Bacterial cellulose mask and preparation method thereof

Abstract

The invention relates to a bacterial cellulose mask and a preparation method thereof.The bacterial cellulose mask is prepared by soaking a bacterial cellulose wet film in a mixed essence solution, and the mixed essence solution is prepared from, by mass, 1%-1.5% of astaxanthin, 1%-1.5% of arbutin, 4%-6% of glycerinum and the balance ultrapure water. According to the method, millet vinegar residues are used as a main raw material, a bacterial cellulose wet film with high yield and excellent performance is safely and conveniently produced and prepared in a microbial fermentation mode, and the bacterial cellulose mask with excellent oxidation resistance is prepared by adsorbing a compound mixed essence solution on the basis of the bacterial cellulose wet film. The method is simple in steps, safe, controllable, high in yield and good in stability and reliability, waste utilization is achieved, and the production cost is greatly reduced.

Description

Technical Field

[0001] The invention relates to the technical field of membrane material preparation, in particular to a method for producing bacterial cellulose using microorganisms and an application of the method in facial masks. Background Art

[0002] Bacterial cellulose is a high molecular polymer composed of D-glucose connected by β-1,4 glycosidic bonds, generally produced by Gram-negative bacteria. It has the advantages of high purity, high water retention, biocompatibility, high crystallinity and strong mechanical properties. It has extensive research value in the field of facial masks, but its low yield and high production cost are still the main reasons limiting its widespread application.

[0003] After searching, it was found that the Chinese invention patent with authorization number CN 109652489B "A one-step method for producing fully fermented bacterial cellulose film containing mushroom fermentation extracts" disclosed a method for producing bacterial cellulose. The method uses lactic acid bacteria, yeast and acetic acid bacteria to jointly ferment a combination of shiitake mushrooms, enoki mushrooms, boletus and abalone mushrooms or a combination of white mushrooms, tea tree mushrooms, oyster mushrooms, hericium erinaceus, matsutake and morels to produce bacterial cellulose. The significant defects of its technical solution are: 1. Its fermentation requires the simultaneous action of lactic acid bacteria, yeast and acetic acid bacteria. The preparation process of the inoculum composed of three or more microorganisms is cumbersome and it is difficult to ensure the quality of the strains. In addition, the simultaneous addition of lactic acid bacteria and yeast during the fermentation process will lead to a mutual growth inhibition effect between the microorganisms; 2. The fermentation raw materials used are mushrooms and coconut water. Some of the mushrooms, such as Hericium erinaceus and Morchella, are expensive, resulting in a high production cost of bacterial cellulose and no actual production value. In addition, the raw materials are highly volatile, resulting in unstable output; 3. It adopts an in-situ synthesis method to prepare bacterial cellulose. The preparation process must be subjected to acid reduction and partial water removal before it can be used as a pharmaceutical excipient and a facial mask substrate. Therefore, it is easy to cause a large amount of active ingredient loss, low raw material utilization, low substrate conversion rate, and serious waste of resources.

[0004] Vinegar dregs are the main by-product produced during the fermentation of vinegar. It is reported that the amount of waste vinegar dregs produced in China each year is as high as 3 million tons, most of which will be directly buried or landfilled, but the acidity of vinegar dregs will pollute the soil and groundwater. Therefore, the research, utilization and redevelopment of vinegar dregs are imminent. Vinegar dregs mainly contain bran, rice bran and fillers containing cellulose and hemicellulose, which can be saccharified into reducing sugars for microbial growth. At present, acid hydrolysis is mostly used in the prior art to hydrolyze vinegar dregs, but acid hydrolysis is difficult to operate and generally requires additional detoxification treatment before it can be used as a carbon source by microorganisms.

[0005] With the development of modern society, the trend of natural and organic facial masks reflects consumers' preference for plant-derived ingredients, so bacterial cellulose masks with stronger breathability and biocompatibility have become a hot topic in current research.

[0006] Therefore, how to use vinegar dregs as the main raw material to produce a bacterial cellulose mask with low raw material cost, high yield, good process controllability and good use effect is of great significance for reducing environmental pollution, reducing the production cost of bacterial cellulose and improving the performance of the mask. Summary of the invention

[0007] The technical purpose of the present invention is to provide a bacterial cellulose mask and a preparation method thereof, which uses millet vinegar dregs as the main raw material, and safely and conveniently produces a bacterial cellulose wet film with high yield and excellent performance by microbial fermentation, and uses the bacterial cellulose wet film as the basis to adsorb a composite mixed essence solution to prepare a bacterial cellulose mask with excellent antioxidant performance. The method itself has simple steps, is safe and controllable, has high yield, and has good stability and reliability, realizes waste utilization, and greatly reduces production costs.

[0008] In order to solve the above technical problems, the technical solution adopted by the present invention is: a bacterial cellulose facial mask, which is made by soaking a bacterial cellulose wet film in a mixed essence solution, wherein the mixed essence solution is composed of the following components in percentage by mass: 1-1.5% astaxanthin, 1-1.5% arbutin, 4-6% glycerol and the remainder ultrapure water.

[0009] Furthermore, the bacterial cellulose wet film needs to be dried to a thickness of 0.75-1.25 mm before being immersed in the mixed essence solution.

[0010] Furthermore, the bacterial cellulose wet film is immersed in the mixed essence solution for 20-40 minutes.

[0011] Furthermore, after the raw material components of the mixed essence solution are evenly mixed, it is necessary to filter and sterilize it.

[0012] Furthermore, the pore size of the filter membrane used in the filtration sterilization treatment is 0.20-0.25 μm.

[0013] Furthermore, the preparation method of the bacterial cellulose wet film is: s1. Preparation of millet vinegar lees enzymatic hydrolysate According to the ratio of solid-liquid ratio of 15-25 mL / g, distilled water and millet vinegar dregs are mixed and stirred at a temperature of 26-30°C for 2.5-3.5 h, then, cellulase is added thereto at an addition amount of 5500-6500 U / g of cellulose, and enzymatic hydrolysis is performed for 8-10 h at a pH of 4.6-5.0 and a temperature of 50-60°C, and after cooling to room temperature, saccharifying enzyme is added thereto at an addition amount of 1500-2500 U / g of starch, and secondary enzymatic hydrolysis is performed for 2.5-3.5 h at a pH of 3.8-4.2 and a temperature of 58-62°C, and after suction filtration, millet vinegar dregs enzymatic hydrolyzate is obtained for standby use; s2. Preparation of fermentation medium Add the millet vinegar lees enzymatic hydrolyzate, nitrogen source, inorganic salt and accelerator obtained in step s1 into distilled water, mix thoroughly, adjust the pH to 2.5-3.5, then sterilize at 115-121° C. for 20-30 min, cool to obtain a fermentation medium, and set aside; s3. Inoculation and culture Under the temperature condition of 25-35°C, inoculate the fermentation medium prepared in step s2 with Bacillus tame SW-1 at a mass ratio of 7-9%, then place it in a shaker at 70-90 r / min and 34-38°C for 6-18 h, then place it in a constant temperature incubator at 34-38°C for static culture for 3-5 d, then inoculate the fermentation medium with Lactobacillus paracasei at a mass ratio of 2-4%, and place it in a constant temperature incubator at 34-38°C for static culture for 4-6 d; s4. Preparation of bacterial cellulose wet film The bacterial cellulose membrane on the upper layer of the fermentation liquid obtained after the inoculation culture in step s3 is taken out, placed in a NaOH solution with a concentration of 0.1 mol / L, and treated in a water bath at a temperature of 75-85°C for 50-70 min. Thereafter, it is transferred to a glacial acetic acid solution with a concentration of 0.1 mol / L for soaking for 50-70 min. After completion, it is taken out and repeatedly rinsed with distilled water until the pH value is neutral to obtain a finished bacterial cellulose wet membrane.

[0014] Furthermore, in step s2, the nitrogen source is yeast powder and corn steep liquor; the inorganic salt is KH2PO4 and MgSO4; and the promoter is citric acid and anhydrous ethanol.

[0015] Furthermore, in the fermentation medium, the amount of each component added per liter is 180-220 mL of millet vinegar lees enzymatic hydrolyzate, 8-12 g of yeast powder, 2-3 g of corn steep liquor, 4-6 g of KH2PO4, 8-12 g of MgSO4, 18-22 g of citric acid, 20-40 mL of anhydrous ethanol, and the balance is distilled water.

[0016] Further, in step s3, the tame bacillus SW-1 is classified and named Komagataeibacterobeodiens SW-1 was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on May 17, 2021, with the deposit number CGMCC NO.22548.

[0017] Furthermore, in step s3, the preparation method of the seed liquid of Bacillus tame SW-1 and Lactobacillus paracasei is as follows: first, the original strain preserved in glycerol is inoculated into a liquid culture medium, and cultured at a temperature of 28-32°C for 36-60 hours to obtain a primary seed liquid of the strain; then, the primary seed liquid is inoculated into a basic culture medium, and cultured at a temperature of 28-32°C for 36-60 hours to obtain a strain with a concentration of (1-5)×10 7 CFU / mL of bacterial seed solution can be used for inoculation.

[0018] Beneficial effects: 1. A method for preparing a bacterial cellulose facial mask of the present invention uses industrial waste, millet vinegar dregs, as the main raw material, and produces a bacterial cellulose wet film with rich internal pore structure, strong adsorption performance, good stability, good air permeability, and excellent biocompatibility safely and conveniently through the synergistic action and compound fermentation of two microorganisms, Bacillus scolopendra SW-1 and Lactobacillus paracasei. The wet film of bacterial cellulose is then used as a basis to prepare a bacterial cellulose facial mask with excellent antioxidant properties by adsorbing a composite mixed essence solution. The method itself has simple steps, is safe and controllable, has high yield, and has good stability and reliability, realizes waste utilization, and greatly reduces production costs.

[0019] 2. A bacterial cellulose facial mask of the present invention uses the porous and breathable bacterial cellulose as the membrane substrate, and is compounded with functional components such as astaxanthin and arbutin to prepare a facial mask with antioxidant, moisturizing and whitening functions. The method itself uses enzymatically hydrolyzed waste vinegar grains as fermentation additives to produce bacterial cellulose, which not only can achieve waste utilization, but also has the advantages of simple operation and low production cost, greatly improves the yield of bacterial cellulose, and can ensure stable and controllable output, and has excellent practical application value and development prospects.

[0020] 3. A bacterial cellulose facial mask of the present invention adopts a combined fermentation method of Bacillus docile and Lactobacillus paracasei to produce bacterial cellulose during the preparation of bacterial cellulose wet film. The process is simple, the conditions are mild, and the output is high. The productivity can reach 86.78 g / Kg of culture. The prepared bacterial cellulose is safe after treatment and can be widely used in food, cosmetics, medical excipients, etc., and has promotion value in the field of bacterial cellulose production technology.

[0021] 4. The bacterial cellulose mask of the present invention utilizes a double enzymatic hydrolysis method to hydrolyze waste millet vinegar dregs, and uses the enzymatic hydrolyzate as the main fermentation substrate to ferment and produce bacterial cellulose. The fermentation method is safe and convenient, and not only has a high yield and is stable and controllable, and has high production efficiency, but also greatly reduces production costs, rationally utilizes vinegar dregs, and increases the added value of by-products after deep processing of millet.

[0022] 5. The preparation method of a bacterial cellulose facial mask of the present invention adopts a non-in situ synthesis method to prepare a finished bacterial cellulose facial mask. Astaxanthin and arbutin in the facial mask ingredients can modify the structure of the bacterial cellulose, so that the facial mask not only has antioxidant properties but also has certain whitening ability. Compared with the in situ synthesis method, this method can efficiently utilize functional substances and greatly reduce the waste of functional substances. The prepared facial mask is more comfortable and safe for the skin. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 This is a schematic photograph of the structure of Example 1 of the present invention after inoculation and culture in a fermenter; Figure 2 This is a photo of the bacterial cellulose wet film prepared in Example 1 of the present invention; Figure 3 This is a scanning electron microscope photograph of the bacterial cellulose dry film prepared in Example 1 of the present invention; Figure 4 This is a distribution diagram of the diameters of cellulose filaments in the bacterial cellulose dry film prepared in Example 1 of the present invention; Figure 5 This is a characterization diagram of the dry resistance of the bacterial cellulose wet film prepared in Example 1 of the present invention in a 37° C. constant temperature incubator; Figure 6 This is a characterization diagram of the rehydration performance of the bacterial cellulose dry film prepared in Example 1 of the present invention; Figure 7 This is a thermal stability characterization diagram of the bacterial cellulose wet film prepared in Example 1 of the present invention; Figure 8 This is a photo of the bacterial cellulose facial mask prepared in Example 1 of the present invention. DETAILED DESCRIPTION

[0024] The specific implementation of the present invention is described in detail below in conjunction with the accompanying drawings and examples, but it should be understood that the protection scope of the present invention is not limited by the specific implementation. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work belong to the scope of protection of the present invention. The experimental methods in the embodiments of the present invention are conventional methods unless otherwise specified.

[0025] The first purpose of the present invention is to provide a method for producing bacterial cellulose by using millet vinegar residues using microorganisms. The method uses an enzymatic method to prepare millet vinegar residue hydrolyzate as an alternative carbon source, and ferments microorganisms to produce bacterial cellulose. The fermentation method is not only safe and convenient, but also ensures stable and reliable output while improving the yield, fully achieving waste utilization, and greatly reducing production costs.

[0026] The second object is to provide a bacterial cellulose film obtained by the above method.

[0027] The third purpose is to provide a facial mask prepared using the bacterial cellulose film obtained above.

[0028] To achieve the above objectives, the inventors of this application have provided the technical solution of the present invention after long-term research and extensive practice. The specific technical contents are as follows: A bacterial cellulose facial mask is prepared by soaking a wet bacterial cellulose film dried in advance to a thickness of 0.75-1.25 mm in a mixed essence solution for 20-40 minutes. The mixed essence solution consists of the following components in percentage by mass: 1-1.5% astaxanthin, 1-1.5% arbutin, 4-6% glycerol and the remainder ultrapure water. After the raw material components of the mixed essence solution are uniformly mixed, it is necessary to filter and sterilize it using a filter membrane with a pore size of 0.20-0.25 μm.

[0029] A method for preparing a bacterial cellulose facial mask is as follows: s1. Preparation of millet vinegar lees enzymatic hydrolysate According to the solid-liquid ratio of 15-25 mL / g, distilled water and millet vinegar dregs are mixed and stirred at a temperature of 26-30°C for 2.5-3.5 h, and then cellulase is added thereto in an amount of 5500-6500 U / g of cellulose, and enzymatic hydrolysis is carried out at a pH of 4.6-5.0 and a temperature of 50-60°C for 8-10 h. After cooling to room temperature, saccharifying enzyme is added thereto in an amount of 1500-2500 U / g of starch, and secondary enzymatic hydrolysis is carried out at a pH of 3.8-4.2 and a temperature of 58-62°C for 2.5-3.5 h. After filtration, the millet vinegar dregs enzymatic hydrolyzate is obtained and set aside.

[0030] s2. Preparation of fermentation medium Add the millet vinegar lees enzymatic hydrolyzate, nitrogen source, inorganic salt and accelerator prepared in step s1 into distilled water, mix thoroughly, adjust the pH to 2.5-3.5, then sterilize at 115-121° C. for 20-30 min, cool to room temperature to obtain a fermentation medium for producing bacterial cellulose, and set aside; The nitrogen source is yeast powder and corn steep liquor; the inorganic salt is KH2PO4 and MgSO4; the promoter is citric acid and anhydrous ethanol. The amount of each component added per liter of fermentation medium is 180-220 mL of millet vinegar residue enzymatic hydrolyzate, 8-12 g of yeast powder, 2-3 g of corn steep liquor, 4-6 g of KH2PO4, 8-12 g of MgSO4, 18-22 g of citric acid, 20-40 mL of anhydrous ethanol, and the balance is distilled water.

[0031] s3. Inoculation and culture When the fermentation medium is cooled to 25-35° C., the fermentation medium prepared in step s2 is inoculated with Bacillus tame SW-1 at a mass ratio of 7-9%, and then the medium is first placed in a shaker at 70-90 r / min and 34-38° C. for 6-18 h, and then placed in a constant temperature incubator at 34-38° C. for static culture for 3-5 d, and then the fermentation medium is inoculated with Lactobacillus paracasei at a mass ratio of 2-4%, and placed in a constant temperature incubator at 34-38° C. for static culture for 4-6 d; Among them, the tame colliform bacillus SW-1 is classified and named Komagataeibacterobeodiens SW-1 was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on May 17, 2021, with the deposit number CGMCC NO.22548.

[0032] The preparation method of the seed liquid of Bacillus tame SW-1 and Lactobacillus paracasei is as follows: firstly, the original strain preserved in glycerol is inoculated into a liquid culture medium, and cultured for 36-60 hours at a temperature of 28-32°C to obtain a first-level seed liquid of the strain; then, the first-level seed liquid is inoculated into a basic culture medium, and cultured for 36-60 hours at a temperature of 28-32°C to obtain a strain with a concentration of (1-5)×10 7 CFU / mL of bacterial seed solution can be used for inoculation.

[0033] s4. Preparation of bacterial cellulose wet film The bacterial cellulose membrane on the upper layer of the fermentation liquid obtained after the inoculation culture in step s3 is taken out, placed in a NaOH solution with a concentration of 0.1 mol / L, and treated in a water bath at a temperature of 75-85°C for 50-70 min. Thereafter, it is transferred to a glacial acetic acid solution with a concentration of 0.1 mol / L for soaking for 50-70 min. After completion, it is taken out and repeatedly rinsed with distilled water until the pH value is neutral to obtain a finished bacterial cellulose wet membrane.

[0034] s5. Preparation of bacterial cellulose dry film The wet bacterial cellulose film obtained in step s4 is placed in an oven at 60-70° C. and dried to a constant weight to obtain a dry bacterial cellulose film.

[0035] s6. Preparation of bacterial cellulose facial mask Ultrapure water, astaxanthin, arbutin and glycerin are used as raw materials, and are evenly mixed to prepare a mixed essence solution with the mass percentage of 1-1.5% astaxanthin, 1-1.5% arbutin, 4-6% glycerin and the remainder ultrapure water, and the mixed essence solution is filtered and sterilized using a filter membrane with a pore size of 0.20-0.25 μm; then, the bacterial cellulose wet film obtained in step s4 is dried to a thickness of 0.75-1.25 mm, cut into shape using a cutting machine to obtain a mask cloth, and then the mask cloth is immersed in the mixed essence solution for 20-40 min to obtain a finished bacterial cellulose mask.

[0036] A bacterial cellulose mask and a preparation method thereof of the present invention, using millet vinegar dregs as the main raw material to produce a bacterial cellulose mask with low raw material cost, high yield, good process controllability and good use effect, which can not only utilize waste, but also has the advantages of simple operation and low production cost, greatly improves the yield of bacterial cellulose and can ensure stable and controllable output, which is of great significance for reducing environmental pollution, reducing the production cost of bacterial cellulose, and improving the performance of the mask. Astaxanthin and arbutin in the mixed essence solution have strong antioxidant properties, and they should be introduced into bacterial cellulose as active ingredients to prepare a mask with strong antioxidant ability, achieve better skin care effects, and better supplement the mask market. The prepared finished bacterial cellulose mask not only has antioxidant ability, but also has higher porosity, and can effectively deliver active ingredients to different skin layers under controllable conditions, and has excellent practical application value and development prospects.

[0037] Example 1 A bacterial cellulose facial mask of this embodiment specifically comprises the following steps: (1) Activation of Bacillus SW-1 and Lactobacillus paracasei The original species of Bacillus tame SW-1 and Lactobacillus paracasei were inoculated into the liquid culture medium of Bacillus tame to obtain the primary seed liquid; (2) Expansion culture of Bacillus SW-1 and Lactobacillus paracasei The first-level seed solution obtained in step (1) was inoculated into the liquid culture medium of Bacillus docileus and the liquid culture medium of MRS to obtain the second-level seed solution, so that the bacterial concentration of each strain reached 2.5×10 7 CFU / mL.

[0038] (3) Preparation of millet vinegar lees enzymatic hydrolysate The vinegar lees and distilled water were mixed at a solid-liquid ratio of 1:20 (g:mL), stirred at 28°C for 3 h, then cellulase was added at a content of 6000 U / g cellulose, and enzymatic hydrolysis was carried out at pH 4.8 and 55°C for 9 h. After cooling to room temperature, saccharifying enzyme was added at a content of 2000 U / g starch, and enzymatic hydrolysis was carried out at pH 4.0 and 60°C for 3 h. The millet vinegar lees enzymatic hydrolysate was obtained by filtration.

[0039] (4) Preparation of bacterial cellulose fermentation medium The addition amount per liter is 10 g of yeast powder, 2.5 g of corn pulp, 20 g of citric acid, 5 g of KH2PO4, 10 g of MgSO4, 30 mL of anhydrous ethanol, 200 mL of millet vinegar residue enzymatic hydrolyzate, and the rest is distilled water. Mix them to make a fermentation medium for producing bacterial cellulose, and adjust the pH value of the medium to 3.0, then sterilize it at 115°C for 30 min.

[0040] (5) Fermentation of bacterial cellulose When the fermentation medium was cooled to 30°C, it was inoculated with Bacillus tame SW-1 at a ratio of 8%. The inoculated culture solution was cultured in a shaker at 80 r / min and 36°C for 12 h, and then placed in a 36°C constant temperature incubator for static culture for 4 d. Thereafter, it was inoculated with Lactobacillus paracasei at an inoculation ratio of 3%, and then placed in a 36°C constant temperature incubator for static culture for 5 d.

[0041] (6) Preparation of bacterial cellulose The bacterial cellulose film on the upper surface of the bacterial cellulose fermentation liquid obtained in step (5) was taken out, and a 0.1 mol / L NaOH solution was added, and the film was placed in a water bath at 80°C for 60 min, and then soaked in a 0.1 mol / L glacial acetic acid solution for 60 min, and then repeatedly rinsed with distilled water until the pH value was neutral to obtain a bacterial cellulose wet film, which was then dried in an oven at 65°C to constant weight to obtain a bacterial cellulose dry film.

[0042] (7) Prepare a mixed essence solution containing 1.25% astaxanthin, 1.25% arbutin and 5% glycerol with ultrapure water, and filter and sterilize with a 0.22 μm filter membrane. Dry the wet bacterial cellulose film obtained in step (6) to a thickness of 1 mm, and soak it in the mixed essence solution for 30 minutes to obtain a finished bacterial cellulose facial mask.

[0043] It was determined that the bacterial cellulose dry film prepared in step (6) of this example had a bacterial cellulose yield (dry weight) of 7.81 g / L and a yield of 86.78 g / kg culture.

[0044] At the same time, the cellulose diameter of the bacterial cellulose membrane obtained by this method is about 84.17 nm. The nano-level cellulose membrane can make the mask fit well and fully contact the skin. At the same time, the porosity is improved, which not only has strong dry resistance, but also can maintain excellent rehydration performance and high thermal stability. At the same time, due to the presence of pores, the bacterial cellulose mask has a certain absorption capacity of functional ingredients, which is suitable as a base material for the mask.

[0045] Example 2 A bacterial cellulose facial mask of this embodiment specifically comprises the following steps: Wherein, steps (1), (2), (4), (5), (6) and (7) are the same as those in Example 1.

[0046] (3) Preparation of millet vinegar lees enzymatic hydrolysate V1: Mix vinegar lees and distilled water at a solid-liquid ratio of 1:15 (g:mL), stir at 30℃ for 2.5 h, then add cellulase at a content of 6500 U / g cellulose, and hydrolyze at pH 5.0 and 50℃ for 8 h. After cooling to room temperature, add saccharifying enzyme at a content of 1500 U / g starch, and hydrolyze at pH 3.8 and 62℃ for 3.5 h. Filter and obtain millet vinegar lees hydrolyzate.

[0047] V2: The vinegar lees were mixed with distilled water at a solid-liquid ratio of 1:25 (g:mL), stirred at 26°C for 2.5 h, then cellulase was added at a content of 6500 U / g cellulose, and enzymatic hydrolysis was carried out at pH 5.0 and 60°C for 8 h. After cooling to room temperature, saccharifying enzyme was added at a content of 1500 U / g starch, and enzymatic hydrolysis was carried out at pH 4.2 and 58°C for 3 h. The millet vinegar lees enzymatic hydrolysate was obtained by filtration.

[0048] V3: Mix vinegar lees and distilled water at a solid-liquid ratio of 1:25 (g:mL), stir at 26°C for 3.5 h, then add cellulase at a content of 5500 U / g cellulose, and hydrolyze at pH 4.6 and 50°C for 10 h. After cooling to room temperature, add saccharifying enzyme at a content of 2500 U / g starch, and hydrolyze at pH 4.2 and 62°C for 3 h. Filter and obtain millet vinegar lees hydrolyzate.

[0049] V4: The vinasse was mixed with distilled water at a solid-liquid ratio of 1:20 (g:mL), stirred at 28°C for 3 h, then cellulase was added at a content of 6000 U / g cellulose, and enzymatic hydrolysis was performed at pH 4.8 and 55°C for 9 h. After cooling to room temperature, saccharifying enzyme was added at a content of 2000 U / g starch, and enzymatic hydrolysis was performed at pH 4.0 and 60°C for 3 h. The vinasse enzymatic hydrolyzate was obtained by filtration.

[0050] After measurement, the test results are shown in Table 1 below.

[0051] Table 1 Example 3 A bacterial cellulose facial mask of this embodiment specifically comprises the following steps: Wherein, steps (1), (2), (3), (6) and (7) are the same as those in Example 1.

[0052] (4) Preparation of bacterial cellulose fermentation medium H1: Each liter of culture medium includes 8 g yeast powder, 2 g corn steep liquor, 18 g citric acid, 6 g KH2PO4, 12 g MgSO4, 40 mL anhydrous ethanol, 220 mL millet vinegar residue enzymatic hydrolyzate, and the rest is distilled water. The pH is adjusted to 2.5 and sterilized at 115°C for 30 min.

[0053] H2: Each liter of culture medium includes 12 g yeast powder, 3 g corn steep liquor, 22 g citric acid, 4 g KH2PO4, 8 g MgSO4, 20 mL anhydrous ethanol, 180 mL millet vinegar lees enzymatic hydrolyzate, and the rest is distilled water. The pH is adjusted to 3.5 and sterilized at 115°C for 30 min.

[0054] H3: Each liter of culture medium includes 10 g of peptone, 2.5 g of corn steep liquor, 15 g of tartaric acid, 5 g of KH2PO4, 14 g of MgSO4, 30 mL of lactic acid, 200 mL of distiller's grains enzymatic hydrolyzate, and the rest is distilled water. The pH is adjusted to 3 and sterilized at 115°C for 30 min.

[0055] H4: Each liter of culture medium includes 10 g yeast powder, 2.5 g beef extract, 15 g citric acid, 5 g Na2HPO4, 14 g CaCO3, 30 mL anhydrous ethanol, 200 mL millet vinegar hydrolysate, and the rest is distilled water. The pH is adjusted to 3 and sterilized at 115°C for 30 min.

[0056] H5: Each liter of culture medium includes 10 g yeast powder, 15 g citric acid, 5 g KH2PO4, 10 g MgSO4, 30 mL anhydrous ethanol, 200 mL millet vinegar lees enzymatic hydrolyzate, and the rest is distilled water. The pH is adjusted to 3 and sterilized at 115°C for 30 min.

[0057] H6: Each liter of culture medium includes 10 g yeast powder, 5 g KH2PO4, 10 g MgSO4, 30 mL anhydrous ethanol, 200 mL millet vinegar lees enzymatic hydrolyzate, and the rest is distilled water. The pH is adjusted to 3 and sterilized at 115°C for 30 min.

[0058] H7: Each liter of culture medium includes 10 g yeast powder, 35 g sucrose, 2.5 g corn steep liquor, 15 g citric acid, 5 g KH2PO4, 10 g MgSO4, 30 mL anhydrous ethanol, 200 mL millet vinegar lees enzymatic hydrolyzate, and the rest is distilled water. The pH is adjusted to 3 and sterilized at 115°C for 30 min.

[0059] H8: Each liter of culture medium includes 10 g yeast powder, 35 g mannitol, 2.5 g corn steep liquor, 15 g citric acid, 5 g KH2PO4, 10 g MgSO4, 30 mL anhydrous ethanol, 200 mL millet vinegar lees enzymatic hydrolyzate, and the rest is distilled water. The pH is adjusted to 3 and sterilized at 115°C for 30 min.

[0060] (5) Fermentation of bacterial cellulose The docile Bacillus fossa obtained in step (2) is inoculated into the bacterial cellulose fermentation medium H1, 3, 5, 7 in step (4) at an inoculation rate of 7%, and then the inoculated culture solution is cultured in a shaking incubator at 90 r / min and 38°C for 6 h, and then placed in a 38°C constant temperature incubator for static culture for 3 days, and then inoculated with Lactobacillus paracasei at an inoculation rate of 4%, and then placed in a 38°C constant temperature incubator for static culture for 6 days.

[0061] The docile Bacillus obtained in step (2) is inoculated into the H2, 4, 6, 8 bacterial cellulose fermentation medium in step (4) at an inoculation rate of 9%, and then the inoculated culture solution is cultured in a shaker at 70 r / min and 34°C for 18 h, and then placed in a 34°C constant temperature incubator for static culture for 5 days, and then inoculated with Lactobacillus paracasei at an inoculation rate of 2%, and then placed in a 34°C constant temperature incubator for static culture for 4 days.

[0062] After measurement, the test results are shown in Table 2 below.

[0063] Table 2 As shown in Table 2, the yield of the bacterial cellulose dry film obtained in Example 1 can reach 7.81 g / L, and the yield can reach 86.78 g / kg culture.

[0064] Example 4 A bacterial cellulose facial mask of this embodiment specifically comprises the following steps: S0: Referring to the mask making method described in Example 1, the obtained composite mask was tested, and the antioxidant capacity DPPH free radical scavenging rate could reach 52.50%.

[0065] M1: Take the wet bacterial cellulose film obtained in step (6) of Example 1 and dry it to a thickness of 1 mm, then cut it into shape using a cutting machine to obtain a facial mask cloth; use ultrapure water to prepare a mixed essence solution containing 1.25% astaxanthin, 1.25% arbutin, 5% glycerol, and the remainder ultrapure water, and filter and sterilize it using a filter membrane with a pore size of 0.22 μm, and soak the prepared facial mask cloth in the mixed essence solution for 40 minutes to obtain a finished bacterial cellulose facial mask. After testing, the antioxidant capacity of the facial mask, the DPPH free radical scavenging rate can reach 45.45%.

[0066] M2: The wet bacterial cellulose film obtained in step (6) of Example 1 was dried to a thickness of 1 mm, and then cut into shape using a cutting machine to obtain a facial mask cloth; a mixed essence solution containing 1.5% astaxanthin, 1.0% arbutin, and 6% glycerol, as well as the remainder of ultrapure water, was prepared using ultrapure water, and the solution was filtered and sterilized using a filter membrane with a pore size of 0.25 μm, and the prepared facial mask cloth was soaked in the mixed essence solution for 20 minutes to obtain a finished bacterial cellulose facial mask. After testing, the antioxidant capacity of the facial mask, the DPPH free radical scavenging rate, can reach 43.18%.

[0067] Test determination Figure 1 This is a schematic structural photograph of Example 1 of the method of the present invention after inoculation and culture in a fermenter; from Figure 1It can be seen that the bacterial cellulose wet film prepared by the method of the present invention is mainly distributed on the gas-liquid contact surface.

[0068] Figure 2 This is a morphological diagram of the bacterial cellulose wet film prepared in Example 1 of the method of the present invention; Depend on Figure 2 It can be seen that the bacterial cellulose wet film prepared by the method of the present invention has a smooth surface, is evenly distributed, and is of moderate size, and can be used as a substrate for a facial mask.

[0069] Figure 3 This is a scanning electron microscope image of the bacterial cellulose dry film prepared in Example 1 of the present invention; Depend on Figure 3 It can be seen that the bacterial cellulose prepared by the method of the present invention is a three-dimensional network structure that is cross-linked with each other and has abundant pores inside, wherein the pores can create an optimal microenvironment for effectively delivering active skin care ingredients.

[0070] Figure 4 This is a distribution diagram of the diameters of cellulose filaments in the bacterial cellulose dry film prepared in Example 1 of the present invention; Depend on Figure 4 It can be seen that the diameter of the bacterial cellulose prepared by the method of the present invention is about 84.17 nm, which is consistent with the common diameter of bacterial cellulose and is evenly distributed.

[0071] Figure 5 This is a characterization diagram of the dry resistance of the bacterial cellulose wet film prepared in Example 1 of the method of the present invention in a 37° C. constant temperature incubator; Depend on Figure 5 It can be seen that the bacterial cellulose prepared by the method of the present invention has good dryness resistance. After being placed in a constant temperature incubator at 37°C for 300 min, the moisture content is completely reduced to 0%.

[0072] Figure 6 This is a characterization diagram of the rehydration performance of the bacterial cellulose wet film prepared in Example 1 of the method of the present invention after being pressed to different initial moisture contents; Depend on Figure 6 It can be seen that the bacterial cellulose prepared by the method of the present invention has good rehydration performance. When the initial moisture content is above 30%, the bacterial cellulose wet film can be rehydrated to 100%, and when the initial moisture content is 15%, it can still be rehydrated to about 60%.

[0073] Figure 7 This is a thermal stability characterization diagram of the bacterial cellulose wet film prepared in Example 1 of the method of the present invention; Depend on Figure 7 It can be seen that the bacterial cellulose prepared by the method of the present invention has good thermal stability and can be used after high temperature sterilization.

[0074] Figure 8 This is a photo of a facial mask using a bacterial cellulose film as a substrate, prepared in Example 1 of the method of the present invention; Depend on Figure 8 It can be seen that the bacterial cellulose facial mask prepared by the method of the present invention has basic facial mask characteristics and can be used as a facial mask substrate.

Claims

1. A bacterial cellulose facial mask, characterized in that: The facial mask is prepared by soaking a bacterial cellulose wet film in a mixed essence solution, wherein the mixed essence solution is composed of the following components in percentage by mass: 1-1.5% astaxanthin, 1-1.5% arbutin, 4-6% glycerol and the remainder ultrapure water.

2. A bacterial cellulose facial mask according to claim 1, characterized in that: The bacterial cellulose wet film needs to be dried to a thickness of 0.75-1.25 mm before being immersed in the mixed essence solution.

3. A bacterial cellulose facial mask according to claim 1 or 2, characterized in that: The bacterial cellulose wet film is immersed in the mixed essence solution for 20-40 minutes.

4. A bacterial cellulose facial mask according to claim 1, characterized in that: After the raw material components of the mixed essence solution are uniformly mixed, it needs to be filtered and sterilized.

5. A bacterial cellulose facial mask according to claim 4, characterized in that: The pore size of the filter membrane used for filtration sterilization is 0.20-0.25 μm.

6. A bacterial cellulose facial mask according to claim 1, characterized in that: The preparation method of the bacterial cellulose wet film is: s1. Preparation of millet vinegar lees enzymatic hydrolysate According to the ratio of solid-liquid ratio of 15-25 mL / g, distilled water and millet vinegar dregs are mixed and stirred at a temperature of 26-30°C for 2.5-3.5 h, then, cellulase is added thereto at an addition amount of 5500-6500 U / g of cellulose, and enzymatic hydrolysis is performed for 8-10 h at a pH of 4.6-5.0 and a temperature of 50-60°C, and after cooling to room temperature, saccharifying enzyme is added thereto at an addition amount of 1500-2500 U / g of starch, and secondary enzymatic hydrolysis is performed for 2.5-3.5 h at a pH of 3.8-4.2 and a temperature of 58-62°C, and after suction filtration, millet vinegar dregs enzymatic hydrolyzate is obtained for standby use; s2. Preparation of fermentation medium Add the millet vinegar lees enzymatic hydrolyzate, nitrogen source, inorganic salt and accelerator obtained in step s1 into distilled water, mix thoroughly, adjust the pH to 2.5-3.5, then sterilize at 115-121° C. for 20-30 min, cool to obtain a fermentation medium, and set aside; s3. Inoculation and culture Under the temperature condition of 25-35°C, inoculate the fermentation medium prepared in step s2 with Bacillus tame SW-1 at a mass ratio of 7-9%, then place it in a shaker at 70-90 r / min and 34-38°C for 6-18 h, then place it in a constant temperature incubator at 34-38°C for static culture for 3-5 d, then inoculate the fermentation medium with Lactobacillus paracasei at a mass ratio of 2-4%, and place it in a constant temperature incubator at 34-38°C for static culture for 4-6 d; s4. Preparation of bacterial cellulose wet film The bacterial cellulose membrane on the upper layer of the fermentation liquid obtained after the inoculation culture in step s3 is taken out, placed in a NaOH solution with a concentration of 0.1 mol / L, and treated in a water bath at a temperature of 75-85°C for 50-70 min. Thereafter, it is transferred to a glacial acetic acid solution with a concentration of 0.1 mol / L for soaking for 50-70 min. After completion, it is taken out and repeatedly rinsed with distilled water until the pH value is neutral to obtain a finished bacterial cellulose wet membrane.

7. A bacterial cellulose facial mask according to claim 1, characterized in that: In step s2, the nitrogen sources are yeast powder and corn steep liquor; the inorganic salts are KH2PO4 and MgSO4; and the accelerators are citric acid and anhydrous ethanol.

8. A bacterial cellulose facial mask according to claim 7, characterized in that: In the fermentation medium, the addition amount of each component per liter is 180-220 mL of millet vinegar residue enzymatic hydrolyzate, 8-12 g of yeast powder, 2-3 g of corn steep liquor, 4-6 g of KH2PO4, 8-12 g of MgSO4, 18-22 g of citric acid, 20-40 mL of anhydrous ethanol, and the balance is distilled water.

9. A bacterial cellulose facial mask according to claim 1, characterized in that: In step s3, the tame bacillus SW-1 is classified as Komagataeibacter obeodiens SW-1 was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on May 17, 2021, with the deposit number CGMCC NO.22548.

10. A bacterial cellulose facial mask according to claim 1, characterized in that: In step s3, the preparation method of the seed liquid of Bacillus tame SW-1 and Lactobacillus paracasei is as follows: first, the original strain preserved in glycerol is inoculated into a liquid culture medium, and cultured at a temperature of 28-32°C for 36-60 h to obtain a first-level seed liquid of the strain; then, the first-level seed liquid is inoculated into a basic culture medium, and cultured at a temperature of 28-32°C for 36-60 h to obtain a strain with a concentration of (1-5)×10 7 CFU / mL of bacterial seed solution can be used for inoculation.

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