Tooth whitening preparation based on compound enzyme synergistic effect and application thereof

Abstract

The invention relates to a tooth whitening preparation based on a compound enzyme synergistic effect and application thereof, and belongs to the field of tooth whitening preparations, the tooth whitening preparation comprises the following substances by weight: 15-20 parts of esterase, 15-25 parts of lipase, 10-15 parts of papain, 10-15 parts of dextranase, and 25-50 parts of maltodextrin. Based on the characteristic that the oral pigment has a'protein-lipid-polyphenol 'composite structure, a quaternary synergistic effect mechanism is provided, protein, lipid-soluble and polyphenol pigments can be removed, and safe and efficient whitening is achieved in the oral environment with the pH of 6.0-7.5; the lipase, the esterase and the papain cooperate with one another, so that pigment matrixes can be decomposed; the dextranase directly acts to decompose water-insoluble glucan, carbohydrate pigments are reduced, decomposition of dental plaque and pigments thereof is accelerated, the lipase, the esterase, the papain, the dextranase and the laccase do not damage teeth, and teeth are not damaged while tooth whitening is achieved.

Description

Technical Field

[0001] This invention belongs to the field of teeth whitening preparations, specifically a teeth whitening preparation based on the synergistic effect of compound enzymes and its application. Background Technology

[0002] In today's society, the demand for teeth whitening is increasing. Traditional teeth whitening methods often have drawbacks such as short-lasting effects and damage to teeth. Currently, the most common teeth whitening procedure involves adding teeth whitening agents to toothpaste. Common additives include chemical bleaching agents and enzyme preparations. However, chemical bleaching agents (such as hydrogen peroxide) can easily cause dentin sensitivity and gum irritation, while enzyme preparations are mostly single enzymes (such as lysozyme) and cannot remove fat-soluble pigments. Furthermore, physical abrasive products can easily damage tooth enamel. Therefore, there is a need for a teeth whitening agent that can effectively whiten teeth without damaging them. Summary of the Invention

[0003] This invention provides a teeth whitening preparation based on the synergistic effect of compound enzymes and its application, in order to overcome the deficiencies in the prior art.

[0004] This invention is achieved through the following technical solution: A teeth whitening preparation based on the synergistic effect of complex enzymes comprises the following substances in parts by weight: 15-20 parts esterase, 15-25 parts lipase, 10-15 parts papain, 10-15 parts dextranase, and 25-50 parts maltodextrin.

[0005] The teeth whitening preparation based on the synergistic effect of complex enzymes as described above, wherein the esterase has an enzyme activity ≥500U / g; the lipase has an enzyme activity ≥1000U / g; the papain has an enzyme activity ≥10000U / g; and the dextranase has an enzyme activity ≥10000U / g.

[0006] The teeth whitening preparation based on the synergistic effect of complex enzymes as described above comprises the following substances in parts by weight: 15-20 parts esterase, 15-25 parts lipase, 10-15 parts papain, 10-15 parts dextranase, 10-15 parts laccase, and 25-50 parts maltodextrin.

[0007] The teeth whitening preparation based on the synergistic effect of complex enzymes as described above, wherein the esterase has an enzyme activity ≥500U / g; the lipase has an enzyme activity ≥1000U / g; the papain has an enzyme activity ≥10000U / g; the dextranase has an enzyme activity ≥10000U / g; and the laccase has an enzyme activity ≥1000U / g.

[0008] The teeth whitening agent based on the synergistic effect of a complex enzyme, as described above, includes the following steps in its preparation method of the esterase: Step 1: Activation of Aspergillus niger strain The well-preserved Aspergillus niger slant culture was inoculated into slant culture medium and incubated at 30℃ for 42h to activate the culture. This activation was repeated 3 times. The composition of the Aspergillus niger slant culture medium is as follows: 200g potato (boiled juice), 20g glucose, 20g agar, 15g traditional Chinese medicine powder, 1000mL distilled water, pH 5.8, sterilized at 121℃ for 20min. Step 2: Liquid seed culture of Aspergillus niger Step ①: Primary seed culture: Wash the activated Aspergillus niger slant culture with sterile water to obtain spores, inoculate into a 500 ml shake flask, add 100 ml of liquid seed culture medium, and culture at 30℃ on a shaker at 100 rpm for 42 h. Step 2: Secondary seed culture: Inoculate 10% of the primary seeds into a 500 ml secondary seed shake flask, and culture under the same conditions as the primary seeds; Step ③: Tertiary seed culture: Inoculate the secondary seeds at an 8% inoculation rate into a 5000 ml tertiary seed shake flask, add 1000 ml of liquid culture medium, and culture at 30℃ on a shaker at 100 rpm for 42 h. Step 4: Primary Seed Tank Cultivation: Inoculate the tertiary seeds at an 8% inoculation rate into a primary seed tank with a total volume of 150L. Fill the fermentation medium with 100L, control the pH value to 6, the cultivation temperature to 30℃, the stirring speed to 300rpm, the aeration rate (V / V) to 1:1, the cultivation time to 42h, and the dissolved oxygen to 20%. The primary, secondary, and tertiary seed culture media for Aspergillus niger consist of: 200g potato (boiled juice), 20g glucose, 18g traditional Chinese medicine powder, 20g trehalose, 1000mL distilled water, pH 5.5, sterilized at 121℃ for 20min. The weight proportions of the primary seed culture medium for Aspergillus niger are as follows: 55g corn flour, 20g soybean flour, 12g wheat bran, 20g trehalose, 2g ammonium sulfate, 2g dipotassium hydrogen phosphate, 2g potassium dihydrogen phosphate, 1000mL purified water, pH 6, sterilized at 121℃ for 20min. The concentration of Aspergillus niger fermentation broth in the primary seed tank is 7.5 x 10⁻⁶ cells / mL. 8 pcs / ml; Step 3: Microbial fermentation The secondary seed tank's symbiotic liquid seeds were inoculated into the fermenter at a 6% inoculum, and cultured at 30℃ with a stirring speed of 350 rpm and an aeration rate (V / V) of 1:2 for 12 hours. The temperature was then slowly lowered to 12℃ at a rate of 2℃ / h and maintained at this temperature for 18 hours. The temperature was then further slowly lowered to 4℃ at a rate of 2℃ / h, at which point the secondary seed tank's symbiotic liquid seeds were added back into the fermenter at a 4% inoculum, and cultured at this temperature for 25 hours. Finally, the temperature was slowly increased to 10-15℃ at a rate of 2℃ / h and cultured at this temperature for 15-20 hours. The temperature was then further slowly increased to 33℃ at a rate of 2℃ / h and cultured at this temperature for 18 hours. Dissolved oxygen control: Dissolved oxygen is controlled at 20% by adjusting the stirring speed and ventilation volume; pH control: The pH value is maintained at 5.2 during fermentation by supplementing with ammonia or dilute phosphoric acid. Feeding control: When the reducing sugar content in the fermentation broth drops to 3mg / ml-8mg / ml, start adding feed medium, and the amount of feed should be enough to maintain the reducing sugar content in the fermentation broth at 2mg / ml-5mg / ml; Standard for placement in the tank: 70% of the bacterial cells are aged and autolyzed, and enzyme activity increases slowly; The fermentation medium consists of: 100g maltodextrin, 55g corn flour, 20g soybean flour, 12g wheat bran, 35g trehalose, 6g yeast powder, 3g corn steep liquor, 2g ammonium sulfate, 2g dipotassium hydrogen phosphate, 2g potassium dihydrogen phosphate, 1000mL purified water, pH 5.2, sterilized at 121℃ for 20min. The feed culture medium has the following weight percentages: maltodextrin 25%, corn flour 15%, soybean flour 20%, traditional Chinese medicine powder 8%, and purified water to make up any shortfall. The pH value is 5.2, and the medium is sterilized at 121℃ for 40 minutes. Step 4: The fermentation broth is filtered through plate and frame filter, finely filtered through filter paper, separated and purified by chromatographic column, finely filtered through filter paper, and spray-dried to obtain solid esterase.

[0009] The teeth whitening agent based on the synergistic effect of complex enzymes, as described above, includes the following steps in its preparation method of the lipase: Step 1: Activation of Aspergillus niger strain The well-preserved Aspergillus niger slant culture was inoculated into slant culture medium and incubated at 30℃ for 42h to activate the culture. This activation was repeated 3 times. The composition of the Aspergillus niger slant culture medium is as follows: 200g potato (boiled juice), 20g glucose, 20g agar, 15g traditional Chinese medicine powder, 1000mL distilled water, pH 5.8, sterilized at 121℃ for 20min. Step 2: Liquid seed culture of Aspergillus niger Step ①: Primary seed culture: Wash the activated Aspergillus niger slant culture with sterile water to obtain spores, inoculate into a 500 ml shake flask, add 100 ml of liquid seed culture medium, and culture at 30℃ on a shaker at 100 rpm for 42 h. Step 2: Secondary seed culture: Inoculate 10% of the primary seeds into a 500 ml secondary seed shake flask, and culture under the same conditions as the primary seeds; Step ③: Tertiary seed culture: Inoculate the secondary seeds at an 8% inoculation rate into a 5000 ml tertiary seed shake flask, add 1000 ml of liquid culture medium, and culture at 30℃ on a shaker at 100 rpm for 42 h. Step 4: Primary Seed Tank Cultivation: Inoculate the tertiary seeds at an 8% inoculation rate into a primary seed tank with a total volume of 150L. Fill the fermentation medium with 100L, control the pH value to 6, the cultivation temperature to 30℃, the stirring speed to 300rpm, the aeration rate (V / V) to 1:1, the cultivation time to 42h, and the dissolved oxygen to 20%. The primary, secondary, and tertiary seed culture media for Aspergillus niger consist of: 200g potato (boiled juice), 20g glucose, 18g traditional Chinese medicine powder, 20g trehalose, 1000mL distilled water, pH 5.5, sterilized at 121℃ for 20min. The weight proportions of the primary seed culture medium for Aspergillus niger are as follows: 55g corn flour, 20g soybean flour, 12g wheat bran, 20g trehalose, 2g ammonium sulfate, 2g dipotassium hydrogen phosphate, 2g potassium dihydrogen phosphate, 1000mL purified water, pH 6, sterilized at 121℃ for 20min. The concentration of Aspergillus niger fermentation broth in the primary seed tank is 7.5 x 10⁻⁶ cells / mL. 8 pcs / ml; Step 3: Microbial fermentation The secondary seed tank's symbiotic liquid seeds were inoculated into the fermenter at a 6% inoculum, and cultured at 30℃ with a stirring speed of 350 rpm and an aeration rate (V / V) of 1:2 for 12 hours. The temperature was then slowly lowered to 12℃ at a rate of 2℃ / h and maintained at this temperature for 18 hours. The temperature was then further slowly lowered to 4℃ at a rate of 2℃ / h, at which point the secondary seed tank's symbiotic liquid seeds were added back into the fermenter at a 4% inoculum, and cultured at this temperature for 25 hours. Finally, the temperature was slowly increased to 10-15℃ at a rate of 2℃ / h and cultured at this temperature for 15-20 hours. The temperature was then further slowly increased to 33℃ at a rate of 2℃ / h and cultured at this temperature for 18 hours. Dissolved oxygen control: Dissolved oxygen is controlled at 20% by adjusting the stirring speed and ventilation volume; pH control: The pH value is maintained at 5.2 during fermentation by supplementing with ammonia or dilute phosphoric acid. Feeding control: When the reducing sugar content in the fermentation broth drops to 3mg / ml-8mg / ml, start adding feed medium, and the amount of feed should be enough to maintain the reducing sugar content in the fermentation broth at 2mg / ml-5mg / ml; Standard for placement in the tank: 70% of the bacterial cells are aged and autolyzed, and enzyme activity increases slowly; The fermentation medium consists of: 100g maltodextrin, 55g corn flour, 20g soybean flour, 12g wheat bran, 35g trehalose, 6g yeast powder, 3g corn steep liquor, 2g ammonium sulfate, 2g dipotassium hydrogen phosphate, 2g potassium dihydrogen phosphate, 1000mL purified water, pH 5.2, sterilized at 121℃ for 20min. The feed culture medium has the following weight percentages: maltodextrin 25%, corn flour 15%, soybean flour 20%, traditional Chinese medicine powder 8%, and purified water to make up any shortfall. The pH value is 5.2, and the medium is sterilized at 121℃ for 40 minutes. Step 4: The fermentation broth is filtered through plate and frame filter, finely filtered through filter paper, separated and purified by chromatographic column, finely filtered through filter paper, and spray-dried to obtain solid lipase.

[0010] The teeth whitening agent based on the synergistic effect of a complex enzyme, as described above, includes the following steps in its preparation method of dextranase: Step 1: Activation of Amorphous Chaetomium strains The well-preserved amorphous Chaetomium slant culture was inoculated into the slant culture medium and incubated at 30℃ for 48h to activate the culture. This activation was repeated 3 times. The composition of the amorphous Chaetomium slant culture medium is as follows: 200g potato, 20g glucose, 20g agar, 1000mL distilled water, pH 5.8, sterilized at 121℃ for 20min. Step 2: Liquid seed culture of amorphous Chaetomium Step ①: Primary seed culture: The activated amorphous chaetotrichum slant culture was inoculated into a 500 ml shake flask with sterile aquatic cells, and 100 ml of liquid seed culture medium was added. The culture was carried out at 30℃ and shaken at 160 rpm for 48 h. Step 2: Secondary seed culture: Inoculate 10% of the primary seeds into a 500 ml secondary seed shake flask, and culture under the same conditions as the primary seeds; Step ③: Tertiary seed culture: Inoculate the secondary seeds at an 8% inoculation rate into a 5000 ml tertiary seed shake flask, add 1000 ml of liquid culture medium, and culture at 30℃ on a shaker at 160 rpm for 48 h. Step 4: Primary Seed Tank Cultivation: Inoculate the tertiary seeds at an 8% inoculation rate into a primary seed tank with a total volume of 150L. Fill the fermentation medium with 100L, control the pH value to 6, the cultivation temperature to 30℃, the stirring speed to 200rpm, the aeration rate to 1:1, the cultivation time to 40h, and the dissolved oxygen to 20%. The primary, secondary, and tertiary seed culture media for the amorphous chaetotrichum are composed of: 200g potato, 20g glucose, 20g trehalose, 1000mL distilled water, pH 5.5, sterilized at 121℃ for 20min. The weight proportions of the amorphous chaetotrichum primary seed tank culture medium are as follows: 55g corn flour, 20g soybean flour, 12g wheat bran, 20g trehalose, 2g ammonium sulfate, 2g dipotassium hydrogen phosphate, 2g potassium dihydrogen phosphate, 1000mL purified water, pH 6, sterilized at 121℃ for 20min. The concentration of the amorphous Chaetomium primary seed tank fermentation broth is 8 x 10⁻⁶ cells / mL. 8 pcs / ml; Step 3: Microbial fermentation The primary seed was inoculated at a rate of 10% into a 500 mL shake flask containing secondary seed culture medium (100 mL). The mixture was incubated at 30°C on a shaker at 160 rpm for 48 hours. After incubation, the secondary seed tank's alternating liquid seed was inoculated into a fermenter at a rate of 6%, incubated at 30°C with a stirring speed of 300 rpm and an aeration ratio of 1:2 for 12 hours. The temperature was then slowly lowered to 12°C at a rate of 2°C / h and maintained at this temperature for 18 hours. The temperature was then further slowly lowered to 4°C at a rate of 2°C / h, at which point the secondary seed tank's alternating liquid seed was added back into the fermenter at a rate of 4%, and the mixture was maintained at this temperature for 25 hours. Finally, the temperature was slowly increased to 10-15°C at a rate of 2°C / h and maintained at this temperature for 15-20 hours. The temperature was then slowly increased to 33°C at a rate of 2°C / h and maintained at this temperature for 18 hours. Dissolved oxygen control: Dissolved oxygen is controlled at 30% by adjusting the stirring speed and ventilation volume; pH control: The pH value is maintained at 5.2 during fermentation by supplementing with ammonia or dilute phosphoric acid. Feeding control: When the reducing sugar content in the fermentation broth drops to 5mg / ml-10mg / ml, start adding feed medium, and the amount of feed should be enough to maintain the reducing sugar content in the fermentation broth at 3mg / ml-6mg / ml; Standard for placement in the tank: 70% of the bacterial cells are aged and autolyzed, protein content does not increase, and enzyme activity increases slowly; The fermentation medium consists of: 100g maltodextrin, 55g corn flour, 20g soybean flour, 12g wheat bran, 35g trehalose, 6g yeast powder, 3g corn steep liquor, 2g ammonium sulfate, 2g dipotassium hydrogen phosphate, 2g potassium dihydrogen phosphate, 1000mL purified water, pH 5.2, sterilized at 121℃ for 20min. The feed culture medium has the following weight percentages: maltodextrin 25%, corn flour 15%, soybean flour 20%, with purified water to make up any shortfall, pH 5.2, and sterilized at 121℃ for 40 minutes. Step 4: The fermentation broth is filtered through plate and frame filter, fine filtration through filter paper, separation and purification by chromatographic column, fine filtration through filter paper, and spray drying to obtain solid dextranase.

[0011] The teeth whitening agent based on the synergistic effect of a complex enzyme, as described above, includes the following steps in its preparation method of the laccase: Step 1: Activation of Aspergillus oryzae strain The well-preserved Aspergillus oryzae slant culture was inoculated into slant culture medium and incubated at 28°C for 24 hours to activate the culture. This activation was repeated 3 times. The Aspergillus oryzae slant culture medium consists of: 200g potato, 20g glucose, 20g agar, 1000mL distilled water, pH 5.8, sterilized at 121℃ for 20min. Step 2: Liquid seed culture of Aspergillus oryzae Step ①: Primary seed culture: Wash the activated Aspergillus oryzae slant culture with sterile water to obtain spores, inoculate into a 500 ml shake flask, add 100 ml of liquid seed culture medium, and culture at 28℃ on a shaker at 120 rpm for 32 h. Step 2: Secondary seed culture: Inoculate 10% of the primary seeds into a 500 ml secondary seed shake flask, and culture under the same conditions as the primary seeds; Step ③: Tertiary seed culture: Inoculate the secondary seeds at an 8% inoculation rate into a 5000 ml tertiary seed shake flask, add 1000 ml of liquid culture medium, and culture at 30℃ on a shaker at 120 rpm for 32 h. Step 4: Primary Seed Tank Cultivation: Inoculate the tertiary seeds at an 8% inoculation rate into a primary seed tank with a total volume of 150L. Fill the fermentation medium with 100L, control the pH value to 6, the cultivation temperature to 30℃, the stirring speed to 200rpm, the aeration rate to 1:1.5, the cultivation time to 32h, and the dissolved oxygen to 25%. The primary, secondary, and tertiary seed culture media for Aspergillus oryzae are composed of: 200g potato, 20g glucose, 20g trehalose, 1000mL distilled water, pH 5.5, sterilized at 121℃ for 20min. The weight proportions of the Aspergillus oryzae primary seed culture medium are as follows: 55g corn flour, 20g soybean flour, 12g wheat bran, 20g trehalose, 2g ammonium sulfate, 2g dipotassium hydrogen phosphate, 2g potassium dihydrogen phosphate, 1000mL purified water, pH 6, sterilized at 121℃ for 20min. The Aspergillus oryzae primary seed tank fermentation broth cell concentration is 6 x 10⁻⁶. 8 pcs / ml; Step 3: Microbial fermentation The primary seed was inoculated at a rate of 10% into a 500 mL shake flask containing secondary seed culture medium (100 mL). The flask was incubated at 30°C for 32 h on a shaker at 120 rpm. After incubation, the secondary seed tank's alternating liquid seed was inoculated into a fermenter at a rate of 6%, incubated at 28°C with a stirring speed of 300 rpm and an aeration ratio of 1:2 for 12 h. The temperature was then slowly lowered to 12°C at a rate of 2°C / h and incubated at this temperature for 18 h. The temperature was then further slowly lowered to 4°C at a rate of 2°C / h, at which point the secondary seed tank's alternating liquid seed was added back into the fermenter at a rate of 4%, and incubated at this temperature for 25 h. Finally, the temperature was slowly increased to 10-15°C at a rate of 2°C / h and incubated at this temperature for 15-20 h. The temperature was then slowly increased to 28°C at a rate of 2°C / h and incubated at this temperature for 20 h. Dissolved oxygen control: Dissolved oxygen is controlled at 25% by adjusting the stirring speed and ventilation volume; pH control: The pH value is maintained at 5.5 during fermentation by adding ammonia or dilute phosphoric acid. Feeding control: When the reducing sugar content in the fermentation broth drops to 5mg / ml-10mg / ml, start adding feed medium, and the amount of feed should be enough to maintain the reducing sugar content in the fermentation broth at 5mg / ml-8mg / ml; Standard for placement in the tank: 70% of the bacterial cells are aged and autolyzed, protein does not increase, and enzyme activity increases slowly; The fermentation medium consists of: 100g maltodextrin, 55g corn flour, 20g soybean flour, 12g wheat bran, 35g trehalose, 6g yeast powder, 3g corn steep liquor, 2g ammonium sulfate, 2g dipotassium hydrogen phosphate, 2g potassium dihydrogen phosphate, 1000mL purified water, pH 5.2, sterilized at 121℃ for 20min. The feed culture medium has the following weight percentages: maltodextrin 25%, corn flour 15%, soybean flour 20%, traditional Chinese medicine powder 8%, and purified water to make up any shortfall. The pH value is 5.2, and the medium is sterilized at 121℃ for 40 minutes. Step 4: The fermentation broth is filtered through plate and frame filter, finely filtered through filter paper, separated and purified by chromatographic column, finely filtered through filter paper, and spray-dried to obtain solid laccase.

[0012] Application of a teeth whitening agent based on the synergistic effect of a complex enzyme, wherein the teeth whitening agent based on the synergistic effect of the complex enzyme is added to toothpaste for use.

[0013] The application of a teeth whitening agent based on the synergistic effect of a compound enzyme as described above, wherein the amount of the teeth whitening agent with the synergistic effect of the compound enzyme added to the toothpaste is 3g / kg.

[0014] The advantages of this invention are as follows: Based on the characteristic of oral pigments having a "protein-lipid-polyphenol" complex structure, this invention proposes a quaternary synergistic mechanism that can remove protein-based, lipid-soluble, and polyphenolic pigments, achieving safe and efficient whitening in an oral environment of pH 6.0-7.5; In this invention, lipase, esterase, and papain work synergistically to decompose the pigment matrix; In this invention, dextranase directly decomposes water-insoluble dextran (dextran), reducing sugar pigments and accelerating the decomposition of dental plaque and its pigments. Moreover, lipase, esterase, papain, dextranase, and laccase do not damage teeth, thus achieving teeth whitening without damaging teeth. Attached Figure Description

[0015] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0016] Figure 1 This is a schematic diagram of the application of tea-stained teeth model to Example 1, the control group, and the blank group of the present invention (left image is blank group, middle image is control group, right image is Example 1); Figure 2 This is a schematic diagram of the application of coffee-stained tooth models to Example 1, the control group, and the blank group of the present invention (left image is blank group, middle image is control group, right image is Example 1). Figure 3 This is a schematic diagram of the application of red wine-stained tooth model to Example 1, the control group, and the blank group of the present invention (left image is blank group, middle image is control group, right image is Example 1). Figure 4 This is a schematic diagram of the application of tea-stained tooth model to Embodiment 2, the control group, and the blank group of the present invention (left image is blank group, middle image is control group, right image is Embodiment 2); Figure 5 This is a schematic diagram of the application of coffee-stained tooth models to Embodiment 2, the control group, and the blank group of the present invention (left image is blank group, middle image is control group, right image is Embodiment 2); Figure 6 This is a schematic diagram of the application of red wine-stained tooth model to Embodiment 2, the control group, and the blank group of the present invention (left image is blank group, middle image is control group, right image is Embodiment 2); Figure 7 This is a schematic diagram of the application of tea-stained teeth model to Example 3, the control group, and the blank group of the present invention (left image is blank group, middle image is control group, right image is Example 3); Figure 8This is a schematic diagram of the application of coffee-stained tooth models to Example 3, the control group, and the blank group of the present invention (left image is blank group, middle image is control group, right image is Example 3). Figure 9 This is a schematic diagram of the application of the present invention in Example 3, the control group, and the blank group to a tooth model stained with red wine (left image is blank group, middle image is control group, right image is Example 3). Figure 10 This is a schematic diagram of the application of tea-stained teeth model to Example 4, the control group, and the blank group of the present invention (left image is blank group, middle image is control group, right image is Example 4). Figure 11 This is a schematic diagram of the application of coffee-stained tooth models to Example 4, the control group, and the blank group of the present invention (left image is blank group, middle image is control group, right image is Example 4). Figure 12 This is a schematic diagram of the application of red wine-stained tooth model to Example 4, the control group, and the blank group of the present invention (left image is blank group, middle image is control group, right image is Example 4). Figure 13 This is a schematic diagram of the application of tea-stained tooth model to Example 5, the control group, and the blank group of the present invention (left image is blank group, middle image is control group, right image is Example 5); Figure 14 This is a schematic diagram of the application of coffee-stained tooth models to Example 5, the control group, and the blank group of the present invention (left image is blank group, middle image is control group, right image is Example 5); Figure 15 This is a schematic diagram of the application of red wine-stained tooth model to Example 5, the control group, and the blank group of the present invention (left image is blank group, middle image is control group, right image is Example 5); Figure 16 This is a schematic diagram of the application of tea-stained teeth model to Example 6, the control group, and the blank group of the present invention (left image is blank group, middle image is control group, right image is Example 6). Figure 17 This is a schematic diagram of the application of coffee-stained tooth models to Example 6, the control group, and the blank group of the present invention (left image is blank group, middle image is control group, right image is Example 6). Figure 18 This is a schematic diagram of the application of red wine-stained tooth model to Example 6, the control group, and the blank group of the present invention (left image is blank group, middle image is control group, right image is Example 6). Detailed Implementation

[0017] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0018] Example 1 Step 1: Weigh out 15 parts of esterase (enzyme activity ≥ 500 U / g), 15 parts of lipase (enzyme activity ≥ 1000 U / g), 10 parts of papain lipase (enzyme activity ≥ 10000 U / g), 10 parts of dextran lipase (enzyme activity ≥ 1000 U / g), and 25 parts of maltodextrin according to the specified ratio. Step 2: Crush the raw materials weighed in Step 1 and mix them evenly to obtain a teeth whitening preparation with synergistic effects of compound enzymes; Step 3: Add the teeth whitening agent with synergistic effect of the compound enzyme obtained in Step 2 to the toothpaste base at an addition rate of 3g / kg and stir evenly to obtain the finished toothpaste. The method for preparing the esterase includes the following steps: Step 1: Activation of Aspergillus niger strain The well-preserved Aspergillus niger slant culture was inoculated into slant culture medium and incubated at 30℃ for 42h to activate the culture. This activation was repeated 3 times. The composition of the Aspergillus niger slant culture medium is as follows: 200g potato, 20g glucose, 20g agar, 15g traditional Chinese medicine powder, 1000mL distilled water, pH 5.8, sterilized at 121℃ for 20min. Step 2: Liquid seed culture of Aspergillus niger Step ①: Primary seed culture: Wash the activated Aspergillus niger slant culture with sterile water to obtain spores, inoculate into a 500 ml shake flask, add 100 ml of liquid seed culture medium, and culture at 30℃ on a shaker at 100 rpm for 42 h. Step 2: Secondary seed culture: Inoculate 10% of the primary seeds into a 500 ml secondary seed shake flask, and culture under the same conditions as the primary seeds; Step ③: Tertiary seed culture: Inoculate the secondary seeds at an 8% inoculation rate into a 5000 ml tertiary seed shake flask, add 1000 ml of liquid culture medium, and culture at 30℃ on a shaker at 100 rpm for 42 h. Step 4: Primary Seed Tank Cultivation: Inoculate the tertiary seeds at an 8% inoculation rate into a primary seed tank with a total volume of 150L. Fill the fermentation medium with 100L, control the pH value to 6, the cultivation temperature to 30℃, the stirring speed to 300rpm, the aeration rate to 1:1, the cultivation time to 42h, and the dissolved oxygen to 20%. The primary, secondary, and tertiary seed culture media for Aspergillus niger consist of: 200g potato, 20g glucose, 18g traditional Chinese medicine powder, 20g trehalose, 1000mL distilled water, pH 5.5, sterilized at 121℃ for 20min. The weight proportions of the primary seed culture medium for Aspergillus niger are as follows: 55g corn flour, 20g soybean flour, 12g wheat bran, 20g trehalose, 2g ammonium sulfate, 2g dipotassium hydrogen phosphate, 2g potassium dihydrogen phosphate, 1000mL purified water, pH 6, sterilized at 121℃ for 20min. The concentration of Aspergillus niger fermentation broth in the primary seed tank is 7.5 x 10⁻⁶ cells / mL. 8 pcs / ml; Step 3: Microbial fermentation The primary seed was inoculated at a rate of 10% into a 500 mL shake flask containing secondary seed culture medium (100 mL). The mixture was incubated at 30°C on a shaker at 100 rpm for 42 h. After incubation, the secondary seed tank's alternating liquid seed was inoculated into a fermenter at a rate of 6%, incubated at 30°C with a stirring speed of 350 rpm and an aeration ratio of 1:2 for 12 h. The temperature was then slowly lowered to 12°C at a rate of 2°C / h and incubated at this temperature for 18 h. The temperature was then further slowly lowered to 4°C, at which point the secondary seed tank's alternating liquid seed was added back into the fermenter at a rate of 4%, and incubated at this temperature for 25 h. Finally, the temperature was slowly increased to 10-15°C at a rate of 2°C / h and incubated at this temperature for 15-20 h. The temperature was then slowly increased to 33°C at a rate of 2°C / h and incubated at this temperature for 18 h. Dissolved oxygen control: Dissolved oxygen is controlled at 20% by adjusting the stirring speed and ventilation volume; pH control: The pH value is maintained at 5.2 during fermentation by supplementing with ammonia or dilute phosphoric acid. Feeding control: When the reducing sugar content in the fermentation broth drops to 3mg / ml-8mg / ml, start adding feed medium, and the amount of feed should be enough to maintain the reducing sugar content in the fermentation broth at 2mg / ml-5mg / ml; Standard for placement in the tank: 70% of the bacterial cells are aged and autolyzed, and enzyme activity increases slowly; The fermentation medium consists of: 100g maltodextrin, 55g corn flour, 20g soybean flour, 12g wheat bran, 35g trehalose, 6g yeast powder, 3g corn steep liquor, 2g ammonium sulfate, 2g dipotassium hydrogen phosphate, 2g potassium dihydrogen phosphate, 1000mL purified water, pH 5.2, sterilized at 121℃ for 20min. The feed culture medium has the following weight percentages: maltodextrin 25%, corn flour 15%, soybean flour 20%, traditional Chinese medicine powder 8%, and purified water to make up any shortfall. The pH value is 5.2, and the medium is sterilized at 121℃ for 40 minutes. Step 4: The fermentation broth is filtered through plate and frame filter, finely filtered through filter paper, separated and purified by chromatographic column, finely filtered through filter paper, and spray-dried to obtain solid esterase; The method for preparing the lipase includes the following steps: Step 1: Activation of Aspergillus niger strain The well-preserved Aspergillus niger slant culture was inoculated into slant culture medium and incubated at 30℃ for 42h to activate the culture. This activation was repeated 3 times. The composition of the Aspergillus niger slant culture medium is as follows: 200g potato, 20g glucose, 20g agar, 15g traditional Chinese medicine powder, 1000mL distilled water, pH 5.8, sterilized at 121℃ for 20min. Step 2: Liquid seed culture of Aspergillus niger Step ①: Primary seed culture: Wash the activated Aspergillus niger slant culture with sterile water to obtain spores, inoculate into a 500 ml shake flask, add 100 ml of liquid seed culture medium, and culture at 30℃ on a shaker at 100 rpm for 42 h. Step 2: Secondary seed culture: Inoculate 10% of the primary seeds into a 500 ml secondary seed shake flask, and culture under the same conditions as the primary seeds; Step ③: Tertiary seed culture: Inoculate the secondary seeds at an 8% inoculation rate into a 5000 ml tertiary seed shake flask, add 1000 ml of liquid culture medium, and culture at 30℃ on a shaker at 100 rpm for 42 h. Step 4: Primary Seed Tank Cultivation: Inoculate the tertiary seeds at an 8% inoculation rate into a primary seed tank with a total volume of 150L. Fill the fermentation medium with 100L, control the pH value to 6, the cultivation temperature to 30℃, the stirring speed to 300rpm, the aeration rate to 1:1, the cultivation time to 42h, and the dissolved oxygen to 20%. The primary, secondary, and tertiary seed culture media for Aspergillus niger consist of: 200g potato, 20g glucose, 18g traditional Chinese medicine powder, 20g trehalose, 1000mL distilled water, pH 5.5, sterilized at 121℃ for 20min. The weight proportions of the primary seed culture medium for Aspergillus niger are as follows: 55g corn flour, 20g soybean flour, 12g wheat bran, 20g trehalose, 2g ammonium sulfate, 2g dipotassium hydrogen phosphate, 2g potassium dihydrogen phosphate, 1000mL purified water, pH 6, sterilized at 121℃ for 20min. The concentration of Aspergillus niger fermentation broth in the primary seed tank is 7.5 x 10⁻⁶ cells / mL. 8 pcs / ml; Step 3: Microbial fermentation The primary seed was inoculated at a rate of 10% into a 500 mL shake flask containing secondary seed culture medium (100 mL). The mixture was incubated at 30°C on a shaker at 100 rpm for 42 h. After incubation, the secondary seed tank's alternating liquid seed was inoculated into a fermenter at a rate of 6%, incubated at 30°C with a stirring speed of 350 rpm and an aeration ratio of 1:2 for 12 h. The temperature was then slowly lowered to 12°C at a rate of 2°C / h and incubated at this temperature for 18 h. The temperature was then further slowly lowered to 4°C, at which point the secondary seed tank's alternating liquid seed was added back into the fermenter at a rate of 4%, and incubated at this temperature for 25 h. Finally, the temperature was slowly increased to 10-15°C at a rate of 2°C / h and incubated at this temperature for 15-20 h. The temperature was then slowly increased to 33°C at a rate of 2°C / h and incubated at this temperature for 18 h. Dissolved oxygen control: Dissolved oxygen is controlled at 20% by adjusting the stirring speed and ventilation volume; pH control: The pH value is maintained at 5.2 during fermentation by supplementing with ammonia or dilute phosphoric acid. Feeding control: When the reducing sugar content in the fermentation broth drops to 3mg / ml-8mg / ml, start adding feed medium, and the amount of feed should be enough to maintain the reducing sugar content in the fermentation broth at 2mg / ml-5mg / ml; Standard for placement in the tank: 70% of the bacterial cells are aged and autolyzed, and enzyme activity increases slowly; The fermentation medium consists of: 100g maltodextrin, 55g corn flour, 20g soybean flour, 12g wheat bran, 35g trehalose, 6g yeast powder, 3g corn steep liquor, 2g ammonium sulfate, 2g dipotassium hydrogen phosphate, 2g potassium dihydrogen phosphate, 1000mL purified water, pH 5.2, sterilized at 121℃ for 20min. The feed culture medium has the following weight percentages: maltodextrin 25%, corn flour 15%, soybean flour 20%, traditional Chinese medicine powder 8%, and purified water to make up any shortfall. The pH value is 5.2, and the medium is sterilized at 121℃ for 40 minutes. Step 4: The fermentation broth is filtered through plate and frame filter, finely filtered through filter paper, separated and purified by chromatographic column, finely filtered through filter paper, and spray-dried to obtain solid lipase.

[0019] The method for preparing dextranase includes the following steps: Step 1: Activation of Amorphous Chaetomium strains The well-preserved amorphous Chaetomium slant culture was inoculated into the slant culture medium and incubated at 30℃ for 48h to activate the culture. This activation was repeated 3 times. The composition of the amorphous Chaetomium slant culture medium is as follows: 200g potato, 20g glucose, 20g agar, 1000mL distilled water, pH 5.8, sterilized at 121℃ for 20min. Step 2: Liquid seed culture of amorphous Chaetomium Step ①: Primary seed culture: The activated amorphous chaetotrichum slant culture was inoculated into a 500 ml shake flask with sterile aquatic cells, and 100 ml of liquid seed culture medium was added. The culture was carried out at 30℃ and shaken at 160 rpm for 48 h. Step 2: Secondary seed culture: Inoculate 10% of the primary seeds into a 500 ml secondary seed shake flask, and culture under the same conditions as the primary seeds; Step ③: Tertiary seed culture: Inoculate the secondary seeds at an 8% inoculation rate into a 5000 ml tertiary seed shake flask, add 1000 ml of liquid culture medium, and culture at 30℃ on a shaker at 160 rpm for 48 h. Step 4: Primary Seed Tank Cultivation: Inoculate the tertiary seeds at an 8% inoculation rate into a primary seed tank with a total volume of 150L. Fill the fermentation medium with 100L, control the pH value to 6, the cultivation temperature to 30℃, the stirring speed to 200rpm, the aeration rate to 1:1, the cultivation time to 40h, and the dissolved oxygen to 20%. The primary, secondary, and tertiary seed culture media for the amorphous chaetotrichum are composed of: 200g potato, 20g glucose, 20g trehalose, 1000mL distilled water, pH 5.5, sterilized at 121℃ for 20min. The weight proportions of the amorphous chaetotrichum primary seed tank culture medium are as follows: 55g corn flour, 20g soybean flour, 12g wheat bran, 20g trehalose, 2g ammonium sulfate, 2g dipotassium hydrogen phosphate, 2g potassium dihydrogen phosphate, 1000mL purified water, pH 6, sterilized at 121℃ for 20min. The concentration of the amorphous Chaetomium primary seed tank fermentation broth is 8 x 10⁻⁶ cells / mL. 8 pcs / ml; Step 3: Microbial fermentation The primary seed was inoculated at a rate of 10% into a 500 mL shake flask containing secondary seed culture medium (100 mL). The mixture was incubated at 30°C on a shaker at 160 rpm for 48 hours. After incubation, the secondary seed tank's alternating liquid seed was inoculated into a fermenter at a rate of 6%, incubated at 30°C with a stirring speed of 300 rpm and an aeration ratio of 1:2 for 12 hours. The temperature was then slowly lowered to 12°C at a rate of 2°C / h and maintained at this temperature for 18 hours. The temperature was then further slowly lowered to 4°C at a rate of 2°C / h, at which point the secondary seed tank's alternating liquid seed was added back into the fermenter at a rate of 4%, and the mixture was maintained at this temperature for 25 hours. Finally, the temperature was slowly increased to 10-15°C at a rate of 2°C / h and maintained at this temperature for 15-20 hours. The temperature was then slowly increased to 33°C at a rate of 2°C / h and maintained at this temperature for 18 hours. Dissolved oxygen control: Dissolved oxygen is controlled at 30% by adjusting the stirring speed and ventilation volume; pH control: The pH value is maintained at 5.2 during fermentation by supplementing with ammonia or dilute phosphoric acid. Feeding control: When the reducing sugar content in the fermentation broth drops to 5mg / ml-10mg / ml, start adding feed medium, and the amount of feed should be enough to maintain the reducing sugar content in the fermentation broth at 3mg / ml-6mg / ml; Standard for placement in the tank: 70% of the bacterial cells are aged and autolyzed, protein content does not increase, and enzyme activity increases slowly; The fermentation medium consists of: 100g maltodextrin, 55g corn flour, 20g soybean flour, 12g wheat bran, 35g trehalose, 6g yeast powder, 3g corn steep liquor, 2g ammonium sulfate, 2g dipotassium hydrogen phosphate, 2g potassium dihydrogen phosphate, 1000mL purified water, pH 5.2, sterilized at 121℃ for 20min. The feed culture medium has the following weight percentages: maltodextrin 25%, corn flour 15%, soybean flour 20%, with purified water to make up any shortfall, pH 5.2, and sterilized at 121℃ for 40 minutes. Step 4: The fermentation broth is filtered through plate and frame filter, fine filtration through filter paper, separation and purification by chromatographic column, fine filtration through filter paper, and spray drying to obtain solid dextranase.

[0020] Example 2 Step 1: Weigh out 20 parts of esterase (enzyme activity ≥ 500 U / g), 25 parts of lipase (enzyme activity ≥ 1000 U / g), 15 parts of papain lipase (enzyme activity ≥ 1000 U / g), 15 parts of dextran lipase (enzyme activity ≥ 1000 U / g), and 50 parts of maltodextrin according to the specified ratio. Step 2: After crushing the raw materials weighed in Step 1, mix them evenly to obtain a teeth whitening preparation with synergistic effects of compound enzymes. Step 3: Add the teeth whitening agent with synergistic effect of the compound enzyme obtained in Step 2 to the toothpaste base at an addition rate of 3g / kg and stir evenly to obtain the finished toothpaste. The preparation methods of the esterase, lipase and dextran lipase are the same as in Example 1.

[0021] Example 3 Step 1: Weigh out the following proportions: 17 parts of esterase (enzyme activity ≥ 500 U / g), 17 parts of lipase (enzyme activity ≥ 1000 U / g), 12.5 parts of papain lipase (enzyme activity ≥ 10000 U / g), 12.5 parts of dextranase (enzyme activity ≥ 10000 U / g), and 41 parts of maltodextrin. Step 2: After crushing the raw materials weighed in Step 1, mix them evenly to obtain a teeth whitening preparation with synergistic effects of compound enzymes. Step 3: Add the teeth whitening agent with synergistic effect of the compound enzyme obtained in Step 2 to the toothpaste base at an addition rate of 3g / kg and stir evenly to obtain the finished toothpaste. The preparation methods of the esterase, lipase and dextran lipase are the same as in Example 1.

[0022] Example 4 Step 1: Weigh out the following ingredients according to the specified ratio: 15 parts of esterase (enzyme activity ≥ 500 U / g), 15 parts of lipase (enzyme activity ≥ 1000 U / g), 10 parts of papain lipase (enzyme activity ≥ 1000 U / g), 10 parts of dextran lipase (enzyme activity ≥ 1000 U / g), 10 parts of laccase (enzyme activity ≥ 1000 U / g), and 25 parts of maltodextrin. Step 2: After crushing the raw materials weighed in Step 1, mix them evenly to obtain a teeth whitening preparation with synergistic effects of compound enzymes. Step 3: Add the teeth whitening agent with synergistic effect of the compound enzyme obtained in Step 2 to the toothpaste base at an addition rate of 3g / kg and stir evenly to obtain the finished toothpaste. The preparation methods of the esterase, lipase and dextran lipase are the same as in Example 1; The method for preparing laccase includes the following steps: Step 1: Activation of Aspergillus oryzae strain The well-preserved Aspergillus oryzae slant culture was inoculated into slant culture medium and incubated at 28°C for 24 hours to activate the culture. This activation was repeated 3 times. The Aspergillus oryzae slant culture medium consists of: 200g potato, 20g glucose, 20g agar, 1000mL distilled water, pH 5.8, sterilized at 121℃ for 20min. Step 2: Liquid seed culture of Aspergillus oryzae Step ①: Primary seed culture: Wash the activated Aspergillus oryzae slant culture with sterile water to obtain spores, inoculate into a 500 ml shake flask, add 100 ml of liquid seed culture medium, and culture at 28℃ on a shaker at 120 rpm for 32 h. Step 2: Secondary seed culture: Inoculate 10% of the primary seeds into a 500 ml secondary seed shake flask, and culture under the same conditions as the primary seeds; Step ③: Tertiary seed culture: Inoculate the secondary seeds at an 8% inoculation rate into a 5000 ml tertiary seed shake flask, add 1000 ml of liquid culture medium, and culture at 30℃ on a shaker at 120 rpm for 32 h. Step 4: Primary Seed Tank Cultivation: Inoculate the tertiary seeds at an 8% inoculation rate into a primary seed tank with a total volume of 150L. Fill the fermentation medium with 100L, control the pH value to 6, the cultivation temperature to 30℃, the stirring speed to 200rpm, the aeration rate to 1:1.5, the cultivation time to 32h, and the dissolved oxygen to 25%. The primary, secondary, and tertiary seed culture media for Aspergillus oryzae are composed of: 200g potato, 20g glucose, 20g trehalose, 1000mL distilled water, pH 5.5, sterilized at 121℃ for 20min. The weight proportions of the Aspergillus oryzae primary seed culture medium are as follows: 55g corn flour, 20g soybean flour, 12g wheat bran, 20g trehalose, 2g ammonium sulfate, 2g dipotassium hydrogen phosphate, 2g potassium dihydrogen phosphate, 1000mL purified water, pH 6, sterilized at 121℃ for 20min. The Aspergillus oryzae primary seed tank fermentation broth cell concentration is 6 x 10⁻⁶. 8 pcs / ml; Step 3: Microbial fermentation The primary seed was inoculated at a rate of 10% into a 500 mL shake flask containing secondary seed culture medium (100 mL). The flask was incubated at 30°C for 32 h on a shaker at 120 rpm. After incubation, the secondary seed tank's alternating liquid seed was inoculated into a fermenter at a rate of 6%, incubated at 28°C with a stirring speed of 300 rpm and an aeration ratio of 1:2 for 12 h. The temperature was then slowly lowered to 12°C at a rate of 2°C / h and incubated at this temperature for 18 h. The temperature was then further slowly lowered to 4°C at a rate of 2°C / h, at which point the secondary seed tank's alternating liquid seed was added back into the fermenter at a rate of 4%, and incubated at this temperature for 25 h. Finally, the temperature was slowly increased to 10-15°C at a rate of 2°C / h and incubated at this temperature for 15-20 h. The temperature was then slowly increased to 28°C at a rate of 2°C / h and incubated at this temperature for 20 h. Dissolved oxygen control: Dissolved oxygen is controlled at 25% by adjusting the stirring speed and ventilation volume; pH control: The pH value is maintained at 5.5 during fermentation by adding ammonia or dilute phosphoric acid. Feeding control: When the reducing sugar content in the fermentation broth drops to 5mg / ml-10mg / ml, start adding feed medium, and the amount of feed should be enough to maintain the reducing sugar content in the fermentation broth at 5mg / ml-8mg / ml; Standard for placement in the tank: 70% of the bacterial cells are aged and autolyzed, protein does not increase, and enzyme activity increases slowly; The fermentation medium consists of: 100g maltodextrin, 55g corn flour, 20g soybean flour, 12g wheat bran, 35g trehalose, 6g yeast powder, 3g corn steep liquor, 2g ammonium sulfate, 2g dipotassium hydrogen phosphate, 2g potassium dihydrogen phosphate, 1000mL purified water, pH 5.2, sterilized at 121℃ for 20min. The feed culture medium has the following weight percentages: maltodextrin 25%, corn flour 15%, soybean flour 20%, traditional Chinese medicine powder 8%, and purified water to make up any shortfall. The pH value is 5.2, and the medium is sterilized at 121℃ for 40 minutes. Step 4: The fermentation broth is filtered through plate and frame filter, finely filtered through filter paper, separated and purified by chromatographic column, finely filtered through filter paper, and spray-dried to obtain solid laccase.

[0023] Example 5 Step 1: Weigh out the following ingredients according to the specified ratio: 20 parts of esterase (enzyme activity ≥ 500 U / g), 25 parts of lipase (enzyme activity ≥ 1000 U / g), 15 parts of papain lipase (enzyme activity ≥ 1000 U / g), 15 parts of dextran lipase (enzyme activity ≥ 1000 U / g), 15 parts of laccase (enzyme activity ≥ 1000 U / g), and 50 parts of maltodextrin. Step 2: After crushing the raw materials weighed in Step 1, mix them evenly to obtain a teeth whitening preparation with synergistic effects of compound enzymes. Step 3: Add the teeth whitening agent with synergistic effect of the compound enzyme obtained in Step 2 to the toothpaste base at an addition rate of 3g / kg and stir evenly to obtain the finished toothpaste. The preparation methods of the esterase, lipase, and dextran lipase are the same as in Example 1; the preparation method of the laccase is the same as in Example 4.

[0024] Example 6 Step 1: Weigh out the following proportions: 17 parts of esterase (enzyme activity ≥ 500 U / g), 17 parts of lipase (enzyme activity ≥ 1000 U / g), 12.5 parts of papain lipase (enzyme activity ≥ 1000 U / g), 12.5 parts of dextran lipase (enzyme activity ≥ 1000 U / g), 10 parts of laccase (enzyme activity ≥ 1000 U / g), and 31 parts of maltodextrin. Step 2: After crushing the raw materials weighed in Step 1, mix them evenly to obtain a teeth whitening preparation with synergistic effects of compound enzymes. Step 3: Add the teeth whitening agent with synergistic effect of the compound enzyme obtained in Step 2 to the toothpaste base at an addition rate of 3g / kg and stir evenly to obtain the finished toothpaste. The preparation methods of the esterase, lipase, and dextran lipase are the same as in Example 1; the preparation method of the laccase is the same as in Example 4.

[0025] Verification test Teeth models with similar conditions were selected and evenly divided into three groups. These groups were soaked in strong tea, strong coffee, and red wine, respectively, at 30°C for 3 days. The resulting tooth models were then examined for tea stains, coffee stains, and red wine stains, respectively. Brushing tests were then conducted using the toothpaste prepared in Examples 1-6. Existing whitening toothpaste was used as a control group, and water was used as a blank control group. The test results are as follows: Figure 1-18 As shown, the whitening toothpaste prepared by comparative examples 1-18 has a significantly better whitening effect than the control group and the blank group, and does not cause damage to teeth.

[0026] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A teeth whitening agent based on the synergistic effect of compound enzymes, characterized in that: It includes the following substances in parts by weight: 15-20 parts esterase, 15-25 parts lipase, 10-15 parts papain, 10-15 parts dextranase, and 25-50 parts maltodextrin.

2. The teeth whitening agent based on the synergistic effect of compound enzymes according to claim 1, characterized in that: The esterase has an enzyme activity ≥500 U / g; the lipase has an enzyme activity ≥1000 U / g; the papain has an enzyme activity ≥10000 U / g; and the dextranase has an enzyme activity ≥10000 U / g.

3. The teeth whitening preparation based on the synergistic effect of compound enzymes according to claim 1, characterized in that: It includes the following substances in parts by weight: 15-20 parts esterase, 15-25 parts lipase, 10-15 parts papain, 10-15 parts dextranase, 10-15 parts laccase, and 25-50 parts maltodextrin.

4. A teeth whitening preparation based on the synergistic effect of compound enzymes according to claim 1, characterized in that: The esterase has an enzyme activity ≥500 U / g; the lipase has an enzyme activity ≥1000 U / g; the papain has an enzyme activity ≥10000 U / g; the dextranase has an enzyme activity ≥10000 U / g; and the laccase has an enzyme activity ≥1000 U / g.

5. A teeth whitening preparation based on the synergistic effect of a complex enzyme according to claim 1 or 3, characterized in that: The method for preparing the esterase includes the following steps: Step 1: Activation of Aspergillus niger strain The well-preserved Aspergillus niger slant culture was inoculated into slant culture medium and incubated at 30℃ for 42h to activate the culture. This activation was repeated 3 times. The composition of the Aspergillus niger slant culture medium is as follows: 200g potato, 20g glucose, 20g agar, 1000mL distilled water, pH 5.8, sterilized at 121℃ for 20min. Step 2: Liquid seed culture of Aspergillus niger Step ①: Primary seed culture: Wash the activated Aspergillus niger slant culture with sterile water to obtain spores, inoculate into a 500 ml shake flask, add 100 ml of liquid seed culture medium, and culture at 30℃ on a shaker at 160 rpm for 42 h. Step 2: Secondary seed culture: Inoculate 10% of the primary seeds into a 500 ml secondary seed shake flask, and culture under the same conditions as the primary seeds; Step ③: Tertiary seed culture: Inoculate the secondary seeds at an 8% inoculation rate into a 5000 ml tertiary seed shake flask, add 1000 ml of liquid culture medium, and culture at 30℃ on a shaker at 100 rpm for 42 h. Step 4: Primary Seed Tank Cultivation: Inoculate the tertiary seeds at an 8% inoculation rate into a primary seed tank with a total volume of 150L. Fill the fermentation medium with 100L, control the pH value to 6, the cultivation temperature to 30℃, the stirring speed to 300rpm, the aeration rate to 1:1, the cultivation time to 42h, and the dissolved oxygen to 20%. The primary, secondary, and tertiary seed culture media for Aspergillus niger consist of: 200g potato, 20g glucose, 20g trehalose, 1000mL distilled water, pH 5.5, sterilized at 121℃ for 20min. The weight proportions of the primary seed culture medium for Aspergillus niger are as follows: 55g corn flour, 20g soybean flour, 12g wheat bran, 20g trehalose, 2g ammonium sulfate, 2g dipotassium hydrogen phosphate, 2g potassium dihydrogen phosphate, 1000mL purified water, pH 6, sterilized at 121℃ for 20min. The concentration of Aspergillus niger fermentation broth in the primary seed tank is 7.5 x 10⁻⁶ cells / mL. 8 pcs / ml; Step 3: Microbial fermentation The primary seed was inoculated at a rate of 10% into a 500 mL shake flask containing secondary seed culture medium (100 mL). The mixture was incubated at 30°C on a shaker at 100 rpm for 42 h. After incubation, the secondary seed tank's alternating liquid seed was inoculated into a fermenter at a rate of 6%, incubated at 30°C with a stirring speed of 350 rpm and an aeration ratio of 1:2 for 12 h. The temperature was then slowly lowered to 12°C at a rate of 2°C / h and incubated at this temperature for 18 h. The temperature was then further slowly lowered to 4°C, at which point the secondary seed tank's alternating liquid seed was added back into the fermenter at a rate of 4%, and incubated at this temperature for 25 h. Finally, the temperature was slowly increased to 10-15°C at a rate of 2°C / h and incubated at this temperature for 15-20 h. The temperature was then slowly increased to 33°C at a rate of 2°C / h and incubated at this temperature for 18 h. Dissolved oxygen control: Dissolved oxygen is controlled at 20% by adjusting the stirring speed and ventilation volume; pH control: The pH value is maintained at 5.2 during fermentation by supplementing with ammonia or dilute phosphoric acid. Feeding control: When the reducing sugar content in the fermentation broth drops to 3mg / ml-8mg / ml, start adding feed medium, and the amount of feed should be enough to maintain the reducing sugar content in the fermentation broth at 2mg / ml-5mg / ml; Standard for placement in the tank: 70% of the bacterial cells are aged and autolyzed, and enzyme activity increases slowly; The fermentation medium consists of: 100g maltodextrin, 55g corn flour, 20g soybean flour, 12g wheat bran, 35g trehalose, 6g yeast powder, 3g corn steep liquor, 2g ammonium sulfate, 2g dipotassium hydrogen phosphate, 2g potassium dihydrogen phosphate, 1000mL purified water, pH 5.2, sterilized at 121℃ for 20min. The feed culture medium has the following weight percentages: maltodextrin 25%, corn flour 15%, soybean flour 20%, traditional Chinese medicine powder 8%, and purified water to make up any shortfall. The pH value is 5.2, and the medium is sterilized at 121℃ for 20 minutes. Step 4: The fermentation broth is filtered through plate and frame filter, finely filtered through filter paper, separated and purified by chromatographic column, and spray-dried to obtain solid esterase.

6. A teeth whitening preparation based on the synergistic effect of a complex enzyme according to claim 1 or 3, characterized in that: The method for preparing the lipase includes the following steps: Step 1: Activation of Aspergillus niger strain The well-preserved Aspergillus niger slant culture was inoculated into slant culture medium and incubated at 30℃ for 42h to activate the culture. This activation was repeated 3 times. The composition of the Aspergillus niger slant culture medium is as follows: 200g potato, 20g glucose, 20g agar, 1000mL distilled water, pH 5.8, sterilized at 121℃ for 20min. Step 2: Liquid seed culture of Aspergillus niger Step ①: Primary seed culture: Wash the activated Aspergillus niger slant culture with sterile water to obtain spores, inoculate into a 500 ml shake flask, add 100 ml of liquid seed culture medium, and culture at 30℃ on a shaker at 160 rpm for 42 h. Step 2: Secondary seed culture: Inoculate 10% of the primary seeds into a 500 ml secondary seed shake flask, and culture under the same conditions as the primary seeds; Step ③: Tertiary seed culture: Inoculate the secondary seeds at an 8% inoculation rate into a 5000 ml tertiary seed shake flask, add 1000 ml of liquid culture medium, and culture at 30℃ on a shaker at 100 rpm for 42 h. Step 4: Primary Seed Tank Cultivation: Inoculate the tertiary seeds at an 8% inoculation rate into a primary seed tank with a total volume of 150L. Fill the fermentation medium with 100L, control the pH value to 6, the cultivation temperature to 30℃, the stirring speed to 300rpm, the aeration rate to 1:1, the cultivation time to 42h, and the dissolved oxygen to 20%. The primary, secondary, and tertiary seed culture media for Aspergillus niger consist of: 200g potato, 20g glucose, 20g trehalose, 1000mL distilled water, pH 5.5, sterilized at 121℃ for 20min. The weight proportions of the primary seed culture medium for Aspergillus niger are as follows: 55g corn flour, 20g soybean flour, 12g wheat bran, 20g trehalose, 2g ammonium sulfate, 2g dipotassium hydrogen phosphate, 2g potassium dihydrogen phosphate, 1000mL purified water, pH 6, sterilized at 121℃ for 20min. The concentration of Aspergillus niger fermentation broth in the primary seed tank is 7.5 x 10⁻⁶ cells / mL. 8 pcs / ml; Step 3: Microbial fermentation The primary seed was inoculated at a rate of 10% into a 500 mL shake flask containing secondary seed culture medium (100 mL). The mixture was incubated at 30°C on a shaker at 100 rpm for 42 h. After incubation, the secondary seed tank's alternating liquid seed was inoculated into a fermenter at a rate of 6%, incubated at 30°C with a stirring speed of 350 rpm and an aeration ratio of 1:2 for 12 h. The temperature was then slowly lowered to 12°C at a rate of 2°C / h and incubated at this temperature for 18 h. The temperature was then further slowly lowered to 4°C, at which point the secondary seed tank's alternating liquid seed was added back into the fermenter at a rate of 4%, and incubated at this temperature for 25 h. Finally, the temperature was slowly increased to 10-15°C at a rate of 2°C / h and incubated at this temperature for 15-20 h. The temperature was then slowly increased to 33°C at a rate of 2°C / h and incubated at this temperature for 18 h. Dissolved oxygen control: Dissolved oxygen is controlled at 20% by adjusting the stirring speed and ventilation volume; pH control: The pH value is maintained at 5.2 during fermentation by supplementing with ammonia or dilute phosphoric acid. Feeding control: When the reducing sugar content in the fermentation broth drops to 3mg / ml-8mg / ml, start adding feed medium, and the amount of feed should be enough to maintain the reducing sugar content in the fermentation broth at 2mg / ml-5mg / ml; Standard for placement in the tank: 70% of the bacterial cells are aged and autolyzed, and enzyme activity increases slowly; The fermentation medium consists of: 100g maltodextrin, 55g corn flour, 20g soybean flour, 12g wheat bran, 35g trehalose, 6g yeast powder, 3g corn steep liquor, 2g ammonium sulfate, 2g dipotassium hydrogen phosphate, 2g potassium dihydrogen phosphate, 1000mL purified water, pH 5.2, sterilized at 121℃ for 20min. The feed culture medium has the following weight percentages: maltodextrin 25%, corn flour 15%, soybean flour 20%, traditional Chinese medicine powder 8%, and purified water to make up any shortfall. The pH value is 5.2, and the medium is sterilized at 121℃ for 40 minutes. Step 4: The fermentation broth is filtered through plate and frame filter, finely filtered through filter paper, separated and purified by chromatographic column, finely filtered through filter paper, and spray-dried to obtain solid lipase.

7. A teeth whitening preparation based on the synergistic effect of a complex enzyme according to claim 1 or 3, characterized in that: The method for preparing dextranase includes the following steps: Step 1: Activation of Amorphous Chaetomium strains Cut a 0.5 × 0.5 cm piece. 2 The slant cultured bacterial blocks were inoculated into activation medium (liquid medium containing 1–1.5% soluble starch and 0.4–0.6% yeast extract) and cultured at 30°C and 200 rpm with shaking until the bacterial solution became turbid (approximately 24–48 hours). The composition of the amorphous Chaetomium slant culture medium is as follows: 200g potato, 20g glucose, 20g agar, 1000mL distilled water, pH 6.5, sterilized at 121℃ for 20min. Step 2: Liquid seed culture of amorphous Chaetomium Step ①: Primary seed culture: Inoculate the activated strain into PD liquid medium (20% potato, 2% glucose), and culture at 30℃ and 180 rpm for 7 days to form mycelial balls or mycelial fragments. Then, inoculate into a 500 ml shake flask with 100 ml of liquid seed medium and culture at 30℃ and 160 rpm for 48 hours. Step 2: Secondary seed culture: Inoculate 10% of the primary seeds into a 500 ml secondary seed shake flask, and culture under the same conditions as the primary seeds; Step 3: Tertiary seed culture: Inoculate the secondary seeds at an 8% inoculation rate into a 5000 ml tertiary seed shake flask, add 1000 ml of liquid culture medium, and culture at 30℃ on a shaker at 180 rpm for 48 h. Step 4: Primary Seed Tank Cultivation: Inoculate the tertiary seeds at an 8% inoculation rate into a primary seed tank with a total volume of 150L. Fill the fermentation medium with 100L, control the pH value at 6.5, the cultivation temperature at 30℃, the stirring speed at 180rpm, the aeration rate at 1:1, the cultivation time at 40h, and the dissolved oxygen at >20%. The primary, secondary, and tertiary seed culture media for amorphous chaetotrichum consist of: 200g potato, 20g glucose, 20g trehalose, 1000mL distilled water, pH 6.5, sterilized at 121℃ for 20min. The concentration of the amorphous Chaetomium primary seed tank fermentation broth is 4 x 10⁻⁶ cells / mL. 8 cells / mL; Step 3: Microbial fermentation The primary seeds were inoculated at a rate of 10% into 500 ml of secondary seed culture medium in a shake flask, and cultured at 30°C for 48 h on a shaker at 180 rpm. After the culture was completed, the secondary seed tank's alternating liquid seeds were inoculated into a fermenter at a rate of 6%, and cultured at 30°C with a stirring speed of 300 rpm and an aeration rate of 1:2 for 36 h. The secondary seed tank's alternating liquid seeds were then added back into the fermenter at a rate of 4%, and cultured at 30°C for 42 h. Dissolved oxygen control: Dissolved oxygen is controlled to be >20% by adjusting the stirring speed and ventilation volume; pH control: The pH value is maintained at 6.5 during fermentation by adding ammonia or dilute phosphoric acid. Feeding control: When the reducing sugar content in the fermentation broth drops to 3mg / ml-6mg / ml, start adding feed medium, and the amount of feed should be enough to maintain the reducing sugar content in the fermentation broth at 5mg / ml-10mg / ml; Standard for placement in the tank: 70% of the bacterial cells are aged and autolyzed, protein content does not increase, and enzyme activity increases slowly; The fermentation medium consists of: 100g maltodextrin, 55g corn flour, 20g soybean flour, 12g wheat bran, 35g trehalose, 6g yeast powder, 3g corn steep liquor, 2g ammonium sulfate, 2g dipotassium hydrogen phosphate, 2g potassium dihydrogen phosphate, 1000mL purified water, pH 6.5, sterilized at 121℃ for 20min. The feed culture medium has the following weight percentages: maltodextrin 25%, corn flour 15%, soybean flour 20%, with purified water to make up any shortfall, pH 6.5, and sterilized at 121℃ for 20 minutes. Step 4: The fermentation broth is filtered through plate and frame filter, fine filtration through filter paper, separated and purified by chromatographic column, and spray-dried to obtain solid dextranase.

8. A teeth whitening preparation based on the synergistic effect of compound enzymes according to claim 3, characterized in that: The method for preparing laccase includes the following steps: Step 1: Activation of Aspergillus oryzae strain The well-preserved Aspergillus oryzae slant culture was inoculated into slant culture medium and incubated at 30℃ for 48h to activate the culture. This activation was repeated 3 times. The Aspergillus oryzae slant culture medium is composed of: 100g glucose, 6g peptone, 20g agar, 5g KH2PO4, 0.5g MgSO4·7H2O, 0.01g Tween-80, 1000mL distilled water, pH 5.5, sterilized at 121℃ for 30 min. Step 2: Liquid seed culture of Aspergillus oryzae Step ①: Primary seed culture: The activated Aspergillus oryzae slant culture was washed with sterile water to obtain spores, filtered through a double layer of 500-mesh nylon cloth, dispersed by small glass beads, counted using a hemocytometer, and diluted to a concentration of approximately 1×10⁻⁶. 7 The number of cells / mL was added to a 500 mL shake flask, and the liquid seed culture medium was added to 100 mL. The culture was carried out at 30℃ and shaken at 120 rpm for 32 h. Step 2: Secondary seed culture: Inoculate 10% of the primary seeds into a 500 ml secondary seed shake flask, and culture under the same conditions as the primary seeds; Step ③: Tertiary seed culture: Inoculate the secondary seeds at an 8% inoculation rate into a 5000 ml tertiary seed shake flask, add 1000 ml of liquid culture medium, and culture at 30℃ on a shaker at 120 rpm for 32 h. Step 4: Primary Seed Tank Cultivation: Inoculate the tertiary seeds at an 8% inoculation rate into a primary seed tank with a total volume of 150L. Fill the fermentation medium with 100L, control the pH value at 5.5, the cultivation temperature at 30℃, the stirring speed at 200rpm, the aeration rate at 1:1.5, the cultivation time at 32h, and the dissolved oxygen at 26%. The primary, secondary, and tertiary seed culture media for Aspergillus oryzae are composed of: 200g potato, 20g glucose, 20g trehalose, 1000mL distilled water, pH 5.5, sterilized at 121℃ for 20min. The weight proportions of the Aspergillus oryzae primary seed culture medium are as follows: 55g corn flour, 20g soybean flour, 20g trehalose, 2g ammonium sulfate, 2g dipotassium hydrogen phosphate, 2g potassium dihydrogen phosphate, 1000mL purified water, pH 5.5, sterilized at 121℃ for 20min. The Aspergillus oryzae primary seed tank fermentation broth cell concentration is 6 x 10⁻⁶. 8 pcs / ml; Step 3: Microbial fermentation The primary seed was inoculated at a rate of 10% into a 500 mL shake flask containing secondary seed culture medium (100 mL). The flask was incubated at 30°C for 32 h on a shaker at 120 rpm. After incubation, the secondary seed tank's alternating liquid seed was inoculated into a fermenter at a rate of 6%, incubated at 28°C with a stirring speed of 300 rpm and an aeration ratio of 1:2 for 12 h. The temperature was then slowly lowered to 12°C at a rate of 2°C / h and incubated at this temperature for 18 h. The temperature was then further slowly lowered to 4°C, at which point the secondary seed tank's alternating liquid seed was added back into the fermenter at a rate of 4%, and incubated at this temperature for 25 h. Finally, the temperature was slowly increased to 10-15°C at a rate of 2°C / h and incubated at this temperature for 15-20 h. The temperature was then slowly increased to 30°C at a rate of 2°C / h and incubated at this temperature for 20 h. Dissolved oxygen control: Dissolved oxygen was controlled at 26% by adjusting the stirring speed and ventilation volume; pH control: The pH value is maintained at 5.5 during fermentation by adding ammonia or dilute phosphoric acid. Feeding control: When the reducing sugar content in the fermentation broth drops to 5mg / ml-10mg / ml, start adding feed medium, and the amount of feed should be enough to maintain the reducing sugar content in the fermentation broth at 5mg / ml-8mg / ml; Standard for release into the tank: 70% of the bacterial cells have aged and autolyzed, the protein content no longer increases, and the enzyme activity increases slowly; The fermentation medium consists of: 100g maltodextrin, 55g corn flour, 20g soybean flour, 35g trehalose, 6g yeast powder, 3g corn steep liquor, 2g ammonium sulfate, 2g dipotassium hydrogen phosphate, 2g potassium dihydrogen phosphate, 1000mL purified water, pH 5.5, sterilized at 121℃ for 20min. The feed culture medium has the following weight percentages: maltodextrin 25%, corn flour 15%, soybean flour 20%, traditional Chinese medicine powder 8%, and purified water to make up any shortfall. The pH value is 5.5, and the medium is sterilized at 121℃ for 20 minutes. Step 4: The fermentation broth is filtered through plate and frame filter, finely filtered through filter paper, separated and purified by chromatographic column, and spray-dried to obtain solid laccase.

9. The application of a teeth whitening agent based on the synergistic effect of compound enzymes, characterized in that: The teeth whitening agent with the synergistic effect of the compound enzyme is added to toothpaste for use.

10. The application of a teeth whitening agent based on the synergistic effect of a complex enzyme according to claim 9, characterized in that: The amount of the teeth whitening agent with the synergistic effect of the compound enzyme added to the toothpaste is 3g / kg.

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