Process method for extracting active metabolite of fermentation probiotics from vegetable drugs

Abstract

The invention relates to the technical field of probiotic fermentation, in particular to a process method for extracting fermentation probiotic active metabolites from botanical drugs, which comprises the following steps: S1, raw material preparation: selecting traditional Chinese medicinal materials with homology of medicine and food, and pretreating the traditional Chinese medicinal materials; s2, extracting and purifying the traditional Chinese medicinal materials: extracting effective components in the traditional Chinese medicinal materials by using a corresponding solvent and a corresponding method; s3, probiotic fermentation: screening and culturing an excellent probiotic flora; s4, application of a fermentation technology; s5, adding an enzyme for catalytic reaction; s6, application of a control project; s7, extracting and purifying a product; the preparation method has the beneficial effects that effective components in plant medicines can be more effectively extracted and utilized, active metabolites of probiotics can also be generated in the fermentation process, the medicine effect and bioavailability can be further enhanced, the combined development of traditional Chinese medicines and a microbial fermentation technology is further facilitated, and the use requirements can be well met.

Description

Technical Field

[0001] The invention relates to the technical field of probiotic fermentation, in particular to a process for extracting active metabolites of fermented probiotics from botanical medicines. Background Art

[0002] With the improvement of living standards, people's awareness of health has become more and more obvious. As a cultural treasure of my country for many years, traditional Chinese medicine has also kept pace with the times in this development trend. With the change of health concepts and the expansion of the health care market, people are paying more and more attention to natural and safe health products and medical supplies. In this context, the process of modernization of traditional Chinese medicine has also promoted the exploration and research of the combination of traditional Chinese medicine and microbial fermentation technology.

[0003] At present, the existing technology lacks a process method that can effectively extract and utilize the active ingredients in botanical medicines and produce active metabolites of probiotics during the fermentation process, which is not conducive to further enhancing the efficacy and bioavailability, and has an adverse impact on the combined development of traditional Chinese medicine and microbial fermentation technology, and cannot meet the use needs well. Therefore, a process method for extracting active metabolites of probiotics from botanical medicines is proposed. Summary of the invention

[0004] In order to overcome the technical defects in the prior art and effectively solve the technical problems in the background technology, the present invention provides the following technical solutions:

[0005] A process for extracting active metabolites of probiotics from botanical medicines, comprising the following steps:

[0006] S1: Raw material preparation: Select Chinese medicinal materials with the properties of both medicine and food, and pre-process them;

[0007] S2: Extraction and purification of Chinese medicinal materials: Use corresponding solvents and methods to extract the active ingredients in Chinese medicinal materials;

[0008] S3: Probiotic fermentation: screening and cultivating excellent probiotic bacteria;

[0009] S4: Application of fermentation technology: Add beneficial bacteria to Chinese medicinal materials, combine purified Chinese medicinal materials with probiotics, and perform anaerobic fermentation; control fermentation conditions to promote the growth and metabolism of probiotics, and use enzymes produced by microorganisms to react with active ingredients in Chinese medicine to increase the content;

[0010] S5: Add enzyme to catalyze the reaction: Use enzyme to destroy the plant cell wall and release active substances, that is, use the catalytic effect of enzyme to accelerate the completion of the reaction;

[0011] S6: Application of control engineering: Introduce control engineering to automatically control the fermentation process of traditional Chinese medicine, monitor and adjust fermentation parameters, and ensure the stable quality of fermentation products;

[0012] S7: Product extraction and purification: After fermentation, the active metabolites of probiotics are extracted, the extract is purified, impurities are removed, and the final product is obtained.

[0013] As a preferred technical solution of the present invention, the Chinese medicinal materials include 15-25 parts of chicken gizzard lining, 35-45 parts of Poria cocos, 15-25 parts of yam, 15-25 parts of hawthorn, 15-25 parts of malt, 15-20 parts of licorice, and 5-10 parts of tangerine peel.

[0014] As a preferred technical solution of the present invention, the probiotics include Bifidobacterium lactis and Lactobacillus paracasei.

[0015] As a preferred technical solution of the present invention, the steps of the Chinese medicinal material pretreatment and extraction and purification are as follows: selecting Chinese medicinal materials according to the component ratio, washing and cutting, decocting with water twice to obtain a decoction, and concentrating and sterilizing the decoction to obtain a Chinese medicinal extract, wherein the decoction step is specifically as follows:

[0016] (1) Mix water and Chinese medicinal materials in a weight ratio of 3:1, soak for 1 to 2 hours and then boil. Keep boiling for 20 to 25 minutes and filter out the decoction;

[0017] (2) Mix water and the filter residue in step (1) in a weight ratio of 1:1, heat and boil, use ultrasound to assist dispersion during the boiling process, and continue for 20 to 25 minutes, then filter out the decoction;

[0018] (3) Mix the decoctions in step (1) and step (2), and concentrate and dry them.

[0019] As a preferred technical solution of the present invention, the probiotic fermentation and application operation steps are: select probiotics, activate the strains, and then inoculate them on the culture medium, then mix the culture medium with the Chinese medicine extract, and after fermentation and cultivation, centrifuge and concentrate to obtain bacterial mud; mix the protective agent and the bacterial mud, vacuum freeze-dry, crush and mix, and purify and remove impurities to obtain the metabolites.

[0020] As a preferred technical solution of the present invention, the culture medium comprises the following components: 8-10 g / L of peptone, 10-12 g / L of agar, 4-5 g / L of lactose, 3-5 g / L of yeast extract, 3-4 g / L of glucose, 1-2 g / L of sodium acetate, 0.5-1 g / L of dipotassium hydrogen phosphate, and 0.5-1 g / L of magnesium sulfate; the pH of the culture medium is adjusted to 6.5, the Bifidobacterium lactis is inoculated on the culture medium at a volume ratio of 1:10; the Lactobacillus paracasei is inoculated on another culture medium at a volume ratio of 1:5, the culture medium inoculated with Bifidobacterium lactis and the culture medium of Lactobacillus paracasei are mixed at a volume ratio of 1:(1-1.5), and then the mixed culture medium is mixed with a traditional Chinese medicine extract at a weight ratio of 1:(80-100).

[0021] As a preferred technical solution of the present invention, the protective agent comprises the following components in weight percentage: 20-30wt% skim milk powder, 10-20wt% whey protein, 5-15wt% glucose, and the balance is trehalose.

[0022] As a preferred technical solution of the present invention, the vacuum freeze-drying specifically comprises the following steps:

[0023] (1) Precooling: control the temperature to -15~-20℃, the vacuum degree to 27~35Pa, and precool for 4~4.5h;

[0024] (2) Sublimation drying: control the temperature to -50~-40℃, the vacuum degree to 15~25Pa, and the sublimation drying time to 12~16h;

[0025] (3) Desorption and drying: control the temperature to 35-50°C, the vacuum degree to 25-30Pa, and the desorption and drying time to 6-8h.

[0026] As a preferred technical solution of the present invention, the fermentation conditions are room temperature, 55% to 65% humidity, pH 6.2 to 6.8, pressure 0.03 to 0.05 MPa, and time 10 to 12 hours.

[0027] As a preferred technical solution of the present invention, the enzyme catalyzing the reaction is a glycosyl hydrolase.

[0028] Compared with the prior art, the beneficial effects of the present invention are as follows: the process method for extracting active metabolites of probiotics from botanical medicines can be operated through corresponding steps, so that the effective ingredients in the botanical medicines can be more effectively extracted and utilized, and at the same time, active metabolites of probiotics can also be produced during the fermentation process, which is beneficial to further enhance the efficacy and bioavailability, and further beneficial to the combined development of traditional Chinese medicine and microbial fermentation technology, and can well meet the use needs. DETAILED DESCRIPTION

[0029] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution in the embodiment of the present invention will be further clearly and completely described below. Obviously, the described embodiment is only a part of the embodiment of the present invention, not all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0030] The present invention provides a technical solution: a process for extracting active metabolites of probiotics from botanical medicines, comprising the following steps:

[0031] S1: Raw material preparation: Select Chinese medicinal materials with the properties of both medicine and food, and pre-process them;

[0032] S2: Extraction and purification of Chinese medicinal materials: Use corresponding solvents and methods to extract the active ingredients in Chinese medicinal materials;

[0033] S3: Probiotic fermentation: screening and cultivating excellent probiotic bacteria;

[0034] S4: Application of fermentation technology: Add beneficial bacteria to Chinese medicinal materials, combine purified Chinese medicinal materials with probiotics, and perform anaerobic fermentation; control fermentation conditions to promote the growth and metabolism of probiotics, and use enzymes produced by microorganisms to react with active ingredients in Chinese medicine to increase the content. Specifically, ferment Chinese herbal medicines under suitable temperature, humidity, air, moisture and other environments, perform biological transformation on the extracted active ingredients of Chinese medicine, degrade macromolecules into small molecules, and improve the absorption and efficacy of active ingredients;

[0035] S5: Add enzyme catalysis reaction: use enzymes to destroy plant cell walls and release active substances, that is, use the catalytic effect of enzymes to accelerate the completion of reactions. Specifically, enzymes (which can be glycosyl hydrolases) can destroy plant cell walls, promote the release of active substances, and transform the chemical components in traditional Chinese medicine through the metabolic activities of microorganisms, thereby improving the efficacy and bioavailability of drugs;

[0036] S6: Application of control engineering: control engineering is introduced to realize automatic control of the fermentation process of traditional Chinese medicine, monitor and adjust fermentation parameters, reduce the impact of human factors, and thus ensure the stable quality of fermentation products;

[0037] S7: Product extraction and purification: After fermentation, the active metabolites of probiotics are extracted, the extract is purified, impurities are removed, and the final product is obtained.

[0038] Furthermore, the Chinese medicinal materials are 20 parts of chicken gizzard lining, 40 parts of Poria cocos, 20 parts of yam, 20 parts of hawthorn, 20 parts of malt, 18 parts of liquorice, and 8 parts of tangerine peel.

[0039] Furthermore, the probiotics include Bifidobacterium lactis and Lactobacillus paracasei.

[0040] Further, the steps of Chinese medicinal material pretreatment and extraction and purification are as follows: selecting Chinese medicinal materials according to the component ratio, washing and cutting, decocting with water twice to obtain a decoction, and concentrating and sterilizing the decoction to obtain a Chinese medicinal extract, wherein the decoction steps are as follows:

[0041] (1) Mix water and Chinese medicinal materials in a weight ratio of 3:1, soak for 1 to 2 hours and then boil. Keep boiling for 20 to 25 minutes and filter out the decoction;

[0042] (2) Mix water and the filter residue in step (1) in a weight ratio of 1:1, heat and boil, use ultrasound to assist dispersion during the boiling process, and continue for 20 to 25 minutes, then filter out the decoction;

[0043] (3) Mix the decoctions in step (1) and step (2), and concentrate and dry them.

[0044] Furthermore, the probiotic fermentation and application operation steps are as follows: select probiotics, activate the strains, inoculate them onto the culture medium, then mix the culture medium with the Chinese medicine extract, and after fermentation and cultivation, centrifuge and concentrate to obtain bacterial mud; mix the protective agent and the bacterial mud, vacuum freeze-dry, crush and mix, and purify and remove impurities to obtain metabolites.

[0045] Furthermore, the culture medium includes the following components: 8-10 g / L of peptone, 10-12 g / L of agar, 4-5 g / L of lactose, 3-5 g / L of yeast extract, 3-4 g / L of glucose, 1-2 g / L of sodium acetate, 0.5-1 g / L of dipotassium hydrogen phosphate, and 0.5-1 g / L of magnesium sulfate; the pH of the culture medium is adjusted to 6.5, and Bifidobacterium lactis is inoculated on the culture medium at a volume ratio of 1:10; Lactobacillus paracasei is inoculated on another culture medium at a volume ratio of 1:5, and the culture medium inoculated with Bifidobacterium lactis is mixed with the culture medium of Lactobacillus paracasei at a volume ratio of 1:(1-1.5), and then the mixed culture medium is mixed with the traditional Chinese medicine extract at a weight ratio of 1:(80-100).

[0046] Furthermore, the protective agent comprises the following components in weight percentage: 20-30wt% skim milk powder, 10-20wt% whey protein, 5-15wt% glucose, and the balance is trehalose.

[0047] Further, vacuum freeze drying specifically comprises the following steps:

[0048] (1) Precooling: control the temperature to -15~-20℃, the vacuum degree to 27~35Pa, and precool for 4~4.5h;

[0049] (2) Sublimation drying: control the temperature to -50~-40℃, the vacuum degree to 15~25Pa, and the sublimation drying time to 12~16h;

[0050] (3) Desorption and drying: control the temperature to 35-50°C, the vacuum degree to 25-30Pa, and the desorption and drying time to 6-8h.

[0051] Furthermore, the fermentation conditions are room temperature, 55% to 65% humidity, pH 6.2 to 6.8, pressure 0.03 to 0.05 MPa, and time 10 to 12 hours.

[0052] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A process for extracting active metabolites of probiotics from botanicals, characterized in that: The following steps are involved: S1: Raw material preparation: Select Chinese medicinal materials with the properties of both medicine and food, and pre-process them; S2: Extraction and purification of Chinese medicinal materials: Use corresponding solvents and methods to extract the active ingredients in Chinese medicinal materials; S3: Probiotic fermentation: screening and cultivating excellent probiotic bacteria; S4: Application of fermentation technology: Add beneficial bacteria to Chinese medicinal materials, combine purified Chinese medicinal materials with probiotics, and perform anaerobic fermentation; control fermentation conditions to promote the growth and metabolism of probiotics, and use enzymes produced by microorganisms to react with active ingredients in Chinese medicine to increase the content; S5: Add enzyme to catalyze the reaction: Use enzyme to destroy the plant cell wall and release active substances, that is, use the catalytic effect of enzyme to accelerate the completion of the reaction; S6: Application of control engineering: Introduce control engineering to automatically control the fermentation process of traditional Chinese medicine, monitor and adjust fermentation parameters, and ensure the stable quality of fermentation products; S7: Product extraction and purification: After fermentation, the active metabolites of probiotics are extracted, the extract is purified, impurities are removed, and the final product is obtained.

2. The process for extracting active metabolites of probiotics from botanical medicine according to claim 1, characterized in that: The Chinese medicinal materials include 15-25 parts of chicken gizzard lining, 35-45 parts of poria cocos, 15-25 parts of yam, 15-25 parts of hawthorn, 15-25 parts of malt, 15-20 parts of liquorice, and 5-10 parts of tangerine peel.

3. The process for extracting active metabolites of probiotics from botanicals according to claim 1, characterized in that: The probiotics include Bifidobacterium lactis and Lactobacillus paracasei.

4. The process for extracting active metabolites of probiotics from botanical medicine according to claim 2, characterized in that: The steps of the Chinese medicinal material pretreatment and extraction and purification are as follows: selecting Chinese medicinal materials according to the component ratio, washing and cutting, decocting with water twice to obtain a decoction, and concentrating and sterilizing the decoction to obtain a Chinese medicinal extract, wherein the decoction steps are specifically as follows: (1) Mix water and Chinese medicinal materials in a weight ratio of 3:1, soak for 1 to 2 hours and then boil. Keep boiling for 20 to 25 minutes and filter out the decoction; (2) Mix water and the filter residue in step (1) in a weight ratio of 1:1, heat and boil, use ultrasound to assist dispersion during the boiling process, and continue for 20 to 25 minutes, then filter out the decoction; (3) Mix the decoctions in step (1) and step (2), and concentrate and dry them.

5. The process for extracting active metabolites of probiotics from botanical medicine according to claim 3, characterized in that: The probiotic fermentation and application operation steps are as follows: selecting probiotics, activating the strains, and then inoculating them on a culture medium, then mixing the culture medium with a traditional Chinese medicine extract, fermenting and culturing, and centrifuging and concentrating to obtain bacterial mud; mixing a protective agent and the bacterial mud, vacuum freeze-drying, crushing and mixing, and purifying and removing impurities to obtain the metabolites.

6. The process for extracting active metabolites of probiotics from botanical medicine according to claim 5, characterized in that: The culture medium comprises the following components: 8-10 g / L of peptone, 10-12 g / L of agar, 4-5 g / L of lactose, 3-5 g / L of yeast extract, 3-4 g / L of glucose, 1-2 g / L of sodium acetate, 0.5-1 g / L of dipotassium hydrogen phosphate, and 0.5-1 g / L of magnesium sulfate; the pH of the culture medium is adjusted to 6.5, the bifidobacterium lactis is inoculated on the culture medium at a volume ratio of 1:10; the lactobacillus paracasei is inoculated on another culture medium at a volume ratio of 1:5, the culture medium inoculated with the bifidobacterium lactis and the culture medium of the lactobacillus paracasei are mixed at a volume ratio of 1:(1-1.5), and then the mixed culture medium is mixed with a traditional Chinese medicine extract at a weight ratio of 1:(80-100).

7. The process for extracting active metabolites of probiotics from botanical medicine according to claim 5, characterized in that: The protective agent comprises the following components in weight percentage: 20-30wt% skim milk powder, 10-20wt% whey protein, 5-15wt% glucose, and the balance is trehalose.

8. The process for extracting active metabolites of probiotics from botanical medicine according to claim 5, characterized in that: The vacuum freeze drying specifically comprises the following steps: (1) Precooling: control the temperature to -15~-20℃, the vacuum degree to 27~35Pa, and precool for 4~4.5h; (2) Sublimation drying: control the temperature to -50~-40℃, the vacuum degree to 15~25Pa, and the sublimation drying time to 12~16h; (3) Desorption and drying: control the temperature to 35-50°C, the vacuum degree to 25-30Pa, and the desorption and drying time to 6-8h.

9. The process for extracting active metabolites of probiotics from botanical medicine according to claim 5, characterized in that: The fermentation conditions are room temperature, 55% to 65% humidity, pH 6.2 to 6.8, pressure 0.03 to 0.05 MPa, and time 10 to 12 hours.

10. The process for extracting active metabolites of probiotics from botanical medicine according to claim 1, characterized in that: The enzyme catalyzing the reaction is a glycosyl hydrolase.