A protease, composition and use having mild exfoliating function

Abstract

The application discloses a protease with mild keratolysis, a composition and application, and belongs to the technical field of genetic engineering or cosmetics. The application provides a basic heat-stable serine protease TVG06 obtained in an acidic hot spring in Ali uninhabited area, and the amino acid sequence of the basic heat-stable serine protease TVG06 is shown as SEQ ID NO. 1. The serine protease in the application has a degradation rate of enzyme activity of less than 15% under the condition of 45 DEG C for 4 weeks, and the enzyme activity is more than 65% after 8 weeks. Compared with traditional keratolysis raw materials, the composition in the application has better oil control, anti-inflammatory and keratolysis effects, and is more soothing.

Description

Technical Field

[0001] This invention relates to a protease, composition, and application with a gentle exfoliating function, belonging to the fields of genetic engineering technology or cosmetic technology. Background Technology

[0002] With the increasing demand for efficient, gentle, and environmentally friendly exfoliating and oil-controlling products in the beauty, skincare, personal hygiene, and industrial cleaning industries, enzyme preparations, due to their advantages such as specific catalysis, low irritation, and easy biodegradability, are gradually replacing traditional chemical exfoliating ingredients (such as glycolic acid and salicylic acid) and physical exfoliating particles, becoming the core active ingredient in exfoliating and oil-controlling systems. Among them, enzyme preparations (such as proteases, lipases, and keratinases) that degrade keratin in the stratum corneum and lipids in sebum are widely used in skincare products, facial cleansers, and oil-controlling shampoos due to their precise ability to decompose aged keratin and their targeted regulation of excess oil.

[0003] However, existing enzyme preparations for exfoliation and oil control generally suffer from poor stability, severely limiting their application scenarios and product shelf life. On the one hand, during product manufacturing, enzyme preparations are extremely sensitive to temperature. Conventional high-temperature sterilization processes can easily destroy the spatial conformation of enzymes, leading to a significant decrease in enzyme activity or even complete inactivation. This forces companies to adopt low-temperature sterilization or aseptic filling processes, significantly increasing production energy consumption and costs. On the other hand, during product storage and use, enzyme preparations are susceptible to activity decay due to fluctuations in ambient temperature, changes in pH, and the influence of other components in the formulation (such as surfactants and preservatives), making it impossible to guarantee the stability of the exfoliation and oil control effect throughout the product's entire life cycle.

[0004] Meanwhile, the limited functionality of existing exfoliating and oil-controlling enzyme preparations makes it difficult to meet complex application needs. Most products can only achieve a single exfoliation or oil-control function, and they are prone to causing potential damage to the skin barrier when they work—if the enzyme activity is too strong, it may over-decompose healthy keratin; if the activity is insufficient, it cannot achieve the expected exfoliation and oil-control effect. In addition, for the exfoliation and oil-control needs in high-temperature scenarios such as industrial cleaning, existing enzyme preparations are completely unsuitable due to their lack of heat resistance, resulting in this field still relying on highly irritating chemical reagents, posing environmental hazards and safety risks.

[0005] To address the aforementioned technical challenges, there is an urgent need in this field to develop an enzyme preparation that possesses high-temperature resistance, excellent activity stability, and the ability to precisely achieve a synergistic effect of exfoliation and oil control. Summary of the Invention

[0006] To address the shortcomings of the prior art, this invention provides a protease, composition, and application with gentle exfoliation properties. The aim is to solve the technical problem of the lack of an enzyme preparation in the prior art that has high temperature resistance, excellent activity stability, and can accurately achieve the synergistic effect of exfoliation and oil control.

[0007] The first technical solution provided by the present invention is an alkaline heat-stable serine protease TVG06, the amino acid sequence of which is shown in SEQ ID NO.1.

[0008] In one embodiment, the serine protease is capable of degrading keratin.

[0009] In one embodiment, the optimal reaction temperature of the serine protease is 50°C, and it exhibits good thermal stability.

[0010] In one embodiment, the optimal reaction pH for the serine protease is 8, exhibiting good pH stability.

[0011] The second technical solution provided by the present invention is a gene encoding the serine protease TVG06 described in the first technical solution.

[0012] The third technical solution provided by the present invention is a recombinant plasmid carrying the gene described in the second technical solution.

[0013] In one embodiment, the expression vector of the recombinant plasmid is the pET-28a plasmid.

[0014] The fourth technical solution provided by the present invention is a recombinant cell that expresses the serine protease TVG06 described in the first technical solution, or contains the gene described in the second technical solution, or is transformed with the recombinant plasmid described in the third technical solution.

[0015] In one embodiment, the recombinant cells use Escherichia coli BW25113 as the host.

[0016] The fifth technical solution provided by this invention is the application of the gene described in the second technical solution, or the recombinant plasmid described in the third technical solution, or the recombinant cell described in the fourth technical solution in the preparation of serine protease TVG06 or products containing alkaline heat-stable serine protease TVG06.

[0017] The sixth technical solution provided by the present invention is a composition containing the serine protease TVG06 described in the first technical solution.

[0018] In one embodiment, the composition further comprises superoxide dismutase and papain.

[0019] In one embodiment, the composition further comprises glycerol, 1,2-pentanediol, 1,2-hexanediol, and tetrahydromethylpyrimidine carboxylic acid.

[0020] Further, by mass percentage, the composition contains 0.5% alkaline heat-stable serine protease TVG06, 0.2% superoxide dismutase, 0.13% papain, 25% glycerol, 1% 1,2-pentanediol, 2% 1,2-hexanediol, 1% tetrahydromethylpyrimidine carboxylic acid, and 70.17% water.

[0021] The seventh technical solution provided by this invention is the application of the alkaline heat-stable serine protease TVG06 described in the first technical solution or the composition described in the sixth technical solution in the preparation of exfoliating daily chemical products.

[0022] In one embodiment, the daily chemical products include facial cleanser, shower gel, serum, and mouthwash.

[0023] Compared with the prior art, the beneficial effects of the present invention are as follows: (1) The present invention provides an alkaline thermostable serine protease obtained from acidic hot springs in the uninhabited area of ​​Ali, the amino acid sequence of which is shown in SEQ ID NO.1.

[0024] (2) The serine protease in this invention has an enzyme activity degradation rate of less than 15% after 4 weeks at 45°C and an enzyme activity of more than 65% after 8 weeks.

[0025] (3) Compared with traditional exfoliating materials, the composition in this invention has better oil control, anti-inflammatory and exfoliating effects, and is more soothing.

[0026] (4) The composition in this invention has good stability. After being placed at 5℃, 25℃, 45℃ and UV conditions for 8 weeks, its enzyme activity for degrading keratin remains unchanged and maintains 100% activity. Attached Figure Description

[0027] Figure 1 Predicting the structure of serine proteases.

[0028] Figure 2 SDS-PAGE analysis of serine protease TVG06.

[0029] Figure 3 This is the optimal reaction temperature for the serine protease TVG06.

[0030] Figure 4 The thermal stability of the serine protease TVG06.

[0031] Figure 5 The optimal reaction pH for the serine protease TVG06 is [value missing].

[0032] Figure 6 This is to assess the enzyme activity and stability of the serine protease TVG06.

[0033] Figure 7 The change rate of protein stripping after different sample treatments.

[0034] Figure 8 The effect of different treatment groups on the degradation of corneal plugs.

[0035] Figure 9 These are the results of a chicken embryo experiment.

[0036] Figure 10 A comparison of the high-temperature stability of serine protease TVG06 and competing products. Detailed Implementation

[0037] The preferred embodiments of the present invention are described below. It should be understood that the embodiments are for better explanation of the present invention and are not intended to limit the present invention.

[0038] Test method: Enzyme activity detection methods Preparation of test samples: Weigh 4 mg of azure keratin into a 1.5 mL EP tube. The reaction mixture contains 0.2 mL of the test sample and 0.8 mL of 10 mM Tris-HCl buffer (pH 8). Incubate at 50 °C and 220 rpm for 3 days. Centrifuge at 12000 rpm for 10 minutes. Take 200 µL of the supernatant and measure the absorbance at 595 nm using a microplate reader. Record this value as A.

[0039] Preparation of blank control sample: Take 4 mg of azure keratin into a 1.5 mL EP tube. The reaction mixture contains 0.2 mL of water and 0.8 mL of 10 mM Tris-HCl buffer (pH 8). Incubate at 50 °C and 220 rpm for 3 days. Centrifuge at 12000 rpm for 10 minutes. Take 200 µL of supernatant and measure the absorbance at 595 nm using a microplate reader. Record the absorbance as A'.

[0040] Preparation of background test sample (without substrate): Add 0.2 mL of the test sample and 0.8 mL of 10 mM Tris-HCl buffer (pH 8) to a 1.5 mL EP tube. Incubate at 50 °C and 220 rpm for 3 days. Centrifuge at 12000 rpm for 10 minutes. Take 200 µL of supernatant and measure the absorbance at 595 nm using a microplate reader. Record this value as A0.

[0041] Preparation of background blank control sample (without substrate): Add 0.2 mL of water and 0.8 mL of 10 mM Tris-HCl buffer (pH 8) to a 1.5 mL EP tube, and incubate at 50 °C and 220 rpm for 3 days. Centrifuge at 12000 rpm for 10 minutes, take 200 µL of supernatant, and measure the absorbance at 595 nm using a microplate reader. Record this as A'0.

[0042] Calculation formula: Keratinase activity (U / L) = (1) Where A is the absorbance at 595 nm of the test sample; A0 is the absorbance at 595 nm of the background of the test sample; A' is the absorbance at 595 nm of the blank control sample; A'0 is the absorbance at 595 nm of the background of the blank control sample; E is 0.01, according to the definition of enzyme activity: in A 595 For the blank control, each change in absorbance value of 0.01 corresponds to 1 U of keratinase activity; 1000 is the dilution factor.

[0043] Materials used in the examples: LB medium: 1% tryptone (purchased from oxoid), 0.5% yeast extract (purchased from oxoid), 1% sodium chloride (purchased from Xilong Scientific), pH natural (approximately 7.0). Solid medium was prepared with 2.0% (w / v) agar (purchased from oxoid).

[0044] 10mM Tris-HCl (pH 8): Weigh 1.2114g of Tris powder (purchased from Solarbio) into a beaker, add 900mL of distilled water, adjust the pH to 8 using HCl, and finally bring the volume to 1L.

[0045] 1M Tris-HCl buffer (pH 7.4): Weigh 121.14g of Tris powder (purchased from Solarbio) into a beaker, add 900mL of distilled water, adjust the pH to 7.4 using HCl, and finally bring the volume to 1L.

[0046] The superoxide dismutase was sourced from Shenzhen Zhongke Xinyang Biotechnology Co., Ltd., with an enzyme activity of 10 W IU / g.

[0047] Papain was obtained from Henan Anrui Biotechnology Co., Ltd., with an enzyme activity of 10 W IU / g.

[0048] Example 1: Screening and Homology Comparison of Serine Proteases In the microbial gene resource library obtained from acidic hydrothermal vents in the Ali uninhabited area, screening was carried out based on highly conserved motifs of serine protease donor-substrate binding, and the heat resistance of serine proteases was predicted. The amino acid sequence of the final screened protease is shown in SEQ ID NO.1.

[0049] SEQ ID NO.1: MLHHRPGHGGGPGGGGGGEEVSSQITPEGVKRVGQPTDSSNGSGIGVAIADTGIDLAHADLAVGSASFNAFNINSSCQDGHSHGTHVAGIVAAINNTIDVIGVAPDATPYCVKILDDNGSGSDSTLMAGLDWIGGNANSVSPPIRVVNMSLGRAGTLNDNTALRASVYALYE VGIAVVVAAGNDSLKEVSQMVPATYPEVFAIASTTAVDGSNKKCKFFSGTIPADTASYFTTDGEFDPGTGIGITISAPGEDKEDVNRPCFAVSEGILSLKLGGGITRMSGTSMASPHVAGIVARMLQTASYSADASGVENIRSNLRFAADQKNIAPL, named serine protease TVG06.

[0050] Structural prediction results as follows Figure 1 As shown, the catalytic active center is D51H83S284 (shown in bright blue), which contains 4 Cys (C77-C111, C216-C261) forming two pairs of SS bonds (shown in yellow). Five pairs of salt bridges (shown in pink) were identified, which are likely the main reasons for the enzyme's high-temperature resistance.

[0051] Example 2: Expression and purification of serine protease The recombinant plasmid of the serine protease used in this invention was synthesized by Shanghai Diwin Biotechnology Co., Ltd. The gene encoding the serine protease TVG06 (SEQ ID NO.2) was inserted into the pET-28a vector plasmid to obtain the recombinant plasmid pET28a-TVG06. The nucleotide sequence of the gene is shown in SEQ ID NO.2.

[0052] SEQ ID NO.2: ATGCTCCACCACCGCCCGGGCCACGGGGGTGGCCCGGGCGGTGGTGGAGGAGGGGAGGAAGTTTCTTCTCAGATTACCCCCGAGGGGGTGAAGCGGGTCGGTCAGCCTACCGATTCCTCCAACGGGAGCGGAATTGGTGTTGCCATTGCCGATACCGGGATCGACCTGGCGCATGCCGACCTGGCGGTCGGCAGCGCCAGCTTCAACGCCTTTAACATCAATAGTAGTTGTCAGGATGGCCACAGCCACGGTACCCACGTGGCGGGTATCGTGGCGGCCATCAACAACACTATTGATGTTATAGGCGTGGCGCCGGACGCAACGCCGTATTGCGTTAAGATCCTGGACGACAACGGCTCAGGCTCAGACAGCACCCTGATGGCGGGGCTAGACTGGATTGGCGGGAACGCCAATTCAGTCTCGCCCCCCATCAGGGTGGTGAACATGAGCCTGGGCCGCGCCGGGACTCTTAACGACAATACGGCGTTGCGTGCGAGCGTCTACGCTCTCTATGAGGTGGGAATTGCGGTGGTGGTGGCCGCCGGGAACGACTCGCTGAAGGAGGTGTCACAGATGGTTCCAGCCACCTACCCCGAGGTGTTCGCGATAGCGAGCACCACGGCGGTAGATGGCTCGAACAAGAAGTGCAAATTCTTCAGCGGGACGATCCCGGCGGACACCGCCTCGTATTTCACCACCGATGGAGAGTTCGATCCCGGTACCGGCATTGGCATCACCATTTCCGCTCCCGGGGAGGATAAGGAGGACGTGAACCGACCCTGCTTCGCCGTCTCGGAGGGTATTCTGAGTCTGAAGCTTGGGGGCGGCATTACCCGGATGTCGGGGACGTCGATGGCGTCCCCGCACGTCGCGGGTATTGTCGCCCGCATGCTTCAGACTGCGTCGTATAGCGCAGATGCTTCGGGTGTGGAGAACATCCGAAGCAATCTGCGCTTTGCGGCGGACCAGAAGAATATTGCTCCCCTC。

[0053] The recombinant plasmid pET28a-TVG06 was transformed into wild-type Escherichia coli BW25113 and named BW1. The obtained BW1 strain was transferred to LB liquid medium containing 100 μg / mL kanamycin and cultured at 37 ℃ and 200 rpm for 8 h. The inoculum was then transferred at a rate of 1% (v / v) to a test tube containing 200 mL of LB liquid medium, and kanamycin was added to a final concentration of 100 μg / mL. The culture was incubated at 37°C and 200 rpm for 3 h until the OD600 nm reached approximately 0.6-0.8. Subsequently, IPTG was added to a final concentration as an inducer, and the culture was continued at 30°C and 200 rpm for 8 h. The cells were collected by centrifugation at 5000 rpm and 4°C for 10 min, and resuspended in 10 mM Tris-HCl (pH 8) to a final OD / mL concentration of 20 to obtain a recombinant *E. coli* cell suspension. This suspension was then sonicated (30% power, 30 min) and centrifuged (12000 rpm, 30 min) to obtain the supernatant. The supernatant was filtered through a 0.22 μm filter membrane; the filtrate contained TVG06 enzyme. SDS-PAGE results (…) Figure 2 The enzyme activity of the enzyme solution obtained in this example was measured to be 45300 U / L.

[0054] Example 3: Optimal Temperature and Thermal Stability Test of Serine Protease Determination of optimal temperature: Serine protease was subjected to enzymatic reaction in 10mM pH 8.5 Tris-HCl buffer at 20℃, 30℃, 37℃, 40℃, 50℃, 60℃, 65℃, 70℃, 80℃, 90℃ and 100℃ respectively. Temperature stability determination: Serine protease was incubated in 10 mM pH 8 Tris-HCl buffer at 37℃, 50℃ and 80℃ for 5 min, 10 min, 20 min, 30 min and 60 min respectively before enzymatic reaction. Untreated enzyme solution was used as control and enzyme activity was detected.

[0055] The optimal reaction temperature results for serine protease TVG06 are as follows: Figure 3 As shown, the relative enzyme activity is 100% at 50℃, and decreases with increasing or decreasing temperature, indicating that the optimal reaction temperature for the serine protease TVG06 is 50℃; the thermal stability results are as follows. Figure 4 As shown, the serine protease TVG06 maintained relatively stable activity after incubation at 37℃ and 50℃ for 60 min, and after incubation at 80℃ for 60 min, the activity remained above 80%. Figure 4 This indicates that the serine protease TVG06 has strong thermal stability.

[0056] Example 4: Optimal pH Determination of the optimal pH of the enzyme: The serine protease TVG06 was subjected to an enzymatic reaction at a reaction temperature of 50°C in a 10 mM Tris-HCl buffer with a pH of 3-11.

[0057] The optimal reaction pH for serine protease TVG06 is 8. Figure 5 ).

[0058] Example 5: Storage stability of serine protease The serine protease TVG06 was placed in stability chambers at 5℃, 25℃, 45℃, and 40℃ + UV, and tested after 1 week, 2 weeks, 4 weeks, and 8 weeks. After removal from the chambers, the samples were allowed to return to room temperature before enzyme activity assays were performed.

[0059] Table 1. Stability of serine protease activity

[0060] The results of the serine protease TVG06 enzyme activity and stability are shown in Table 1 and Figure 6 As shown, after being placed under conditions of 5℃, 25℃, 45℃, and 40℃+UV for 8 weeks, the activity was maintained at over 65%, indicating that the serine protease TVG06 has excellent enzyme activity stability.

[0061] Example 6: Composition and Efficacy Testing Table 2 Composition of the Composition (Polar Smart Enzyme)

[0062] The composition (trade name: Polar Smart Enzyme) was formulated according to the formulation in Table 3 and its performance was tested.

[0063] 1. Exfoliation effect Different sample groups were set up, all of which were prepared with water and adjusted to pH 5.5. The samples were: 1# 0.5% capryloyl salicylic acid; 2# 2% salicylic acid; 3# 30% lactic acid; 4# 10% lactic acid; 5# 10% lactobionic acid; 6# 2% succinic acid; 7# 10% mandelic acid; 8# 4% of the combination. All samples were prepared with water. A blank control group was set up with the following formulation: 0.2% superoxide dismutase, 0.13% papain, 25% glycerol, 1% 1,2-pentanediol, 2% 1,2-hexanediol, 1% tetrahydromethylpyrimidine carboxylic acid, and 70.67% water. 100 μL of PBS was added to each group. Take fresh pig skin from the back and drop 100 μL of the above sample into a 6 cm * 6 cm area. Process for 10 min, collect cells, lyse the cells, and detect the protein content using the BCA method to measure the relative amount of cells detached.

[0064] The results are as follows Figure 7 As shown, under the conditions set in this experiment (pH 5.5, treatment for 10 min), the exfoliation effects of different components were ranked as follows: 10% lactobionic acid > 30% lactic acid ≈ 4% composition (Polar Smart Enzyme). The 4% composition (Polar Smart Enzyme) achieved a good exfoliation effect with a low addition amount, namely, a change rate of exfoliated protein of 40.3%, which has high application value.

[0065] 2. Degradation effect on corneal plugs Keratin plug samples were collected from individuals at a beauty salon. Samples were treated with 2% and 4% composite solutions for 0, 1, 3, and 5 minutes, respectively, and photographed under a microscope after treatment. Both 2% and 4% composite solutions were prepared using 1M Tris-HCl buffer (pH 7.4). A blank control group was also established, consisting of 0.2% superoxide dismutase, 0.13% papain, 25% glycerol, 1% 1,2-pentanediol, 2% 1,2-hexanediol, 1% tetrahydromethylpyrimidinecarboxylic acid, and 70.67% water. A positive control group consisted of 2% salicylic acid, prepared using 1M Tris-HCl buffer (pH 7.4).

[0066] Experimental results are as follows Figure 8 As shown, both the 2% and 4% compositions exhibited the effect of degrading keratin plugs, with the effect increasing with increasing concentration. At the 4% concentration, the edges of the keratin plugs showed signs of scattering. Furthermore, the composition's effect on degrading keratin plugs was stronger than that of acid, possibly due to the degradation of proteins in the keratin plug tissue by the protease.

[0067] 3. Stimulation test (1) Reagent preparation: Weigh 0.9 g NaCl and dilute to 100 mL of ultrapure water to prepare a 0.9% NaCl solution; weigh 0.0400 g NaOH and dilute to 10 mL of ultrapure water to prepare a 0.1 M NaOH solution; weigh 4 g of the composition (Polar Smart Enzyme) and add to 96 g of pure water, and adjust the pH to 5.5 using 10% citric acid solution.

[0068]

[0069] Table 3 Experimental Design (2) CAM preparation: Purchase 8-day-old chicken embryos and incubate them until they are 9 days old. Inspect and discard defective chicken embryos. Mark the location of the air cell on the surface of the normal chicken embryo eggshell, peel off part of the eggshell to expose the white egg membrane; carefully remove the inner membrane with tweezers, ensuring that the vascular membrane is not damaged.

[0070] (3) The experiment was conducted using the stimulus rating method, with at least 6 chicken embryos in each group. 0.3 mL of the test substance was directly added to the CAM surface, and the CAM reaction was observed. The time of occurrence of each toxic effect within 5 min was recorded, accurate to the second, including three reactions: hemorrhage, coagulation and angiogenesis. The degree of the reaction was also recorded.

[0071] (4) Data processing: The reaction time method was used for the experiment, and the stimulus score (IS) was calculated using Formula 1. The results were rounded to two decimal places. Formula 1 sec H (bleeding time) – the average time it takes for bleeding to begin to occur as observed on the CAM membrane, in seconds (s). sec L (vascular dissolution time) – the average time, in seconds (s), observed on the CAM membrane to begin vascular dissolution. sec C (clotting time) – the average time it takes for clotting to begin to occur as observed on a CAM membrane, measured in seconds (s). The irritancy of the test substance is classified according to the calculated IS value as shown in Table 4.

[0072] Table 4 Evaluation of Stimulus Rating Method Results

[0073] The results are shown in Table 5 and Figure 9 As shown in the figure, according to the judgment criteria of SN / T 2329-2009, under the experimental conditions, the IS score of the 4% Polar Smart Enzyme (m / m) test sample was 1.03, which is considered mildly irritating. The figure also shows that there was no significant difference between the sample group and the negative control group.

[0074] Table 5 Scoring Results

[0075] Example 7: High Temperature Stability Comparison Serine protease TVG06 and papain (Nanning Pangbo Biotechnology Co., Ltd.), a commonly used exfoliating enzyme, were placed in a stability incubator at 45°C. Samples were taken out after 1, 2, 4, and 8 weeks for testing. After removal, the samples were allowed to return to room temperature before enzyme activity assays. Results are as follows: Figure 10 As shown, the serine protease TVG06 exhibits a degradation rate of less than 15% after 4 weeks at 45℃, and its activity remains above 65% after 8 weeks. In contrast, papain shows a 50% activity loss after 4 weeks at 45℃, and an 80% activity loss after 8 weeks.

[0076] Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Anyone skilled in the art can make various modifications and alterations without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be determined by the claims.

Claims

1. A serine protease TVG06, characterized in that, The amino acid sequence of the serine protease TVG06 is shown in SEQ ID NO.

1.

2. The gene encoding the serine protease TVG06 of claim 1.

3. A recombinant plasmid carrying the gene described in claim 2.

4. The recombinant plasmid according to claim 3, characterized in that, The expression vector for the recombinant plasmid is pET-28a plasmid.

5. Recombinant cells expressing the serine protease TVG06 of claim 1, or containing the gene of claim 2, or transformed with the recombinant plasmid of any one of claims 3 to 4.

6. The use of the gene of claim 2, or the recombinant plasmid of claim 3 or 4, or the recombinant cell of claim 5 in the preparation of serine protease TVG06 or products containing serine protease TVG06.

7. A composition, characterized in that, The composition contains the serine protease TVG06 as described in claim 1.

8. The composition according to claim 7, characterized in that, The composition also contains superoxide dismutase and papain.

9. The composition according to claim 8, characterized in that, The composition also contains glycerol, 1,2-pentanediol, 1,2-hexanediol, and tetrahydromethylpyrimidine carboxylic acid.

10. The use of the serine protease TVG06 of claim 1 or the composition of any one of claims 7-8 in the preparation of exfoliating daily chemical products.

Images