CN107772076B - Cordyceps sinensis based feed immunopotentiator and preparation method thereof - Google Patents
Cordyceps sinensis based feed immunopotentiator and preparation method thereof Download PDFInfo
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Abstract
The invention belongs to the field of feed additives, and particularly relates to an immunopotentiator for cordyceps sinensis base feed and a preparation method thereof. Cordyceps militaris with oat as culture medium is cultured for 35-45 days, and the residual Cordyceps militaris medium after fruiting bodies are picked contains special bioactive components, other nutritional components and impurities of Cordyceps militaris. The weight of the bioactive components accounts for 65 percent of the total weight; the weight of the bioactive components is as follows: 29.6 to 30.4 percent of cordyceps polysaccharide, 0.07 to 0.09 percent of cordycepin, 0.015 to 0.025 percent of adenosine, 12.8 to 13 percent of crude protein, 12.8 to 13.6 percent of cordycepic acid, 0.052 to 0.065 percent of SOD enzyme, 8.7 to 9.1 percent of amino acid, 0.022 to 0.028 percent of calcium and 0.031 to 0.037 percent of iron. It can be used as feed additive for aquatic animals (such as Stichopus japonicus) to enhance immunity. The product has no influence on health, and is a pure green powder preparation.
Description
Technical Field
The invention belongs to the field of feed additives, and particularly relates to an immunopotentiator for cordyceps sinensis base feed and a preparation method thereof.
Background
The rapid development of the aquaculture industry at present enables the aquaculture scale to be continuously enlarged and the aquaculture yield to be continuously increased. However, as the culture intensification degree is continuously improved, the stress conditions in the living environment of the aquatic animals are increased, the diseases are more and more serious, and the disease control and healthy culture of the aquatic animals are more and more concerned. Traditional disease control mainly uses chemical drugs such as antibiotics to inhibit or kill germs, but the drug resistance of aquatic animals to antibiotics is continuously increased, the immune function of the aquatic animals is reduced, the problem of drug residues in the aquatic animals is increasingly serious, and the food safety and the human health are affected. Attempts have been made to improve the constitution of aquatic animals immunologically and to increase their resistance to pathogenic microorganisms. Therefore, the development and application of the immunopotentiator for aquatic animals have become a research hotspot.
The immunopotentiator is a substance which is added into feed, improves the nutritional status of aquatic animals by improving the immunity of the aquatic animals, and can remarkably enhance the resistance of the aquatic animals to pathogenic microorganisms. The growth and development of the aquatic animals are influenced by various factors, and the immunopotentiator can enhance the anti-stress and anti-infection capability and improve the production performance of the aquatic animals by improving the non-specific immune function of the aquatic animals; in addition, the substances can enhance the capability of the fish to resist the invasion of external pathogens and relieve the immunosuppressive action of external environmental stress on the fish. The immunopotentiator has the characteristics of stable performance, wide action, safety, high efficiency and the like, and becomes one of the important ways for solving the diseases of the aquatic animals at present. The immunopotentiator mainly comprises saccharides, vitamins, trace elements, nucleotides, microecologics, Chinese herbal medicines and the like.
The cordyceps militaris (also known as cordyceps militaris and cordyceps militaris) contains rich nutrient components, is a precious Chinese herbal medicine with a nourishing function, and has the effects of regulating immunity, preventing cancer and aging, protecting lung and tonifying kidney, protecting heart and strengthening brain, improving organ functions, resisting fatigue, protecting skin and beautifying, inhibiting bacteria, resisting tumors and the like. The fruiting bodies of China are mainly used as health care products and medicines, while the residual culture medium (called cordyceps sinensis base) after the fruiting bodies are picked is generally discarded as waste, and particularly after large-scale production, a large amount of cordyceps sinensis base waste materials are randomly discarded, so that huge waste is caused, and the environment is polluted. The cordyceps substrate still contains a large amount of hypha for propagation, and the cordyceps substrate also contains functional components such as protein, amino acid, fat, trace elements and the like besides the special active secondary metabolites of cordyceps such as cordycepin, cordycepic acid, adenosine, polysaccharide, SOD and the like. Has obvious effect on the aspects of regulating the functions of the immune system of animals and the like, and is a natural, safe and efficient feed immunopotentiator.
At present, there are many patents on immunopotentiators for cordyceps based feeding. For example, CN101066090A patent adopts short-term fermentation of mixed strain liquid, the period is only 2-5 days, but hypha is not completely developed, and only crude polysaccharide is a nutritional index, the nutritional value is not sufficient, and the immunity of the most effective nutritional components of Cordyceps militaris can not be embodied; patent CN103892052A adopts a solid culture medium, and after freeze drying, the superfine grinding technology is carried out to prepare the cordyceps immune enhancer, although the nutrient content is sufficient, the content of each component is not high, and the high-efficiency dissolution and utilization of medicinal active ingredients and nutrient ingredients are not achieved.
In conclusion, the development of the immune enhancer for the cordyceps sinensis base feed, which is rich in nutrition and remarkable in immune effect, is particularly important.
Disclosure of Invention
In order to solve the problems, the invention provides an immunopotentiator for cordyceps sinensis base feed and a preparation method thereof. The immunopotentiator has the advantages of remarkable immunization effect, simple preparation method and environmental protection.
In order to achieve the aim, the invention provides an immunopotentiator for a cordyceps substrate feed, wherein the cordyceps substrate is cordyceps militaris with oat as a culture medium, and the rest cordyceps substrate is obtained after fruiting bodies are picked up after 35-45 days of culture; the immunopotentiator contains special bioactive components of Cordyceps militaris, other nutritional components and impurities.
The bioactive components are as follows: 29.6-30.4% of cordyceps polysaccharide, 0.07-0.09% of cordycepin, 0.015-0.025% of adenosine, 12.8-13% of crude protein, 12.8-13.6% of cordycepic acid, 0.052-0.065% of SOD enzyme, 8.7-9.1% of amino acid, 0.022-0.028% of calcium and 0.031-0.037% of iron, wherein the weight of the bioactive components accounts for 65% of the weight of the pretreated cordyceps substrate.
The weight of other nutrient components accounts for 27 percent of the weight of the pretreated cordyceps sinensis base, and the nutrient components are as follows: 2.21-2.25% of alkaloid, 5.63-5.65% of terpenoid, 8.28-8.3% of sterol, 9.2-9.3% of glycoside and 1.52-1.59% of vitamin.
The preferred scheme is as follows: 2.25% of alkaloid, 5.65% of terpenoid, 8.3% of sterol, 9.21% of glycosides and 1.59% of vitamin.
The balance being impurities.
The amino acid is selected from glycine, valine, leucine, isoleucine, proline, methionine, arginine or histidine.
The alkaloid is selected from pyridine, isoquinoline, indole or purine, specifically nicotine, caffeine or vincristine.
The terpenoids are selected from lauraceae, geraniol or marigold ketone.
The glycosides are as follows: adenine, deoxyuric acid, guanosine, deoxythymidine, hypoxanthine, uridine, N6-methyladenosine or O6-acetyl cordycepin.
The sterol is as follows: ergosterol, beta-sitosterol or allose alcohol.
The vitamins are as follows:vitamin A, vitamin B12Vitamin B6Vitamin B1Vitamin B2A plurality of vitamin C, vitamin D or vitamin E.
In order to realize the aim, the invention also provides a preparation method of the immune enhancer for the cordyceps sinensis-based feed, which comprises the following steps.
(1) Pretreatment of cordyceps sinensis base powder: culturing Cordyceps militaris for 35-45 days, picking fruiting body, and weighing Cordyceps militaris base; selecting Cordyceps base, removing impurities without visible impurities, and cleaning with pure water for 2 times; drying at 60 deg.C for 1-3 hr in a drying oven at low temperature; pulverizing the dried Cordyceps militaris base with a superfine pulverizer, and sieving with a 80-mesh sieve to obtain Cordyceps militaris culture medium powder for use.
(2) Mixing materials: placing the pretreated Cordyceps sinensis base powder in an extraction tank, adding distilled water, and stirring with a stirrer.
(3) Carrying out first enzymolysis: adding amylase into the mixed solution after fully and uniformly stirring, adjusting the pH to 5.5 by using dilute hydrochloric acid, and carrying out enzymolysis for 2.5h at the temperature of 65 ℃.
(4) And (3) carrying out second enzymolysis: adding protease into the enzymolysis liquid in the step (3), adjusting the pH to 6.0 by using dilute hydrochloric acid, introducing cooling water, cooling to 55 ℃, and carrying out enzymolysis for 3 hours.
(5) Ultrasonic oscillation treatment: starting an ultrasonic generator, setting the ultrasonic frequency to be 140W, and introducing the enzymolysis liquid into the ultrasonic generator for oscillation treatment for 1 h.
(6) Centrifuging and drying: and (3) introducing the oscillating liquid obtained in the step (5) into a centrifugal tank for centrifugal treatment, introducing the obtained supernatant into a concentration tank, concentrating to 20% of the volume of the mixed liquid obtained in the step (1), rapidly passing through a spray dryer, and performing spray drying for 30-45min at the temperature of 100 ℃ and 120 ℃ to obtain the cordyceps sinensis immunopotentiator.
(7) And (4) preparing the cordyceps immunopotentiator obtained in the step (6) into powder, wherein the particle size reaches 150-200 meshes.
The immune enhancer for the cordyceps sinensis basal feed can be used as a feed additive for aquatic animals (such as sea cucumbers) to enhance the immune capability of the aquatic animals.
Compared with the prior art, the invention has the beneficial effects.
The cordyceps militaris medium used in the invention is a solid medium taking oat as a culture medium, and the residue of the medium also contains the specific active substances of cordyceps, such as cordycepin, cordyceps polysaccharide, adenosine and the like, and also contains the effective components of alkaloid, terpenoid, sterol, glycoside, phenol, enzyme, vitamin, SOD and the like. Therefore, the auxiliary materials under the cordyceps culture medium are effectively developed and utilized to prepare the immunopotentiator for the aquatic animals, 200 g of the immunopotentiator can be prepared per kilogram of the cordyceps culture medium, the market value is about 160 yuan/kg, the production cost of the cordyceps can be reduced, and the overall economic benefit is improved. In the culture production, the pure natural cordyceps immunopotentiator is applied, so that the immunity level of an animal body can be improved, the congenital resistance can be enhanced, and the use of medicaments for improving the animal immunity can be avoided, so that the problem of medicament residue in aquatic animals is solved, and the food safety and the human health are ensured.
The invention prepares the cordyceps immune enhancer by degrading starch and protein in the cordyceps substrate in a mode of combining the compounding of a plurality of enzymes and ultrasonic waves, and the method does not use chemical reagents such as ethyl acetate and the like, only uses a small amount of enzymes, and reduces the cost of raw materials; in the preparation process, firstly, two enzymes are used for carrying out enzymolysis on the cordyceps sinensis base powder aqueous solution, so that nutritional ingredients such as protein in the cordyceps sinensis base powder aqueous solution are subjected to enzymolysis to form a plurality of amino acids, and further organic micromolecules which are easy to absorb by a human body are generated; then, an ultrasonic generator is adopted for oscillation treatment, so that the nutrient components in the solution are completely oscillated, and the extraction of the components of the cordyceps sinensis immunopotentiator is facilitated; the reaction condition is mild, no influence is caused on health, and the product is a pure green powder preparation; the special biological active component content of the cordyceps sinensis immunopotentiator powder prepared by the preparation method is high, and the nutritional value is high.
Detailed Description
The present invention will be further described with reference to the following specific examples.
Example 1.
Cordyceps militaris with oat as culture medium is cultured for 35 days, and the rest Cordyceps militaris medium is taken for pretreatment after fruiting bodies are picked.
The cordyceps sinensis base is pretreated, and the treatment steps are as follows: (1) culturing Cordyceps militaris for 35 days, picking fruiting bodies, and weighing 11kg of Cordyceps militaris base; (2) selecting Cordyceps base, removing impurities without visible impurities to obtain 10kg Cordyceps base, and cleaning with pure water for 2 times; (3) drying at 60 deg.C in a drying oven at low temperature; (4) pulverizing the dried Cordyceps militaris base with a superfine pulverizer, and sieving with a 80-mesh sieve to obtain Cordyceps militaris culture medium powder for use.
The feed immunopotentiator is prepared from the obtained cordyceps militaris culture medium powder, and the preparation method comprises the following steps: (1) 10kg of pretreated cordyceps sinensis base powder is taken and placed in an extraction tank, 50kg of distilled water is added into the extraction tank, a stirrer is started to stir, the stirring speed is 120 r/min, and the stirring is carried out for 0.5-1 h; (2) stirring thoroughly, adding amylase 0.12kg, adjusting pH to 5.5 with 0.5% diluted hydrochloric acid, and performing enzymolysis at 65 deg.C for 2.5 hr; (3) adding protease 0.12kg, adjusting pH to 6.0 with 0.5% diluted hydrochloric acid, cooling to 55 deg.C with cooling water, and performing enzymolysis for 3 hr; (4) starting an ultrasonic generator, setting the ultrasonic frequency to be 140W, introducing the enzymolysis liquid obtained in the step (3) into the ultrasonic generator, carrying out oscillation treatment for 1h, introducing the obtained liquid into a centrifugal tank, carrying out centrifugal treatment for 20 minutes, introducing the supernatant into a concentration tank, concentrating for 5h to 20% of the volume of the step (1), rapidly carrying out spray drying through a spray dryer, drying for 30 minutes, and obtaining the cordyceps sinensis immunopotentiator, wherein the drying temperature is 120 ℃; (5) preparing the cordyceps immunopotentiator obtained in the step (4) into powder, wherein the particle size reaches 150-200 meshes; (6) and (4) packaging the powder obtained in the step (5) into 100 g/bag, 200 g/bag or 500 g/bag to obtain the cordyceps sinensis immunopotentiator product for later use.
The prepared immunopotentiator contains special bioactive components of Cordyceps militaris, other nutritional components and impurities.
The bioactive components are as follows: 30.4% of cordyceps polysaccharide, 0.08% of cordycepin, 0.015% of adenosine, 12.8% of crude protein, 12.9% of cordycepic acid, 0.052% of SOD enzyme, 1.03% of glycine, 0.87% of valine, 0.83% of leucine, 0.92% of isoleucine, 0.95% of proline, 1.08% of methionine, 1.22% of arginine, 1.8% of histidine, 0.022% of calcium and 0.031% of iron, wherein the bioactive components account for 65% of the weight of the pretreated cordyceps substrate.
The other nutrient components account for 27 percent of the weight of the pretreated cordyceps sinensis base, and specifically comprise 0.79 percent of nicotinic acid, 0.73 percent of caffeine, 0.73 percent of vincristine, 2.98 percent of geraniol, 2.67 percent of marigold ketone, 2.42 percent of ergosterol, 3.28 percent of β -sitosterol, 2.6 percent of allose, 3.07 percent of adenine, 2.98 percent of deoxyuric acid, 3.16 percent of guanosine, and 3.16 percent of vitamin B60.36 percent of vitamin B10.35% of vitamin B20.38%, vitamin C0.3%, vitamin D0.2%.
The rest is impurities, and the impurities are unhydrolyzed protein, starch and other substances which do not harm the animal body.
The prepared cordyceps sinensis is used for enhancing immunity and is prepared into a sea cucumber feed additive for enhancing the immunity of the sea cucumber feed additive.
Firstly, detecting an enhancer.
150 sea cucumbers with the same growth environment and no obvious difference in growth conditions are randomly divided into an experimental group, a control group (A) and a control group (B), and each group comprises 50 sea cucumbers. The feed of the experimental group was added with the cordyceps immunopotentiator obtained in example 1 in an amount of 10% by weight of the feed of the sea cucumber, the feed of the control group (a) was added with the common feed additive containing acremonium terricola purchased in the market in an amount of 10% by weight of the feed of the sea cucumber, and the feed of the control group (B) was not added with any immunopotentiator. After feeding for 60 days, the physical conditions of three groups of sea cucumbers are observed, and the detection results of specific indexes are shown in table 1.
Table 1 the results of physical condition examination of three groups of sea cucumbers.
Therefore, the experimental group of the feed additive prepared by the immunopotentiator disclosed by the invention feeds the sea cucumber without antibiotic residues, so that the food safety and the human health are ensured. The proportion of the sea cucumber amino acid to the dry weight of the body wall is 11 percent higher than that of a control group (A) added with the common feed additive containing acremonium terricola and 13 percent higher than that of a control group (B) not added with any immunopotentiator; the weight gain rate of the sea cucumber is 8 percent higher than that of a control group (A) added with the common feed additive containing acremonium terricola and 10 percent higher than that of a control group (B) not added with any immunopotentiator; the survival number of the sea cucumber is 1 more than that of the control group (A) added with the common feed additive containing the acremonium terricola and 6 more than that of the control group (B) not added with any immunopotentiator. The experimental result shows that the immunity and the disease resistance of the sea cucumber can be improved and the survival rate can be improved only by adding the cordyceps immune enhancer because the sea cucumber is very easily infected by pathogenic microorganisms in the feeding process; the immunopotentiator contains nutrient components, so that the amino acid content in the sea cucumber can be increased; by adopting the immunopotentiator disclosed by the invention, the residue of antibiotics in a body is avoided, and the nutritive value of the sea cucumber is improved.
Claims (2)
1. A preparation method of an immunopotentiator for a cordyceps sinensis basal feed comprises the following steps:
(1) pretreatment of cordyceps sinensis base powder: culturing Cordyceps militaris for 35-45 days, picking fruiting body, and weighing Cordyceps militaris base; selecting Cordyceps base, removing impurities without visible impurities, and cleaning with pure water for 2 times; drying at 60 deg.C for 1-3 hr in a drying oven at low temperature; pulverizing the dried Cordyceps militaris base with a superfine pulverizer, and sieving with a 80-mesh sieve to obtain Cordyceps militaris culture medium powder for use;
(2) mixing materials: placing the pretreated cordyceps sinensis base powder in an extraction tank, adding distilled water into the extraction tank, and starting a stirrer to stir;
(3) carrying out first enzymolysis: adding amylase into the mixed solution after fully and uniformly stirring, adjusting the pH to 5.5 by using dilute hydrochloric acid, and performing enzymolysis for 2.5 hours at the temperature of 65 ℃;
(4) and (3) carrying out second enzymolysis: adding protease into the enzymolysis liquid obtained in the step (3), adjusting the pH to 6.0 by using dilute hydrochloric acid, introducing cooling water, cooling to 55 ℃, and performing enzymolysis for 3 hours;
(5) ultrasonic oscillation treatment: starting an ultrasonic generator, setting the ultrasonic frequency to be 140W, and introducing the enzymatic hydrolysate into the ultrasonic generator for oscillation treatment for 1 h;
(6) centrifuging and drying: introducing the obtained liquid into a centrifugal tank for centrifugal treatment, introducing the supernatant into a concentration tank, concentrating to 20% of the volume of the mixed liquid in the step (1), rapidly passing through a spray dryer, and performing spray drying for 30-45min at the temperature of 100-;
(7) preparing the cordyceps immunopotentiator obtained in the step (6) into powder, wherein the particle size reaches 150-200 meshes;
the cordyceps substrate is cordyceps militaris with oat as a culture medium, and the residual cordyceps substrate is obtained after fruiting bodies are picked up after 35-45 days of culture; the immunopotentiator consists of special bioactive components of cordyceps militaris, other nutrient components and impurities;
the weight of the bioactive components accounts for 65 percent of the base weight of the cordyceps sinensis after pretreatment; the weight percentage of the bioactive components is as follows: 29.6 to 30.4 percent of cordyceps polysaccharide, 0.07 to 0.09 percent of cordycepin, 0.015 to 0.025 percent of adenosine, 12.8 to 13 percent of crude protein, 12.8 to 13.6 percent of cordycepic acid, 0.052 to 0.065 percent of SOD enzyme, 8.7 to 9.1 percent of amino acid, 0.022 to 0.028 percent of calcium and 0.031 to 0.037 percent of iron;
the weight of other nutrient components accounts for 27 percent of the weight of the pretreated cordyceps sinensis base, and the weight percentages of the nutrient components are as follows: 2.21-2.25% of alkaloid, 5.63-5.65% of terpenoid, 8.28-8.3% of sterol, 9.2-9.3% of glycoside and 1.52-1.59% of vitamin;
the amino acid is a plurality of glycine, valine, leucine, isoleucine, proline, methionine, arginine or histidine;
the alkaloid is a plurality of pyridines, isoquinolines, indoles or purines;
the terpenoids are various of laurel, geraniol or marigold ketone;
the glycosides are as follows: adenine, deoxyuric acid, guanosine, deoxythymidine, hypoxanthine, uridine, N6-methyladenosine or O6-a plurality of acetyl cordycepin;
the sterol is as follows: a plurality of ergosterol, beta-sitosterol or allose alcohol;
the vitamins are as follows: vitamin A, vitamin B12Vitamin B6Vitamin B1Vitamin B2A plurality of vitamin C, vitamin D or vitamin E;
the immune enhancer for the cordyceps sinensis basal feed is used as an additive of aquatic animal feed to enhance the immune capacity of the aquatic animal feed.
2. The method for preparing an immune enhancer for a cordyceps sinensis-based feed according to claim 1, wherein the alkaloid is a plurality of alkaloid selected from the group consisting of nicotine, caffeine and vincristine.
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| CN109632980B (en) * | 2018-11-15 | 2022-04-29 | 东莞市东阳光冬虫夏草研发有限公司 | Method for simultaneously detecting uridine and ergosterol and application thereof |
| CN111528344A (en) * | 2020-05-08 | 2020-08-14 | 蚌埠学院 | Method for preparing turtle immunity feed by using cordyceps militaris waste culture medium |
| CN112674236A (en) * | 2020-09-24 | 2021-04-20 | 蚌埠学院 | River crab functional feed and preparation method thereof |
| CN113349308A (en) * | 2021-05-10 | 2021-09-07 | 广州清科生物技术有限公司 | Composite zymolyte for pet food and preparation process thereof |
| CN114681496B (en) * | 2022-01-13 | 2023-09-05 | 合肥河川生物医药科技有限公司 | Application of Acremonium terricola culture |
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| CN102870600A (en) * | 2012-10-17 | 2013-01-16 | 山西万海澳生物科技有限责任公司 | Cordyceps militaris fruit body cultivation technology for stabilizing cordycepin content |
| CN103892052A (en) * | 2014-04-18 | 2014-07-02 | 江苏康能生物工程有限公司 | Cordyceps militaris medium immunopotentiator for feeding and production method of immunopotentiator |
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| CN101856062A (en) * | 2009-04-10 | 2010-10-13 | 南京康能生物工程有限公司 | Immune potentiator for wormgrass base feed and production method thereof |
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| CN102870600A (en) * | 2012-10-17 | 2013-01-16 | 山西万海澳生物科技有限责任公司 | Cordyceps militaris fruit body cultivation technology for stabilizing cordycepin content |
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