阅读说明：本技术 乳酸菌生产高含γ-氨基丁酸发酵液及其在化妆品中的应用 (Fermentation liquor with high content of gamma-aminobutyric acid produced by lactic acid bacteria and application of fermentation liquor in cosmetics ) 是由 梁金钟 肖雯娟 林枫翔 于 2019-09-16 设计创作，主要内容包括：本发明涉及发酵技术及化妆品领域,具体涉及乳酸菌生产高含γ-氨基丁酸发酵液及其在化妆品中的应用,所述制备方法包括如下步骤：(1)将糙米和大麦进行发芽,和桑叶混合,粉碎,得混合粉；(2)将混合粉加水浸泡,过滤,得滤液；(3)在滤液中加入谷氨酸或/和谷氨酸盐,接入发酵菌,过滤,即得,具有γ-氨基丁酸含量高、应用于化妆品中使用效果好、无副作用的优点。(The invention relates to the field of fermentation technology and cosmetics, in particular to a fermentation liquor with high content of gamma-aminobutyric acid produced by lactic acid bacteria and application thereof in cosmetics, wherein the preparation method comprises the following steps: (1) germinating brown rice and fructus Hordei vulgaris, mixing with folium Mori, and pulverizing to obtain mixed powder; (2) soaking the mixed powder in water, and filtering to obtain filtrate; (3) adding glutamic acid or/and glutamate into the filtrate, inoculating zymocyte, and filtering to obtain the final product with high content of gamma-aminobutyric acid, good use effect in cosmetic, and no adverse side effect.)

1. A preparation method for producing a fermentation liquor containing high content of gamma-aminobutyric acid by lactic acid bacteria is characterized by comprising the following steps:

(1) germinating brown rice and fructus Hordei vulgaris, mixing with folium Mori, and pulverizing to obtain mixed powder;

(2) soaking the mixed powder in water, and filtering to obtain filtrate;

(3) adding glutamic acid or/and glutamate into the filtrate, inoculating zymocyte, and filtering.

2. The preparation method according to claim 1, wherein the mass ratio of the brown rice, barley and mulberry leaves in step (1) is 1-2:1-2: 0.5-1.5; the mass-volume ratio of the mixed powder to water in the step (2) is 0.25-1.1:1 g/mL; the mass fraction of the glutamic acid or/and glutamate in the step (3) is 0.5-2.5%; the zymocyte comprises at least one of lactobacillus plantarum, lactobacillus acidophilus, lactobacillus casei and lactobacillus rhamnosus.

3. The preparation method according to claim 1, wherein the brown rice and barley germinate in step (1) to 1/3-2/3 with leaf bud length of seed, and the germination is terminated by heating to 40-50 ℃ for 5-15 min.

4. The method according to claim 1, wherein the step (2) comprises the steps of: soaking the mixed powder in water, heating at 55-65 deg.C for 2-6 hr, and filtering to obtain filtrate; the step (3) comprises the following steps: adding glutamic acid or/and glutamate into the filtrate, heating to 105-.

5. The production method according to claim 4, wherein the step (2) comprises the steps of: soaking the mixed powder in water, heating at 55-57 deg.C for 0.5-1h, heating to 60-65 deg.C, maintaining the temperature for 2-5h, and filtering to obtain filtrate.

6. The lactic acid bacteria produced by the method according to any one of claims 1 to 5 produce a fermentation broth with high gamma-aminobutyric acid content.

7. A cosmetic composition comprising the lactic acid bacterium according to claim 6 for producing a fermentation broth rich in gamma-aminobutyric acid.

8. The cosmetic composition according to claim 7, characterized by comprising the following components: lactobacillus produces high gamma-aminobutyric acid broth, DPG, glycerol, Starlub-7, glyceryl polyether-26, A91, xanthan gum, hydroxyethyl cellulose, HA, p-hydroxyacetophenone, betaine, Geobab gum, ERT37, Tridermol Hydrant, Aminose R, DermalRxKBGA, allantoin, acetyl hexapeptide-8, MultiEX Waterin, CalmYang, ceramide, bergamot essential oil, and water.

9. The cosmetic composition according to claim 8, which is characterized by comprising the following components in percentage by mass: lactic acid bacteria production high gamma-aminobutyric acid content fermentation liquor 1.0-5.0%, DPG 2.0-6.0%, glycerol 1.0-4.0%, Starlub-70.5-2.0%, glycerol polyether-260.5-2.5%, A910.1-0.5%, transparent xanthan gum 0.1-0.5%, hydroxyethyl cellulose 0.01-0.05%, HA 0.05-0.5%, p-hydroxyacetophenone 0.01-0.4%, betaine 0.05-3.0%, luba gum 1.0-6.0%, ERT 370.05-2.0%, Tridermol Hydrant 0.05-2.0%, Amitose R0.05-2.0%, Dermalrx KBGA 0.01-1.0%, allantoin 0.05-1.0%, acetyl hexapeptide-80.05-1.0%, Muterin 0.05-2.0%, Yang 2.0.001-2.0%, Californide 0.01-1.0%, and the balance of bergamot oil.

10. Use of the lactobacillus strain of claim 6 for producing a fermentation broth rich in gamma-aminobutyric acid or the cosmetic composition of claims 7 to 9 in cosmetics, preferably, the cosmetics are facial mask essence or skin lotion.

Technical Field

The invention relates to the field of fermentation technology and cosmetics, in particular to fermentation liquor with high gamma-aminobutyric acid content produced by lactic acid bacteria and application of the fermentation liquor in cosmetics.

Background

Gamma-aminobutyric acid (GABA), English name gamma-aminobutyric acid, chemical name: 4-aminobutyric acid, alternative name: aminobutyric acid and pipecolic acid are widely distributed in animals and plants. GABA is contained in seeds, rhizomes and tissue fluid of plants such as beans, ginseng, Chinese herbal medicines, etc. In animals, GABA is present almost exclusively in nerve tissue, with brain tissue at levels of about 0.1-0.6 mg/g tissue. GABA is an important neurotransmitter, has various physiological activity functions, and has physiological effects of physiological repair, cell activation, skin tendering and beautifying and the like on human epidermal cells. GABA is also closely related to the quality of sleep and is called the brain's natural sedative. In addition, it also has effects of lowering blood pressure and reducing blood ammonia. GABA can promote oxygen and glucose metabolism in brain tissue, increase brain blood flow, and improve headache, heaviness of head, fatigue, dizziness, tinnitus, hypomnesis, and hypovolemia. GABA can relieve physical and psychological discomfort symptoms, and symptoms such as anxiety, uneasiness, fatigue, insomnia, dreaminess, intolerance to pain, low resistance and the like can be caused when GABA in a human body is insufficient, and GABA should be supplemented in time at the moment. Generally, people with high stress, such as competitive athletes, office workers, students with high learning pressure, and the like, are easy to lack GABA, so that GABA needs to be supplemented to relieve emotion.

The preparation method of the gamma-aminobutyric acid mainly comprises a chemical synthesis method and a biological synthesis method.

The chemical synthesis method is reported in patent documents, has higher cost and lower yield, and uses dangerous solvents, even toxic solvents, in the production process. Therefore, gamma-aminobutyric acid prepared by a chemical synthesis method cannot be used for food, and cannot be considered as a natural food additive.

Compared with the biosynthesis method, the method is safe and low in cost. In the early research, the fermentation method for producing gamma-aminobutyric acid takes escherichia coli as a production bacterium, and a fermentation medium comprises bran hydrolysate, corn steep liquor, peptone, minerals and the like. In the fermentation process, the L-glutamic acid is converted into gamma-aminobutyric acid by the action of escherichia coli decarboxylase, and then the GABA product is obtained through separation and purification. However, the use of Escherichia coli has various problems in terms of safety as long as food development is attempted. According to the latest research reports and patent documents, some high-safety microorganisms such as lactic acid bacteria, yeast and aspergillus have been applied to the preparation of gamma-aminobutyric acid foods, so that the biosynthetic gamma-aminobutyric acid products can be used as ingredients of high-grade functional health-care foods.

The microbial fermentation method mainly utilizes the decarboxylation of a fermentation bacterium, namely Glutamic Acid Decarboxylase (GAD), so that carboxyl on the alpha carbon position of glutamic acid or sodium salt thereof is subjected to decarboxylation reaction, and one carboxyl is removed, thereby converting the glutamic acid or sodium salt thereof into GABA.

Chinese patent CN104388514A discloses a method for preparing gamma-aminobutyric acid by using composite strain fermentation, wherein the fermentation strains comprise lactobacillus plantarum ZSM-002, kast bouquet yeast ZSM-001 and a collection number of CCTCC NO: m2014376 Rhizopus (Rhizopus oryzae) zsm-005 takes grain seeds or embryo-containing byproducts as raw materials, and prepares a high-concentration liquid gamma-aminobutyric acid product through the processes of cleaning, disinfection, gamma-aminobutyric acid enrichment, separation, extraction, refining, purification and the like, and prepares a high-concentration solid gamma-aminobutyric acid product through the drying process of the liquid gamma-aminobutyric acid. The patent adopts multi-strain synergistic fermentation, and can improve the capability of enriching gamma-aminobutyric acid of grains or embryo-containing byproducts. However, the types and functions of metabolites after the fermentation of the three types of zymophytes are limited, and the content of an efficacy product gamma-aminobutyric acid is low, so that the application of the efficacy product gamma-aminobutyric acid in cosmetics is limited.

At present, the gamma-aminobutyric acid is mainly applied to food, and the gamma-aminobutyric acid is applied to cosmetics. GABA has weight reducing and skin caring effects, and can be used for decomposing body fat by combustion, increasing and prolonging catabolic process, and burning more fat to achieve weight reducing effect. GABA is a small molecule substance which can directly permeate into the inside of cells, and the function must be performed in a sleeping state, so that beauty and weight loss can be realized by applying a mask containing GABA to sleep. Meanwhile, GABA can relieve great mental stress and eliminate fatigue, anxiety and dull complexion.

However, when the gamma-aminobutyric acid is applied to cosmetics and acts on skin, it is inevitable that uncomfortable symptoms such as tingling, numbness, itching, burning and the like are generated when the concentration of GABA reaches a desired effect, and the side effect has a positive correlation with the concentration, and at present, the content of the gamma-aminobutyric acid in the cosmetics is recommended to be at most 5.0%, and how to reduce the side effect at a lower concentration or at a higher concentration is a problem to be solved urgently.

Therefore, it is necessary to develop a lactobacillus capable of solving the above technical problems to produce a fermentation broth with high content of gamma-aminobutyric acid, and a composition and application thereof.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provide the lactobacillus fermentation liquor with high content of gamma-aminobutyric acid, good using effect when being applied to cosmetics and no side effect, as well as the composition and the application thereof.

The invention is realized by the following technical scheme:

a preparation method for producing fermentation liquor containing high content of gamma-aminobutyric acid by lactic acid bacteria comprises the following steps:

(1) germinating brown rice and fructus Hordei vulgaris, mixing with folium Mori, and pulverizing to obtain mixed powder;

(2) soaking the mixed powder in water, and filtering to obtain filtrate;

(3) adding glutamic acid or/and glutamate into the filtrate, inoculating zymocyte, and filtering.

Preferably, the mass ratio of the brown rice, the barley and the mulberry leaves in the step (1) is 1-2:1-2: 0.5-1.5.

Preferably, the brown rice and barley germinate in step (1) to 1/3-2/3 with the leaf bud length of the seeds.

More preferably, the brown rice and barley germinate in the step (1) to 1/3-2/3 of the length of the leaf bud of the seed, and the germination is terminated.

More preferably, the germination is terminated by heating to 40-50 deg.C for 5-15 min.

More preferably, the step (1) includes the steps of: germinating brown rice and fructus Hordei vulgaris until the leaf bud length is 1/3-2/3 of seed, heating to 40-50 deg.C for 5-15min, stopping germination, mixing with folium Mori at a mass ratio of 1-2:1-2:0.5-1.5, and pulverizing to obtain mixed powder.

Preferably, the mass-volume ratio of the mixed powder to the water in the step (2) is 0.25-1.1:1 g/mL.

Preferably, the mixed powder in the step (2) is soaked in water and then heated at the temperature of 55-65 ℃ for 2-6 h.

More preferably, the heating is as follows: heating at 55-57 deg.C for 0.5-1h, heating to 60-65 deg.C, and maintaining for 2-5 h.

More preferably, the step (2) includes the steps of: soaking the mixed powder in water according to the mass-volume ratio of 0.25-1.1:1g/mL, heating at 55-65 ℃ for 2-6h, and filtering while hot to obtain filtrate.

More preferably, the step (2) includes the steps of: soaking the mixed powder in water according to the mass-volume ratio of 0.25-1.1:1g/mL, heating at 55-57 ℃ for 0.5-1h, then heating to 60-65 ℃, preserving heat for 2-5h, and filtering while hot to obtain a filtrate.

Preferably, the mass fraction of the glutamic acid or/and glutamate in the step (3) is 0.5-2.5%. Namely the mass fraction of the glutamic acid or the mass fraction of the glutamate or the total mass fraction of the glutamic acid and the glutamate is 0.5-2.5%.

Preferably, the fermentation bacteria in step (3) include at least one of Lactobacillus plantarum (Lactobacillus plantarum), Lactobacillus acidophilus (Lactobacillus acidophilus), Lactobacillus casei (Lactobacillus casei), and Lactobacillus rhamnosus (Lactobacillus rhamnosus).

Preferably, the zymophyte is fermented for 6 to 24 hours at 35 to 40 ℃ in the step (3).

Preferably, the amount of the zymocyte in the step (3) is 1.0-10.0%.

More preferably, the step (3) includes the steps of: adding 0.5-2.5% of glutamic acid or/and glutamate by mass percent into the filtrate, heating to 105-plus materials at 121 ℃ for sterilization for 10-30min, inoculating zymocyte, fermenting for 6-24h at 35-40 ℃ with the inoculum size of 1.0-10.0%, and filtering to obtain the filtrate which is the fermentation liquor for producing the high gamma-aminobutyric acid by the lactobacillus.

More preferably, the preparation method comprises the following steps:

(1) germinating brown rice and fructus Hordei vulgaris until the leaf bud length is 1/3-2/3 of seed, heating to 40-50 deg.C for 5-15min, stopping germination, mixing with folium Mori at a mass ratio of 1-2:1-2:0.5-1.5, pulverizing to obtain mixed powder;

(2) soaking the mixed powder in water according to the mass-volume ratio of 0.25-1.1:1g/mL, heating at 55-57 ℃ for 0.5-1h, heating to 60-65 ℃, preserving heat for 2-5h, and filtering while hot to obtain a filtrate;

(3) adding 0.5-2.5% of glutamic acid or/and glutamate by mass percent into the filtrate, heating to 105-plus materials at 121 ℃ for sterilization for 10-30min, inoculating zymocyte, fermenting for 6-24h at 35-40 ℃ with the inoculum size of 1.0-10.0%, and filtering to obtain the filtrate which is the fermentation liquor for producing the high gamma-aminobutyric acid by the lactobacillus.

The invention also relates to a fermentation liquor with high content of gamma-aminobutyric acid produced by the lactobacillus prepared by the preparation method.

The invention also relates to a cosmetic composition which comprises the lactobacillus fermentation liquor for producing the high-content gamma-aminobutyric acid.

The fermentation bacteria of the present invention produce various metabolites through fermentation, and the metabolites include: lactic acid, malic acid, acetic acid, lysine, threonine, phenylalanine, methionine, leucine, cystine, valine, etc., in addition to which many vitamins, exopolysaccharides, small peptides and some biological enzymes are produced.

The organic acid generated by metabolism can make the pH value of the skin to be weakly acidic, thereby achieving the effect of balancing the pH value on the surface of the skin. The amino acids, small molecular peptides, vitamins and biological enzymes generated by metabolism can nourish and whiten skin, repair and maintain epidermal cells, remove pigmentation, prevent pachylosis, make skin bright and elastic, and achieve the purpose of beautifying.

In addition, the zymocyte can also generate a plurality of micromolecule antibacterial and antioxidant substances, can effectively inhibit the growth and reproduction of harmful bacteria on the surface of the skin such as acne bacillus, and has good effects of resisting acne and removing spots.

The invention ferments the zymocyte in a certain culture medium and controls certain conditions, and the zymocyte can ferment and metabolize to generate a plurality of bioactive substances. The fermentation metabolites and some substances beneficial to skin health are scientifically combined and prepared, and various facial masks or other skin care products with the effects of whitening, cleaning, removing wrinkles and moisturizing can be prepared.

Preferably, the cosmetic composition comprises the following components: lactobacillus produces high gamma-aminobutyric acid broth, DPG, glycerol, Starlub-7, glyceryl polyether-26, A91, xanthan gum, hydroxyethyl cellulose, HA, p-hydroxyacetophenone, betaine, Geobab gum, ERT37, Tridermol Hydrant, Aminose R, DermalRx KBGA, allantoin, acetyl hexapeptide-8, MultiEX Waterin, CalmYang, ceramide, bergamot essential oil and water.

Preferably, the cosmetic composition consists of the following components in percentage by mass: lactic acid bacteria production high gamma-aminobutyric acid content fermentation liquor 1.0-5.0%, DPG 2.0-6.0%, glycerol 1.0-4.0%, Starlub-70.5-2.0%, glycerol polyether-260.5-2.5%, A910.1-0.5%, transparent xanthan gum 0.1-0.5%, hydroxyethyl cellulose 0.01-0.05%, HA 0.05-0.5%, p-hydroxyacetophenone 0.01-0.4%, betaine 0.05-3.0%, luba gum 1.0-6.0%, ERT 370.05-2.0%, Tridermol Hydrant 0.05-2.0%, Amitose R0.05-2.0%, Dermalrx KBGA 0.01-1.0%, allantoin 0.05-1.0%, acetyl hexapeptide-80.05-1.0%, MuEX Waterin 0.05-2.0.001-2.0%, Yang 0.01-1.0%, Californide 0.5-0%, and the balance of bergamot oil.

The invention also relates to the application of the lactobacillus in producing the fermentation liquor with high content of gamma-aminobutyric acid or the cosmetic composition in cosmetics.

Preferably, the cosmetic is a mask essence or a skin moisturizing lotion.

The invention has the beneficial effects that:

according to the invention, brown rice, barley and mulberry leaves are used as raw materials of a fermentation medium, and by limiting the germination conditions of the brown rice and the barley and the ratio of the brown rice to the barley, the fermentation bacteria can be metabolized to obtain high-content gamma-aminobutyric acid (up to 4.8-5.5g/100 mL), and various bioactive substances can be obtained.

The invention further improves the gamma-aminobutyric acid content of the final metabolite by further limiting the variety of zymophyte, the preparation of the culture medium and the fermentation conditions.

According to the invention, the fermentation liquor which is produced by lactic acid bacteria and contains high content of gamma-aminobutyric acid is applied to cosmetics and is matched with other components to act, so that the dull face skin is obviously improved, the whiteness of the skin is obviously improved, the elasticity and moisture of the skin are also obviously improved, and wrinkles are obviously reduced. The long-term use can improve the problems of tarnish, dehydration, dryness and wrinkles of the skin and keep the skin tender and white.

Detailed Description

The invention will be further described with reference to specific embodiments, and the advantages and features of the invention will become apparent as the description proceeds. These examples are illustrative only and do not limit the scope of the present invention in any way. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention, and that such changes and modifications may be made without departing from the spirit and scope of the invention.