CN113439838B

CN113439838B - Fermentation method, fermentation product and kit containing fermentation product

Info

Publication number: CN113439838B
Application number: CN202010226008.5A
Authority: CN
Inventors: 彭宁; 张彦; 张海波; 覃敬羽; 杨树金; 冯玖
Original assignee: Angel Nutt Co ltd
Current assignee: Angel Nutt Co ltd
Priority date: 2020-03-26
Filing date: 2020-03-26
Publication date: 2022-09-06
Anticipated expiration: 2040-03-26
Also published as: CN113439838A

Abstract

The invention provides a fermentation method, a fermentation product and a kit containing the fermentation product. The fermentation method comprises the following steps: preparing a to-be-fermented liquid by taking ginseng, polygonatum odoratum, lotus seeds and coix seeds as raw materials, and performing combined fermentation on the to-be-fermented liquid by utilizing a plurality of bacteria to obtain a fermentation product, wherein the plurality of bacteria comprise at least one yeast and at least one lactic acid bacteria. The raw material composition containing ginseng, polygonatum, lotus seed and coix seed is fermented by utilizing the fermentation characteristics of edible safe strains such as saccharomycetes and lactic acid bacteria, the two strains generate various functional substances in the growth process, the enzyme system is complex, the release of different functional components in the raw materials can be promoted, the content of the functional substances in the traditional Chinese medicine is increased, the structure is converted, and the overall effect of the fermentation product is improved.

Description

Fermentation method, fermentation product and kit containing fermentation product

Technical Field

The invention relates to the field of fermentation, and particularly relates to a fermentation method, a fermentation product and a kit containing the fermentation product.

Background

Ginseng, radix Ginseng, sweet in taste and slightly cold in nature, has the effects of nourishing five internal organs, calming mind, stopping palpitation, eliminating pathogenic factors, improving eyesight, and improving intelligence. It can be taken for a long time, and has effects of reducing weight and prolonging life. One for each person and one for each ghost. And (4) growing the valley. Shen nong Ben Cao Jing.

Fragrant solomonseal rhizome, rhizoma Polygonati Odorati, is neutral in nature, sweet in flavor, soft, moist and edible. Compendium of materia Medica.

The lotus is sweet in flavor, astringent in nature and fresh and fragrant in nature, so as to obtain the flavor of crops, and also the spleen. Earth is the mother of original qi, and the mother qi is harmonized with body fluids, so the spirit is self-generated, long-term vision is durable, and so its right is also. Xiren is indicated for disharmony between heart and kidney, impairment of white turbidity due to overstrain, and has Qingxin Lian Yin; it is also called Rui Lian Wan for tonifying heart and kidney, nourishing essence and blood. Compendium of materia Medica.

Yi ren, yang ming herb also can strengthen spleen and tonify stomach. It is indicated for consumptive lung disease and pulmonary abscess because it tonifies its mother herb in case of deficiency. For the diseases of tendons and bones, it is indicated for yang-yin, spasm and wind-arthralgia. For it can generate water and remove dampness, it is indicated for diarrhea and edema- -Ben Cao gang mu.

Ginseng, polygonatum, lotus seed and coix seed which are used as traditional medicinal materials and food have a long-standing utilization tradition in China, the components are also used as additives to be mixed with a plurality of other components to be used as common food in the prior art, or single components in the common food are fermented or combined with other substances to be fermented, however, the functions of the food or the fermented product in the aspects of medicine, eating or skin care are still not effectively developed and utilized.

Disclosure of Invention

The invention mainly aims to provide a fermentation method, a fermentation product and a kit containing the fermentation product, so as to solve the problem that the development and utilization degree of the traditional Chinese medicinal materials and food materials in the aspects of medicine, eating or skin care is still low in the prior art.

In order to achieve the above object, according to one aspect of the present invention, there is provided a fermentation method comprising: preparing a to-be-fermented liquid by using ginseng, polygonatum odoratum, lotus seeds and coix seeds as raw materials, and performing combined fermentation on the to-be-fermented liquid by using a plurality of bacteria to obtain a fermented product, wherein the plurality of bacteria comprise at least one yeast and at least one lactic acid bacteria.

Further, the step of performing combined fermentation on the fermentation liquid to be fermented by adopting a plurality of bacteria to obtain the fermentation product comprises the following steps: mixing various bacteria and a to-be-fermented solution, and performing primary culture to obtain a primary culture; and carrying out anaerobic or aerobic culture on the primary culture to obtain a fermentation product.

Further, the step of performing combined fermentation on the fermentation liquor to be fermented by adopting a plurality of bacteria to obtain a fermentation product comprises the following steps: mixing saccharomycetes and a liquid to be fermented, and then carrying out primary culture to obtain a primary culture; and (4) carrying out anaerobic or aerobic culture on the primary culture by using lactic acid bacteria to obtain a fermentation product.

Further, the step of performing combined fermentation on the fermentation liquor to be fermented by adopting a plurality of bacteria to obtain a fermentation product comprises the following steps: mixing lactic acid bacteria and a liquid to be fermented, and then carrying out primary culture to obtain a primary culture; and (4) carrying out anaerobic or aerobic culture on the primary culture by using saccharomycetes to obtain a fermentation product.

Further, the step of performing combined fermentation on the fermentation liquor to be fermented by adopting a plurality of bacteria to obtain a fermentation product comprises the following steps: mixing lactic acid bacteria and a liquid to be fermented, and then carrying out fermentation culture to obtain a first fermentation culture; mixing saccharomycetes and a to-be-fermented solution, and then performing fermentation culture to obtain a second fermentation culture; mixing the first fermentation culture with the second fermentation culture to obtain a fermentation product; preferably, the first fermentation culture is mixed with the second fermentation culture in equal volumes to yield the fermentation product.

Further, the step of preparing the liquid to be fermented by taking the ginseng, the polygonatum odoratum, the lotus seed and the coix seed as raw materials comprises the following steps: mixing Ginseng radix, rhizoma Polygonati Odorati, semen Nelumbinis and Coicis semen to obtain mixed material; carrying out enzymolysis treatment on the mixed material to obtain an enzymolysis liquid; adding a nitrogen source and a carbon source into the enzymolysis liquid to obtain a liquid to be fermented; wherein the enzymolysis comprises amylase enzymolysis, saccharification enzymolysis and protease enzymolysis; preferably, the mixing ratio of the ginseng, the polygonatum odoratum, the lotus seed and the coix seed in parts by weight is 1-10: 1-10: 1-10: 1-10; preferably, 1-20 parts of mixed material is added into 100 parts of deionized water by weight for enzymolysis treatment; preferably, the nitrogen source is selected from any one or more of ammonium sulfate, ammonium chloride, hydrolyzed soy protein, yeast extract and milk powder; more preferably, the amount of the nitrogen source is 0.5 to 10 parts; preferably, the carbon source is selected from any one or more of glucose, fructose, maltose and hydrolyzed starch; more preferably, the amount of the carbon source added is 0.5 to 20 parts.

Further, before the liquid to be fermented is subjected to combined fermentation, the fermentation method further comprises the step of sterilizing the liquid to be fermented, preferably, the sterilization temperature is 80-130 ℃, and the sterilization time is 3s-240 min.

Further, the initial culture is carried out for 24-96h at 25-30 ℃ by adopting a shaking table, and the rotation speed of the shaking table is preferably 160-200 rpm/min.

Further, the anaerobic culture or aerobic culture comprises the following steps: adding a nitrogen source and a carbon source into the primary culture to obtain primary fermentation liquor; adjusting the pH value of the primary fermentation liquid to 6.5-7.5, and then carrying out anaerobic culture or aerobic culture; preferably, the anaerobic culture or aerobic culture is carried out for 24-240 hours at 35-37.5 ℃.

Further, before the combined fermentation of the fermentation liquid to be fermented by using a plurality of bacteria, the fermentation method further comprises the following steps: respectively carrying out strain activation, strain purification and enlarged culture on at least one yeast and at least one lactic acid bacteria in sequence.

Further, the yeast is selected from Saccharomyces cerevisiae A3.12 with a preservation number of CCTCC NO: M2017781, Pichia pastoris C1.8 with a preservation number of CCTCC NO: M2017780, or Candida candida C1.7 with a preservation number of CCTCC NO: M2017782; preferably, the lactobacillus is Lactobacillus pentosus 001 with the preservation number of CCTCC NO: M2017625, Lactobacillus casei 001 with the preservation number of CCTCC NO: M2017631, Lactobacillus acidophilus 001 with the preservation number of CCTCC NO: M2017628, Lactobacillus plantarum 001 with the preservation number of CCTCC NO: M2017629 or Lactobacillus rhamnosus 001 with the preservation number of CCTCC NO: M2017630.

Further, the multiple bacteria comprise 0.1-5 parts of saccharomycetes and 0.1-5 parts of lactic acid bacteria in parts by weight; preferably, the dosage ratio of the various bacteria to the liquid to be fermented is 0.2-10: 100-200.

In order to achieve the above object, according to a second aspect of the present invention, there is provided a fermentation product, which is fermented by any one of the above fermentation methods.

In order to achieve the above object, according to a third aspect of the present invention, there is provided a fermentation kit, comprising a fermentation raw material, at least one yeast and at least one lactic acid bacterium, wherein the fermentation raw material is selected from the group consisting of ginseng, polygonatum odoratum, lotus seed and coix seed.

Further, the yeast is selected from at least one of saccharomyces cerevisiae A3.12 with a preservation number of CCTCC NO: M2017781, pichia pastoris C1.8 with a preservation number of CCTCC NO: M2017780 and candida albicans C1.7 with a preservation number of CCTCC NO: M2017782, and the lactobacillus is selected from at least one of lactobacillus pentosus 001 with a preservation number of CCTCC NO: M2017625, lactobacillus casei 001 with a preservation number of CCTCC NO: M2017631, lactobacillus acidophilus 001 with a preservation number of CCTCC NO: M2017628, lactobacillus plantarum 001 with a preservation number of CCTCC NO: M2017629 and lactobacillus rhamnosus 001 with a preservation number of CCTCC NO: M2017630.

By applying the technical scheme of the invention, the raw material composition containing ginseng, polygonatum odoratum, lotus seed and coix seed is fermented by utilizing the fermentation characteristics of edible safe strains such as saccharomycetes and lactic acid bacteria, the two strains generate various functional substances in the growth process, the enzyme system is complex, the release of different functional components in the raw materials can be promoted, the improvement of the content of the functional substances in the traditional Chinese medicine and the conversion of the structure are realized, and the overall effect of the fermentation product is improved.

Drawings

The accompanying drawings, which are incorporated in and constitute a part of this application, illustrate embodiments of the invention and, together with the description, serve to explain the invention and not to limit the invention. In the drawings:

fig. 1 shows the moisturizing rates of the complex extract and the complex fermented product according to example 2 of the present invention.

Information on strain preservation

The strains used by the invention are preserved in China Center for Type Culture Collection (CCTCC) at the preservation addresses of: wuhan, Wuhan university, zip code: 430072; telephone: (027) -68754052. The preservation information of the specific strains is as follows:

saccharomyces cerevisiae A3.12(Saccharomyces cerevisiae A3.12) is preserved in China Center for Type Culture Collection (CCTCC) in 2017, 12 months and 11 days, and the preservation number is CCTCC NO: M2017781.

Candida C1.7(Wickerhamomyces anomalus C1.7) is preserved in China Center for Type Culture Collection (CCTCC) in 2017, 12 and 11 months, and the preservation number is CCTCC NO: M2017782.

Pichia pastoris C1.8 (Cyberlindera fabianii C1.8) is preserved in China Center for Type Culture Collection (CCTCC) in 2017, 12 months and 11 days, with the preservation number of CCTCC NO: M2017780.

Lactobacillus pentosus 001(Pediococcus pentosaceus 001) is preserved in China Center for Type Culture Collection (CCTCC) in 2017, 10 and 25 months, and the preservation number is CCTCC NO: M2017625.

Lactobacillus plantarum 001(Lactobacillus plantarum 001) is preserved in the China Center for Type Culture Collection (CCTCC) in 2017, 10 and 25 months, and the preservation number is CCTCC NO: M2017629.

Lactobacillus rhamnosus 001(Lactobacillus rhamnosus 001) is preserved in China Center for Type Culture Collection (CCTCC) in 2017 in 10 and 25 months, with the preservation number of CCTCC NO: M2017630.

Lactobacillus acidophilus 001(Lactobacillus acidophilus 001) is preserved in China Center for Type Culture Collection (CCTCC) in 2017, 10 months and 25 days, and the preservation number is CCTCC NO: M2017628.

Lactobacillus casei 001(Lactobacillus casei 001) is preserved in China Center for Type Culture Collection (CCTCC) in 2017, 10 and 25 months, and the preservation number is CCTCC NO: M2017631.

Detailed Description

It should be noted that the embodiments and features of the embodiments in the present application may be combined with each other without conflict. The present invention will be described in detail with reference to examples.

As mentioned in the background of the invention, the fermented product in the prior art has low efficiency of utilizing the efficacy of traditional herbs or food materials, and in order to improve the current situation, in an exemplary embodiment of the present application, a fermentation method is provided, which includes: preparing a to-be-fermented liquid by taking ginseng, polygonatum odoratum, lotus seeds and coix seeds as raw materials, and performing combined fermentation on the to-be-fermented liquid by utilizing a plurality of bacteria to obtain a fermentation product, wherein the plurality of bacteria comprise at least one yeast and at least one lactic acid bacteria.

The raw materials in the fermentation method are ginseng, polygonatum, lotus seed and coix seed, wherein the ginseng mainly contains ginsenoside, and the polygonatum polysaccharide and flavone are the main substances. The water-soluble polysaccharide, the total flavone and the superoxide dismutase in the lotus seeds are the main three substances. The main active components contained in Coicis semen include Coicis semen ester, triglyceride, fatty acids, lactam, Coicis semen lactone, saccharide, sterol, and triterpene. The raw materials can play a role in synergy among different functional components after being matched and combined, and meanwhile, the four components contain abundant starch, protein, trace elements and the like, so that sufficient raw materials can be provided for microbial fermentation.

By utilizing the fermentation characteristics of edible safe strains such as saccharomycetes and lactic acid bacteria, the raw material composition containing ginseng, polygonatum odoratum, lotus seeds and coix seeds is fermented, the two strains generate various functional substances in the growth process, the enzyme system is complex, the release of different functional components in the raw materials can be promoted, the content of the functional substances in the traditional Chinese medicine is increased, the structure is converted (for example, ginsenoside is converted from common saponin into rare ginsenoside, flavonoid structure changes the water solubility and the like), and the overall effect of the fermentation product is improved.

In a preferred embodiment, the step of performing a combined fermentation on a fermentation broth to obtain a fermentation product comprises: mixing various bacteria with a to-be-fermented solution, and performing primary culture to obtain a primary culture; and carrying out anaerobic or aerobic culture on the primary culture to obtain a fermentation product.

Mixing various bacteria and the liquid to be fermented, and then carrying out primary culture, aiming at obtaining the optimal culture condition of the bacteria and facilitating the subsequent fermentation. Then anaerobic culture or aerobic culture is carried out for fermentation, so that the fermentation is more sufficient, and the utilization rate of the raw materials is high. The aerobic culture here means: introducing sterilized air during the fermentation process.

In the preferred embodiment, the yeast and the lactic acid bacteria are in a competitive and interdependent relationship under symbiotic conditions by mixing a plurality of bacteria of at least one yeast and at least one lactic acid bacteria and then performing combined fermentation of the liquid to be fermented of the raw material, so that the yeast and the lactic acid bacteria compete for nutrients and simultaneously provide the other with substances required by the other. Such as: lactic acid bacteria are auxotrophic strains, yeast provides substances such as amino acid, vitamin pyruvate and the like for the lactic acid bacteria in the fermentation process, and metabolites of the lactic acid bacteria can provide energy sources for the yeast. Therefore, through the initial culture and the subsequent anaerobic or aerobic culture, various enzymes are released in the fermentation process, the release of the functional substances in the raw materials is promoted, and the function is improved, so that the obtained fermentation product has the effects of resisting fatigue, delaying aging and preserving moisture.

In a preferred embodiment, the step of performing combined fermentation on the fermentation broth to obtain the fermentation product by using a plurality of bacteria comprises: mixing saccharomycetes and a liquid to be fermented, and then carrying out primary culture to obtain a primary culture; and (4) carrying out anaerobic or aerobic culture on the primary culture by using lactic acid bacteria to obtain a fermentation product.

In a preferred embodiment, the step of performing combined fermentation on the fermentation broth to obtain the fermentation product by using a plurality of bacteria comprises: mixing lactic acid bacteria and a liquid to be fermented, and performing primary culture to obtain a primary culture; and (4) carrying out anaerobic or aerobic culture on the primary culture by using saccharomycetes to obtain a fermentation product.

In the two preferable embodiments, the two kinds of strains are fermented successively, and substances released by the fermentation of the former strain can provide nutrient substances or energy for the latter strain, so that various functional species in the raw materials are fully released, and the overall efficacy of the fermentation product is improved.

In a preferred embodiment, the step of performing combined fermentation on the fermentation broth to obtain the fermentation product by using a plurality of bacteria comprises: mixing lactic acid bacteria and a liquid to be fermented, and then carrying out fermentation culture to obtain a first fermentation culture; mixing saccharomycetes and a to-be-fermented solution, and then performing fermentation culture to obtain a second fermentation culture; mixing the first fermentation culture with the second fermentation culture to obtain a fermentation product; preferably, the first fermentation culture is mixed with the second fermentation culture in equal volumes to give the fermentation product.

In the preferred embodiment described above, the four raw materials are fermented by the two types of bacterial species, respectively, and then the fermentation cultures of the two types of bacteria are mixed so that the fermentation product includes the fermentation products of the two types of bacteria. Compared with the fermentation product of single type of bacteria, the two types of bacteria are richer in the fermentation products of the four raw materials and better in overall effect.

In the above various combined fermentation schemes, after the fermentation product is obtained, the step of filtering the fermentation product to obtain the supernatant may be further included. The filtering step removes solid residues in the fermentation product, so that the performance of the fermentation product is more excellent. The specific filtering mode is not limited, and any filtering mode which is favorable for solid-liquid separation is suitable for the application. In the present application, centrifugation and plate-and-frame filtration are preferably used for filtration to obtain the supernatant.

In the fermentation method, the raw materials are also required to be prepared into environmental conditions suitable for bacterial fermentation, for example, enzymolysis is firstly carried out to form enzymolysis liquid, then nitrogen sources and carbon sources required by bacterial growth and propagation are added, and the specific preparation process and the proportion can be reasonably adjusted according to actual requirements.

In a preferred embodiment, the step of preparing the liquid to be fermented using ginseng, polygonatum odoratum, lotus seed and coix seed as raw materials comprises: mixing Ginseng radix, rhizoma Polygonati Odorati, semen Nelumbinis and Coicis semen to obtain mixed material; carrying out enzymolysis treatment on the mixed material to obtain an enzymolysis liquid; adding a nitrogen source and a carbon source into the enzymolysis liquid to obtain a liquid to be fermented; wherein the enzymolysis comprises amylase enzymolysis, saccharification enzyme enzymolysis and protease enzymolysis; preferably, the mixing ratio of the ginseng, the polygonatum odoratum, the lotus seed and the coix seed is 1-10: 1-10: 1-10: 1-10; preferably, 1-20 parts of mixed material is added into 100 parts of deionized water by weight for enzymolysis treatment; preferably, the nitrogen source is selected from any one or more of ammonium sulfate, ammonium chloride, hydrolyzed soy protein, yeast extract and milk powder; more preferably, the amount of the nitrogen source is 0.5 to 10 parts; preferably, the carbon source is selected from any one or more of glucose, fructose, maltose and hydrolyzed starch; more preferably, the amount of the carbon source added is 0.5 to 20 parts.

In the preferred embodiment, the raw materials are subjected to enzymolysis according to enzymolysis conditions of different enzymes respectively, so that various nutrient components are released, the raw materials are fully utilized by the zymophyte for growth and propagation, and a better fermentation product is obtained. The four raw materials are mixed according to the proportion, so that the effective components of the raw materials are conveniently and uniformly utilized. The nitrogen source and the carbon source are nutrient substances required by the growth and the propagation of the zymophyte. The raw material proportion of the liquid to be fermented has the beneficial effects of rich and balanced nutrition and being beneficial to efficient fermentation of various bacteria. The nitrogen source adopts the compounds of the above categories, which has the advantages of inorganic nitrogen source and organic nitrogen source combination and is suitable for yeast and lactobacillus, and the carbon source adopts monosaccharide and polysaccharide combination, which can meet different requirements of multiple strains in different periods.

In order to obtain a fermentation product with better efficacy, the pollution of mixed bacteria is avoided in the fermentation process, and any components possibly brought into the mixed bacteria need to be sterilized. In a preferred embodiment, before the liquid to be fermented is subjected to combined fermentation, the fermentation method further comprises the step of sterilizing the liquid to be fermented, preferably, the sterilization temperature is 80-130 ℃, and the sterilization time is 3s-240 min.

The specific conditions of the initial culture can be obtained by reasonably optimizing according to different types of strains used. In a preferred embodiment, the initial culture is performed at 25-30 ℃ for 24-96h by using a shaking table, and the rotation speed of the shaking table is preferably 160-200 rpm/min.

Shaking culture is a common culture method for strain culture condition screening. According to the different mixed strains, the temperature of 25-30 ℃ is selected for culture, which is beneficial to the growth and propagation of the strains. The higher the rotating speed of the shaking table is, the more frequent the contact between the fermentation solution and the strain is, and the relatively higher the growth speed of the strain is; conversely, the slower the rotation speed, the lower the contact frequency between the fermentation solution and the strain, and the relatively slower the growth rate of the strain. Therefore, the culture time is relatively short as the rotation speed is higher, whereas the culture time is relatively long as the rotation speed is lower, for obtaining a certain number of cells.

The above-mentioned pressure oxygen culture or aerobic culture is carried out after relatively optimum culture conditions are obtained in the initial culture, and therefore, the culture may be carried out according to the obtained optimum culture conditions. In a preferred embodiment, the step of anaerobic or aerobic culturing is: adding a nitrogen source and a carbon source into the primary culture to obtain primary fermentation liquor; adjusting the pH value of the primary fermentation liquid to 6.5-7.5, and then carrying out anaerobic culture or aerobic culture; preferably, the anaerobic culture or aerobic culture is carried out for 24-240 hours at 35-37.5 ℃.

In the preferred embodiment, after the optimal culture conditions are obtained in the initial culture stage, the fermentation stage is usually transferred to a fermentation tank, and the growth environment in the fermentation tank is closer to the ideal environment (including temperature, pH value and the like), so that the biomass grows more vigorously, the nutrient consumption is faster, the nutrient is insufficient, and a nitrogen source and a carbon source are supplemented to ensure that the fermentation reaches the optimal state, so that the biomass grows and breeds quickly until the production capacity is reduced.

In the above fermentation method, the plurality of bacteria include at least one yeast and at least one lactic acid bacteria, and the amount of the bacteria used is suitable for the subsequent fermentation. In a preferred embodiment, before performing the combined fermentation on the fermentation broth to be fermented by using a plurality of bacteria, the fermentation method further comprises: respectively carrying out strain activation, strain purification and enlarged culture on at least one yeast and at least one lactic acid bacteria in sequence.

The steps of sequentially activating, purifying and expanding the strains are conventional steps. Specifically, in certain preferred embodiments of the present application, the step of activating the yeast and lactic acid bacteria species is: the colony of the candida (or other yeast strains) is put into a YPD liquid culture medium in a ring mode, and the strain is activated in a shaking table. The lactobacillus plantarum (or other lactobacillus) colony is picked and put into MRS liquid culture medium in a ring mode, and the culture medium is activated in a shaking table.

In other preferred embodiments, the step of purifying the bacterial species is: and diluting and plating the activated two bacteria liquid in a gradient manner to obtain a single colony.

In certain preferred embodiments, the yeast species is grown by the steps of: selecting single yeast colony in the plate, inoculating into YPD liquid culture medium, shake culturing at 28 deg.C, and obtaining zymocyte liquid (the strain is in log phase and has concentration of about 1 × 10) when OD value is 1.0 ⁸ CFU/ml), and the thalli are collected by centrifugation.

In certain preferred embodiments, the step of expanding the lactic acid bacterial species is: selecting Lactobacillus plantarum (or other lactic acid bacteria as above)Seed) single colony is placed into MRS liquid culture medium, anaerobic or aerobic culture is carried out at 37 deg.C, when OD value is above 0.8, lactobacillus zymocyte liquid (strain is in logarithmic phase, concentration is about 1 × 10) ⁸ CFU/ml), and then centrifuged to obtain the cells.

In the above fermentation method, yeast and lactobacillus can be selected from existing strains. In a preferred embodiment, the yeast is selected from Saccharomyces cerevisiae A3.12 with the preservation number of CCTCC NO: M2017781, Pichia pastoris C1.8 with the preservation number of CCTCC NO: M2017780, or Candida albicans C1.7 with the preservation number of CCTCC NO: M2017782; preferably, the lactobacillus is selected from lactobacillus pentosus 001 with the preservation number of CCTCC NO: M2017625, lactobacillus casei 001 with the preservation number of CCTCC NO: M2017631, lactobacillus acidophilus 001 with the preservation number of CCTCC NO: M2017628, lactobacillus plantarum 001 with the preservation number of CCTCC NO: M2017629 or lactobacillus rhamnosus 001 with the preservation number of CCTCC NO: M2017630.

In the application, the yeast strains of the above types and the lactobacillus strains of the above types are preferably selected for combined fermentation, and the strains have multiple types of generated functional substances and complex enzyme systems in the growth process, and have the beneficial effects of promoting the release of raw material components in the liquid to be fermented and improving the efficacy.

In the above fermentation method, the amount of the bacteria used for fermentation is different depending on the ratio of the raw materials and the species of the bacteria. In a preferred embodiment, the plurality of bacteria comprises 0.1-5 parts by weight of yeast and 0.1-5 parts by weight of lactic acid bacteria; preferably, the dosage ratio of the various bacteria to the liquid to be fermented is 0.2-10: 100-200. The proportion of the two types of bacteria is controlled within the range, which is beneficial to shortening the fermentation process and improving the fermentation conversion efficiency. The dosage ratio of the various bacteria and the liquid to be fermented is controlled within the range, which is beneficial to more thorough fermentation of the raw materials and higher efficacy of the fermentation product.

In a more preferred embodiment, when yeast and lactic acid bacteria are mixed and then fermented with the liquid to be fermented, 0.1-5 parts of yeast and 0.1-5 parts of lactic acid bacteria are used to ferment 100 parts of the liquid to be fermented. When the two types of bacteria are adopted to ferment the fermentation liquor in sequence, the fermentation is carried out according to the dosage ratio. When the fermentation is carried out by respectively adopting the method of respectively fermenting and then mixing the two types of bacteria to the fermentation liquid, the using amount of each type of bacteria is 0.1-5 parts, and 100 parts of the fermentation liquid to be fermented are fermented.

In a second exemplary embodiment of the present application, a fermentation product is also provided, which is obtained by fermentation using any one of the fermentation methods described above. According to the application, the two edible fungi of saccharomycetes and lactic acid bacteria are fermented by different processes, so that the structural upgrade of functional substances in the efficacy of the fermented product is realized, and the effective components are fully released, so that the fermented product has the effects of resisting fatigue, delaying senescence and preserving moisture. In addition, the fermentation product is also combined with abundant microbial metabolites, so that the obtained fermentation product has pleasant flavor. Compared with the existing fermented product, the fermented product has good overall effect and has the effects of resisting fatigue, delaying aging and preserving moisture, so that the fermented product can be applied to various industries of food, health-care food and skin care products.

In a third exemplary embodiment of the present application, a fermentation kit is further provided, which comprises a fermentation raw material, at least one yeast and at least one lactic acid bacteria, wherein the fermentation raw material is ginseng, polygonatum odoratum, lotus seed and coix seed. When the kit containing the mixed bacteria of at least one yeast and at least one lactic acid bacteria is used for carrying out combined fermentation on the liquid to be fermented of the applied four Chinese medicinal material raw materials, the yeast and the lactic acid bacteria have a competitive and interdependent relationship under symbiotic conditions, and can provide substances required by the other party while competing for nutrient substances. When the fermentation is carried out respectively or sequentially, different enzymes are released respectively, so that the release of functional substances in the raw materials is promoted, and the function is upgraded, so that the obtained fermentation product has the effects of resisting fatigue, delaying senescence and preserving moisture.

In a preferred embodiment, the yeast is at least one selected from saccharomyces cerevisiae A3.12 with the preservation number of CCTCC NO: M2017781, pichia pastoris C1.8 with the preservation number of CCTCC NO: M2017780 and candida albicans C1.7 with the preservation number of CCTCC NO: M2017782, and the lactobacillus is at least one selected from lactobacillus pentosus 001 with the preservation number of CCTCC NO: M2017625, lactobacillus casei 001 with the preservation number of CCTCC NO: M2017631, lactobacillus acidophilus 001 with the preservation number of CCTCC NO: M2017628, lactobacillus plantarum 001 with the preservation number of CCTCC NO: M2017629 and lactobacillus rhamnosus 001 with the preservation number of CCTCC NO: M2017630. The yeast strains and the lactobacillus strains are preferably selected, so that various functional substances are generated in the growth process, the enzyme system is complex, and the beneficial effects of promoting the release of different functional components in the raw materials and improving the overall effect are achieved.

In a most preferred embodiment of the present application, the fermentation process of the fermentation product of the present application comprises the steps of:

1) activation of yeast and lactobacillus strains: the Candida (or other yeast species mentioned above) is picked and placed in YPD liquid medium in a ring-by-ring manner, and then placed in a shaking table to activate the species. And (3) picking bacterial colonies of the lactobacillus plantarum (or other lactobacillus strains) and putting the bacterial colonies into an MRS liquid culture medium in a ring mode, and activating in a shaking table.

2) And (3) strain purification: and (4) diluting and plating the two activated bacteria solutions in a gradient manner to obtain a single colony.

3) And (3) yeast strain amplification culture: selecting single yeast colony from the plate, inoculating to YPD liquid culture medium, shake culturing at 28 deg.C, and obtaining zymocyte liquid (the strain is in log phase and has concentration of about 1 × 10) when OD value is 1.0 ⁸ CFU/ml), and the thalli are collected by centrifugation.

4) And (3) carrying out enlarged culture on the lactobacillus strain: selecting Lactobacillus plantarum (or other lactobacillus strains), placing into MRS liquid culture medium, performing anaerobic or aerobic culture at 37 deg.C, and obtaining lactobacillus zymocyte liquid (the strain is in logarithmic phase and has concentration of about 1 × 10) when OD value is above 0.8 ⁸ CFU/ml), and then centrifuged to obtain the cells.

5) Treating ginseng, polygonatum, lotus seed and coix seed: and (3) crushing the dried raw materials, and sieving the crushed raw materials with a sieve of 80-200 meshes for later use.

6) Raw material enzymolysis and batching: ginseng, polygonatum, lotus seed and coix seed, wherein the proportion is as follows: 1-10: 1-10: 1-10: 1-10, adding the mixed powder according to the proportion of 1-20 in 100 parts of deionized water, and performing hydrolysis treatment according to the optimized treatment process of amylase, saccharifying enzyme and protease. Adding 0.5-20 parts of glucose or fructose or maltose or hydrolyzed starch, 0.5-10 parts of ammonium sulfate or ammonium chloride or hydrolyzed soybean protein or yeast extract or milk powder, and sterilizing at 80-130 deg.C for 3s-240 min.

7) The fermentation method 1: adding 0.1-5 parts of candida thalli and 0.1-5 parts of lactobacillus plantarum thalli into 100 parts of the raw material mixed solution cooled in the last step, carrying out shake culture at 25-30 ℃ for 24-96h (shaking table rotation speed 160-200rpm/min), adding 0.5-10 parts of fructose, glucose, sucrose, maltose or hydrolyzed starch, 0.5-10 parts of ammonium sulfate, ammonium chloride, soybean protein hydrolysate, milk powder or yeast extract and the like, and then regulating the pH to be between 6.5-7.5 and carrying out anaerobic or oxygen-consuming culture fermentation at 37 ℃ for 24-240 h.

The fermentation method 2: adding 0.1-5 parts of candida thalli into 100 parts of the raw material mixed solution cooled in the last step, performing shake culture at 25-28 ℃ for 24-120h (the rotation speed of a shaking table is 160-. Adding sterilized fructose, glucose, sucrose, maltose or hydrolyzed starch 0.5-10 parts, ammonium sulfate or ammonium chloride or soybean protein hydrolysate or milk powder or yeast extract 0.5-10 parts, and adjusting pH to 6.5-7.5. Then 0.1-5 parts of lactobacillus plantarum thallus is added again, and the lactobacillus plantarum thallus is cultured and fermented for 24-240 hours under anaerobic or oxygen-consuming conditions at 35-37 ℃. Or fermenting lactobacillus first and then yeast.

The fermentation method 3: 0.1-5 parts of yeast thallus is added into 100 parts of the raw material mixed solution cooled in the last step, and the mixture is cultured for 24-96h by a shaking table at the temperature of 25-28 ℃ (the rotation speed of the shaking table is 160-200 rpm/min). Simultaneously, 0.1 to 5 parts of lactobacillus plantarum thalli are added into 100 parts of the raw material mixed liquor cooled in the last step, and anaerobic or oxygen-consuming culture fermentation is carried out for 24 to 240 hours at the temperature of between 35 and 37 ℃. After the two-step fermentation is finished respectively, equal-volume mixing is carried out.

8) Clarification treatment: and (3) treating the fermented product by centrifuging and plate-frame filtering to obtain supernatant, namely the composite fermented product.

The advantageous effects of the present application will be further described with reference to specific examples.

The raw materials of the following examples are ginseng, polygonatum odoratum, lotus seed, coix seed, nitrogen sources (including but not limited to ammonium sulfate, ammonium chloride, hydrolyzed soy protein, milk powder, yeast extract), carbon sources (including but not limited to glucose, fructose, sucrose, maltose, hydrolyzed starch, etc.), yeasts (including but not limited to saccharomyces cerevisiae, pichia, candida), lactic acid bacteria (including but not limited to lactobacillus pentosus, lactobacillus casei, lactobacillus acidophilus, lactobacillus plantarum, lactobacillus rhamnosus), and the raw materials can be purchased from the market except for the strains having the above preservation information.

Example 1

Yeast and lactobacillus strain activation: a ring of saccharomyces cerevisiae colonies is picked and put into YPD liquid culture medium, and then put into a shaking table to activate strains. And (3) picking lactobacillus acidophilus colonies, putting the lactobacillus acidophilus colonies into an MRS liquid culture medium in a ring mode, and activating in a shaking table.

And (3) strain purification: and diluting and plating the two activated bacteria liquid in a gradient manner to obtain a single colony.

And (3) yeast strain amplification culture: and (3) selecting a single colony in the plate in the previous step, inoculating the single colony into an YPD liquid culture medium, carrying out shake culture at 28 ℃, obtaining a zymophyte liquid when the OD value is 1.0, and centrifugally collecting thalli.

Carrying out amplification culture on lactobacillus strains: and (3) selecting lactobacillus acidophilus colonies, putting the lactobacillus acidophilus colonies into an MRS liquid culture medium in a loop mode, performing purification and amplification culture, performing anaerobic fermentation at 37 ℃, obtaining a zymophyte liquid when the OD value is 1.0, and centrifuging to collect thalli.

Treating ginseng, polygonatum, lotus seed and coix seed: pulverizing dried raw materials, processing, and sieving with 100 mesh sieve.

Post-treatment and compounding of raw materials: ginseng, polygonatum, lotus seed and coix seed, wherein the proportion is as follows: 5: 2: 3: 5, mixing, adding 10 parts of the mixed powder into 100 parts of deionized water, and hydrolyzing by using amylase, saccharifying enzyme and protease. After the hydrolysis is finished (obtaining the composite extract), 10 parts of glucose and 5 parts of ammonium sulfate are added, and the mixture is sterilized for 30min at 90 ℃.

Mixing and synchronously fermenting: 5 parts of yeast thallus after enlarged culture and 5 parts of lactobacillus acidophilus thallus are added into the raw material mixed liquid cooled in the previous step, and shaking culture is carried out for 96 hours at 25 ℃ (the rotation speed of a shaking table is 180 rpm/min). And 5 parts of sterilized glucose and 3 parts of soybean protein hydrolysate are added again, the ph is adjusted to be 6.5, and the fermentation is carried out for 48 hours at 37 ℃ in an anaerobic or aerobic culture mode.

Clarification treatment: after the fermentation is finished, the centrifuge centrifuges the mixture for 10min at high speed of 5000r/min to obtain supernatant, namely the fermentation product.

And (3) sterilization: sterilizing the fermented product at 60 deg.C for 0.5 h.

Example 2

Yeast and lactobacillus strain activation: one ring of candida colony is picked and put into YPD liquid culture medium, and the strain is activated in a shaking table. The lactobacillus plantarum colony is picked and placed in MRS liquid culture medium in a ring mode, and the lactobacillus plantarum colony is activated in a shaking table.

And (3) yeast strain amplification culture: and (3) selecting a single colony in the plate in the previous step, inoculating the single colony into an YPD liquid culture medium, carrying out shake culture at 28 ℃, obtaining zymocyte liquid when the OD value is more than or equal to 1.0, and centrifugally collecting thalli.

And (3) carrying out enlarged culture on the lactobacillus strain: and (3) selecting lactobacillus casei bacterial colonies, putting the lactobacillus casei bacterial colonies into an MRS liquid culture medium, performing purification and amplification culture, performing anaerobic fermentation at 37 ℃, obtaining zymocyte liquid when the OD value is 1.0, and centrifuging to collect thalli.

Treating ginseng, polygonatum, lotus seed and coix seed: pulverizing the dried raw materials, and sieving with 200 mesh sieve.

Raw material treatment and compounding: ginseng, polygonatum, lotus seed and coix seed, wherein the proportion is 7: 3: 7: 3, mixing, adding 5 parts of the mixed powder into 100 parts of deionized water, and hydrolyzing by using amylase, saccharifying enzyme and protease. After the hydrolysis (to obtain the composite extract), 2 parts of maltose and 2 parts of milk powder are added, and the mixture is sterilized for 30min at 90 ℃.

Respectively fermenting yeast and lactobacillus: adding 1 part of yeast thallus into 100 parts of the raw material mixed solution cooled in the previous step, and performing shake cultivation at 25 ℃ for 72h (the rotation speed of a shaking table is 200 rpm/min). Meanwhile, 1 part of lactobacillus pentosus thallus is added into 100 parts of the raw material mixed solution cooled in the previous step, and anaerobic culture and fermentation are carried out for 96 hours at the temperature of 35-37 ℃. After the two-step fermentation is respectively finished, the equal volume mixing is carried out.

And (3) sterilization: sterilizing the fermented product at 130 deg.C for 15 s.

Example 3

Yeast and lactobacillus strain activation: and (3) picking a ring of pichia pastoris bacterial colony, putting the ring into a YPD liquid culture medium, and putting the YPD liquid culture medium into a shaking table to activate the strain. And (3) picking a colony of the lactobacillus pentosus, putting the colony into an MRS liquid culture medium in a ring mode, and activating in a shaking table.

And (3) yeast strain amplification culture: and (3) selecting a single colony in the flat plate in the previous step, inoculating the single colony into an YPD liquid culture medium, performing shake culture at 28 ℃, obtaining zymocyte liquid when the OD value is more than or equal to 1.0, and performing centrifugation to collect thalli.

And (3) carrying out enlarged culture on the lactobacillus strain: and (3) selecting a colony of the lactobacillus pentosus, putting the colony into an MRS liquid culture medium, performing purification and amplification culture, performing oxygen-consuming fermentation at 35 ℃, obtaining a zymocyte liquid when the OD value is 1.0, and centrifuging to collect thalli.

Raw material treatment and compounding: ginseng, polygonatum, lotus seed and coix seed, wherein the proportion is as follows: 2: 5: 7: 5, mixing, adding 10 parts of the mixed powder into 100 parts of deionized water, and hydrolyzing by using amylase, saccharifying enzyme and protease. After the hydrolysis (obtaining the composite extract), 1 part of hydrolyzed starch sugar, 1 part of ammonium sulfate and 1 part of yeast extract are added, and the mixture is sterilized for 20min at 120 ℃.

Step-by-step fermentation of yeast and lactic acid bacteria: adding 5 parts of yeast thallus into 100 parts of the raw material mixed solution cooled in the previous step, performing shake culture at 28 ℃ for 24h (shaking table rotation speed of 200rpm/min), and heating for inactivation. The sterilized 7 parts of sucrose and 3 parts of soy protein hydrolysate were added again, and then ph was adjusted to 6.5 to 7.5. And adding 0.1 part of lactobacillus pentosus thallus again, and performing anaerobic culture and fermentation at 37 ℃ for 96 hours.

Clarification treatment: and filtering by using a plate and frame filter after the fermentation is finished to obtain supernatant, namely the composite fermentation product.

And (3) sterilization: sterilizing the fermented product at 130 deg.C for 5 s.

Examples 4 to 9

Examples 4 to 9 Using the starting materials shown in tables 1 and 2 and the conditions shown in Table 3, the same procedures as in example 1 were carried out to obtain fermentation products of the respective examples.

Comparative example 1

The fermentation is carried out by adopting a single zymophyte fermentation mode, and the method comprises the following specific steps:

the starting material from example 7 was used and 10% pichia pastoris seed solution (containing 5 x 10 pichia pastoris per ml seed solution) was added ⁷ CFU), stirring uniformly, fermenting for 15 days at the temperature of 28 ℃, and stopping fermentation when the pH of the fermentation system reaches 4.0 to obtain the pichia pastoris fermentation liquor.

Clarification treatment: and filtering by using a plate and frame filter after the fermentation is finished to obtain supernatant, namely the fermentation product.

And (3) sterilization: sterilizing the fermentation product at 120 deg.C for 15 min.

Comparative example 2

Using the starting material obtained in example 7, 10% of a seed solution of Lactobacillus casei (containing 5 x 10 Lactobacillus casei per ml of seed solution) was added ⁸ CFU), stirring uniformly, fermenting at 37 deg.C for 15 days, and stopping fermentation when pH of the fermentation system reaches 4.3 to obtain lactobacillus fermentation liquid.

Comparative example 3

Using the material obtained in example 7, 10% of Lactobacillus casei seed solution (containing 5 x 10 Lactobacillus casei per ml of seed solution) was added ⁸ CFU), stirring, fermenting at 37 deg.C for 15 days, and stopping fermentation when pH of the fermentation system reaches 4.3And obtaining the lactobacillus fermentation liquor.

Adding 8% Lactobacillus plantarum (4.5 x 10 Lactobacillus plantarum per ml seed solution) into lactobacillus fermentation broth ⁷ CFU), stirring uniformly, and fermenting at 37 ℃ for 60 days to obtain lactobacillus plantarum fermentation liquor.

Table 1: raw materials for examples and comparative examples

Table 2:

item	The mass ratio of ginseng, polygonatum, lotus seed and coix seed	Particle size of raw material
			Example 1	5：2：3：5	100 mesh
Example 2	7：3：7：3	200 mesh
			Example 3	2：5：7：5	200 mesh
Example 4	1:1:1:1	90 mesh
			Example 5	10:1:1:1	120 mesh
Example 6	1:10:1:1	180 mesh
			Example 7	1:1:10:1	180 mesh
Example 8	1:1:1:10	210 mesh
			Example 9	10:0.5:1:10	180 mesh
Comparative example 1	1:1:10:1	180 mesh
			Comparative example 2	1:1:10:1	180 mesh
Comparative example 3	1:1:10:1	180 mesh

Attached: the Saccharomyces cerevisiae and Lactobacillus bulgaricus used in example 9 were all commercially available products, and the Saccharomyces cerevisiae was obtained from low-sugar high-activity dry yeast of Angel Yeast, Inc., with production lot number of 2018021201. The lactobacillus was obtained from Lactobacillus bulgaricus of Shanghai purple stone Biotechnology Ltd, and its production lot was 20171225.

Table 3:

and (3) testing:

(1) weight bearing swimming experiment

The swimming exercise is carried out after the experimental mice are initially selected, 30 mice with the standard deviation of the load measurement basic time less than 10min are selected and randomly divided into 3 groups, namely a control group and an experimental group, wherein the experimental group comprises a compound extract group (namely a product obtained by hydrolyzing a raw material before fermentation) and a compound fermentation liquid group (namely a supernatant obtained finally after fermentation) of each embodiment and each comparative example. The experimental group was administered 10g/kg weight concentration of the complex extract and the complex fermented product, respectively, and the control group was administered 10g/kg daily of stomach-feeding sterile distilled water. The mice are subjected to continuous 30d intragastric administration and a weight swimming test 2h after 30d intragastric administration, lead blocks with the weight of 5% of the mice are tied at the tail part near the heart end 1/2 and are placed in a water depth of 50cm and a volume of about 1m ³ And (3) in a water tank with the water temperature of 28 +/-2 ℃, the mice sink for 10s and cannot float to mark exhaustion, and the swimming exhaustion time of the mice is measured and recorded. The test results are shown in Table 4.

Table 4: influence of the Complex fermentation group and Complex extraction group on the outcome of swimming with load (n-10)

The mouse weight-bearing swimming test is the most direct method for investigating the anti-fatigue capability, and the swimming time is positively correlated with the animal exercise fatigue degree. As can be seen from table 4, the swimming exhaustion time of the compound extract group and the compound fermentation group of each example is significantly longer than that of the control group, and the compound fermentation group is higher than that of the compound extract group, which indicates that the compound fermentation group has more significant anti-fatigue and physical performance enhancing effects.

(2) Comparison of proliferation effects of composite extract and composite fermentation product on human fibroblasts under ultraviolet irradiation damage

Human dermal fibroblasts (HSF) were divided into a control group, a model group and a sample group. Culturing the control group in normal culture solution without UVA irradiation; model group and sample group: removing culture solution before UVA irradiation, washing with PBS once, adding small amount of PBS to cover the upper layer of irradiated cell, and irradiating with UVA light source at irradiation dose of 10J/cm ² . Model group cells were plated at 4X 10 ⁴ Inoculating each cell/well in 96-well plate, adding the compound extract and compound fermentation product into culture medium after cell adherence, respectively, adding equal amount of culture solution as control to blank group and model, and standing at 37 deg.C and 5% CO ₂ After the culture is continued for 24h in the incubator, the culture solution is carefully removed by aspiration, then 100 μ L of 5mg/mL tetramethylazozolium (MTT) solution is added to each well, after the culture is continued for 4h, the supernatant is carefully removed by aspiration, 150 μ L of DMSO is added to each well, shaking is performed for 15min to fully dissolve the precipitate, the absorbance at the wavelength of 570nm is measured by using a microplate reader, and the cell survival rate of each group is calculated (the calculation formula is as follows), and the detection results of each group are shown in table 5:

table 5: the effect of the complex extract and the complex fermentation product on the proliferation activity of HSF cells after UVA irradiation (n-3,

)

as shown in Table 5 above, the test piece passed 10J/cm in comparison with the control group ² UVA irradiationThe light absorption value and the survival rate of the model group cells after the model is made are reduced to 0.432 +/-0.02 and 57.1 percent respectively; compared with the model group, the cell proliferation activity of the added compound extract group and the compound fermentation group is obviously recovered, the compound fermentation and the compound extract have the effect of promoting the proliferation of human fibroblasts, the effect of the compound fermentation is better than that of the compound extract, compared with the experimental group, the fermentation group of the comparison group is also better than that of the extraction group, but the increase effect is not as obvious as that of the experimental group.

(3) In vitro moisture retention test

Selecting a culture dish with the constant weight, respectively pasting a layer of 3M medical breathable adhesive tape on the inner side of the culture dish, respectively sucking 500 mu L of the compound extracting solution in the embodiment 2 and the compound fermentation liquor in the embodiment 2, respectively, dropping the compound extracting solution and the compound fermentation liquor on the breathable adhesive tape, weighing the compound extracting solution and the compound fermentation liquor in a constant-temperature constant-humidity dryer with the relative humidity of 60 percent at the temperature of 25 ℃ for 8 hours, weighing the compound extracting solution and the compound fermentation liquor once every 1 hour, and calculating the moisturizing rate according to the difference between the initial weight:

X(％)＝Mt/M0×100％；

note: m0 is initial weight in g; mt is the weight of the sampling point and is given in g; t is the sampling time, h.

The results are shown in FIG. 1. As can be seen from fig. 1, both the complex extract and the complex fermented product have moisturizing effects, and the difference is small within 3 hours in the initial period of the experiment, and the moisturizing rate of the normal complex fermented product is higher than that of the complex extract over time.

From the above description, it can be seen that the above-described embodiments of the present invention achieve the following technical effects: the raw materials of the application are ginseng, polygonatum, lotus seed and coix seed, wherein ginsenoside is the main component playing various effects in the ginseng, and polygonatum polysaccharide and flavone are the main substances in the polygonatum. The water-soluble polysaccharide, total flavone and superoxide dismutase in lotus seeds are the main three substances. The main active components contained in Coicis semen include Coicis semen ester, triglyceride, fatty acids, lactam, Coicis semen lactone, saccharide, sterol, and triterpene. The raw materials can play a synergistic interaction role among different functional components after being matched and combined, and meanwhile, the four raw materials contain abundant starch, protein, trace elements and the like, so that sufficient raw material substances can be provided for microbial fermentation.

By utilizing the fermentation characteristics of edible safe strains such as saccharomycetes and lactic acid bacteria, the raw material composition containing ginseng, polygonatum odoratum, lotus seeds and coix seeds is fermented, the two strains generate various functional substances in the growth process, the enzyme system is complex, the release of different functional components in the raw materials can be promoted, the content of the functional substances in the traditional Chinese medicine is increased, the structure is converted (for example, ginsenoside is converted from common saponin into rare ginsenoside, flavonoid structure changes the water solubility and the like), and the overall effect of the fermentation product is improved. Has effects in relieving fatigue, delaying aging and keeping moisture

The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A fermentation process, comprising:

preparing a liquid to be fermented by taking ginseng, polygonatum, lotus seed and coix seed as raw materials,

performing combined fermentation on the liquid to be fermented by using various bacteria to obtain a fermentation product,

wherein the plurality of bacteria comprises at least one yeast and at least one lactic acid bacteria;

the yeast is selected from Saccharomyces cerevisiae A3.12 with the preservation number of CCTCC NO: M2017781, Pichia pastoris C1.8 with the preservation number of CCTCC NO: M2017780, or Candida albicans C1.7 with the preservation number of CCTCC NO: M2017782;

the lactobacillus selects lactobacillus pentosus 001 with the preservation number of CCTCC NO: M2017625, lactobacillus casei 001 with the preservation number of CCTCC NO: M2017631, lactobacillus acidophilus 001 with the preservation number of CCTCC NO: M2017628, lactobacillus plantarum 001 with the preservation number of CCTCC NO: M2017629 or lactobacillus rhamnosus 001 with the preservation number of M2017630.

2. The fermentation method according to claim 1, wherein the step of performing combined fermentation on the liquid to be fermented by using a plurality of bacteria to obtain the fermentation product comprises:

mixing the various bacteria with the liquid to be fermented and then carrying out primary culture to obtain a primary culture;

and carrying out anaerobic or aerobic culture on the primary culture to obtain the fermentation product.

3. The fermentation method according to claim 1, wherein the step of performing combined fermentation on the liquid to be fermented by using a plurality of bacteria to obtain the fermentation product comprises:

mixing the saccharomycetes and the liquid to be fermented and then carrying out primary culture to obtain a primary culture;

and (3) carrying out anaerobic or aerobic culture on the primary culture by using the lactic acid bacteria to obtain the fermentation product.

4. The fermentation method according to claim 1, wherein the step of performing combined fermentation on the liquid to be fermented by using a plurality of bacteria to obtain the fermentation product comprises:

mixing the lactic acid bacteria with the liquid to be fermented and then carrying out primary culture to obtain a primary culture;

and carrying out anaerobic or aerobic culture on the primary culture by using the microzyme to obtain the fermentation product.

5. The fermentation method according to claim 1, wherein the step of performing combined fermentation on the liquid to be fermented by using a plurality of bacteria to obtain the fermentation product comprises:

mixing the lactic acid bacteria with the liquid to be fermented, and then carrying out fermentation culture to obtain a first fermentation culture;

mixing the yeast and the liquid to be fermented, and then carrying out fermentation culture to obtain a second fermentation culture;

mixing the first fermentation culture with the second fermentation culture to obtain the fermentation product.

6. The fermentation process of claim 5, wherein the first fermentation culture and the second fermentation culture are mixed in equal volumes to provide the fermentation product.

7. The fermentation method of claim 1, wherein the step of preparing the liquid to be fermented using ginseng, polygonatum odoratum, lotus seed and coix seed as raw materials comprises:

mixing Ginseng radix, rhizoma Polygonati Odorati, semen Nelumbinis and Coicis semen to obtain mixed material;

carrying out enzymolysis treatment on the mixed material to obtain an enzymolysis liquid;

adding a nitrogen source and a carbon source into the enzymolysis liquid to obtain the liquid to be fermented;

wherein the enzymolysis comprises amylase enzymolysis, saccharification enzyme enzymolysis and protease enzymolysis.

8. The fermentation method according to claim 7, wherein the mixing ratio of the ginseng, the polygonatum odoratum, the lotus seed and the coix seed is 1-10: 1-10: 1-10: 1 to 10.

9. The fermentation method according to claim 7, wherein 1 to 20 parts by weight of the mixed material is added to 100 parts by weight of deionized water to perform the enzymatic hydrolysis treatment.

10. The fermentation method according to claim 7, wherein the nitrogen source is selected from any one or more of ammonium sulfate, ammonium chloride, hydrolyzed soybean protein, yeast extract, and milk powder.

11. The fermentation method according to claim 10, wherein the nitrogen source is added in an amount of 0.5 to 10 parts.

12. The fermentation process of claim 7, wherein the carbon source is selected from any one or more of glucose, fructose, maltose and hydrolyzed starch.

13. The fermentation method according to claim 12, wherein the carbon source is added in an amount of 0.5 to 20 parts.

14. The fermentation process of claim 7, wherein the fermentation process further comprises the step of sterilizing the liquid to be fermented before the combined fermentation of the liquid to be fermented.

15. The fermentation method according to claim 14, wherein the sterilization temperature is 80 ℃ to 130 ℃, and the sterilization time is 3s to 240 min.

16. The fermentation method according to any one of claims 2 to 4, wherein the primary culture is performed at 25 ℃ to 30 ℃ for 24 to 96 hours by using a shaker.

17. The fermentation process of claim 16, wherein the rotation speed of the rocking platforms is 160 to 200 rpm/min.

18. The fermentation process of claim 16, wherein the step of anaerobic or aerobic culturing is:

adding a nitrogen source and a carbon source into the primary culture to obtain primary fermentation liquor;

and adjusting the pH value of the primary fermentation liquid to 6.5-7.5, and then carrying out anaerobic culture or aerobic culture.

19. The fermentation method according to claim 18, wherein the anaerobic culture or aerobic culture is carried out for 24-240 hours at a temperature of 35-37.5 ℃.

20. The fermentation method of claim 1, wherein prior to the combined fermentation of the liquid to be fermented with a plurality of bacteria, the fermentation method further comprises:

respectively carrying out strain activation, strain purification and enlarged culture on at least one kind of the microzyme and at least one kind of the lactic acid bacteria in sequence.

21. The fermentation method according to any one of claims 1 to 15, wherein the plurality of bacteria includes 0.1 to 5 parts by weight of the yeast and 0.1 to 5 parts by weight of the lactic acid bacteria.

22. The fermentation method according to claim 21, wherein the ratio of the plurality of bacteria to the liquid to be fermented is 0.2-10: 100-200.

23. A fermentation product obtained by fermentation using the fermentation process of any one of claims 1 to 22.