing their condition and leading to greater fabric defects than the transglutaminase-containing detergent (Table 6).
Table 6. Evaluation of effectiveness of restoration of the structure of fibers based
on protein substances after 5 and 10 washes using the transglutaminase-containing
detergent
| Tested sample | Structure restoration | |||
| Detergent base | Composition with 0.0025% transglutaminase | |||
| After 5 washes | After 10 washes | After 5 washes | After 10 washes | |
| Sample 1 Silk | 2.5 | 2.0 | 3.0 | 4.5∗ |
| Sample 2 Wool | 2.0 | 1.0 | 4.5∗ | 4.5∗ |
| ∗statistically significant result (p<0.05) | ||||
Example 5.
[0079] A laboratory study was carried out to determine changes in strength characteristics of samples of selected delicate fabrics after wash with protease and transglutaminase. The study was required to prove the hypothesis of improvement of strength characteristics after wash of delicate fabrics with transglutaminase. The liquid concentrated delicate laundry detergent given in Table 1 was used as the base for the experiment.
[0080] During wear textile is subjected to numerous stresses: stretching, compression, bending, friction. Fabric durability and maintaining of consumer properties depend on the ability of material to withstand such impacts. The mechanical properties of textile mean resistance to factors that worsen the structure as a result of continuous use. The ability of textile to withstand breaking strain is its tensile strength.
[0081] The test method is based on physical and chemical methods, determination of breaking strain and elongation of fabrics. The tests were carried out at the ambient temperature of 21.6°C, relative humidity of 60%, pressure of 97.3 kPa. This method enables evaluating effectiveness of impact after one wash with the concentrated delicate laundry detergent with addition of enzymes of the invention on the structure of fabric fibers, strength and elasticity of garments. The test principle is based on the steel ball method that consists in the following: a fabric sample of a certain area is pressed in the fixed base circle, while a round spherical upper rod is vertically pressed to the sample with a constant movement speed in such a way that the sample is deformed until it breaks and the tensile strength is measured. Usually the instrument used to measure fabric strength by the steel ball method is an electronic bullet-type machine for measuring fabric strength or a tensile tester equipped with a steel-ball tool. This is use of steel spherical surfaces for fabric tear.
[0082] The tests were carried out with the use of 18 fabric samples sized 50×100 mm washed with the concentrated delicate laundry detergent with and without transglutaminase of the invention. The samples were washed once in a Miele washing machine. The following requirements were selected as the test conditions: temperature of 30°C, water hardness 10.9° dH and "delicate fabrics" mode. The concentration of the liquid concentrated delicate laundry detergent was 5.0 g/l per standard washing machine load of 2-4 kg. The washed fabric samples were tested on a universal test machine Shimadzu AG 10kNX in longitudinal and transverse directions. The fabric strips are fixed on the universal tensile tester and are stretched with the impact of a force stop. Warp and weft data are recorded and the arithmetic mean of tensile strength is determined. At the same time, tear strength and elongation of fabric are determined. There are plots of changes of strength characteristics of pair samples in "force (N) - elongation (mm)" coordinates. The mean test results with the data of changes of each pair of samples in relation to the original fabric (samples 1-2) for each fabric type (wool+cashmere, silk) are given in Table 10.
[0083] The tensile load and elongation were determined on a universal tester Shimadzu AG 10kNX with the use of wedge-shaped clamps. Silk (blue washed silk twill Max Mara SVM-M40- 1808212; composition 100% silk with density ~60 g/m2) was tested at the rate of 20 mm/min. Wool (berry-red wool with cashmere M-U30 18082147; composition 80% wool and 20% cashmere with density ~400 g/m2) was tested at the rate of 50 mm/min. The values of tensile load and elongation were determined for the fabric samples. The sizes and mean values of test series results for the fabric samples are given in the table below.
Results.
[0084] The liquid concentrated delicate laundry detergent with biotransglutaminase enables increasing strength and elasticity of fibers of selected delicate fabrics, which speaks of regulation of elasticity of delicate fibers and correspondingly regeneration of microdamage. The impact of 0.0025% transglutaminase after treatment of fabrics with the liquid concentrated delicate laundry detergent containing 0.0025 % protease gives better results of strength characteristics for silk than initial effect on the structure of fabrics with transglutaminase and then protease. The effect of 0.0025% transglutaminase before treatment of fabric with the liquid concentrated delicate laundry detergent containing 0.0025 % protease gives better results of strength characteristics for wool than initial effect on the structure of fabrics with protease and then transglutaminase (Table 7).
Table 7. Evaluation of strength characteristics of fibers based on protein substances
after wash of wool and silk with transglutaminase
| Ingredients | Fabric | Tensile strength, N | Elongation, mm | Difference, % Strength | Difference, % Elasticity |
| Alternate treatment with transglutaminase and protease | |||||
| 1 wash with protease 0.0025%, then TG 0.0025% | Silk | 298.2 | 7.8 | +9.03∗ preventative fabric strengthening before use of protease | +9.30∗ preventative fabric strengthening before use of protease |
| 1 wash with TG 0.0025%, then protease 0.0025% | Silk | 327.8* | 8.6* | ||
| 1 wash with protease 0.0025%, then TG 0.0025% | Wool | 146.5* | 28.5 | +20.18∗ post-restoration of fibers damaged by protease | +16.18∗ preventative fabric strengthening before use of protease |
| 1 wash with TG 0.0025%, then protease 0.0025% | Wool | 121.9 | 34.0 * | ||
| ∗significant result | |||||
[0085] The samples of silk fabrics washed with the liquid laundry detergent with 0.0025% biotransglutaminase and then treated with 0.0025% bioprotease demonstrated an increase of tensile load for the fabric in Newtons by 9.03% and an increase in elongation of the fabric in mm by 9.3%, which speaks of an increase of strength and elasticity of garment fibers and regeneration of microdamage with transglutaminase. The samples of wool fabrics washed with the liquid laundry detergent with 0.0025% biotransglutaminase, before wash with 0.0025% bioprotease, demonstrated an increase of tensile load for the fabric in Newtons by +20.18% and an increase in elongation of the fabric in mm by +16.18%, which speaks of an increase of strength and elasticity of garment fibers and effectiveness of transglutaminase enzyme impact on the fabric structure.
Example 6.
[0086] A study of changes in strength characteristics of samples of selected delicate fabrics after one and multiple washes with transglutaminase of the composition and with the liquid laundry detergent base was carried out (Table 1). The study was required to prove the hypothesis of improvement of strength characteristics after multiple washes of delicate fabrics with the composition of the invention. The liquid concentrated delicate laundry detergent given in Table 1 was used as the base for the experiment.
[0087] The study was carried out using the method described in Example 5 in the same conditions to prove effectiveness of transglutaminase after multiple use in washing of delicate fabrics. Transglutaminase was added in concentrations of 0.001%, 0.0025%, 0.005% (expressed as active substance). The duration of the cycle of exposure on the fabric structure made up 40 minutes, which corresponds to automatic wash mode "delicate fabrics".
[0088] Upon completion of the study of tensile strength, tensile elongation, constant elongation load the selected transglutaminase concentrations were evaluated after multiple use in the delicate laundry detergent (up to 5 washes).
Results.
[0089] Based on the results of evaluation of strength and elasticity of structure of delicate fabrics it was established that the analysed composition with addition of the optimal transglutaminase concentration from 0.001 % to 0.005 % in the liquid delicate laundry detergent has a pronounced effect. After multiple use a delicate garment does not lose its original shape as a result of fiber deformation or microdamage. The experiment demonstrated an increase in the tensile load in Newtons and elongation in mm with the use of Shimadzu AG 10kNX with establishment of the optimal transglutaminase concentration within the range from 0.001% to 0.005% (Table 8).
Table 8. Evaluation of strength characteristics of the wool and silk fabric surface
after numerous washes with transglutaminase
| Ingredients | Fabric | Tensile strength, N | Elongation, mm | Strain, N/mm | |||
| Wool:cashmere | |||||||
| Reference | Wool | 150.5 | 20.5 | 3.0 | |||
| After 5 washes with the detergent base w/o TG | Wool | 264.0 | - | 31.9 | - | 3.9 | - |
| After 5 washes with 0.001% TG | Wool | 401.9* | +67.1% | 51.7 | +62.1% | 6.0* | +53.8% |
| After 5 washes with 0.005% TG | Wool | 376.2* | +42.5% | 49.1 * | +53.9% | 5.7* | +46.2% |
| Silk | |||||||
| Reference | Silk | 501.3 | 11.4 | 66.8 | |||
| After 5 washes with the detergent base w/o TG | Silk | 203.5 | - | 11.4 | - | 16.9 | - |
| After 5 washes with 0.001% TG | Silk | 537.1* | +163.9% | 14.8 | +29.8% | 44.8* | +165.1% |
| After 5 washes with 0.0025% TG | Silk | 315.8* | +55.2% | 17.1 * | +50.0% | 52.5* | +210.6% |
| After 5 washes with 0.005% TG | Silk | 648.8* | +218.8% | 17.2* | +50.9% | 59.8* | +253.8% |
[0090] The data obtained by means of the tensile test can determine elastic limit, elongation, elastic modulus, proportional limit, area reduction, ultimate tensile strength, tear strength and other indicators of material strength. This enables precise determination of structural and mechanical properties of delicate fabrics mediated by regeneration of microdamage.
[0091] Force required for tear of delicate fibers increases with addition of transglutaminase of the invention. For silk the breaking force measured in Newtons increases from 203.5 N to 537.1 N at enzyme concentration of 0.001% and up to 648.8 N at enzyme concentration of 0.005% after 5 washes, which speaks of good restoration and strengthening of silk fibers. Breaking force depends on the thickness of fibers and yarn, therefore for fibers of wool: cashmere the tensile load increases from 264.0 N to 401.9 N (+67.1% to the indicator) at enzyme concentration of 0.001% and to 376.2 N (+42.5% to the indicator) at enzyme concentration of 0.005% after 5 washes, which speaks of good restoration of microdamage and strengthening of wool fibers.
[0092] Elongation at tensile break is called elongation at break and it indicates the ability of textile material to withstand tensile deformation. Addition of transglutaminase enzyme of the invention results in tensile elongation increase. For wool the percentage of elongation in relation to the initial length increased from 31.9 mm to 51.7 mm (+62.1% to the indicator) at enzyme concentration of 0.001% and to 49.1 mm (+53.9% to the indicator) at enzyme concentration of 0.005% after 5 washes. For silk fiber elongation increased from 11.4 mm to 14.8 (+29.8% to the indicator) at enzyme concentration of 0.001% and to 17.2 mm (+50.9% to the indicator) at enzyme concentration of 0.005% after 5 washes. An increase in the tensile elongation value is accompanied with an increase in force as fabric becomes more elastic and it gets more difficult to tear it, which positively influences the structural and mechanical characteristics of fiber, its wear resistance.
[0093] Relative fabric strength is characterised with maximum tension value per fiber or yarn thinness unit. Addition of transglutaminase enzyme of the invention contributes to increasing of tension of fabrics consisting of protein fibers. For silk the stress increases from 16.9 N/mm to 44.8 N/mm (+165.1% to the indicator) at enzyme concentration of 0.001% and to 59.8 N/mm (+253.8% to the indicator) at enzyme concentration of 0.005%. For wool the stress increases from 3.9 N/mm to 6.0 N/mm (+53,8% to the indicator)) at enzyme concentration of 0.001 % and to 5.7 N/mm (+46.2% to the indicator) at enzyme concentration of 0.005%.
[0094] Material properties from the point of view of deformation and damage under impact of tensile forces can be measured using the tensile plot, which is one of the most fundamental and important concepts in material mechanics. The horizontal curve coordinate is strain, while the vertical coordinate is stress. The curve form reflects different deformation processes that take place in material under the impact of external forces. It is shown on the curves representing the relation between load and elongation of textile material under tension (see Figures 4a, 4b and 4c) that transglutaminase reduces significant elongation increase at significant increase of resistance of acting load, which speaks of strengthening of fabric after its use.
[0095] Changing of sizes of textile materials under continuous action of a load of a value much less than the breaking one (i.e. creep and stress relaxation) at deformation stop as well as deformation relaxation (at rest) are important characteristics of mechanical properties having great importance in use of garments. Relaxation characteristics depend on the fibrous composition and texture of materials and greatly determine properties of materials.
[0096] Elastic deformation arises because under the impact of an external force there are slight changes in mean distances between adjacent chains and atoms in macromolecules of fibers making delicate fabrics. Elastic deformation cannot be high: with separation of particles to great distances the bond between them is destroyed, cracks and tears arise.
[0097] σe is the elastic limit of material and it represents maximum tension at which material remains elastically deformed. In the elastic phase there is a special linear segment wherein there is a linear relation between σ and ε. This is called a proportional phase, also known as the linear elastic phase.
[0098] When the load exceeds σe value to a certain value, the linear dependence between tension and deformation is broken and deformation significantly increases, uniform plastic deformation arises. This phenomenon of increasing resistance to plastic deformation alongside plastic deformation increase is known as strain hardening.
[0099] The phase of uniform sample deformation ends when strain reaches σb. This maximum strain σb is called breaking stress or ultimate tensile strength, which indicates material resistance to maximum uniform plastic deformation, i.e. maximum strain that material can withstand up to tensile break. (Table 9). After reaching the tension value σb the sample starts to deform non-uniformly and form necking, strain decreases and, finally, the sample is broken (torn), when the strain reaches σc. σc - is the fracture strength of material, which is the limit resistance of material to plasticity. As a whole, indicators of plastic properties of materials are elongation and area reduction.
[0100] Addition of transglutaminase enzyme to the composition of the invention in concentrations from 0.0025 to 0.005% gives the most optimal values of ultimate tensile strength σb for both delicate fabrics. The force applied for maximum tensile stress for silk 380 N and for wool 245 N at transglutaminase concentration of 0.0025% elongates the fabric by 20 mm for silk and by 34 mm for wool. Addition of 0.005% transglutaminase to a concentrated delicate laundry detergent increases the applied force for maximum tensile strength for silk up to 535 N and 455 N for wool and elongates the fabric by 17 mm for silk and by 46 mm for wool. Higher transglutaminase concentrations have greater influence on the tensile strength and elongation, which speaks of strengthening of delicate fabrics, improvement of their elasticity and strength at the same time.
Example 7.
[0101] A study of measuring of color difference between samples of selected fabrics (wool:cashmere and silk) after a single wash with protease and transglutaminase of the invention was carried out. The study aimed at proving the hypothesis of improvement of color preservation of delicate fabrics after a single wash with the new biotransglutaminase enzyme. The analysed liquid laundry detergent was the liquid concentrated delicate laundry detergent specified in Table 1.
[0102] Fading or change of color of dyed fabrics can be evaluated by means of the color stability test. Transition of a part of the dye from initial fibers to other nearby fabrics is called coloration or dye transfer. Coloration depends on how much sample color was applied on standard adjacent fabric or how much the adjacent fabric changed its color.
[0103] The test method is based on generally accepted recommendations of the international standard ISO 7724-3:1984 "Paints and varnishes - Colorimetry - Part 3: Calculation of colour differences", to determine differences between samples by color coordinates. The International Commission recommended two calculation formulae for general use. One of them in the system of color coordinates L∗, a∗, b∗, V (Cl ELAB system) is recommended to determine color differences. The possibility of its use for colorimetric evaluation of color differences was proven in practice.
[0104] This test enables evaluating effectiveness of the composition by the degree of preservation of color of protein materials, after actual wash conditions. The fabrics were washed in a washing machine Bosch WAB 24272 CE. The following recommendations were selected as the test conditions: temperature of 30°C, water hardness 10.9 ° dH and automatic wash mode (delicate fabrics, 40 minutes). The concentration of the liquid concentrated delicate laundry detergent was 4.5 g/l per standard washing machine load of 2-4 kg. After wash of the selected fabric samples with the composition containing biotransglutaminase and with the base of the liquid concentrated delicate laundry detergent, differences in color, lightness, color tone and color purity of the test fabric sample and reference by their color coordinates (L ∗, a∗, b∗) using a colorimeter Konica Minolta Chroma Meters CR-400 with the measurement area of 8 mm, were determined. There were three repetitions (n=3), the error was max. 0.1% for each sample. The measuring time, minimum interval between the measurements was 1-3 sec. The values are based on indicators of total color difference ΔE∗ab between two colors - geometric distance between two color space points (L∗, a∗, b∗), which are calculated using the formula: ΔE∗ab =[(ΔL∗)2 +(Δa∗)2 + (Δb∗)2]1/2. The method has a lot of advantages, in particular, high accuracy, reproducibility, a low error (less than 5%). The effective and statistically significant difference is 2 and more units ΔE∗ab.
Results.
[0105] Based on the results of evaluation of fabric color preservation it was established that the tested transglutaminase-based composition included in the liquid concentrated delicate fabric laundry detergent has a pronounced effect in relation to color tone preservation, saturation, color purity and total color difference comparing to the reference not containing the composition of the invention (Table 10).
[0106] Washing delicate fabrics with the liquid delicate laundry detergent first with protease and then with transglutaminase contributes to partial dye washout from the fabric, silk samples become lighter than the reference as the obtained value ΔL is positive, with "+" sign. For wool, on the opposite, it contributes to more intense coloring as the value ΔL is negative, with "-" sign. The obtained values ΔL for the selected protein fibers in Experiment 2 demonstrated better results. For silk the value ΔL was lower, and for wool the value ΔL was higher.
[0107] Washing delicate fabrics with the liquid concentrated delicate laundry detergent first with transglutaminase and then with protease contributes to preservation of color purity for silk as the value ΔC is positive, with "+" sign, for wool the sample color is slightly dirtier than the reference as the value ΔC is negative, with "-" sign. The obtained ΔC values for the selected delicate fibers in Experiment 2 demonstrated better results. For silk there was a greater positive value ΔC, while for wool there was a lower negative value ΔC.
[0108] Washing delicate fabrics with the liquid concentrated delicate laundry detergent first with protease and then with transglutaminase contributes to a lower difference in color tone of the initial fabric sample, the color tone of the reference remains as the obtained values Δh in Experiment 1 for wool and silk are nearer to zero. Use of the transglutaminase-based composition contributes to regeneration of microdamage of fibers of delicate fabrics and, as a result, preservation of brightness and purity of color.
[0109] A less significant value of differences by total color tone ΔE for the selected protein fabrics was observed in Experiment 3, which proves the hypothesis that addition of biotransglutaminase enzyme to the composition of the liquid concentrated delicate laundry detergent contributes not only to regeneration of fiber microdamage, but also to lesser color washout from delicate fabrics. Transglutaminase contributes to dye fixation and its lesser washout due to restoration of fibers, formation of a protective layer on the surface of delicate fibers without use of additional agents.
Example 8.
[0110] A study was carried out to measure color difference between samples of selected protein-based fabrics after a single and multiple washes with the enzyme cross-linking protein bonds in fibers. The study aimed at proving the hypothesis of preservation of color of delicate fabrics after multiple washes with the new biotransglutaminase enzyme. The analysed liquid laundry detergent was the liquid concentrated delicate laundry detergent specified in Table 1.
[0111] The study was carried out using the method described in Example 7 in the same conditions to prove performance of transglutaminase enzyme after multiple use in washing of delicate fabrics. Transglutaminase was added at the concentration of 0.0025%. The duration of the cycle of exposure on the fabric structure made up 40 minutes, which corresponds to hand wash and automatic wash mode "delicate fabrics".
[0112] Upon completion of the study color difference was measured by means of a spectrophotometer in color coordinates L, a, b in three different areas of the fabric sample treated with transglutaminase included in the laundry detergent after multiple washes (up to 10 washes).
Results.
[0113] Based on the results of evaluation of color difference, lightness, color tone and color purity of the dyes it was established that the analysed composition based on 0.0025% transglutaminase included in the liquid concentrated delicate laundry detergent has a pronounced effect. It was established that after multiple use the garment does not lose its original color because of the impact on the structure of fibers of the composition of the invention. Repeatability of ΔE∗ab is within 0.6 (at 12 measurements of ceramic references BCRA II comparing to the result of the reference instrument).
[0114] Experimentally and visually, preservation of color characteristics of delicate fabrics was demonstrated by means of a standalone portable instrument Konica Minolta Chroma Meters CR-400 with the use of 0.0025% transglutaminase (Table 11).
Table 11. Evaluation of preservation of color characteristics of protein fabrics after
numerous washes with biotransglutaminase
| Parameters | Reference of samples for test with the detergent base | After 1 wash with 0.0025% TG | After 5 washes with 0.0025% TG | After 10 washes with 0.0025% TG | After 5 washes with detergent base | After 10 washes with detergent base | ||||||
| Silk | Wool | Silk | Wool | Silk | Wool | Silk | Wool | Silk | Wool | Silk | Wool | |
| Lmean1 | 82.08 | 24.83 | 23.15 | 24.60 | 23.02 | 24.20 | 23.10 | 24.06 | 81.23 | 26.46 | 80.80 | 24.62 |
| amean | -1.00 | 25.67 | 10.17 | 37.50 | 9.84 | 37.43 | 9.98 | 38.32 | 0.04 | 26.20 | -0.24 | 25.34 |
| bmean | 7.03 | 6.81 | -44.37 | 10.78 | -43.06 | 10.12 | -43.85 | 10.36 | 7.18 | 8.53 | 7.33 | 8.03 |
| h | -81.90 | 14.86 | -77.09 | 16.04 | -77.13 | 15.13 | -77.18 | 15.13 | -89.68 | 18.03 | -88.15 | 17.58 |
| C | 7.10 | 26.55 | 45.52 | 39.02 | 44.17 | 38.77 | 44.97 | 39.70 | 7.18 | 27.55 | 7.33 | 26.58 |
| ΔC | - | - | 20.65 | -6.50 | 19.30 | -6.75 | 20.10 | -5.82 | 0.08 | 0.99 | 0.23 | 0.03 |
| Δ h | - | - | -0.61 | 1.37 | -0.65 | 0.46 | -0.70 | 0.46 | -7.78 | 3.17 | -6.25 | 2.72 |
| Δ Lmean | -9.85 | -67.10 | 0.97 | -0.73 | 0.84 | -1.13 | 0.92 | -1.27 | -0.85 | 1.63 | -1.28 | -0.21 |
| Δ Emean | 10.76 | 71.59 | 20.68 | 6.69 | 19.33 | 6.86 | 20.13 | 5.98 | 7.82 | 3.70 | 6.38 | 2.72 |
[0115] The study results demonstrated an accumulative effect of increase of brightness, saturation, general color tone of tested delicate fabrics after multiple use of the composition of the invention, which spoke of preservation of the original color of clothes (Table 12).
[0116] At the end of the laboratory study pronounced changes of evaluated indicators concerning preservation of color characteristics were observed. According to the dynamics of the indicators, addition of highly-effective biotransglutaminase as part of the liquid concentrated delicate laundry detergent enabled preserving color of delicate fabrics and inhibiting transfer of dyes. The composition with 0.0025% biotransglutaminase made it possible to protect color against fading by 97% for silk and by 89% for wool after the 10th wash, which is a statistically significant result for preservation of color of fabrics after multiple washes (p <0.05). The base sample without the composition of the invention demonstrated weak prevention of fiber fading, which speaks of insufficient effectiveness of surfactants and other ingredients for creation of a protective layer around clothes and protection of clothes shine against fading.
Example 9.
[0117] A study to determine dye washout from samples of delicate fibers after a single wash with protease and transglutaminase of the invention was carried out. The study aimed at proving the hypothesis of improvement of color preservation of delicate fabrics after a single wash with the new biotransglutaminase enzyme. The analysed liquid laundry detergent was the universal liquid concentrated delicate laundry detergent specified in Table 1.
[0118] The analysis method was based on measuring of the quantity of the dye washed out by means of electromagnetic emission in a certain narrow wavelength band. Spectrophotometry in the ultraviolet band is used for quantification of the dye washed out from fabric to the solution. The method sensitivity is determined mainly by the ability of the substance to absorb and is expressed with the molar absorption coefficient. Limit concentrations of substances analysed by means of spectrophotometry are, as a rule, lower than in standard wash conditions. The main condition for quantitative analysis is observance of the Bouguer-Lambert-Beer law: relative optical density is directly proportional to the dye molecule concentration.
[0119] Solutions of different dyed compounds are characterised by means of their absorption spectra - light absorption plots that determine the dependence of optical density D or molar absorption coefficient ε on wavelength λ or frequency n. The spectral region where intensive emission absorption is observed is called the absorption band. Of the greatest interest for analysis are the following spectrum characteristics: the number of peaks (absorption bands), their position on the wavelength scale, peak height (value of molar coefficient e in absorption peak), absorption band intensity, band width and shape. The wavelength at max absorption point is called λmax and it determines the color of compound, lgε determines intensity of coloring. The width of absorption band at the height middle dropped from max absorption point determines dye brightness or purity. The wider the absorption band is, the dirtier the dye color is. The narrower the absorption band is, the brighter and purer the dye color is. Light absorption is measured in the optical spectrum range in the ultraviolet (185-400 nm) and visible (400-760 nm) spectrum regions. The light absorption peak of colored substances, in most cases, is located in the visible spectrum region (λ= 500 nm). The light absorption plots of analysed solutions for quantitative analysis of dye release from fibers was drawn by means of a UV-spectrometer Thermo Scientific Evolution 300. To measure the spectral directional transmittance coefficient, optical density and optical density change rate of transparent liquid prepared solutions and to determine concentration of substances in solutions after washing fabric samples with the composition of the invention, a photoelectric photometer KFK-3-ZOMZ, factory No. 1970693, was used. Test conditions: temperature 21.6°C, relative humidity 60%, pressure 97.3 kPa. Then spectral absorption plots - plots of dependence of absorption intensity (lgε) on wavelength (λ) - were drawn.
Results.
[0120] Based on the results of evaluation of dye/pigment washout from delicate fabric fibers after washing it was established that the tested composition of the invention has a high performance in terms of delicate fabric color preservation. The composition of the invention as part of the delicate laundry detergent makes it possible to achieve a high degree of preservation of color characteristics at a minimum concentration of the component. The absorption spectra (see Figure 5) show that transglutaminase at concentration of 0.0025% reduces dye release 3.5 times, i.e. it fixes color in delicate fabric fibers, makes it strong by strengthing of fabric fibers. Pre-treatment of fibers with transglutaminase reduces dye release 3.5 times comparing to the fabric washed with protease, which speaks of regeneration of microdamage of fibers and, as a result, dye retention in delicate fabric microfibers.
[0121] Color change in a sequence corresponding to the absorption peak shift (λmax) to the long-wavelength spectrum part is called a bathochromic shift, when the color deepens. Color change in the reverse direction corresponding to the absorption peak shift to the short-wavelength spectrum part is called a hypsochromic shift. Washing of delicate fibers with the concentrated detergent first with 0.0025% transglutaminase, then with 0.0025% protease demonstrates insignificant optical density A on the level of 0.025-0.04, which speaks of slight dye washout from delicate fabric fibers. Addition of 0.0025% protease first and then 0.0025% transglutaminase leads to a bathochromic shift and reaching of optical density A on the level of 0.14-0.15, color deepens, which speaks of much greater dye release from the structure of delicate fibers to the analysed solution.
[0122] The position of spectral absorption plot peak on y-axis (lgε-y) characterises absorption intensity, coloring intensity. Washing with a concentrated delicate laundry detergent first with 0.0025% transglutaminase and then with 0.0025% protease leads to a hypochromic effect, absorption intensity decreases (A=0.04 at λmax), which speaks of lesser fabric damage and dye washout from delicate fabric fibers. Washing with the detergent first with 0.0025% protease and then with 0.0025% transglutaminase leads to a hyperchromic effect, absorption intensity increases 3.5 times (A= 0.14 at λmax), which speaks of much greater dye release from fiber structure to the analysed solution and, correspondingly, damage of delicate fabrics. Washing with the concentrated delicate laundry detergent first with 0.0025 % protease and then with 0.0025% transglutaminase influences the absorption band width, namely, it becomes narrow, the absorption peak becomes more obvious, which speaks of a high dye concentration and active color washout.
[0123] After pre-treatment with protease the scale layer of wool fiber is partially damaged, revealing more active groups such as the amino group, primary hydroxyl group and phenol group, mainly, existing in serine and tyrosine residues. Addition of transglutaminase enzyme contributes to regeneration of microdamage and improvement of structure of delicate fibers, preventing further washout of dye and pigments from delicate fabrics.
Example 10.
[0124] A study was carried out to evaluate the softening effect after washing delicate fabrics with detergent samples. The study was required to prove the hypothesis of having a softening effect on fabrics, making them more pleasant to touch after washing and gentle impact on hand skin during hand washing with the new biotransglutaminase enzyme. The analysed liquid laundry detergent was the liquid concentrated delicate laundry detergent specified in Table 1.
[0125] The analysis method was based on determination of delicate fabric softness after washing. The softening effect was determined organoleptically on wool and silk fabric samples. Softness of initially soft fabric samples sized 30×50 cm was compared by touch initially and after 1 wash with the tested detergent. The samples of selected fabrics were washed in a washing machine Bosch WAB 24272 CE, hand wash cycle at 40°C, water hardness 11.2° dH, detergent consumption of 50 g per 1 wash. Upon completion of the wash cycle the fabrics were dried in horizontal position in a room where constant temperature and humidity were maintained. After complete drying the softening effect was evaluated in scores: 0 - no difference; 1 - probably, softer, 2 - surely, softer; 3 - much softer.
Results.
[0126] Based on the results of evaluation of the softening effect after washing it was established that the tested composition of the invention has a high softening effect on delicate fabrics. Biotransglutaminase included in the delicate laundry detergent enables achieving a high softening effect, removing roughness from surfaces by regeneration of microfibers at minimum transglutaminase concentration (Table 13).
Table 13. Determination of transglutaminase impact on softening effect
| Tested sample | Softening effect, scores | ||
| Before wash | After 1 wash | ||
| Wool | Universal delicate laundry detergent base w/o TG | 0 | 1 |
| Universal delicate laundry detergent with 0.0025% TG | 0 | 2 | |
| Silk | Universal delicate laundry detergent base w/o TG | 0 | 1 |
| Universal delicate laundry detergent with 0.0025% TG | 0 | 2 | |
[0127] Water containing salts, high wash temperature, fabric peculiarities cause excessive stiffness of fabrics, specifically, delicate fabrics. Contact between stiff rough material and skin is unpleasant. Addition of transglutaminase enzyme of the invention to the concentrated delicate laundry detergent enables softening fibers of delicate fabrics (silk, wool) and has a gentle impact on body skin even during daily wear of such fabrics.
Example 11.
[0128] Skin diagnostic (dermatoscopy) was carried out with ASW (Aramo Smart Wisard) magnification using accessories (ASW) designed for visual skin diagnostic (dermatoscopy). Skin diagnostic was used to determine skin hydration and elasticity after a single hand wash with the liquid concentrated delicate laundry detergent base with and without the composition of the invention containing 0.0025% transglutaminase (Table 1). Hand wash imitated a short-term contact with hand skin during hand wash of delicate fabrics to evaluate the impact on main skin indicators and the possibility of use by people with sensitive skin.
[0129] The analysis method was based on the skin surface epiluminescence microscopy. This kind of examination, in distinction to standard light microscope, enables not only examining skin surface relief, but also visualising interskin morphological structures sized 0.2 µm and more located in the epidermis and papillary dermis. The skin humidity and elasticity indicators were measured using a dermatoscope Aramo Smart Wizard (ASW-300). The skin condition in the test area - hands - was evaluated before and after wash with the laundry detergent. 7 people with different hand skin hydration degrees participated in the study. Each measurement was taken three times (n=3), the mean of these three measurements, with account for the standard deviation (SD), was considered. Measurement for each participant was taken in the same hand skin area, specifically, on wrist skin.
Results.
[0130] Based on the results of evaluation of the influence of the detergent with transglutaminase and alpha-amylase of the invention on hydration of the skin surface layer and skin elasticity after a single hand wash it was established that the analysed composition of the invention does not impair physiological hand skin parameters such as hydration and elasticity comparing to the liquid concentrated delicate laundry detergent not containing transglutaminase and alpha-amylase. Skin surface daily experiences the influence of negative external factors: people regularly wet hands, submerge them in water, wash clothes by hand, subjecting hands to the impact of chemicals included in laundry detergents. The skin hydration and elasticity indicators after washing hands with the delicate laundry detergent base decrease by 44%, almost twice than before hand washing, which speaks of the negative influence on the skin barrier and significant moisture loss after a single contact (p<0.05). Use of the delicate laundry detergent with biotransglutaminase does not lead to formation of local dry areas, is not accompanied with the feeling of skin tightness, skin does not itch, crack, exfoliate and has no discomfort feeling. Thus, the composition of the invention protects the skin surface forming a protective barrier that guards it against negative influence of laundry detergent components and skin hydration and elasticity reduction (Table 14).
Table 14. Determination of the effect of delicate laundry detergents on skin hydration
and elasticity after a single contact
| Sample | Hydration before hand washing | Elasticity before hand washing | Hydration after hand washing | Elasticity after hand washing |
| Scores, M±SD | ||||
| Basic universal delicate laundry detergent w/o TG | 13.2±5.4 | 13.0±5.6 | 7.1±3.3 | 7.0±3.8 |
| Universal delicate laundry detergent with 0.0025% TG and with 0.01% alpha-amylase | 16.7±11.8 | 15.0±8.8 | 15.2±9.3 | 14.3±8.7 |
[0131] Hydrated skin looks fresh and beautiful, effectively fulfils its protective function and effectively copes with negative influence of the environment. Hydration is the first and main stage in basic skin care as it is very important that hand laundry detergents are non-aggressive and have a gentle impact on sensitive hand skin. Addition of the composition of the invention to the liquid concentrated delicate laundry detergent and further washing enable maintaining hand skin smoothness, elasticity and hydration without causing dryness and tightness. But after a single hand wash with the liquid concentrated delicate laundry detergent not containing the composition of the invention it causes first signs of hand skin hydration loss and reduction of elasticity. The skin hydration and elasticity indicators decrease by more than 44% after use of the laundry detergent without the composition of the invention. Use of the concentrated delicate laundry detergent with transglutaminase and alpha-amylase will make it possible to maintain skin hydration and elasticity on the previous level, with no harm for skin health (sse the Plots as per Figures 6 and 7).
[0132] Hand wash with the liquid concentrated delicate laundry detergent base reduces hand skin hydration and elasticity, which negatively affects skin health in the long run. The components of the composition of the invention, namely transglutaminase and alpha-amylase, take care not only of delicate fabric fiber structure, but also of hand skin during hand wash and daily contact with detergents.
Example 12.
[0133] A laboratory study of washing performance of components of the composition of the invention included in the liquid concentrated delicate laundry detergent specified in Example 1 was carried out.
[0134] The test method was based on generally recognized recommendations of the European Association A.I.S.E. [A.I.S.E. Laundry Detergent Testing Guidelines Minimum requirements for comparative detergents testing https://www.aise.eu/documents/document/20180625164030-laundry _detergent _testing guidelines_v_5_2 june_2018_.pdf] to determine washing performance with account for the stain removal degree (SRI) and L,a,b values. The SRI index was calculated using the formula from the ASTM D 4265 Standard guide for evaluating stain removal performance in home laundering [DOI: 10.1520/D4265-14]. The present guide was prepared by the Committee D-12 for soaps and other detergents of the American Society for Testing Materials (ASTM) and was published in 2014. This test enables evaluating performance of the composition by the degree of removal of fresh and old stains of different origin from dense surfaces, specifically, from cotton and synthetic materials imitating actual wash conditions. The fabrics were washed in a washing machine Bosch WAB 24272 CE. The selected test conditions were popular recommendations in the territory of EU and RF, specifically, temperature 40°C, water hardness 11.2° dH and standard wash cycle (mixed fabrics). The detergent consumption was 50 g per 1 wash.
[0135] The test was carried out with the use of stubborn stains of different origin, including those containing starch compounds based on amylose and amylopectine (Table 15). The selected stubborn biostains enabled evaluating general washing performance of the formula and performance for individual soils due to presence of transglutaminase and alpha-amylase of the invention.
Table 15. Tested biostains and biosoils from A.I.S.E.
| Name | Ingredients | Type |
| EMPA 111 | Blood | Protein, enzymatic |
| EMPA 164 | Grass | Protein, mixed, enzymatic and bleachable |
| CFT CS-44 | Chocolate | Mixed∗ |
| CFT CS-27 | Amylase starch-based | Starch, enzymatic∗ |
| ∗contain compounds based on starch or its derivatives | ||
[0136] Also there was evaluation of removal of stains based on starch from different vegetable sources: CS-28/Rice starch on cotton and CS-29/Tapioca Starch Colored (Table 16). The selected stubborn starch-based stains enabled evaluating general washing performance of the formula upon presence of transglutaminase and alpha-amylase of the invention. The test was carried out in a TOM laboratory washing machine at the temperature of 30°C, wash duration 20 minutes, wringing rate 120 PRM, water hardness 15°dH, detergent consumption 3.33 g/l. The test was carried out twice (n=2). SRI was measured at Rem 460 nm.
Table 16. Tested starch biostains from A.I.S.E.
| Name | Ingredients | Type |
| CS-28/Rice starch on cotton | Rice starch | Enzymatic |
| CS-29/Tapioca Starch Colored | Tapioca Starch | Enzymatic |
[0137] SRI is the quantitative index of removal of soils and stains in conventional units. The values are based on indices of whiteness L and color a,b, calculated mathematically using the special formula from ASTM D 4265-14 standard [DOI: 10.1520/D4265-14.]:
SRI=100-[(Lc-Lw)2+(ac-aw)2+(bc-bw)2]1/2,
where
where: L is reflectance, a is red/yellow ratio, b is yellow/blue ratio, c is unsoiled fabric washed in wash conditions, w is washed soiled fabric.
[0138] The method has a lot of advantages, specifically, high accuracy, reproducibility, a low error (below 5%), it takes into account colourity of fabrics and its change in the washing process, different types of fabrics and it is closest to actual visual perception. Besides, it levels out the effect of optical dyes widely spread in the composition of laundry detergents. The effective and statistically significant difference is 2 and more SRI units. According to practical experience, 1 SRI unit corresponds to 5% of additional washing performance and contributes to the total performance of the composition.
Results.
[0139] Based on the results of effectiveness and rate of removal of stubborn stains based on starch and other organic compounds, it was established that the combination of transglutaminase and alpha-amylase in the composition of the delicate laundry detergent has a pronounced stain-removing effect with respect to stubborn stains comparing to the reference containing no transglutaminase and no alpha-amylase of the invention.
[0140] Upon completion of the study there were obvious changes in the tested indicator of performance of removal of stubborn stains based on various organic compounds, including starch compounds. According to the indicator dynamics, addition of highly-effective alpha-amylase and transglutaminase increased the degree of removal of stubborn stains of various origin. Addition of 0.0025% transglutaminase in combination with 0.01% alpha-amylase enabled increasing the SRI index by 62.8 units (+23.4%) comparing to wash with the liquid delicate laundry detergent without the composition of the invention. Universal liquid delicate laundry detergent with transglutaminase and alpha-amylase with respect to stubborn stains of blood, grass, chocolate drink and starch. Alpha-amylase in combination with transglutaminase made it possible to significantly increase stain-removal performance when included in the composition of the universal liquid delicate laundry detergent. The sample of the detergent base without the composition did not demonstrate very high performance of removal of selected stains and soils, which speaks of insufficient performance of surfactants in removal of stains and soils in the wash process and the need in pre-treatment of delicate fabrics (Table 17).
[0141] According to the indicator dynamics, addition of highly-effective alpha-amylase in combination with transglutaminase together with selected composition components increased the degree of removal of stubborn starch-based soils. Addition of 0.0025% transglutaminase and 0.01% alpha-amylase made it possible to significantly increase the SRI indicator in relation to the detergent base with respect of fixed starch stains. The composition with 0.0025% transglutaminase and 0.01% alpha-amylase increased the SRI index by 24.92 units (+25.25%), which demonstrates synergism between the components and is a statistically significant result in removal of biostains during washing (p<0.05). The sample of the base without the composition of the invention did not demonstrate high removal of selected biosoils, which speaks of insufficient performance of surfactants in removal of these soils during washing (Table 18).
Table 18. Evaluation of starch-based stain removal performance
| Tested sample | Composition components | SRI index | ||
| CS-28 Rice starch | CS-29 Tapioca starch | SRI total by 2 stains | ||
| Sample 1 Base | - | 59.82 | 38.87 | 98.69 |
| Sample 2 Base + composition components | Transglutaminase 0.0025% | 67.59 | 56.02 | 123.61 |
| Alpha-amylase 0.01% | ||||
| ∗mean significant difference, statistically significant (p<0.05), increase of the standard
washing performance by 10-20% ∗∗best significant difference, statistically significant (p<0.05), increase of the standard washing performance by more than 20% |
Example 13.
[0142] In addition, a laboratory study of washing performance of components of the composition of the invention included in the liquid concentrated delicate laundry detergent in the form of capsules specified in Table 19 was carried out.
Table 19. Ingredients of the concentrated delicate laundry detergent in the form of
capsules with the claimed composition
| No. | Component | Content, % wt. |
| 1 | Purified water | up to 100.00 |
| 2 | Propylene Glycol | 15.0-35.0 |
| 3 | Glycerin | 5.0-15.0 |
| 5 | Sorbitol | 0.01-1.0 |
| 6 | MEA-Cocoate | 15.0-35.0 |
| 7 | PEG-7 Glyceryl Cocoate | 15.0-25.0 |
| 8 | Sodium Laureth sulfate | 5.0-15.0 |
| 9 | Sodium diethylenetriamine pentamethylene phosphonate | 0.1-2.0 |
| 10 | Protease (subtilisin) | 0.01-0.3 |
| 10 | Amylase - expressed as active substance | 0.005-0.3 |
| 11 | Transglutaminase - expressed as active substance | 0.005-0.3 |
| 12 | Lipase - expressed as active substance | 0.005-0.3 |
| 13 | Sodium formate | 0.01-0.3 |
| 14 | Denatonium benzoate | 0.005-0.3 |
| 15 | Calcium chloride | 0.00001-0.1 |
| 16 | Polyvinyl alcohol | - |
[0143] The study was carried out based on the methods described in Example 12 to prove the performance and synergism of alpha-amylase and transglutaminase in the composition of the laundry detergent with basic medium (pH 8.5-10.5 for 20% detergent solution depending on the composition).
[0144] The test was carried out with the use of stubborn starch compounds based on amylose and amylopectine (Table 20). The selected stubborn biostains enabled evaluating general washing performance of the formula and performance for starch soils upon presence of transglutaminase and alpha-amylase of the invention.
Table 20. Tested starch biostains from A.I.S.E.
| Name | Ingredients | Type |
| CFT CS-27 | Dyed starch | Enzymatic |
| CFT CS-02 | Cocoa / chocolate drink | Enzymatic |
| Results. |
[0145] Based on the results of performance and rate of removal of stubborn starch-based stains, it was established that the combination of transglutaminase and alpha-amylase in the composition of the concentrated delicate laundry detergent in the form of capsules has a pronounced stain-removing effect with respect to stubborn stains comparing to the reference containing no transglutaminase and no alpha-amylase of the invention.
[0146] Upon completion of the study there were obvious changes of the analysed indicator of performance of removal of stubborn stains based on starch compounds that are strongly held in delicate fabrics. According to the indicator dynamics, addition of highly-effective alpha-amylase and transglutaminase increased the degree of removal of stubborn starch-based soils. Addition of 0.0025% transglutaminase and 0.01% alpha-amylase made it possible to significantly increase the SRI index in relation to the detergent base with respect to fixed starch stains by 21.15% comparing to the detergent without these components. The composition with 0.0025% transglutaminase and 0.01% alpha-amylase increased the SRI index by 27.7 units (+21.14%), which demonstrates synergism between the components and is a statistically significant result in removal of biostains during washing (p<0.05). The sample of the base without the composition of the invention did not demonstrate high removal of the selected biosoil, which speaks of insufficient performance of surfactants and basic ingredients in removal of these soils during washing (Table 21).
Table 21. Evaluation of starch-based stain removal performance
| Tested sample | Composition components | SRI index | ||
| CFT CS-27 Starchy dye | CFT CS-02 Cocoa / chocolate drink | SRI total by 2 stains | ||
| Sample 1 Base of capsules | - | 55.1 | 75.9 | 131.0 |
| Sample 2 Base of capsules + composition components | Alpha-amylase 0.02% | 78.1 | 77.6 | 155.7** |
| Sample 2 Base of capsules + composition components | Transglutaminase 0.0025% | 78.2 | 80.5 | 158.7** |
| Alpha-amylase 0.02% | ||||
| ∗mean significant difference, statistically significant (p<0.05), increase of the standard
washing performance by 10-20% ∗∗best significant difference, statistically significant (p<0.05), increase of the standard washing performance by more than 20% |
1. A composition comprising the following components:
A) transglutaminase; and
B) α-amylase, and
wherein the mass ratio of said components is preferably the following:| A | : | B |
| 0.0010-0.1000 | 0.0025-0.0500 |
2. The composition of any of the preceding claims, wherein said transglutaminase is further
defined by at least one of the following:
i) is produced biotechnologically from a microorganism, preferably from bacteria or yeast-like fungi, further preferably from protease-resistant bacteria or yeast-like fungi;
ii) has a biological activity of at least 100 U/g at 37°C and/or a viscosity of 0-1000 sPas at 37 °C;
iii) has the CAS number 80146-85-6.
3. The composition of any of the preceding claims, wherein said α-amylase is further
defined by at least one of the following:
i) is produced biotechnologically from a microorganism;
ii) has a biological activity of at least 100 U/g at 37°C and/or a viscosity of 0-1000 sPas at 37 °C.
4. The composition of any of the preceding claims, wherein the amount of transglutaminase
in said mass ratio of transglutaminase to α-amylase (0.001-0.1):(0.0025-0.05) is 0.001,
0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.01, 0.02, 0.03, 0.04, 0.05,
0.06, 0.07, 0.08, 0.09 or 0.10.
5. The composition of any of the preceding claims, wherein the amount of α-amylase in
said mass ratio of transglutaminase to α-amylase (0.001-0.1):(0.0025-0.05) is 0.0025,
0.0030, 0.0035, 0.0040, 0.0045, 0.0050, 0.0055, 0.0060, 0.0065, 0.0070, 0.0075, 0.0080,
0.0085, 0.0090, 0.0095, 0.01, 0.02, 0.03, 0.04 or 0.05.
6. The composition of any of the preceding claims, wherein at least one of components
A) and B) are in a solution selected from the following: water-glycerol, water-sorbitol,
water-glycerol-sorbitol, water-propylene glycol, water-glycerol-maltitol.
7. The composition of any of the preceding claims, wherein said composition comprises
- component A) as per 0.001-0.1 wt.% in said composition, preferably as per the following wt.% or ranges defined by the following wt.%: 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10; and/or
- component B) as per 0.0025-0.05 wt.% in said composition.
8. A laundry washing and/or conditioning composition comprising the composition of any
of the preceding claims, wherein said laundry washing and/or conditioning composition
preferably comprises 0.10-25.50 wt.%, further preferably 0.10-1.0 wt.% of said composition
of claims 1-7.
9. The composition of any of the preceding claims, wherein said composition is a formulation
selected from the following: solution, paste, gel, powder, granulate, wherein said
formulation is preferably a solution, further preferably a solution selected from
the following: water-glycerol, water-sorbitol, water-glycerol-sorbitol, water-propylene
glycol, water-glycerol-maltitol.
10. The composition of any of the preceding claims assembled as a single composition comprising
both components A) and B) or assembled as a kit-of-parts, wherein said kit-of-parts
preferably separates components A) and B).
11. Use of the composition of any of the preceding claims for at least one of the following
uses:
a) as hand cleaning washing composition, preferably in combination with any of the uses as per below items b)-e);
b) for removal of starch and/or polysaccharide residue-based stains;
c) for fabric care, including brightness care of colored fabrics;
d) for fabric fibre microdamage repair or for fabric fibre regeneration and/or for fabric fibre elasticity regulation, including increasing of elasticity of said fabric fibres, wherein preferably said microdamage and/or elasticity loss is caused by previous protease-based treatments of said fabric fibres;
e) for laundry care and/or laundry washing, wherein said laundry is preferably a fabric
i) selected from cotton, synthetic, membrane, wool, silk, cashmere, merino, down, feather and mixtures thereof; or
ii) comprising or consisting of animal protein-based textile fibres, further preferably hair, including wool, cashmere, merino, and/or silk; and/or said fabric comprises at least one of the following components to be treated: skin sebum, blood, sauce, grass, juice, jam, tea, cacao, coffee, wine, beer, chocolate, vinegar, wherein said components are preferably contaminations on surfaces of said fabric.
12. The composition or use of the composition of any of the preceding claims, wherein
i) said composition has a pH of 7.0 to 10.5, preferably selected from the following pH sub-ranges: 7.5-10.0, 7.5-9.5, 7.0-9.0, 8.5-10.5, and combinations thereof; and/or
ii) said use of the composition is for use at
- a pH of 7.0 to 10.5, preferably selected from the following pH sub-ranges: 7.5-10.0, 7.5-9.5, 7.0-9.0, 8.5-10.5, and combinations thereof, and/or
- a temperature range of +15 °C to +60 °C, preferably selected from the following temperature sub-ranges: +20 °C to +60 °C, +20 °C to +55 °C, +30 °C to +55 °C, +40 °C to +55 °C, and combinations thereof.
13. The composition or use of the composition of any of the preceding claims, said composition
- additionally comprises added proteins derived from hair, including wool, cashmere, merino, and/or silk; or
- not comprises added proteins derived from hair, including wool, cashmere, merino, and/or silk.
14. The composition or use of the composition of any of the preceding claims, said composition
not comprising surfactants.
15. A method for cleaning fabrics and/or for fabric care, said method comprising:
a) solving the composition of any of claims 1-10 in water for obtaining an aqueous solution;
b) optionally, adjusting
- the pH of said aqueous solution to a pH of a pH of 7.0 to 10.5, preferably selected from the following pH sub-ranges: 7.5-10.0, 7.5-9.5, 7.0-9.0, 8.5-10.5, and combinations thereof, and
- a water hardness of 0-15 °dH; and
c) applying said aqueous solution to said fabric, preferably at a temperature range of 15 °C to 60 °C, further preferably selected from the following temperature sub-ranges: +20 °C to +60 °C, +20 °C to +55 °C, +30 °C to +55 °C, +40 °C to +55 °C, and combinations thereof.
REFERENCES CITED IN THE DESCRIPTION
This list of references cited by the applicant is for the reader's convenience only. It does not form part of the European patent document. Even though great care has been taken in compiling the references, errors or omissions cannot be excluded and the EPO disclaims all liability in this regard.
Non-patent literature cited in the description
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