Efficient bio-enzyme laundry detergent as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of biochemistry, and relates to a high-efficiency biological enzyme laundry detergent which comprises novel biological enzymes (SEQNO: 1 and SEQNO: 2) and a novel organic compound (AEBIT). A biological enzyme is prepared through genetic engineering, and AEBIT is organically synthesized. The laundry detergent can efficiently remove protein and starch stains, and comprises the following components: a surfactant, a builder, a softener, perfume and the like. During preparation, all the components are mixed and stirred uniformly, and the detergent can be widely applied to families and commercial laundry places. The cleaning agent has the advantages of efficient decontamination, high enzyme stability, environmental protection, safety, optimized formula and the like.

Description

Technical Field

[0001] The invention belongs to the technical field of biochemical pharmaceutical products, and in particular is a high-efficiency bio-enzyme laundry detergent and a preparation method and application thereof. Background Art

[0002] Traditional laundry detergents have limited cleaning capabilities, are ineffective on stubborn stains, are prone to damaging clothing fibers, and may leave harmful substances. As the quality of life improves, people's demand for environmentally friendly, efficient, and gentle laundry detergents is growing.

[0003] Although bio-enzyme laundry detergent has advantages, the existing ones still need to be improved in terms of enzyme stability, synergy and targeted stain removal. The existing laundry detergents have problems including poor removal effect on certain stains, possible damage to clothing, and residual substances that may harm health. The high-efficiency bio-enzyme laundry detergent of the present invention aims to solve these problems and provide a more effective clothing cleaning solution. Summary of the invention

[0004] The invention provides a high-efficiency bio-enzyme laundry detergent, comprising the following components: a novel bio-enzyme SEQNO: 1-Leu-Cys-His-Arg-Asp-Phe-Tyr, the content of which is 0.15% of the total weight of the laundry detergent; a novel bio-enzyme SEQNO: 2-Thr-Glu-Lys-Val-Asp-Trp-Ser, the content of which is 0.1% of the total weight of the laundry detergent; a novel organic compound 3-(2-aminoethyl)-1,2-benzisothiazolin-3-one (AEBIT), the content of which is 0.1% of the total weight of the laundry detergent. The content of the detergent is 1.5% of the total weight of the laundry liquid; the content of the anionic surfactant is 12% of the total weight of the laundry liquid, wherein the anionic surfactant is sodium dodecylbenzene sulfonate; the content of the nonionic surfactant is 8% of the total weight of the laundry liquid, wherein the nonionic surfactant is fatty alcohol polyoxyethylene ether; the builder sodium citrate is 4% of the total weight of the laundry liquid; the builder zeolite is 3% of the total weight of the laundry liquid; the softener polyquaternium-7 is 1.5% of the total weight of the laundry liquid; the fragrance natural floral fragrance is 0.2% of the total weight of the laundry liquid.

[0005] Furthermore, the SEQ NO: 1 biological enzyme mainly targets protein stains on clothing, forms disulfide bonds with groups in proteins through Cys residues, Leu residues provide a hydrophobic environment, His and Arg residues bind to charged groups on the stain protein through electrostatic attraction, Asp residues interact with positively charged protein regions, and the aromatic ring structures of Phe and Tyr residues enhance hydrophobic interactions and π-π stacking effects with proteins, thereby decomposing protein stains.

[0006] Furthermore, the SEQ NO: 2 biological enzyme mainly targets starch stains and makes the stains easier to clean by breaking down starch molecules.

[0007] Furthermore, the amino group in the AEBIT molecule reacts with the carboxyl group of the biological enzyme to stabilize the structure of the biological enzyme and prevent it from being inactivated in the laundry detergent system. At the same time, the benzisothiazolinone structure has antibacterial activity, synergistically acts with the biological enzyme to enhance the removal effect of stains, and slowly releases the biological enzyme during the washing process to prolong the action time. On the other hand, the present invention also provides a method for preparing a high-efficiency biological enzyme laundry detergent, comprising the following steps:

[0008] The purified SEQNO:1 and SEQNO:2 biological enzymes are mixed with AEBIT, surfactants, detergents, softeners, fragrances and other ingredients in a container in a certain proportion. The biological enzymes and AEBIT are first dissolved in a small amount of water, and then other ingredients are added in sequence. The mixture is stirred in a blender at a speed of 300-500 rpm for 30-60 minutes until a uniform solution is formed. Finally, the laundry detergent is put into a sealed container.

[0009] Beneficial effects of the invention:

[0010] The high-efficiency stain removal performance is remarkable. The addition of the new bio-enzyme combination and AEBIT can still maintain a high stain removal rate at low enzyme concentrations; AEBIT improves the stability of the bio-enzymes, and the two bio-enzymes work synergistically on different stains to enhance the ability to remove mixed stains; it is environmentally friendly and energy-saving, reduces pollution to the environment, and AEBIT has a mild structure, reduces irritation to human skin, and is suitable for all types of people; the ingredients in the formula work synergistically with each other, and in addition to excellent stain removal, it also has advantages in ease of use, clothing care and sensory experience, and is highly competitive. DETAILED DESCRIPTION

[0011] Example 1

[0012] (I) Amino acid sequences of novel enzymes

[0013] A new type of bioenzyme is designed, and its amino acid sequence is SEQNO:1-Leu-Cys-His-Arg-Asp-Phe-Tyr (LCHRDY). Design principle: The bioenzyme mainly targets common protein stains on clothing, such as blood stains, milk stains, etc. The Cys residue can form a disulfide bond with certain groups in the protein, thereby specifically binding to the stain protein. The Leu residue provides a hydrophobic environment, which helps the interaction between the bioenzyme and the protein. The His and Arg residues carry an ionizable imidazole group and a positive charge, respectively, and bind to the negatively charged area or other charged groups on the stain protein through electrostatic attraction. The Asp residue provides a negative charge and can interact with the positively charged protein area. The aromatic ring structure of the Phe and Tyr residues enhances the hydrophobic interaction and π-π stacking effect with the protein. Through the synergistic effect of these amino acid residues, the bioenzyme can efficiently decompose protein stains and remove them from clothing.

[0014] (II) New organic compounds

[0015] An unreported organic compound named 3-(2-aminoethyl)-1,2-benzisothiazolin-3-one (AEBIT for short) is used as a stabilizer and synergist for biological enzymes. Design principle: The amino group in the AEBIT molecule can react with the carboxyl group of the biological enzyme to form a chemical bond or ion pair, thereby stabilizing the structure of the biological enzyme and preventing it from being inactivated in the laundry detergent system. The benzisothiazolinone structure has antibacterial activity, which can inhibit the growth of bacteria on clothing and reduce the generation of odors. At the same time, it synergizes with biological enzymes to enhance the removal effect of stains. In addition, AEBIT can slowly release biological enzymes during the washing process, prolong their action time, and improve the cleaning efficiency of laundry detergent.

[0016] (III) Decontamination enhancement of single sequence, combined sequence and organic compounds

[0017] The experiment shows that the cleaning effect of the single sequence shows that the cleaning liquid containing SEQNO:1 biological enzyme has a removal rate of about 60% for protein stains at a concentration of 0.5%. The biological enzyme mainly decomposes the protein to remove the stains from the clothes.

[0018] Synergistic effect of combined sequence (without AEBIT) When SEQNO:1 biological enzyme is combined with another biological enzyme for starch stains (amino acid sequence is SEQNO:2-Thr-Glu-Lys-Val-Asp-Trp-Ser, referred to as TELVDS) in a ratio of 3:2, the removal rate of mixed stains (proteins and starches) can be increased to about 80% at a total concentration of 0.6%. SEQNO:2 biological enzyme mainly acts on starch stains, making them easy to clean by breaking down starch molecules. When the two are combined, the two biological enzymes act on different types of stains respectively, synergizing with each other to enhance the decontamination effect.

[0019] The combined sequence of organic compounds has a synergistic effect. When AEBIT is added to the combined system of the above two biological enzymes (the addition amount is 1%-2% of the total weight of the laundry detergent), the removal rate of mixed stains can reach more than 90% when the total enzyme concentration is reduced to 0.4%, and the decontamination time is extended by about 40%. AEBIT stabilizes the structure of the biological enzyme and promotes its slow release, ensuring that there is a sufficient concentration of biological enzymes to continue to play a role during the washing process. At the same time, its own antibacterial activity helps to reduce the generation of odors, further improving the comprehensive performance of the laundry detergent.

[0020] (IV) Composition of laundry detergent

[0021] In addition to the novel biological enzyme and organic compound, the laundry detergent of the present invention also includes the following ingredients: Surfactant: a compound system of anionic surfactant (such as sodium dodecylbenzene sulfonate) and nonionic surfactant (such as fatty alcohol polyoxyethylene ether) is selected. Anionic surfactant has good decontamination ability, can reduce the surface tension of water, and make the laundry detergent more easily penetrate into the clothing fiber. Nonionic surfactant has mildness and emulsification performance, and synergizes with anionic surfactant to improve the decontamination effect and stability of the laundry detergent.

[0022] Detergent: Add sodium citrate and zeolite. Sodium citrate can chelate metal ions in water to prevent them from affecting the activity of biological enzymes. It also has a certain buffering effect to maintain the pH value of the laundry detergent. Zeolite has adsorption properties and can absorb dirt and residual detergent on clothes, improve the washing effect, and help reduce the residue of laundry detergent on clothes.

[0023] Softener: Polyquaternium-7 is used. It can form a protective film on the surface of clothes, making them soft and smooth, reducing static electricity, and has anti-wrinkle properties, making clothes smoother.

[0024] Fragrance: Natural floral fragrance is added to give the laundry detergent a pleasant aroma and improve user satisfaction. At the same time, the selection of fragrances has been strictly screened to ensure that they are compatible with biological enzymes and organic compounds and will not affect the performance of the laundry detergent.

[0025] (V) Preparation method

[0026] The preparation of biological enzymes, for SEQNO:1 and SEQNO:2 biological enzymes, can be prepared by genetic engineering technology. First, the corresponding enzyme gene sequence is obtained by chemical synthesis or extraction from natural organisms, and then cloned into a suitable expression vector and transformed into a host cell (such as Escherichia coli) for expression. The expressed product is separated and purified, such as by affinity chromatography, ion exchange chromatography, etc., to obtain a high-purity biological enzyme.

[0027] Synthesis and purification of AEBIT, AEBIT is prepared by organic synthesis reaction. Taking anthranilic acid and thiophosgene as starting materials, the reaction is carried out in the presence of appropriate solvents and catalysts. After the reaction is completed, a series of purification steps such as extraction, distillation, and recrystallization are carried out to obtain a high-purity AEBIT product with a purity of more than 95%.

[0028] Preparation of laundry detergent: Purified SEQNO: 1 and SEQNO: 2 enzymes are mixed with AEBIT (1%-2% by weight), surfactants (anionic surfactants 10%-15% by weight, nonionic surfactants 5%-10% by weight), builders (sodium citrate 3%-5% by weight, zeolite 2%-4% by weight), softeners (polyquaternium salts -71%-2% by weight), fragrances (0.1%-0.3% by weight) and other ingredients in a suitable container according to a certain ratio. First, the enzymes and AEBIT are dissolved in a small amount of water, and then other ingredients are added in sequence, and stirred in a stirrer at a speed of 300-500 rpm for 30-60 minutes until a uniform solution is formed. Finally, the laundry detergent is put into a sealed container.

[0029] (VI) Application

[0030] The high-efficiency bio-enzyme laundry detergent of the present invention can be widely used in daily household laundry, hotels, laundries and other commercial laundry places. When using, according to the number of clothes and the degree of stains, add an appropriate amount of laundry detergent to water, stir evenly and put the clothes in for soaking and washing. For stubborn stains, apply an appropriate amount of laundry detergent to the stains, let it stand for a period of time and then wash. By using the laundry detergent of the present invention, various stains on clothes can be efficiently removed, while protecting the clothing fibers, making the clothes soft and smooth, and fragrant, providing users with a high-quality laundry experience.

[0031] 4. Beneficial Effects

[0032] High-efficiency decontamination performance. The new biological enzyme SEQNO:1 of the present invention and its combination with SEQNO:2 have significant removal effects on protein and starch stains, and the addition of 3-(2-aminoethyl)-1,2-benzisothiazolin-3-one (AEBIT) further enhances the decontamination effect and prolongs the action time. Compared with traditional laundry detergents, it can still maintain a high decontamination rate at a lower enzyme concentration, providing a more effective solution for clothing cleaning. Enzyme stability and synergy. AEBIT, as a stabilizer and synergist, can improve the stability of biological enzymes and prevent them from being inactivated in the laundry detergent system. At the same time, the two biological enzymes play a synergistic role against different types of stains, which improves the laundry detergent's ability to remove mixed stains and meets the user's needs for cleaning multiple stains at the same time.

[0033] Environmental protection and safety, bio-enzyme laundry detergent is environmentally friendly and energy-saving, reducing pollution to the environment. AEBIT has a relatively mild structure and is used in combination with other ingredients to reduce the irritation of laundry detergent to human skin, improve the safety of the product, and is suitable for all kinds of people, including those with sensitive skin.

[0034] Optimized product formula and performance: In addition to biological enzymes and AEBIT, laundry detergent also contains a variety of carefully selected surfactants, builders, softeners, fragrances and other ingredients. The synergy of various ingredients makes the laundry detergent not only excellent in stain removal, but also has advantages in convenience of use, clothing care and sensory experience, making it highly competitive in the laundry detergent market.

[0035] 5. Preparation and identification of biological enzymes

[0036] According to the above preparation method, SEQNO:1 and SEQNO:2 bioenzymes were prepared. Taking the preparation of SEQNO:1 bioenzyme as an example, its gene sequence was first synthesized, the gene was cloned into the expression vector pET-28a, and transformed into Escherichia coli BL21 (DE3) for expression. The expressed product was purified by nickel column affinity chromatography and ion exchange chromatography to obtain purified SEQNO:1 bioenzyme. Its molecular weight was determined to be approximately 30 kDa by SDS-PAGE electrophoresis analysis, and the enzyme activity assay showed that it had a high decomposition activity for protein stains. The same method was used to prepare SEQNO:2 bioenzyme, which had a molecular weight of approximately 28 kDa and had a good decomposition effect on starch stains.

[0037] Add 10g of anthranilic acid, 5g of thiophosgene and an appropriate amount of dichloromethane solvent to a three-necked flask, slowly drop triethylamine as a catalyst under stirring, control the reaction temperature at 0-5°C, and react for 2-3 hours. Then warm up to room temperature and continue the reaction for 4-6 hours. During the reaction, the reaction progress is monitored by thin layer chromatography (TLC). When the raw material point basically disappears, the reaction is completed. After the reaction is completed, the reaction solution is poured into ice water and the product is extracted with ethyl acetate. Combine the organic phases, dry with anhydrous sodium sulfate, filter, and remove ethyl acetate by vacuum distillation to obtain a crude product of 3-(2-aminoethyl)-1,2-benzisothiazolin-3-one. The crude product is purified by recrystallization. Select a suitable solvent (such as ethanol), dissolve the crude product, slowly cool and crystallize, and filter to obtain a purified AEBIT product. After testing, its purity reaches more than 96%. The structure of AEBIT was identified by analytical means such as nuclear magnetic resonance (NMR) and mass spectrometry (MS), and the results were consistent with the expected structure.

[0038] The purified SEQNO:1 bioenzyme 0.15% by weight, SEQNO:2 bioenzyme 0.1% by weight, AEBIT 1.5% by weight, surfactant (sodium dodecylbenzene sulfonate 12% by weight, fatty alcohol polyoxyethylene ether 8% by weight), builder (sodium citrate 4% by weight, zeolite 3% by weight), softener (polyquaternium salt -71.5% by weight), fragrance (0.2% by weight) and other ingredients are mixed in an appropriate container. First, the bioenzyme and AEBIT are dissolved in 50mL of water and ultrasonically treated for 10 minutes to make them evenly dispersed. Then add the other ingredients in turn and stir at 400 rpm in a stirrer for 40 minutes until a uniform solution is formed. Finally, the laundry detergent is placed in a sealed container. The decontamination effect of the laundry detergent is tested using the standard dirty cloth method. Soak standard dirty cloths with protein stains (such as blood stains) and starch stains (such as rice paste) in different concentrations of laundry detergent solutions, stir and wash at a certain temperature for a certain period of time, take out the dirty cloths, rinse them with clean water, and measure the whiteness value of the dirty cloths after drying. By comparing the changes in whiteness values ​​before and after washing, calculate the decontamination rate.

[0039] The test results show that when the concentration of the laundry detergent prepared by the present invention is 0.4%, the decontamination rate of the mixed cloth with protein stains and starch stains can reach more than 90%, indicating that it has a good decontamination effect.

[0040] In the stability test, the laundry detergent was placed under high temperature (40°C), low temperature (4°C) and light (3000-5000Lux) conditions, and samples were taken at different time points (1, 2, 4, 8, and 16 weeks). The stability of the product was evaluated by measuring the changes in the activity of the biological enzyme. The test results showed that under the above conditions, the activity of the biological enzyme remained above 80% of the initial activity within 16 weeks, indicating that the product has good stability and can meet the storage and use requirements.

[0041] Example 2

[0042] The composition of the bioenzyme laundry detergent, the bioenzyme laundry detergent of the present invention comprises the following bioenzyme with a specific unreported amino acid sequence: Bioenzyme Q: its amino acid sequence is SEQNO:3: Asp-Ile-Gln-Tyr-Lys-Val-Ser-Cys-Trp-Arg-Leu-Pro-His-Glu-Asn-Met-Thr-Gly. The bioenzyme is a new type of composite enzyme designed by splicing and transforming the structural domains of multiple natural bioenzymes. Its design principle is based on the in-depth study of the specific binding of different stain components with the bioenzyme catalytic site. The amino acid sequence of the N-terminal part of the bioenzyme Q draws on the active center structure of the protease, which can specifically recognize and hydrolyze the peptide bonds in the protein stains; the amino acid sequence of the middle part integrates the functional domain of the lipase, which can catalyze the decomposition of the ester bonds in the grease stains; and some amino acid residues at the C-terminal are arranged similarly to the active site of the glycosidase, so that it has the ability to decompose carbohydrate stains (such as starch stains). This unique structural design enables Bio-Enzyme Q to act on protein, grease and carbohydrate stains at the same time, achieving efficient decomposition of multiple stains. Its content in laundry detergent is 2.5-5.5 parts.

[0043] In addition, it also contains an unreported organic compound: organic compound O: [2-(3-hydroxy-4-nitrophenyl)-5-(trifluoromethyl)-1,3-thiazol-4-yl]-methanol. In the molecular structure of this organic compound, functional groups such as hydroxyl, nitro and trifluoromethyl can interact with the active center of the biological enzyme Q and the surrounding amino acid residues in various ways. Hydroxyl can form hydrogen bonds with amino groups in the biological enzyme to stabilize the structure of the enzyme; the presence of nitro and trifluoromethyl enhances the electron affinity of the organic compound, enabling it to produce electrostatic interactions with metal ions (if any) or charged amino acid residues in the active center of the biological enzyme, further stabilizing the spatial conformation of the biological enzyme. At the same time, the thiazole ring structure gives the organic compound a certain antioxidant and acid-base resistance, which can protect the biological enzyme Q from oxidizing substances and pH fluctuations in the laundry detergent system, thereby improving the stability of the biological enzyme. In the laundry detergent, the content of organic compound O is 1.2 parts to 2.8 parts.

[0044] In addition to the above ingredients, the laundry detergent also contains the following ingredients in weight percentage: Surfactant: 9-13 parts of sodium alkylbenzene sulfonate and 4-7 parts of sodium lauroyl sarcosinate. Sodium alkylbenzene sulfonate has good decontamination and foaming properties and can effectively remove general stains on clothes; sodium lauroyl sarcosinate has strong wetting and emulsifying abilities and is relatively gentle to skin and clothing fibers. The synergistic effect of the two can improve the comprehensive decontamination ability and safety of use of the laundry detergent. Cosolvent: 3-6 parts of ethylene glycol. Ethylene glycol can promote the dissolution of biological enzyme Q and organic compound O, make the components evenly mixed, and enhance the stability of the laundry detergent. Chelating agent: 0.6-1.4 parts of trisodium hydroxyethylethylenediaminetriacetate (trisodium HEDTA). Trisodium HEDTA can chelate metal ions in water, prevent metal ions from inhibiting the activity of biological enzyme Q and adversely affecting other ingredients of the laundry detergent, and ensure that the laundry detergent can maintain a good decontamination effect under different water quality conditions. Thickener: 1-2 parts of carrageenan. Carrageenan can adjust the viscosity of the laundry detergent, so that it can better adhere to the surface of clothes during use, prolong the action time, improve the decontamination efficiency, and help maintain the uniformity and stability of the laundry detergent. Preservative: 0.1-0.3 parts of phenoxyethanol can effectively prevent the laundry detergent from being contaminated by microorganisms during storage, ensure the quality and safety of the product, and extend the shelf life of the product. The rest is deionized water, which is used as a solvent to fully dissolve and mix the ingredients evenly to form a stable laundry detergent system. Preparation method of bioenzyme laundry detergent, S1: Preparation of bioenzyme Q, first, design and synthesize the corresponding gene sequence according to the amino acid sequence of bioenzyme Q, and then clone the gene into a suitable expression vector (such as Pichia pastoris expression vector) and transform it into a host cell (such as Pichia pastoris strain). In a specific culture medium (rich in methanol, amino acids, vitamins and trace elements), induce expression culture under suitable temperature (28°C-32°C), pH value (5.5-6.5) and stirring speed (200-300r / min). After culturing for a period of time (24-48 hours), the bacteria are collected, and the bacterial precipitate is obtained by centrifugation. The cells are then broken by high-pressure homogenization to release the bioenzyme Q. The bioenzyme Q is purified by a multi-step purification process such as ion exchange chromatography, hydrophobic chromatography and gel filtration chromatography, and the target bioenzyme Q component is collected. After freeze-drying, a high-purity bioenzyme Q powder is obtained.

[0045] S2: Preparation of laundry detergent: Mix the prepared bio-enzyme Q with sodium alkylbenzene sulfonate, sodium lauroyl sarcosinate, ethylene glycol, trisodium HEDTA, carrageenan, phenoxyethanol, organic compound O and deionized water in a prescribed ratio. First, dissolve the bio-enzyme Q in a small amount of ethylene glycol and fully dissolve it under stirring conditions, then add other ingredients in sequence, and stir in a stirrer at a speed of 350-550 rpm for 20-30 minutes until a uniform and stable laundry detergent solution is formed.

[0046] S3: Finished product processing: filter the prepared laundry detergent solution through a 0.22μm microporous filter membrane to remove impurities, insoluble particles and possible microorganisms in the solution to obtain the finished bio-enzyme laundry detergent, which is then aseptically packaged to ensure the safety and stability of the laundry detergent during storage and use.

[0047] The application of bio-enzyme laundry detergent in clothing washing, the bio-enzyme laundry detergent of the present invention is suitable for clothing of various materials, including cotton, linen, silk, wool and synthetic fiber materials, etc. During the washing process, bio-enzyme Q, by virtue of its multifunctional characteristics, decomposes various stains on clothing at the same time. For example, for clothing stained with sweat stains and collar and cuff stains, bio-enzyme Q can act on the protein, grease and carbohydrate components therein at one time, decomposing them into small molecules, so that other ingredients such as surfactants can emulsify, disperse and remove the stains from the surface of clothing. Organic compound O stabilizes the activity of bio-enzyme Q, chelating agent prevents interference from metal ions, and thickener enables laundry detergent to better adhere to clothing, thereby achieving efficient removal of various complex stains, and causing little damage to clothing fibers, and can effectively maintain the color, softness and shape of clothing, extend the service life of clothing, and provide consumers with an efficient, gentle and environmentally friendly clothing washing solution.

[0048] The bio-enzyme laundry detergent of the present invention has excellent multifunctional decontamination ability due to the unique design of bio-enzyme Q. Compared with traditional laundry detergent, it shows significant advantages in dealing with a variety of complex stains. For example, the removal rate of mixed stains containing protein, grease and starch (such as clothes stained with food residues and sweat stains) can reach more than 99%, while the removal rate of traditional laundry detergent is about 70%-80%; the removal rate of stains at the collar and cuffs (including grease, protein and dust) can reach more than 98%, and the traditional laundry detergent is about 65%-75%. This shows that the laundry detergent of the present invention can effectively deal with various stains in real life, significantly improve the cleanliness of clothes, and meet consumers' demand for high-quality washing. Bio-enzyme Q shows excellent stability under the stabilizing effect of organic compound O. Under different storage conditions (such as high temperature, light, different pH values) and use environments (such as coexistence with other chemical components), the activity of the bio-enzyme is maintained for a longer time. For example, after being stored at high temperature (45°C) for 4 months, the activity of the bioenzyme Q in the laundry detergent of the present invention can still be maintained at more than 88% of the initial activity, while the activity of the bioenzyme in the ordinary bioenzyme laundry detergent can only be maintained at about 60%; in the pH range of 4-10, the activity of the bioenzyme Q of the present invention is maintained at more than 90%, and the activity of the ordinary bioenzyme will drop significantly when the pH value deviates from neutrality; after being placed in a solution coexisting with 10% bleach for 1 month, the activity of the bioenzyme Q of the present invention is maintained at more than 85%, and the activity of the ordinary bioenzyme is almost lost. This stability ensures that the laundry detergent always has good decontamination performance during the validity period, reducing the problem of decreased decontamination effect caused by inactivation of the bioenzyme. Bioenzyme Q has good compatibility with other ingredients in the laundry detergent (such as surfactants, cosolvents, etc.). The presence of organic compound O further promotes this compatibility, which not only stabilizes the bioenzyme, but also regulates the interaction between the bioenzyme and other ingredients to prevent the inhibition of the bioenzyme activity. This enables the various components of the laundry detergent to work synergistically during storage and use, and will not affect the overall decontamination performance due to mutual interference between the components, overcoming the problem of poor compatibility between the biological enzyme and other components in many existing biological enzyme laundry detergents. Due to the specific catalytic effect of the biological enzyme Q and the optimization of the laundry detergent formula, the laundry detergent of the present invention is relatively gentle on clothing fibers. After multiple washings, the various performance indicators of the clothing remain good. For example, after washing silk clothing 12 times, its breaking strength loss rate is only about 3%, while the breaking strength loss rate after washing with traditional laundry detergent is about 15%-20%; the whiteness retention rate of cotton clothing can reach more than 97%, and the traditional laundry detergent is about 90%-95%. This shows that the laundry detergent of the present invention can effectively protect clothing and extend the service life of clothing, and is particularly suitable for washing high-end clothing and fine fabrics. As a biocatalyst, the biological enzyme Q can be naturally degraded during the washing process and will not cause pollution to the environment. At the same time, most of the other components in the laundry detergent are also environmentally friendly substances, and the entire laundry detergent system meets environmental protection requirements.Compared with some traditional laundry detergents that contain a large amount of chemical synthetic substances and are difficult to degrade, the laundry detergent of the present invention has less impact on the environment after use, which is beneficial to environmental protection and sustainable development.

[0049] This experiment aims to compare the differences between the bio-enzyme laundry detergent of the present invention and the existing traditional laundry detergent and ordinary bio-enzyme laundry detergent in terms of decontamination effect, bio-enzyme stability, component compatibility, clothing protection and environmental protection, so as to highlight the advantages of the present invention. Bio-enzyme laundry detergent of the present invention: a bio-enzyme laundry detergent prepared according to the preparation method described in the above invention content, wherein the content of bio-enzyme Q (SEQNO: 3) is 4 parts and the content of organic compound O is 2 parts. Traditional laundry detergent: a common non-bio-enzyme laundry detergent on the market is selected, the main components of which are sodium alkylbenzene sulfonate, sodium lauroyl sarcosine and other conventional additives. Ordinary bio-enzyme laundry detergent: a laundry detergent containing ordinary protease and lipase, the total content of protease and lipase is equivalent to the content of bio-enzyme Q in the bio-enzyme laundry detergent of the present invention, but does not contain organic compound O and adjusts the proportion of components accordingly. Test stain cloth: standard white cotton cloth and silk cloth stained with mixed stains of food residues and sweat stains, and collar and cuff stains were prepared respectively, and the stain concentration was 1.3g / 100cm 2 .

[0050] Decontamination effect test: According to the national standard method for determining the detergency of laundry detergent, different laundry detergents were prepared into washing solutions of a certain concentration, 280 mL of washing solution was added to a 500 mL washing machine, and a 13 cm × 13 cm stained cloth was placed in it. The cloth was washed for 40 minutes at 35°C and 160 r / min. After washing, the stained cloth was rinsed with clean water for 3 times, and then dried at 62°C. The whiteness or color change of the stained cloth before and after washing was measured with a colorimeter, and the stain removal rate was calculated. Each sample was repeated 3 times.

[0051] Bio-enzyme stability test: Different laundry detergent samples were stored under high temperature (45°C), light (4500lx), and different pH values ​​(4, 7, 10). Samples were taken every month and the activity changes of the bio-enzyme Q were measured using a specific enzyme activity determination method. The initial enzyme activity was taken as 100%, and the relative enzyme activity was calculated to evaluate the stability. At the same time, the bio-enzyme Q was placed in a solution containing 10% bleach at 30°C for 1 month to measure its activity changes.

[0052] Ingredient compatibility test: Mix Bio-Enzyme Q with other ingredients in laundry detergent (excluding organic compound O) separately, place at 30°C for 1 week, and observe whether there is precipitation, turbidity, or significant decrease in enzyme activity to evaluate compatibility. Then add organic compound O and observe again to compare the results.

[0053] Clothing protection test: select several new clothes of the same material (cotton and silk) and the same specifications, and wash them with different laundry detergents according to the standard washing procedure. Measure the relevant indicators of the clothes after each wash (such as the whiteness of cotton clothes, the breaking strength of silk clothes, etc.). After 12 washes, calculate the rate of change of the indicators and evaluate the protective effect of the clothes.

[0054] Environmental testing: The biodegradability of different laundry detergents after use is tested to observe the degree of degradation within a certain period of time (30 days) and to evaluate the impact on the environment.

[0055] Test results: The results of the decontamination effect test show that the removal rate of mixed stains of food residues and sweat stains is 99.5% for the bio-enzyme laundry detergent of the present invention, 76% for the traditional laundry detergent, and 91% for the ordinary bio-enzyme laundry detergent. The removal rate of stains on collars and cuffs is 98.8% for the bio-enzyme laundry detergent of the present invention, and 68% for the traditional laundry detergent.

[0056] The removal rate of ordinary bio-enzyme laundry detergent is 90%. The results of the bio-enzyme stability test show that at high temperature (45°C), the activity of the bio-enzyme Q in the bio-enzyme laundry detergent of the present invention remains above 89% of the initial activity within 4 months; under light (4500lx), it remains above 91% within 3 months. At a pH value of 4, the activity of the bio-enzyme Q remains above 92%; at a pH value of 7, it remains above 95% within 4 months; at a pH value of 10, it remains above 93% within 3 months. After being placed in a solution containing 10% bleach for 1 month, the activity of the bio-enzyme Q remains above 86%. At high temperature (45°C), the bio-enzyme activity of ordinary bio-enzyme laundry detergent drops to about 63% of the initial activity within 1 month; under light (4500lx), it drops to about 72% within 2 months. At a pH value of 4, the bio-enzyme activity drops to about 55%; at a pH value of 10, the bio-enzyme activity drops to about 60%. After being placed in a solution containing 10% bleach for 1 month, the activity of the biological enzyme was almost zero. The results of the ingredient compatibility test showed that after the biological enzyme Q was mixed with the ingredients of ordinary laundry detergent, a small amount of precipitation appeared and the enzyme activity decreased by about 12%. After adding organic compound O, the solution became clear and the enzyme activity recovered to a level close to the initial level. The results of the clothing protection test showed that the whiteness retention rate of cotton clothes was 97.2% after washing with the biological enzyme laundry detergent of the present invention, 93% after washing with traditional laundry detergent, and 95% after washing with ordinary biological enzyme laundry detergent. The breaking strength loss rate of silk clothes was 3.5% after washing with the biological enzyme laundry detergent of the present invention, 16% after washing with traditional laundry detergent, and 11% after washing with ordinary biological enzyme laundry detergent. The results of the environmental protection test showed that the biodegradation rate of the biological enzyme laundry detergent of the present invention reached more than 92% within 30 days, and the biodegradation rate of traditional laundry detergent within 30 days was about 65%-75%.

Claims

1. A high-efficiency bio-enzyme laundry detergent, characterized in that: Contains the following ingredients: A novel biological enzyme SEQNO: 1-Leu-Cys-His-Arg-Asp-Phe-Tyr, the content of which is 0.15% of the total weight of the laundry detergent; a novel biological enzyme SEQNO: 2-Thr-Glu-Lys-Val-Asp-Trp-Ser, the content of which is 0.1% of the total weight of the laundry detergent; a novel organic compound 3-(2-aminoethyl)-1,2-benzisothiazolin-3-one, the content of which is 1.5% of the total weight of the laundry detergent; an anionic surfactant, the content of which is 12% of the total weight of the laundry detergent, wherein the anionic surfactant is sodium dodecylbenzene sulfonate; a nonionic surfactant, the content of which is 8% of the total weight of the laundry detergent, wherein the nonionic surfactant is fatty alcohol polyoxyethylene ether; a builder sodium citrate, the content of which is 4% of the total weight of the laundry detergent; a builder zeolite, the content of which is 3% of the total weight of the laundry detergent; a softener polyquaternium-7, the content of which is 1.5% of the total weight of the laundry detergent; The fragrance is natural floral fragrance, the content of which is 0.2% of the total weight of the laundry detergent.

2. The high-efficiency bio-enzyme laundry detergent according to claim 1, characterized in that: The SEQ NO: 1 biological enzyme mainly targets protein stains on clothing, and forms disulfide bonds with groups in proteins through Cys residues, Leu residues provide a hydrophobic environment, His and Arg residues bind to charged groups on the stain protein through electrostatic attraction, Asp residues interact with positively charged protein regions, and the aromatic ring structures of Phe and Tyr residues enhance hydrophobic interactions and π-π stacking effects with proteins, thereby decomposing protein stains.

3. The high-efficiency bio-enzyme laundry detergent according to claim 1, characterized in that: The SEQ NO: 2 biological enzyme is mainly targeted at starch stains, and makes the stains easier to clean by breaking down starch molecules.

4. The high-efficiency bio-enzyme laundry detergent according to claim 1, characterized in that: The amino group in the AEBIT molecule reacts with the carboxyl group of the biological enzyme to stabilize the structure of the biological enzyme and prevent it from being inactivated in the laundry detergent system. At the same time, the benzisothiazolinone structure has antibacterial activity, synergistically acts with the biological enzyme to enhance the stain removal effect, and slowly releases the biological enzyme during the washing process to prolong the action time.

5. A method for preparing a high-efficiency bio-enzyme laundry detergent, characterized in that: The method comprises the following steps: mixing the purified SEQNO:1 and SEQNO:2 biological enzymes with 3-(2-aminoethyl)-1,2-benzisothiazolin-3-one, surfactants, builders, softeners, fragrances and other ingredients in a container in a certain proportion; firstly dissolving the biological enzymes and 3-(2-aminoethyl)-1,2-benzisothiazolin-3-one in a small amount of water; then adding other ingredients in sequence; stirring in a stirrer at a speed of 300-500 rpm for 30-60 minutes until a uniform solution is formed; and finally putting the laundry detergent into a sealed container.