CN108324633B

CN108324633B - Preparation method of brown rice fermentation stock solution for cosmetics and product thereof

Info

Publication number: CN108324633B
Application number: CN201810171483.XA
Authority: CN
Inventors: 陆震; 魏玉洁; 孙元军; 石艳丽
Original assignee: Bloomage Biotech Co Ltd; Shandong Bloomage Hyinc Biopharm Co Ltd
Current assignee: Huaxi Biotechnology Tianjin Co ltd; Bloomage Biotech Co Ltd
Priority date: 2018-03-01
Filing date: 2018-03-01
Publication date: 2021-08-13
Anticipated expiration: 2038-03-01
Also published as: CN108324633A

Abstract

The invention provides a preparation method of brown rice fermentation stock solution for cosmetics. The preparation method provided by the invention can efficiently enrich the active substances in the brown rice in a very short time by controlling the fermentation process, has the advantages of short period, controllable process, easily-amplified process, stable product, high content of the active substances, good effect, easy absorption, safety and health, and meets the requirements of cosmetic raw materials.

Description

Preparation method of brown rice fermentation stock solution for cosmetics and product thereof

Technical Field

The invention relates to the field of cosmetic raw materials, and in particular relates to a preparation method of a high-activity brown rice fermentation stock solution for cosmetics.

Background

China is a large country for rice production and consumption, the annual output is more than 2 hundred million tons, the brown rice is different from common polished rice, namely the brown rice has no glume, the brown rice not only contains endosperm parts, but also retains a bran layer removed in the whitening process, the bran layer is a main nutrient component enrichment part in the rice, and the bran layer contains more active components, such as vitamins, phytosterol, phenolic acid, gamma-aminobutyric acid and the like, wherein the content of the gamma-aminobutyric acid is highest. Gamma-aminobutyric acid is an inhibitory neurotransmitter in the central nervous system, has extremely important physiological functions, has good effects of resisting oxidation, whitening skin and the like on human skin, is mainly applied to the field of foods at present, rarely relates to the field of cosmetics, and is widely commented on products which are sold as cosmetic raw materials by rice at home, particularly in Japan, such as face cream, facial mask and the like, so that the development of the cosmetic raw materials of rice, particularly brown rice, has very important significance.

The rice extract is prepared by gelatinizing and saccharifying raw materials, adding stabilizer and emulsifier, and extracting with water. The method is low in efficiency, active ingredients of the rice are seriously damaged in a high-temperature environment, the rice is difficult to extract and retain efficiently, the content of gamma-aminobutyric acid serving as a main functional ingredient is low, most of the active ingredients exist in the brown rice, however, most of raw material products of rice extracts in the market mainly adopt the method, and how to release the active ingredients by using a mild method is the key to successfully utilize the brown rice, and the requirement can be met by microbial fermentation. Part of tissues of the brown rice can be fermented to fully degrade polysaccharide components with simple structures, so that a carbon source required by growth of microorganisms is provided, and active substances existing on the polysaccharide in a binding state in the brown rice are released, so that the fermentation liquor has functional activity. Based on the fermentation metabolism regulation theory, the fermentation period can be shortened by adopting a yeast cell conversion mode, the content of an active ingredient gamma-aminobutyric acid can be efficiently improved by adding coenzyme, in addition, the yeast can also metabolize other various active products in the growth process, such as various amino acids, sterol, phytic acid, vitamins and the like which are necessary for small molecules, the nutrient density and the physiological activity are greatly improved, so that the nutrient substances of the brown rice after fermentation are further concentrated, and the value of the brown rice is far higher than that of the brown rice.

So far, many studies on fermented brown rice have been made, for example, chinese patent CN104000266A discloses "a method for preparing a rice fermented beverage", which is characterized in that rice is soaked, sieved and gelatinized, then enzymatic hydrolysate is added for enzymolysis, and then lactobacillus is used for fermentation to obtain a finished product. The disadvantage is that lactic acid bacteria are fermented, the final product has high acidity, and the lactic acid bacteria can be applied to the food field, but if the lactic acid bacteria are used as cosmetic raw materials in a formula, the lactic acid bacteria easily cause damage to the skin epidermis.

Chinese patent CN104212698 discloses a method for preparing fermented brown rice vinegar, which comprises carrying out enzymolysis on germinated brown rice, carrying out liquid fermentation by using yeast, and carrying out solid fermentation by using acetic acid bacteria liquid to obtain the fermented brown rice vinegar with high nutritional activity. The disadvantages of the method include (1) the seed liquid is utilized to ferment, the fermentation period is longer, the amplification is not easy, and the large-scale production is difficult to realize; (2) the metabolism is difficult to accurately control the fermentation metabolic pathway by utilizing the two-step fermentation process, and the product stability is difficult to ensure; (3) the second fermentation step using acetic acid bacteria also has the problem of over-high acidity.

Chinese patent CN1957752A discloses a brown rice fermentation extract, wherein brown rice is fermented by yeast twice to obtain fermentation liquor, and the disadvantages of the brown rice fermentation are that yeast is not inoculated in the first fermentation step, fermentation metabolism control is lacked, pathogenic bacteria are easy to infect, and safety can not be ensured.

Chinese patent CN1386423A discloses a rice fermented beverage and a preparation method thereof, rice is gelatinized and inoculated with microorganisms for fermentation for 1-5 days, and then the rice is filtered and sterilized to obtain a finished product, which has the defects that the raw material is peeled rice, rice bran is wasted, and the active ingredients of the fermented product do not relate to the content of gamma-aminobutyric acid, which is related to the fact that the fermentation strain is purchased from the market and has no conversion capability on the gamma-aminobutyric acid; meanwhile, the fermentation period is longer, and can reach 5 days.

Chinese patent CN103705436 discloses a germinated brown rice herbal skin care product and a preparation method thereof, brown rice powder is gelatinized and enzymolyzed and then compounded with herbal essence to obtain a finished product, and the disadvantage is that the extraction rate of active ingredients is not high by adopting a common extraction method.

In summary, the rice raw materials in the current market mainly have the following disadvantages:

1. the main source of the raw material is peeled rice, and rich phytosterol rice bran is directly discarded.

2. The rice fermentation raw material is mainly applied to the field of foods, and the field of cosmetics relates to a few;

3. most of rice fermentation products are usually evaluated in the food field only by taking physical characteristics such as appearance states as guidance, and the most important fermentation process is guided by taking active ingredients of the products as guidance, particularly the content of gamma-aminobutyric acid, is ignored, and meanwhile, product efficacy data are lacked, which are very important factors applied to the cosmetic field;

4. the existing rice fermentation patents lack intermediate control of fermentation, and mostly adopt multi-step fermentation, and the defects of the multi-step fermentation are that the intermediate process is difficult to control, so that the process is difficult to amplify, the product stability is difficult to ensure, and on the other hand, the fermentation period is generally longer and the efficiency is not high.

Disclosure of Invention

Aiming at the problems in the prior art, the invention provides the preparation method of the brown rice fermentation stock solution for cosmetics, which has the advantages of short production period, stable product, high active substance content, safety and health, and capability of meeting the requirements of cosmetic raw materials.

In order to achieve the purpose, the invention adopts the following technical scheme.

A method for preparing brown rice fermentation stock solution for cosmetics comprises the following steps:

(1) soaking brown rice in water for accelerating germination, drying, and pulverizing into brown rice powder;

(2) adding water into brown rice powder, mixing to obtain rice slurry, adding enzyme, and performing enzymolysis to obtain rice slurry enzymatic hydrolysate;

(3) inoculating yeast cells into a culture solution containing rice pulp enzymatic hydrolysate, adding coenzyme, and fermenting to the end point to obtain a fermentation solution;

(4) removing impurities from the fermentation liquor, and sterilizing to obtain the finished product.

In the step (1), the soaking amount is 1.5-2 times of the mass of the brown rice, and the germination accelerating time is 12-24 h.

In the step (1), the drying temperature is 60-80 ℃, and the drying time is 5-10 h.

In the step (1), the moisture content of the brown rice powder is less than 10%; the fineness is that the mixture is sieved by a sieve of 80-200 meshes.

In the step (2), the brown rice powder in the rice milk accounts for 5-10% by mass.

In the step (2), the adding amount of the enzyme is 0.002-0.2% of the mass of the rice milk.

In the step (2), the enzyme is amylase and saccharifying enzyme, preferably alpha-amylase and alpha-saccharifying enzyme. Preferably, the enzyme activity is preferably 100-150U/g and 300-400U/g respectively; the ratio of the two is preferably 1: 1-2.

In the step (2), the enzymolysis temperature is 20-50 ℃, and the enzymolysis time is 0.5-3 h.

In the step (2), the glucose content at the enzymolysis end point at intervals of 0.5h is not changed any more. The glucose content can be determined by a conventional chemical method such as DNS method, Fehling's reagent method, indirect iodometric method, and optical rotation method.

In the step (3), the culture solution further contains a nitrogen source and inorganic salts. The nitrogen source is common nitrogen source for microorganism culture, such as beef extract, peptone, yeast powder, and soybean cake powder. The inorganic salt is preferably dipotassium hydrogen phosphate.

In the step (3), the addition amount of the yeast cells is 0.5-1% wt.

In the step (3), the yeast cells are obtained by: the yeast strain is inoculated in a sterile seed culture solution containing 4-8 wt% of carbon source, 0.5-1 wt% of nitrogen source and 0.1-0.5 wt% of inorganic salt, the rotation speed of a shaking table is 150-200rpm, the culture is carried out at the temperature of 25-35 ℃ until the logarithmic phase, and the strain is obtained by centrifuging and discarding the supernatant. The centrifugal speed is 8000-10000 rpm, and the centrifugal time is 30-120 min. The carbon source is a common carbon source for microbial culture, such as sucrose and glucose. The nitrogen source is common nitrogen source for microorganism culture, such as beef extract, peptone, yeast powder, and soybean cake powder. The inorganic salt is preferably dipotassium hydrogen phosphate.

In the step (3), the yeast is selected from safe strains applicable to the field of cosmetics; preferably Saccharomyces cerevisiae (Saccharomyces cerevisiae) Grape juice yeast (1)Saccharomyces uvarum) Rhodosporidium toruloides (A)Rhodospordium spp.) Beer yeast (Saccharomyces cerevisiaeHansen), Hansenula sporogenes (B.sp.), (B.sporum)Hanseniaspora spp.) Pichia pastoris (A), (B)Pichia spp.）。

In the step (3), the fermentation temperature is 25-35 ℃; the fermentation time is 15-20 h.

In the step (3), the pH value is controlled to be 5-5.5 in the fermentation process. As an optimization, pH was controlled by addition of glutamic acid.

In the step (3), the fermentation end point is that the fermentation liquor has no glucose. The glucose content can be determined by a conventional chemical method such as DNS method, Fehling's reagent method, indirect iodometric method, or optical rotation method.

In the step (3), the coenzyme is vitamin and derivatives thereof; preferably one or more of nicotinic acid, biotin, tetrahydrofolic acid or thiamine pyrophosphate; the addition amount is preferably 0.0001 to 0.001% by weight.

In the step (4), the impurity removal process is to remove the precipitate by centrifugation at 1000-.

In the step (4), the sterilization process is performed by filtering and sterilizing a filter element with the aperture of 0.22 mu m.

A brown rice fermentation stock solution obtained by the above preparation method is provided. The mass percentage content of the gamma-aminobutyric acid in the brown rice fermentation stock solution is preferably 0.15-0.25%.

An application of the brown rice fermentation stock solution in cosmetics is provided. The brown rice fermentation stock solution has the effects of resisting oxidation, whitening and repairing damage in cosmetics.

The invention has the following advantages:

(1) according to the invention, the brown rice is subjected to a germination accelerating mode before fermentation, so that the nutrient substances in the brown rice can be further enriched; (2) according to the invention, the brown rice is subjected to enzymolysis, so that nutrients existing in the brown rice in a combined state can be primarily converted into a free state, and the subsequent fermentation efficiency is improved, such as macromolecular starch is degraded into oligosaccharide or monosaccharide and the like; (3) the invention combines the metabolism regulation and control means, and the thallus cell conversion mode is adopted in the fermentation process, so that the fermentation efficiency can be effectively improved, and the fermentation period is shortened; (4) according to the invention, coenzyme is added in the fermentation process to improve the enzyme activity, so that the conversion efficiency of gamma-aminobutyric acid is further improved; (5) according to the invention, glutamic acid is supplemented to adjust the pH in the fermentation process, so that the fermentation process can be controlled in the optimum pH range of enzyme activity, and the yield of gamma-aminobutyric acid can be remarkably improved.

Drawings

FIG. 1 is an HPLC chromatogram for determining the content of gamma-aminobutyric acid in different samples;

FIG. 2 is a graph showing the results of promoting cell repair.

Detailed Description

The present invention will be further described with reference to the following examples and drawings, but the present invention is not limited to the following examples.

Weighing 100-150U/g alpha-amylase and 300-400U/g alpha-diastase to respectively prepare enzyme solutions, wherein the activities are both 500U/mL, and mixing the enzyme solutions for use in the examples and the comparative examples. Both alpha-amylase and alpha-saccharifying enzymes are commercially available.

Example 1 fresh cell preparation of Yeast

Selecting Hanseng spore yeast strain from a test tube, inoculating the Hanseng spore yeast strain into 500mL of sterile seed culture solution containing 4-8% wt of glucose, 0.5-1% wt of peptone and 0.1-0.5% wt of dipotassium hydrogen phosphate, culturing at the rotating speed of a shaking table of 150rpm at 30 ℃ for 13h, then, reaching the logarithmic phase, inoculating and expanding the strain into 5L of seed culture medium again, culturing for 10h for the second time until the logarithmic phase, centrifuging at 10000 rpm for 30min, and then, removing the supernatant to obtain the Hanseng spore yeast strain.

Example 2

(1) Weighing 10kg of brown rice, soaking the brown rice into 20kg of water for accelerating germination for 24h, drying the brown rice for 10h at the temperature of 60 ℃, and crushing the brown rice into brown rice powder which passes through 100 meshes completely, wherein the water content is 5.2%;

(2) taking 8.5kg of brown rice powder in the step (1), adding 100L of water, mixing to obtain rice slurry, adding 100mL of alpha-amylase liquid and alpha-diastase liquid, performing enzymolysis, measuring the glucose content every 0.5h, and obtaining rice slurry enzymolysis liquid when the glucose content is not changed any more, wherein the residual sugar content is 58.5 g/L;

(3) adding 5g/L of bean cake powder as a nitrogen source and 1g/L of dipotassium hydrogen phosphate as a culture solution into the rice pulp enzymatic hydrolysate, inoculating the saccharomycete thallus in the example 1, adding 1g of biotin at the same time, supplementing 55g of glutamic acid powder in the middle to maintain the pH between 5.0 and 5.5, fermenting at 30 ℃ for 20 hours, and after no residual sugar exists, ending the fermentation to obtain a fermentation liquid;

(4) centrifuging the fermentation liquid at 3000rpm to remove impurities, filtering the supernatant with 0.22 μm polyethersulfone filter element to obtain brown rice fermentation stock solution S1 with relative density of 1.01.

Example 3

(1) Weighing 3kg of brown rice, soaking the brown rice into 4.5kg of water for accelerating germination for 18h, drying the brown rice for 5h at 80 ℃, and crushing the brown rice into brown rice powder which passes through 150 meshes completely, wherein the water content is 6.2%;

(2) taking 2.2kg of brown rice powder in the step (1), adding 30L of water, mixing to obtain rice slurry, adding 25ml of alpha-amylase liquid and alpha-diastase liquid, performing enzymolysis, measuring the glucose content every 0.5h, and obtaining rice slurry enzymolysis liquid when the glucose content is not changed any more, wherein the residual sugar content is 63.1 g/L;

(3) adding 5g/L yeast powder as a nitrogen source and 1g/L dipotassium hydrogen phosphate as a culture solution into the rice pulp enzymatic hydrolysate, inoculating the yeast thallus in the embodiment 1, simultaneously adding 0.3g of nicotinic acid, supplementing 15g of glutamic acid powder in the middle to maintain the pH between 5.0 and 5.5, fermenting for 16 hours at 32 ℃, and after no residual sugar exists, finishing fermentation to obtain a fermentation liquid;

(4) centrifuging the fermentation liquid at 5000rpm to remove impurities, filtering the supernatant with 0.22 μm polyethersulfone filter element to obtain brown rice fermentation stock solution S2 with relative density of 1.01.

Comparative example 1

(1) Weighing 1kg of polished rice, baking at 80 deg.C for 30min, sufficiently drying to water content of 3.5%, pulverizing dried polished rice with a mill pulverizer, sieving with 40 mesh sieve to obtain about 860g of rice flour;

(2) adding about 8.5L of purified water, adding alpha-amylase to make the addition amount of the alpha-amylase be 0.01%, reacting at 90 ℃ for 10min, adding alpha-diastase to make the addition amount of the alpha-diastase be 0.05%, reacting at 60 ℃ for 1h, heating the hydrolysate to 100 ℃, keeping the temperature for 10min, and performing enzyme deactivation treatment to obtain an enzymatic hydrolysate;

(3) centrifuging the enzymolysis solution at 5000rpm to remove impurities, and collecting supernatant as rice hydrolysis extract C1.

Comparative example 2

The method steps are the same as example 2, except that no coenzyme is added in the yeast fermentation process, and brown rice fermentation liquor C2 is obtained.

Comparative example 3

The procedure of the method was the same as in example 2, except that citric acid was added to adjust the pH, to obtain brown rice fermentation broth C3.

Example 4 physicochemical Properties examination

1. Amino acid content

The amino acid content was measured using S1 sample in example 2 and C1 sample in comparative example 1 as experimental samples, and the results are shown in table 1 using an automatic amino acid analyzer.

TABLE 1 comparison of amino acid content in different samples

As can be seen from the table, the content of the small molecular amino acid in the brown rice fermentation broth S1 prepared by the method is obviously higher than that of the sample C1 obtained by the traditional extraction method.

2. Content of gamma-aminobutyric acid

The content of gamma-aminobutyric acid is determined by using an o-phthalaldehyde pre-column derivatization method by taking the sample S1 in example 2, the sample S2 in example 3 and the sample C1 in comparative example 1, the sample C2 in comparative example 2 and the sample C3 in comparative example 3 as experimental samples, and the HPLC conditions are as follows: a chromatographic column: hypersil ODS C₁₈Column temperature: 40 ℃; mobile phase: 0.75% sodium acetate acetonitrile (75: 25), flow rate: 1.0 mL/min, detection wavelength: 338 nm; the chromatogram is shown in FIG. 1, wherein a) is a standard substance (the content concentration of gamma-aminobutyric acid is 2 g/L), b) is S1, C) is S2, d) is C1, e) is C2, and f) is C3; the results are shown in Table 2.

TABLE 2 comparison of the content of gamma-aminobutyric acid in the conventional germinated brown rice

	S1	S2	C1	C2	C3
						Content (g/L)	1.91	2.15	0.051	0.41	0.54

As can be seen from the data in Table 2, the content of gamma-aminobutyric acid in the samples S1 and S2 is obviously higher than that in C1, C2 and C3, so that the active ingredient content in the product obtained by the process method is obviously higher than that in the conventional rice enzymolysis method, and the content of gamma-aminobutyric acid in the product can be effectively increased by adding coenzyme and supplementing glutamic acid to adjust the pH during fermentation.

3. Antioxidant activity

Accurately measuring 5.0mL of DPPH solution and 5.0mL of S1 sample solution with different concentrations respectively, placing the solutions in test tubes with plugs, and mixing the solutions uniformly. The blank is a mixed solution of water and 95% ethanol with equal volume. The mixture was left at room temperature for 30 minutes, and the absorbance values of the solutions were measured at 523nm, respectively. Another set of the two solutions was prepared by precisely measuring 5.0mL of DPPH solution and 5.0mL of purified water, and mixing the two solutions as described above. The calculation method is as follows:

clearance (%) =1-

The results are shown in Table 3. As can be seen from the data in the table, the sample S1 has higher ability of scavenging free radicals under the condition of adding the concentration gradient of 0.5-3%, and can reach 49% under the condition of 3% concentration, thus having better oxidation resistance.

TABLE 3 DPPH radical scavenging Activity

Concentration of stock solution (%)	0.5	1	2	3
					Clearance (%)	10	17	33	49

4. Whitening effect

Cell culture solution: 1640 culture medium containing 10% FBS; NaOH lysate: preparing lysis solution with the concentration of 0.1mol/L by using 10% DMSO solution; monkey head extract solution: preparing 20 mu M of a mao monkey hormone solution by using a cell culture solution; sample solution: preparing the brown rice extract into 4% mother liquor with a mao monkey extract solution, and diluting to a target concentration before use; MTT: and (3) preparing an MTT (methyl thiazolyl tetrazolium) acting solution by using PBS (phosphate buffer solution), filtering and sterilizing by using a filter membrane of 0.22 mu m, and storing for later use in a refrigerator at 4 ℃.

B16 cells were cultured at 1X 10⁵The cells were inoculated in 6-well plates at a density of one/mL and cultured at 37 ℃ under 5% CO2 for 24 hours. Adding the sample, reacting for 72h, cracking the cells with NaOH lysate, heating at 80 ℃ for 30min, and detecting the absorbance by an enzyme-labeling instrument. Simultaneously inoculating cells with the same volume density in a 96-well plate as a proliferation control, and inspecting sample pairsEffect of melanin level in unit cell.

The results are shown in Table 4. As can be seen from the data in the table, the sample S1 has a better melanin inhibition rate at a concentration of 1% -4%, and reaches about 80% at 4%, and most of melanin is inhibited.

TABLE 4 influence of brown rice fermentation extract on the level of melanin in unit cell

Concentration of brown rice (%)	Melanin inhibition per cell (%)
		1.0	34.46
2.0	68.05
		4.0	80.83

5. Promoting cell repair effect

Cell culture solution: DMEM medium containing 10% FBS. Sample solution: the brown rice extract is prepared into 4 percent mother liquor by using cell culture solution.

HaCaT cells were cultured at 5X 10⁴Inoculating to 24-well plate at 37 deg.C and 5% CO₂Culturing for 24h under the condition. On the near-confluent monolayer cells, a 200 μ L gun head was used to vertically scribe lines in each well of the 24-well plate. The liquid in the hole is discarded, the sample solution (the final serum content is 2.5 percent) is added, and the culture is continued for 24 hours and 48 hours, and then the photographing observation is carried out.

The results are shown in fig. 2, where a is the C1 treatment group initial, b is the C1 treatment group 24h, C is the C1 treatment group 48h, d is the S1 treatment group initial, e is the S1 treatment group 24h, and f is the S1 treatment group 48 h. Compared with C1, S1 at a concentration of 4.0% contacted with damaged cells for 48h can better promote cell migration and self-repair, and has a mostly intact morphology and a long spindle shape.

Example 5

(1) Weighing 5kg of brown rice, soaking the brown rice into 8.5kg of water for accelerating germination for 20h, drying the brown rice for 8h at 70 ℃, and crushing the brown rice into brown rice powder which is completely sieved by 200 meshes, wherein the water content is 5.7%;

(2) taking 5kg of brown rice powder in the step (1), adding 100L of water, mixing to obtain rice slurry, adding 100mL of alpha-amylase liquid and alpha-diastase liquid, performing enzymolysis, measuring the glucose content every 0.5h, and obtaining rice slurry enzymolysis liquid when the glucose content is not changed any more, wherein the residual sugar content is 59.4 g/L;

(3) adding 5g/L of bean cake powder as a nitrogen source and 1g/L of dipotassium hydrogen phosphate as a culture solution into the rice pulp enzymolysis solution, inoculating the saccharomycete thallus in the embodiment 1, simultaneously adding 0.8g of tetrahydrofolic acid, supplementing 38g of glutamic acid powder in the middle to maintain the pH between 5.0 and 5.5, fermenting for 20 hours at 25 ℃, and after no residual sugar exists, finishing fermentation to obtain a fermentation solution;

(4) centrifuging the fermentation liquor at 1500rpm to remove impurities, filtering the supernatant with 0.22 μm polyethersulfone filter element to obtain brown rice fermentation stock solution with relative density of 1.01 and gamma-aminobutyric acid content of 1.71g/L, removing impurities, and sterilizing.

Example 6

(1) Weighing 6kg of brown rice, soaking the brown rice into 10kg of water for accelerating germination for 12h, drying the brown rice for 10h at 80 ℃, and crushing the brown rice into brown rice powder which passes through 100 meshes and has the water content of 5.0 percent;

(2) taking 5kg of brown rice powder in the step (1), adding 50L of water, mixing to obtain rice slurry, then adding 50mL of alpha-amylase liquid and alpha-diastase liquid, performing enzymolysis, measuring the glucose content every 0.5h, and obtaining rice slurry enzymolysis liquid when the glucose content is not changed any more, wherein the residual sugar content is 64.8 g/L;

(3) adding 5g/L yeast powder as a nitrogen source and 1g/L dipotassium phosphate as a culture solution into the rice pulp enzymatic hydrolysate, inoculating the yeast thallus in the embodiment 1, simultaneously adding 0.5g of thiamine pyrophosphate, supplementing 46g of glutamic acid powder in the middle to maintain the pH value between 5.0 and 5.5, fermenting for 15h at 35 ℃, and after no residual sugar exists, ending the fermentation to obtain a fermentation liquid;

(4) centrifuging the fermentation liquor at 5000rpm to remove impurities, filtering the supernatant with 0.22 μm polyethersulfone filter element to obtain brown rice fermentation stock solution with relative density of 1.02 and gamma-aminobutyric acid content of 1.98g/L, removing impurities, and sterilizing.

Claims

1. A preparation method of brown rice fermentation stock solution for cosmetics is characterized by comprising the following steps:

(4) removing impurities from the fermentation liquor, and sterilizing to obtain a finished product;

in the step (2), the enzyme is alpha-amylase and alpha-saccharifying enzyme;

in the step (3), the coenzyme is one or more of nicotinic acid, biotin, tetrahydrofolic acid or thiamine pyrophosphate.

2. The method according to claim 1, wherein the brown rice flour is contained in the rice slurry in an amount of 5 to 10% by mass in the step (2).

3. The preparation method according to claim 1, wherein in the step (2), the enzyme is added in an amount of 0.002% -0.2% by mass of the rice pulp.

4. The method according to claim 1, wherein in the step (2), the mass ratio of the α -amylase to the α -saccharifying enzyme is 1: 1-2.

5. The method according to claim 1, wherein in the step (3), yeast cells are addedThe amount is 0.5-1% wt of the culture solution; the yeast is saccharomyces cerevisiae (Saccharomyces cerevisiae) Grape juice yeast (1)Saccharomyces uvarum) Rhodosporidium toruloides (A)Rhodospordium spp.) Beer yeast (Saccharomycescerevisiae Hansen) Hanseng spore yeast (A)Hanseniaspora spp.) Or Pichia pastoris (Pichia spp.）。

6. The method according to claim 1, wherein in the step (3), the pH during the fermentation is controlled to 5 to 5.5.

7. The process according to claim 6, wherein in the step (3), pH during the fermentation is controlled by adding glutamic acid to control pH.

8. The method according to claim 1, wherein the coenzyme in the step (3) is added in an amount of 0.0001 to 0.001% wt based on the culture broth.

9. The preparation method according to claim 1, wherein in the step (2), the glucose content at the enzymolysis end point is not changed any more at intervals of 0.5 h; in the step (3), the end point of the fermentation is that the fermentation liquor does not contain glucose.

10. A brown rice fermentation stock for cosmetics obtained by the production method according to any one of claims 1 to 9.

11. Use of the brown rice fermentation liquor of claim 10 in cosmetics.