CN110591925A - Eurotium cristatum production fermentation method - Google Patents
Eurotium cristatum production fermentation method Download PDFInfo
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Abstract
The invention relates to the field of biochemical engineering, in particular to a production and fermentation method of eurotium cristatum; mixing tea leaves and active additive components in proportion to obtain a raw material, processing the raw material to obtain a pre-filtrate and a first filter residue, processing the first filter residue to obtain fermented tea residues, processing the fermented tea residues to obtain a post-filtrate, mixing part of the pre-filtrate and the post-filtrate to obtain a eurotium cristatum spore suspension, mixing the rest of the pre-filtrate and carbon source nutrient substances to obtain a sterile culture solution, inoculating the eurotium cristatum spore suspension into the sterile culture solution, and performing fermentation treatment to obtain eurotium cristatum; the method prepares the fermentation culture solution by five steps of preparing pre-filtrate, post-filtrate, sterile culture solution, eurotium cristatum suspension and eurotium cristatum liquid and taking common tea leaves and pure natural substances such as medlar, red dates, American ginseng and the like as additives. The invention has simpler production raw materials, low raw material cost and controllable production process, and can completely reach the industrialized production.
Description
Technical Field
The invention relates to the field of biochemical engineering, in particular to a production and fermentation method of eurotium cristatum.
Background
In daily life, medlar, red dates and the like are often used for making tea or soaking medicinal liquor. Researches show that the medlar contains various amino acids, and contains special nutritional ingredients such as betaine, zeaxanthin, and baccatin, and has very good health care effect. The Chinese wolfberry has the effects of regulating immunity, resisting aging, resisting tumors, resisting fatigue, resisting radiation damage, regulating blood fat, reducing blood sugar, reducing blood pressure, protecting reproductive system, improving vision, improving respiratory disease resistance, maintaining beauty, keeping young, moistening skin, protecting liver, enhancing hematopoietic function and the like. The fructus Jujubae can improve immunity and inhibit cancer cell growth, and rutin contained therein can soften blood vessel. The American ginseng is taken as a first-selected medicine for tonifying qi and health care, can improve the immunity of the organism and inhibit the growth of cancer cells; the saponin in the American ginseng can effectively enhance the function of the central nervous system; long-term administration of radix Panacis Quinquefolii can protect cardiovascular system and promote blood activity; meanwhile, the medicine has the functions of reducing blood sugar and regulating insulin secretion.
Tea is a plant rich in polyphenols, and contains more than 700 kinds of compounds. The tea beverage using tea leaf extract, concentrated powder, tea powder, etc. as main material has the refreshing taste of tea leaf and contains effective components of tea polyphenol, catechin, caffeine, etc. The existing research results show that the effective components in the tea can directly eliminate free radicals and avoid oxidative damage. Therefore, the effective components can act on a plurality of diseases related to free radicals, such as atherosclerosis, diabetes, body aging and the like, and have a plurality of pharmacological actions of preventing cardiovascular and cerebrovascular diseases, resisting bacteria and viruses and the like.
Among six basic tea types in China, the Fuzhuan tea is one of compressed black tea, and is reprocessed tea prepared by taking raw dark green tea or crude old green tea as a raw material through a compression process. In the 'flower growing' process of the Fuzhuan tea, a dominant flora, namely eurotium cristatum exists, and the dominant flora can promote the growth and the propagation of the eurotium cristatum by controlling external conditions, so that a large amount of macroscopic golden yellow capsule shells, commonly called 'golden flowers', are grown on the surface of the Fuzhuan tea. Due to the existence of eurotium cristatum, the Fuzhuan tea is endowed with unique quality and flavor. Therefore, people often judge the quality of the Fuzhuan tea by the quantity and quality of the golden flowers. Research finds that the Eurotium cristatum has the effects, and the Eurotium cristatum can absorb nutrient substances in tea leaves in the growth process in order to meet the growth and development of the Eurotium cristatum, and secrete a plurality of metabolites such as antioxidant metabolites, lipid-lowering metabolites, enzyme metabolites, aroma metabolites, amino acid metabolites, antibacterial metabolites and antitumor metabolites while utilizing and converting the nutrient substances. Taking antibacterial and anti-tumor metabolites as an example, in the 'flower growing' process of the Fuzhuan tea, the mass growth and propagation of eurotium cristatum can obviously inhibit and interfere the obvious growth of other bacteria and moulds. Buting et al found that the Eurotium cristatum liquid fermentation broth has an antibacterial effect on both Escherichia coli and Staphylococcus aureus in 2012. The inhibition effect of the extracellular polysaccharide of the eurotium cristatum on tumor model cells is researched by dangming (high-throughput screening test for bioactivity of extracellular polysaccharide of the eurotium cristatum, 2007) and the like, and the result proves that the extracellular polysaccharide of the eurotium cristatum has the anti-tumor cell activity. Therefore, the metabolic activity of eurotium cristatum not only promotes various components in the tea leaves to be obviously changed, but also forms the unique color, fragrance and taste of the Fuzhuan tea to a great extent, and brings about the magical effect of benefiting the human health. It is these unique life activities that make the research and development of Eurotium cristatum very important. When the eurotium cristatum is cultured in a solid state, the eurotium cristatum grows vigorously, but is not suitable for large-scale production; when the liquid fermentation technology is used for culturing eurotium cristatum, the production period is short, the cost is low, the yield is high, and when the mycelium of the eurotium cristatum is cultured by liquid, the growth and metabolism of the mycelium are more active, so that the accumulation of active ingredients is more facilitated. Therefore, the modern liquid fermentation technology can be used for realizing the large-scale culture and production of eurotium cristatum.
At present, most of studied eurotium cristatum fermentation generally has the defects of simple medium components, insufficient and single effects of related health care products.
Disclosure of Invention
In order to solve the technical problems, the invention provides a production and fermentation method of eurotium cristatum;
the invention is realized by the following steps:
a fermentation method for producing eurotium cristatum comprises the steps of mixing tea leaves and active additive components in proportion to obtain raw materials, processing the raw materials to obtain a pre-filtrate and a first filter residue, processing the first filter residue to obtain fermented tea residues, processing the fermented tea residues to obtain a post-filtrate, mixing part of the pre-filtrate and the post-filtrate to obtain eurotium cristatum spore suspension, mixing the rest of the pre-filtrate and carbon source nutrient substances to obtain a sterile culture solution, inoculating the eurotium cristatum spore suspension into the sterile culture solution, and performing fermentation treatment to obtain the eurotium cristatum.
Further, the method for treating the pre-filtrate comprises the following steps: mixing the tea leaves and the active additive components according to the weight ratio of 100: 4-8: 0.5-3, and decocting for 1-2 h.
Further, the tea leaves comprise a mixture of dark tea and poria cocos tea in a weight ratio of 4-20: 1, and the active additive ingredients comprise one or more of wolfberry, red dates and American ginseng.
Further, the preparation method of the fermented tea leaves comprises the following steps: adding vinegar into the first filter residue, adjusting the pH to 5.5-6.5, then airing until the humidity is 60-75%, placing into a container, sealing, and storing for 15-45 days at 15-20 ℃.
Further, the preparation method of the post-filtrate comprises the following steps: and adding water into the fermented tea residues, decocting for 30-60 min, filtering and collecting filtrate to obtain the final filtrate.
Further, the mass ratio of the water to the fermented tea leaves is 5-10: 1.
Further, the preparation method of the sterile culture solution comprises the following steps: mixing the rest pre-filtrate and carbon source nutrient substances according to the volume mass ratio of 100: 4-8 to prepare a solution with the pH of 5.0-5.5, and continuously sterilizing for 20-30min under the conditions of the temperature of 115 ℃ and the temperature of 121 ℃ and the pressure of 0.1-0.15 MPa.
Further, the carbon source nutrient is any one of sucrose or glucose.
Furthermore, the inoculation amount of the eurotium cristatum spore suspension is 5-8% of the mass of the sterile culture solution.
Further, the fermentation environment of the Eurotium cristatum is 500r/min at the rotation speed of 100-.
The beneficial effect of above-mentioned scheme:
the method prepares the fermentation culture solution by five steps of preparing pre-filtrate, post-filtrate, sterile culture solution, eurotium cristatum suspension and eurotium cristatum liquid and taking common tea leaves and pure natural substances such as medlar, red dates, American ginseng and the like as additives. The invention has simpler production raw materials, low raw material cost and controllable production process, and can completely reach the industrialized production.
Detailed Description
A fermentation method for producing eurotium cristatum comprises the steps of mixing tea leaves and active additive components in proportion to obtain raw materials, processing the raw materials to obtain a pre-filtrate and a first filter residue, processing the first filter residue to obtain fermented tea residues, processing the fermented tea residues to obtain a post-filtrate, mixing part of the pre-filtrate and the post-filtrate to obtain eurotium cristatum spore suspension, mixing the rest of the pre-filtrate and carbon source nutrient substances to obtain a sterile culture solution, inoculating the eurotium cristatum spore suspension into the sterile culture solution, and performing fermentation treatment to obtain the eurotium cristatum.
Example 1
Step 1, weighing 20 parts of dark tea, 60 parts of shinyleaf yellowhorn leaves and 5 parts of poria cocos tea according to parts by weight;
step 2, putting the three raw materials into water, decocting for 1 hour, filtering and collecting filtrate, namely pre-filtrate, adjusting the pH of the pre-filtrate to 5.5-6.5 by using table vinegar, and refrigerating for later use;
step 3, adding table vinegar into the tea leaves obtained in the step 2, adjusting the pH value to 5.5-6.5, then airing the tea leaves until the humidity is 60%, wrapping the tea leaves with three layers of medical gauze, putting the tea leaves into an enamelware, sealing the enamelware, and storing the tea leaves at 15 ℃ for 15 days to obtain fermented tea leaves;
step 4, adding purified water into the fermented tea dregs obtained in the step 3, decocting for 30 minutes, and then filtering and collecting filtrate, namely post-filtrate;
and 5, mixing the post filtrate with the pre filtrate to prepare the eurotium cristatum spore suspension.
And 6, mixing the remaining pre-filtrate Eurotium cristatum suspension prepared in the step 2 with carbon source nutrient substances according to the volume-mass ratio of 100: 4 to prepare a solution, wherein the pH of the prepared solution is 5.0-5.5, and continuously sterilizing for 20min at the temperature of 115 ℃ and the pressure of 0.12MPa to obtain the sterile culture solution.
And 7, inoculating the eurotium cristatum suspension prepared in the step 5 into the sterile culture solution prepared in the step two according to the inoculation amount of 8%, and setting fermentation parameters as follows: the rotation speed is 500r/min, the pH value is 5.5, the dissolved oxygen is 40%, the temperature is 30 ℃, the culture is carried out for 168 hours, and the fermentation is stopped when the color of the fermentation liquor is red and transparent and the hypha amount is not increased any more.
And 8, collecting and concentrating the fermentation liquor obtained in the step 7, and removing water to obtain eurotium cristatum.
Example 2
Step 1, weighing 40 parts of dark tea, 40 parts of shinyleaf yellowhorn leaves and 2 parts of Fu tea according to parts by weight;
step 2, putting the three raw materials into water, decocting for 1.5 hours, filtering and collecting filtrate, namely pre-filtrate, adjusting the pH of the pre-filtrate to 5.5-6.5 by using table vinegar, and refrigerating for later use;
step 3, adding table vinegar into the tea leaves obtained in the step 2, adjusting the pH value to 5.5-6.5, then airing the tea leaves until the humidity is 70%, wrapping the tea leaves with three layers of medical gauze, putting the tea leaves into an enamelware, sealing the enamelware, and storing the tea leaves at 117 ℃ for 30 days to obtain fermented tea leaves;
step 4, adding purified water into the fermented tea dregs obtained in the step 3, decocting for 45 minutes, and then filtering and collecting filtrate, namely post-filtrate;
and 5, mixing the post filtrate with the pre filtrate to prepare the eurotium cristatum spore suspension.
And 6, mixing the remaining pre-filtrate Eurotium cristatum suspension prepared in the step 2 with carbon source nutrient substances according to the volume-mass ratio of 100: 4 to prepare a solution, wherein the pH of the prepared solution is 5.0-5.5, and continuously sterilizing for 20min at the temperature of 115 ℃ and the pressure of 0.12MPa to obtain the sterile culture solution.
And 7, inoculating the eurotium cristatum suspension prepared in the step 5 into the sterile culture solution prepared in the step two according to the inoculation amount of 8%, and setting fermentation parameters as follows: the rotation speed is 500r/min, the pH value is 5.5, the dissolved oxygen is 30%, the temperature is 30 ℃, the culture is carried out for 168 hours, and the fermentation is stopped when the color of the fermentation liquor is red and transparent and the hypha amount is not increased any more.
And 8, collecting and concentrating the fermentation liquor obtained in the step 7, and removing water to obtain eurotium cristatum.
Example 3
Step 1, weighing 35 parts of dark tea, 45 parts of shinyleaf yellowhorn leaves and 4 parts of poria cocos tea according to parts by weight;
step 2, putting the three raw materials into water, decocting for 1-2 hours, filtering and collecting filtrate, namely pre-filtrate, adjusting the pH of the pre-filtrate to 5.5-6.5 by using table vinegar, and refrigerating for later use;
step 3, adding table vinegar into the tea dregs obtained in the step 2, adjusting the pH value to 5.5-6.5, then airing the tea dregs until the humidity is 60-75%, wrapping the tea dregs with three layers of medical gauze, putting the tea dregs into an enamelware, sealing, and storing for 15-45 days at 15-20 ℃ to obtain fermented tea dregs;
step 4, adding purified water into the fermented tea dregs obtained in the step 3, decocting for 30-60 minutes, and then filtering and collecting filtrate, namely post-filtrate;
and 5, mixing the post filtrate with the pre filtrate to prepare the eurotium cristatum spore suspension.
And 6, mixing the remaining pre-filtrate Eurotium cristatum suspension prepared in the step 2 with carbon source nutrient substances according to the volume-mass ratio of 100: 8 to prepare a solution, wherein the pH of the prepared solution is 5.0-5.5, and continuously sterilizing for 30min at the temperature of 125 ℃ and the pressure of 0.15MPa to obtain the sterile culture solution.
And 7, inoculating the eurotium cristatum suspension prepared in the step 5 into the sterile culture solution prepared in the step two according to the inoculation amount of 8%, and setting fermentation parameters as follows: the rotation speed is 300r/min, the pH value is 5.5, the dissolved oxygen is 40%, the temperature is 30 ℃, the culture is carried out for 120h, and the fermentation is stopped when the color of the fermentation liquor is red and transparent and the hypha amount is not increased any more.
And 8, collecting and concentrating the fermentation liquor obtained in the step 7, and removing water to obtain eurotium cristatum.
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 (10)
1. A eurotium cristatum production fermentation method is characterized in that tea leaves and active additive components are mixed in proportion to obtain raw materials, the raw materials are processed to obtain a pre-filtrate and a first filter residue, the first filter residue is processed to obtain fermented tea residues, the fermented tea residues are processed to obtain a post-filtrate, part of the pre-filtrate and part of the post-filtrate are mixed to obtain eurotium cristatum suspension, the rest of the pre-filtrate is mixed with carbon source nutrient substances to obtain sterile culture solution, and the eurotium cristatum suspension is inoculated into the sterile culture solution to be fermented to obtain the eurotium cristatum.
2. The Eurotium cristatum production fermentation method according to claim 1, wherein the pre-filtrate treatment method comprises: mixing the tea leaves and the active additive components according to the weight ratio of 100: 4-8: 0.5-3, and decocting for 1-2 h.
3. The eurotium cristatum production and fermentation method according to claim 2, wherein the tea leaves comprise a mixture of dark tea and poria cocos tea in a weight ratio of 4-20: 1, and the active additive ingredients comprise one or more of lycium barbarum, red dates and American ginseng.
4. The production and fermentation method of eurotium cristatum according to claim 1, wherein the preparation method of the fermented tea leaves comprises: adding vinegar into the first filter residue, adjusting the pH to 5.5-6.5, then airing until the humidity is 60-75%, placing into a container, sealing, and storing for 15-45 days at 15-20 ℃.
5. The Eurotium cristatum production fermentation method according to claim 1, wherein the preparation method of the post-filtrate comprises: and adding water into the fermented tea residues, decocting for 30-60 min, filtering and collecting filtrate to obtain the final filtrate.
6. The Eurotium cristatum production fermentation method according to claim 5, wherein the mass ratio of the added water to the fermented tea leaves is 5-10: 1.
7. The Eurotium cristatum production fermentation method according to claim 1, wherein the sterile culture solution preparation method comprises: mixing the rest pre-filtrate and carbon source nutrient substances according to the volume mass ratio of 100: 4-8 to prepare a solution with the pH of 5.0-5.5, and continuously sterilizing for 20-30min under the conditions of the temperature of 115 ℃ and the temperature of 121 ℃ and the pressure of 0.1-0.15 MPa.
8. The Eurotium cristatum producing fermentation method according to claim 7, wherein the carbon source nutrient is any one of sucrose or glucose.
9. The Eurotium cristatum production fermentation method according to claim 1, wherein the inoculation amount of the Eurotium cristatum spore suspension is 5% -8% of the quality of the sterile culture solution.
10. The fermentation method of Eurotium cristatum as claimed in claim 9, wherein the fermentation environment of Eurotium cristatum is 100-500r/min, pH 5.0-5.5, dissolved oxygen 20% -40%, temperature 28-30 ℃, culture time 120-168 h.
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| CN111803533A (en) * | 2020-07-01 | 2020-10-23 | 陕西巨子生物技术有限公司 | Composition for reducing blood sugar and blood fat, preparation method and application thereof |
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