阅读说明：本技术 一种蛹虫草发酵提取液的制备方法及所得产品和应用 (Preparation method of cordyceps militaris fermentation extract, product obtained by preparation method and application of product ) 是由 孙劭靖 杨旭 陆震 毛华 马良 温亮亮 石艳丽 郭学平 于 2020-07-07 设计创作，主要内容包括：本发明公开了一种蛹虫草发酵提取液的制备方法及所得产品和应用,该方法将乳酸菌在以蛹虫草为氮源的发酵培养基中进行发酵培养,得到蛹虫草发酵提取液。本发明通过选择合适的菌种和培养基,使微生物最大限度的利用分解蛹虫草粉释放有效成分；经微生物分解后的虫草素含量稳定,虫草酸含量与提取法相比提高6-8倍；提取液中既包含蛹虫草的功效成分,又包含乳酸菌发酵产生的一些活性物质,二者结合协同增效,使蛹虫草发酵提取液的营养物质、营养密度及生理活性大幅度提高,其价值远远超过了蛹虫草或乳酸菌发酵提取物本身,是天然优质的护肤产品、食品或保健品原料。(The invention discloses a preparation method of cordyceps militaris fermentation extract, an obtained product and application. The invention makes the microorganism utilize and decompose the cordyceps militaris powder to the maximum extent to release the effective components by selecting proper strains and culture media; the cordycepin content after microbial decomposition is stable, and the cordycepic acid content is improved by 6-8 times compared with that of an extraction method; the extract not only contains the functional components of the cordyceps militaris, but also contains some active substances generated by lactobacillus fermentation, and the two substances are combined for synergistic interaction, so that the nutrient substances, the nutrient density and the physiological activity of the cordyceps militaris fermentation extract are greatly improved, the value of the cordyceps militaris fermentation extract is far higher than that of the cordyceps militaris or lactobacillus fermentation extract, and the cordyceps militaris fermentation extract is a natural high-quality skin care product, food or health care product raw material.)

1. A preparation method of cordyceps militaris fermentation extract is characterized by comprising the following steps: carrying out fermentation culture on lactic acid bacteria in a fermentation culture medium taking cordyceps militaris as a nitrogen source to obtain cordyceps militaris fermentation extract, wherein the fermentation culture medium taking cordyceps militaris as the nitrogen source comprises the following components: 5-10 g/L of cordyceps militaris powder, 5g/L of glucose, 15-20g/L of fructose, 3-8 g/L of sodium acetate and 1-3g/L of magnesium sulfate.

2. The method of claim 1, wherein: the fermentation medium taking cordyceps militaris as a nitrogen source comprises the following components: 8g/L of cordyceps militaris powder, 5g/L of glucose, 18g/L of fructose, 5g/L of sodium acetate and 2g/L of magnesium sulfate.

3. The method according to claim 1 or 2, characterized in that: the method comprises the following steps:

(1) carrying out seed culture on lactobacillus to obtain a seed solution;

(2) inoculating the seed solution into a fermentation medium using cordyceps militaris as a nitrogen source, ventilating, stirring and culturing, and ending fermentation until no residual sugar exists to obtain a fermentation liquid;

(3) and treating the fermentation liquor to obtain cordyceps militaris fermentation extract.

4. The method of claim 1, 2 or 3, wherein: the lactic acid bacteria include lactobacillus, leuconostoc or bifidobacterium, preferably lactobacillus.

5. The method of claim 3, wherein: inoculating the seed liquid into a fermentation culture medium taking cordyceps militaris as a nitrogen source according to the inoculation amount of 5-10%.

6. The method of claim 1, 2 or 3, wherein: in the step (2), the ventilation amount is kept at 0.3-0.6vvm during fermentation culture.

7. The method of claim 3, wherein: in the step (3), sequentially carrying out operations of heating sterilization, pH adjustment and filtration on the fermentation liquor to obtain cordyceps militaris fermentation extract; preferably, the temperature for heat sterilization is 60-70 deg.C, and pH is adjusted to 5.0-6.0.

8. The cordyceps militaris fermentation extract prepared by the preparation method of the cordyceps militaris fermentation extract according to any one of claims 1 to 7; preferably, the cordycepin content is 40-90mg/L, the cordycepic acid content is 14-17g/L, and the total amino acid content is 1-2.5 g/L.

9. The application of the cordyceps militaris fermentation extract liquid as claimed in claim 8 in health care products, foods or cosmetics.

10. A health product, food or cosmetic is characterized in that: comprises the cordyceps militaris fermentation extract liquid as claimed in claim 8 or 9.

Technical Field

The invention relates to a preparation method of a cordyceps militaris fermentation extract, in particular to a method for preparing a cordyceps militaris fermentation extract by fermenting lactic acid bacteria serving as fermentation strains in a fermentation medium taking cordyceps militaris as a unique nitrogen source, and also relates to the cordyceps militaris fermentation extract and application of the cordyceps militaris fermentation extract.

Background

Cordyceps militaris is a model species of Ascomycota, Hypocreales, Clavicipitaceae, Cordyceps. The scientific name is Cordyceppsmilitaris, also called cordyceps militaris and cordyceps militaris, called cordyceps militaris for short, and the cordyceps militaris cultured by living pupa is generally called cordyceps militaris. Cordyceps militaris is distributed worldwide, and the quantity of natural resources is small. The main active ingredients in Cordyceps militaris comprise cordycepin, cordycepic acid and Cordyceps polysaccharide. Cordycepin (cordycepin), also known as cordycepin, 3-deoxyadenosine, is the first nucleoside antibiotic isolated from fungi. The cordycepin contained in the cordyceps militaris can obviously inhibit various inflammatory factors due to the specific antibacterial and antiviral effects of the cordycepin. Cordycepic acid is another important quality index of cordyceps militaris, and has the effects of resisting free radicals, resisting oxidation, promoting urination and dehydration, relieving cough, eliminating phlegm and relieving asthma. Cordyceps polysaccharide has anti-inflammatory and antioxidant effects, and can improve immunity, reduce blood sugar activity, resist tumor, and resist radiation.

Cordyceps militaris has been widely added into cosmetics and health foods, and nowadays, the application method of cordyceps militaris mostly takes cordyceps militaris as a raw material to extract effective components thereof, or takes cordyceps militaris living bodies as strains to obtain cordyceps militaris extracts or single effective components thereof by a fermentation method. For example, patent CN 107602660 a discloses a cordyceps militaris polypeptide product, and a preparation method and application thereof, in the patent, cordyceps militaris raw materials are added with an extracting solution and ground to obtain homogenate, and the homogenate is centrifuged and filtered, and then hydrolyzed by an enzyme preparation to obtain the polypeptide product. The cordyceps militaris polypeptide product prepared by the method is mostly applied to health-care food or beauty and skin care products. However, this method requires a large amount of enzyme. For example, patents CN 109136112A and CN 109294927 a disclose methods for improving cordycepin and cordyceps polysaccharide in cordyceps militaris, and these patents only research methods for improving a certain component and applications thereof.

At present, in the prior art, the utilization of active ingredients in cordyceps militaris is single, the utilization rate is low, and the effect of cordyceps militaris cannot be fully exerted.

Disclosure of Invention

Aiming at the defects in the prior art, the invention provides a preparation method of cordyceps militaris fermentation extract, which can also be called as a utilization method of cordyceps militaris. By the method, the whole Cordyceps militaris is fully utilized, and the fermentation extract is rich in nutrient components.

At present, no relevant report that lactic acid bacteria are used for fermentation culture in a fermentation culture medium with cordyceps militaris as a unique nitrogen source is seen. According to the invention, by selecting proper strains and culture media, the production period is shortened, the utilization rate of cordyceps militaris is improved, and the obtained fermentation extract has higher content of active substances such as cordycepin, cordycepic acid and amino acid; and the product is stable, and is beneficial to later purification.

The specific technical scheme of the invention is as follows:

a preparation method of cordyceps militaris fermentation extract comprises the following steps: carrying out fermentation culture on lactic acid bacteria in a fermentation culture medium taking cordyceps militaris as a nitrogen source to obtain cordyceps militaris fermentation extract, wherein the fermentation culture medium taking cordyceps militaris as the nitrogen source comprises the following components: 5-10 g/L of cordyceps militaris powder, 5g/L of glucose, 15-20g/L of fructose, 3-8 g/L of sodium acetate, 1-3g/L of magnesium sulfate and the balance of water.

Furthermore, in the fermentation medium, the cordyceps militaris powder is 8-10 g/L and the fructose is 18g/L, so that the effect is best. When the content of sodium acetate is 3 to 8g/L, the content of the effective component in the extract tends to increase with the increase of the content of sodium acetate, but the effect is equivalent when the content of sodium acetate is 5 to 8g/L, and therefore, 5g/L of sodium acetate is preferable. When magnesium sulfate is 1 to 3g/L, the content of the effective component in the extract tends to increase with the increase of the content of magnesium sulfate, but the effect is equivalent when the content of magnesium sulfate is 2 to 3g/L, and therefore, sodium acetate is preferably 2 g/L.

Preferably, when the following contents are selected from the fermentation medium taking cordyceps militaris as a nitrogen source, the effect is better: 8-10 g/L of cordyceps militaris powder, 5g/L of glucose, 15-20g/L of fructose, 5-8g/L of sodium acetate, 2-3g/L of magnesium sulfate and the balance of water.

More preferably, when the following contents are selected from the fermentation medium taking cordyceps militaris as a nitrogen source, the effect is best: 8g/L of cordyceps militaris powder, 5g/L of glucose, 18g/L of fructose, 5g/L of sodium acetate, 2g/L of magnesium sulfate and the balance of water.

The preparation method specifically comprises the following steps:

(1) carrying out seed culture on lactobacillus to obtain a seed solution;

(2) inoculating the seed solution into a fermentation medium using cordyceps militaris as a nitrogen source, ventilating, stirring and culturing, and ending fermentation until no residual sugar exists to obtain a fermentation liquid;

(3) and treating the fermentation liquor to obtain cordyceps militaris fermentation extract.

Further, the cordyceps militaris powder is powder formed by drying and crushing natural cordyceps militaris sporocarp, and the particle size of the powder has no special requirement and is generally 40 meshes or below. The Cordyceps militaris raw material contains various active ingredients such as cordycepin, cordycepic acid, Cordyceps polysaccharide and the like.

Further, various lactic acid bacteria species disclosed in the prior art of lactic acid bacteria, such as lactobacillus, leuconostoc, bifidobacterium, and the like, preferably lactobacillus. Each of the lactic acid bacteria can be purchased from the market.

Further, in step (1), the lactic acid bacteria can select a suitable seed culture medium and seed culture conditions from the disclosed prior art to obtain a seed solution required for fermentation culture. The conditions for seed culture, for example, temperature, pH, etc., may be selected to be optimum for each species.

Further, in the step (2), inoculating the seed solution into a fermentation culture medium taking cordyceps militaris as a nitrogen source according to the inoculation amount of 5-10%, performing fermentation culture, and stopping fermentation when residual sugar is exhausted without controlling the pH value in the fermentation process. The fermentation temperature and pH are optimum for each strain, for example, the fermentation temperature of lactobacillus is maintained at about 37 deg.C, pH is maintained at about 6.2-6.6, and aeration stirring is carried out during the culture process, with aeration amount of 0.3-0.6 vvm.

Further, in the step (3), after the fermentation is finished, the fermentation liquid is sequentially subjected to operations of heating sterilization, pH adjustment and filtration to obtain the cordyceps militaris fermentation extract. The sterilization is carried out at 60-70 deg.C and the heating time is about 1-2 h. Adjusting pH to 5.0-6.0. Filtration is generally carried out using a filter membrane having a pore size of 0.45. mu.m.

The invention also protects the cordyceps militaris fermentation extract obtained by the method. The Cordyceps militaris fermentation extract is rich in nutrition, high in activity, and rich in cordycepin and cordycepic acid, and is a good raw material for food, health product and cosmetic.

According to the invention, microbial lactic acid bacteria are adopted as strains, cordyceps militaris powder is utilized for fermentation to prepare cordyceps militaris fermentation extract, and the cordyceps militaris powder is used as a nitrogen source, so that the cordyceps militaris powder is decomposed to the maximum extent by utilizing microorganisms, various active ingredients in the cordyceps militaris powder are released, various active ingredients in the cordyceps militaris are all introduced into the fermentation extract, and the utilization rate of the cordyceps militaris is improved. In addition, after being decomposed by microorganisms, other active products such as various small molecule essential amino acids, lactic acid, sterol, phytic acid, vitamins and the like can be metabolized. The nutrient components in the fermented extract of cordyceps militaris obtained after fermentation are very rich, and the combination of microbial fermentation and cordyceps militaris has a good synergistic interaction effect, so that the nutrient substances, the nutrient density and the physiological activity of the fermented extract of cordyceps militaris are greatly improved, and the value of the fermented extract of cordyceps militaris far exceeds that of cordyceps militaris or microbial fermentation.

Further, the cordyceps militaris fermentation extract obtained by the invention has the following effects: 1. has the function of softening cuticle, can well play the effect of smoothing and tendering skin, and is particularly suitable for removing dandruff of scalp and oily skin; 2. the moisturizing and skin-moistening cream has good effects of moisturizing and moistening skin, can prevent the loss of moisture, and forms a protective film on the surface layer of the skin; 3. has the whitening effect: can inhibit melanin precipitation, effectively improve freckle, mottle, etc., whiten skin more, promote cell metabolism, and rapidly renew skin cells; 4. has antibacterial and antiseptic effects, and can prevent bacteria from invading skin and regulate skin pH. Compared with the cordyceps militaris products reported in the prior art, the cordyceps militaris fermentation extract disclosed by the invention is short in preparation period, low in cost, rich in components, diversified in effect and better in application value.

Furthermore, in the cordyceps militaris fermentation extract, the cordycepin content is 40-90mg/L, the cordycepic acid content is 14-17g/L, and the total amino acid content is 1-2.5 g/L.

Further, the invention also provides application of the cordyceps militaris fermentation extract in health care products, foods or cosmetics.

Furthermore, the invention also provides a health-care product, food or cosmetic, and the health-care product, food or cosmetic contains the cordyceps militaris fermentation extract.

The invention has the following advantages:

1. the invention takes the cordyceps militaris powder as a unique nitrogen source, and makes microorganisms utilize and decompose the cordyceps militaris powder to the maximum extent by selecting a proper strain and a proper fermentation medium formula, so that the effective ingredients in the cordyceps militaris are fully released.

2. The cordycepin content after microbial decomposition is stable and basically consistent with that obtained by an extraction method, and the cordycepic acid content is improved by 6-8 times compared with that obtained by the extraction method.

3. The extract not only contains the functional components of the cordyceps militaris, but also contains some active substances generated by microbial fermentation, and the two are combined for synergistic interaction, so that the nutrient substances, the nutrient density and the physiological activity of the cordyceps militaris fermentation extract are greatly improved, and the value of the cordyceps militaris fermentation extract is far higher than that of the cordyceps militaris or the microbial fermentation extract.

4. The invention has simple process, short production period and convenient operation, can prepare the cordyceps militaris fermentation extract with higher active matter content by simple microbial catabolism and purification process, has stable product quality, and is a high-quality food, health-care product or cosmetic raw material.

Detailed Description

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

In the following examples, the lactic acid bacteria used were commercially available products. The cordyceps militaris powder is obtained by drying and crushing commercially available cordyceps militaris sporocarp, and the particle size is about 40 meshes.