CN102925527A - Method for mixing and fermenting flammulina velutipes and lucid ganoderma - Google Patents
Method for mixing and fermenting flammulina velutipes and lucid ganoderma Download PDFInfo
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- CN102925527A CN102925527A CN201210455815XA CN201210455815A CN102925527A CN 102925527 A CN102925527 A CN 102925527A CN 201210455815X A CN201210455815X A CN 201210455815XA CN 201210455815 A CN201210455815 A CN 201210455815A CN 102925527 A CN102925527 A CN 102925527A
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Abstract
The invention relates to a method for mixing and fermenting flammulina velutipes and lucid ganoderma. The method comprises steps as follows: (1) respectively preparing fermenting seed liquid of flammulina velutipes and the fermenting seed liquid of the lucid ganoderma; (2) preparing mixed fermenting culture medium, wherein the culture medium comprises sucrose, starch, potato, corn and soybean; and (3) inoculating the fermenting seed liquid of flammulina velutipes and the fermenting seed liquid of the lucid ganoderma obtained in step (1) to the mixed fermenting culture medium for deeply culturing together, so as to obtain active products such as polysaccharide and triterpenoids. The method has the advantages that (1) mixed fermenting culturing of the flammulina velutipes and lucid ganoderma is achieved; (2) the culture medium comprises natural components, and is convenient to use; natural food crops are used in mixed fermenting culture medium, and no chemical agent is added, preparation is convenient and simple, and the pH (Potential Of Hydrogen) is not needed to be adjusted during a fermenting process; and (3) compared with individual fermenting of the flammulina velutipes and the lucid ganoderma, mixed fermenting of the flammulina velutipes and the lucid ganoderma has the advantages that outputs of exopolysaccharide and triterpenoids can be increased by 8 to 35%.
Description
Technical field
The present invention relates to a kind of method of mixed fungus fermentation, be specifically related to the method for a kind of needle mushroom, glossy ganoderma mixed fungus fermentation.
Background technology
Needle mushroom (
Flammulina velutipes) be called as " increasing the intelligence mushroom ", contain 18 seed amino acids, the amino acid that comprises 8 kinds of needed by human, be rich in the several mineral materials such as zinc, calcium, phosphorus, iron, have to promote human intelligence, the effect of growing, improving body immunity, especially in enhancing body the effect of the aspects such as the adaptive faculty of exercise load, antifatigue, Ginseng Extract is better than the edible mushroomss such as mushroom, mushroom, auricularia auriculajudae.
Glossy ganoderma (
Ganoderma lucidum) contain ten plurality of active ingredients such as polysaccharide, triterpene compound, polypeptide.Glossy ganoderma is different from general medicine to certain disease and plays therapeutic action; also being different from general nutritive health-care food only replenishes and strengthens the deficiency of nutrient substance in a certain respect; but two-ways regulation function of human body balance on the whole; balance the body metabolism function; improve autoimmunization ability and hypoxia-bearing capability, impel internal organ or organ function normalizing.
Artificial culture needle mushroom, glossy ganoderma since the sporophore growth cycle long, be subjected to season and raw-material restriction, yield poorly, can not satisfy the demand, at present research mainly concentrates on respectively to be carried out on deep layer fermenting process and the active substance function needle mushroom, glossy ganoderma, but generally use during the fermentation some biochemical substances as the nutrient media components of its fermentation, this not only causes the substratum price more expensive, and the after product that ferments simultaneously utilizes the aspect also to have safety issue.
Mixed fungus fermentation refers to adopt the synergy of two or more microorganisms jointly to finish a kind of novel fermentation technology of certain fermenting process, it is the new development of pure-blood ferment technology, also be a kind of the needs to carry out the novel fermentation technology that complicated DNA vitro recombination but can obtain similar effect, can improve fermentation efficiency even can form product innovation.The refined treasure of the king of Shanghai Normal University once used glossy ganoderma, agaric pilose mixed fungus fermentation, find mixed fungus fermentation after because collaborative, the synergy between active substance, complementation, and the new active substance that produces, its physiological function effect is better than original single bacterium tunning.Fungi mixed fungus fermentation technology and related products thereof still await further research at present.
Summary of the invention
The objective of the invention is to provide a kind of needle mushroom, glossy ganoderma mixed fungus fermentation method that can promote, improve and change active result quality in the fermented liquid.
The object of the present invention is achieved like this: this mixed fungus fermentation method is: the mixed fungus fermentation substratum is comprised of disaccharide, polysaccharide, starch based grain, protein-based grain, and described mixed fungus fermentation substratum comprises: 10 ~ 20g/L starch, 15 ~ 25g/L potato, 10 ~ 20g/L sucrose, 10 ~ 30g/L corn, 1 ~ 10g/L soya bean; Adopt the liquid submerged fermentation technology, mixed bacterium is cultivated needle mushroom and glossy ganoderma; Take needle mushroom and glossy ganoderma as bacterial classification, prepare single bacterium fermentation seed liquid through potato dextrose agar activation, liquid fermentation and culture, combined inoculation needle mushroom, glossy ganoderma are in the mixed fungus fermentation substratum, inoculative proportion 1:1 ~ 3:1,22 ~ 26 ℃ of temperature, pH 4.5 ~ 5.5, and incubation time is 120 ~ 240h, and mycelia and fermented liquid behind the mixed fungus fermentation can be used for tunning: the extraction and determination of biomass, polysaccharide and triterpene compound.
Concrete steps are as follows:
(1) preparation potato dextrose agar slant medium: according to GB4789.15-2010 " microbiological test of food hygiene Molds and yeasts counting " preparation, be used for recovery and the preservation of bacterial classification;
(2) actication of culture: get needle mushroom No. 5.65 bacterial strains and glossy ganoderma No. 5.786 bacterial strains and be inoculated in respectively and activate on the potato dextrose agar slant medium 1 ~ 2 time;
(3) prepare respectively single bacterium fermentation seed liquid: the bacterium piece of getting after the activation of needle mushroom, glossy ganoderma is inoculated in respectively in the liquid nutrient medium 22 ~ 26 ℃ of fermentation culture 168 ~ 288h;
(4) preparation mixed fungus fermentation substratum: be cut into approximately 1cm behind the peeling potatoes
3It is 100 μ m that fritter, corn, analysis for soybean powder are broken to granular size, and substratum consists of 10 ~ 20g/L starch, 15 ~ 25g/L potato, 10 ~ 20g/L sucrose, 10 ~ 30g/L corn, 1 ~ 10g/L soya bean, adds the water preparation; First potato, corn, soybean with water are boiled 25min, add again dissolving good starch and sucrose, sterilize after adding the water constant volume;
(5) preparation mixed fungus fermentation seed liquor: get needle mushroom, the single bacterium fermentation seed liquid of glossy ganoderma, be inoculated in the mixed fungus fermentation substratum, nutrient solution acidity-basicity ph 4.5 ~ 5.5, the inoculum size proportioning is 1:1 ~ 3:1, amounts to 5%, 22 ~ 26 ℃ of fermentation culture 144 ~ 240h;
(6) separating bio amount: filtering fermentation liquor is collected mycelia, dries to constant mass and claims quality for 60 ℃;
(7) crude extracellular polysaccharide extraction and determination: centrifuging and taking supernatant liquor behind the filtering fermentation liquor, 80 ~ 90 ℃ of water-baths are condensed into 0.2 ~ 0.25 times of original volume, add 4 ~ 5 times of volume 95% ethanol, 4 ℃ leave standstill, and concentrated solution is centrifugal, precipitate twice with absolute ethanol washing, dry to constant weight, get crude extracellular polysaccharide;
(8) extraction of triterpene compound in the born of the same parents: get and filter after 1 g mycelia adds 50 mL methyl alcohol standing over night, add 4 mL, 95% dissolve with ethanol behind the evaporate to dryness;
(9) extraction of the outer triterpene compound of born of the same parents: get fermented liquid 15mL, with the water-saturated n-butanol extraction of 3 times of volumes 3 times, merging n-butanol extracting liquid, evaporated under reduced pressure adds 4mL 95% dissolve with ethanol.
Beneficial effect, owing to adopting such scheme, the mixed fungus fermentation substratum composition that uses is food crop, substratum comprises potato, corn, analysis for soybean powder, starch and sucrose, mixed fungus fermentation substratum of the prior art all uses chemical reagent, the grain substratum more is conducive to obtain green, natural, safe active result and functional product than the chemical reagent Nutrients of culture medium more comprehensively; Promote, improve and changed active result quality in the fermented liquid, reached purpose of the present invention.
Advantage:
(1) the present invention has realized that first the mixed fungus fermentation of needle mushroom, Ganoderma mycelium cultivates;
(2) culture medium prescription is natural, and is easy to use: in mixed fungus fermentation substratum of the present invention, use be natural food crop, without chemical reagent, preparation is convenient, simple, and usually during the fermentation, needn't regulate the pH value;
(3) by needle mushroom, glossy ganoderma mixed fungus fermentation, to compare with needle mushroom, the single bacterium fermentation of glossy ganoderma, exocellular polysaccharide, triterpene compound output all are able to 8% ~ 35% raising.
By mixed fungus fermentation, compare with more single bacterium fermentation, promote, improve and changed active result quality in the fermented liquid.
Embodiment
Below in conjunction with embodiment the present invention is described in further detail:
Embodiment 1:
1, bacterial classification:
Needle mushroom (
Flammulina velutipes): available from Chinese common micro-organisms culture presevation administrative center, be numbered 5.65;
Glossy ganoderma (
Ganoderma lucidum): available from Chinese common micro-organisms culture presevation administrative center, be numbered 5.786, behind mutagenic and breeding, produce the polysaccharide ability and be able to a certain degree raising.
2, substratum:
Mixed fungus fermentation substratum desired raw material comprises sucrose, starch, potato, corn, soya bean, available from the supermarket, is cut into approximately 1cm behind the peeling potatoes
3It is 100 μ m that fritter, corn, analysis for soybean powder are broken to granular size.Consist of 10 ~ 20g/L sucrose, 10 ~ 20g/L starch, 15 ~ 25g/L potato, 10 ~ 30g/L corn, 1 ~ 10g/L soya bean, the backwater preparation; First potato, corn, soybean with water are boiled 25min, add again dissolving good starch and sucrose, sterilize behind the water constant volume.
The extraction and determination of needle mushroom, the cultivation of glossy ganoderma mixed fungus fermentation and tunning
The mixed fungus fermentation method is: the mixed fungus fermentation substratum is comprised of disaccharide, polysaccharide, starch based grain, protein-based grain, adopts the liquid submerged fermentation technology, and mixed bacterium is cultivated needle mushroom and glossy ganoderma; Take needle mushroom and glossy ganoderma as bacterial classification, prepare single bacterium fermentation seed liquid through potato dextrose agar activation, liquid fermentation and culture, combined inoculation needle mushroom, glossy ganoderma are in the mixed fungus fermentation substratum, inoculative proportion 1:1 ~ 3:1,22 ~ 26 ℃ of temperature, pH 4.5 ~ 5.5, and incubation time is 120 ~ 240h, and mycelia and fermented liquid behind the mixed fungus fermentation can be used for tunning: the extraction and determination of biomass, polysaccharide and triterpene compound.
The substratum of described potato dextrose agar for being widely adopted in the laboratory, described mixed fungus fermentation substratum is the grain substratum.
1, zymotechnique (culturing bottle cultivation)
With needle mushroom No. 5.65 bacterial strains and glossy ganoderma No. 5.786 bacterial strains 24 ~ 28 ℃ of lower heat insulating culture activated spawn 6 ~ 9 days, the bacterium piece of getting after the activation is inoculated in the liquid nutrient medium, 25 ℃ of shaking tables were cultivated 7 days, be prepared into needle mushroom, the single bacterium fermentation seed liquid of glossy ganoderma, described liquid nutrient medium is the common liq substratum that is used for cultivating needle mushroom, glossy ganoderma in the laboratory; Be inoculated in the 320mL mixed fungus fermentation substratum 500mL shaking flask according to inoculum size 1:1 ~ 3:1,25 ℃ of shaking tables are cultivated 192h, and behind the filtering fermentation liquor, mycelia is dried to constant weight and gets biomass, and the result is as follows.
Biological accumulation amount (doing g/100mL): needle mushroom: 4 ~ 6; Glossy ganoderma: 3 ~ 5; Mixed bacterium: 4 ~ 6.
The result shows, that adopts that the above-mentioned corn that contains different concns, soya bean mixed fungus fermentation substratum all can success carries out needle mushroom, glossy ganoderma mixed fungus fermentation.
2, zymotechnique (10L fermentor cultivation)
Get needle mushroom, the single bacterium fermentation seed liquid of glossy ganoderma, be inoculated in respectively in the 10L fermentor tank that contains the mixed fungus fermentation substratum and ferment, the inoculum size proportioning is 1:1 ~ 3:1 during mixed bacterium inoculation, amounts to 5%, pH 4.5 ~ 5.5; 24 ℃ of temperature; Time: 144 ~ 168h.Behind the filtering fermentation liquor, mycelia is dried to constant weight and gets biomass, and the result is as follows.
Biological accumulation amount (doing g/100mL): needle mushroom: 5.5 ~ 6.5; Glossy ganoderma: 4 ~ 5; Mixed bacterium: 5 ~ 6.
The result shows, under above-mentioned mixed fungus fermentation culture medium condition, all can carry out needle mushroom, the single bacterium of glossy ganoderma and mixed fungus fermentation tank and cultivate, and incubation time is cultivated to some extent shortening than culturing bottle.
3, the extraction of exocellular polysaccharide
4000r/min centrifuging and taking supernatant liquor behind the filtering fermentation liquor, 80 ℃ of water-baths are condensed into 0.2 times of original volume, add 5 times of volume 95% ethanol, 4 ℃ leave standstill 12h, and concentrated solution 4000r/min is centrifugal, precipitate twice with absolute ethanol washing, dry to constant weight, get crude extracellular polysaccharide for 70 ℃.
4, the extraction of triterpene compound in the born of the same parents
Get 1 g mycelia and add and filter after 50 mL methyl alcohol leave standstill 12h, add 4 mL, 95% dissolve with ethanol behind the evaporate to dryness.
5, the extraction of the outer triterpene compound of born of the same parents
Get fermented liquid 15mL, with the water-saturated n-butanol of 3 times of volumes extraction 3 times, merge n-butanol extracting liquid, evaporated under reduced pressure adds 4mL 95% dissolve with ethanol.
6, activeconstituents accumulation
(1) accumulation of exocellular polysaccharide
Exocellular polysaccharide (g/L): needle mushroom: 37 ~ 39; Glossy ganoderma: 27 ~ 29; Mixed bacterium: 41 ~ 43.
The result shows, mixed fungus fermentation exocellular polysaccharide content all is improved than needle mushroom, the single bacterium fermentation of glossy ganoderma.
(2) accumulation of triterpene compound
Triterpene output (* 10 in the born of the same parents
-2Mg/g): needle mushroom: 64 ~ 68; Glossy ganoderma: 72 ~ 78; Mixed bacterium: 86 ~ 90.
The outer triterpene output (* 10 of born of the same parents
-2G/L): needle mushroom: 66 ~ 70; Glossy ganoderma: 71 ~ 74; Mixed bacterium: 80 ~ 84.
The result shows, behind the mixed fungus fermentation in the born of the same parents, the outer triterpene output of born of the same parents all is improved than needle mushroom, the single bacterium fermentation of glossy ganoderma.
The above results shows, adopt the mixed fungus fermentation substratum to carry out needle mushroom, glossy ganoderma mixed fungus fermentation after, exocellular polysaccharide, triterpene compound all are able to 8% ~ 35% raising.
Claims (1)
1. the method for a needle mushroom, glossy ganoderma mixed fungus fermentation, it is characterized in that: this mixed fungus fermentation method is: the mixed fungus fermentation substratum is comprised of disaccharide, polysaccharide, starch based grain, protein-based grain, and described mixed fungus fermentation substratum comprises: 10 ~ 20g/L starch, 15 ~ 25g/L potato, 10 ~ 20g/L sucrose, 10 ~ 30g/L corn, 1 ~ 10g/L soya bean; Adopt the liquid submerged fermentation technology, mixed bacterium is cultivated needle mushroom and glossy ganoderma; Take needle mushroom and glossy ganoderma as bacterial classification, prepare single bacterium fermentation seed liquid through potato dextrose agar activation, liquid fermentation and culture, combined inoculation needle mushroom, glossy ganoderma are in the mixed fungus fermentation substratum, inoculative proportion 1:1 ~ 3:1,22 ~ 26 ℃ of temperature, pH 4.5 ~ 5.5, and incubation time is 120 ~ 240h, and mycelia and fermented liquid behind the mixed fungus fermentation can be used for tunning: the extraction and determination of biomass, polysaccharide and triterpene compound;
Concrete steps are as follows:
(1) preparation potato dextrose agar slant medium: according to GB4789.15-2010 " microbiological test of food hygiene Molds and yeasts counting " preparation, be used for recovery and the preservation of bacterial classification;
(2) actication of culture: get needle mushroom No. 5.65 bacterial strains and glossy ganoderma No. 5.786 bacterial strains and be inoculated in respectively and activate on the potato dextrose agar slant medium 1 ~ 2 time;
(3) prepare respectively single bacterium fermentation seed liquid: the bacterium piece of getting after the activation of needle mushroom, glossy ganoderma is inoculated in respectively in the liquid nutrient medium 22 ~ 26 ℃ of fermentation culture 168 ~ 288h;
(4) preparation mixed fungus fermentation substratum: be cut into approximately 1cm behind the peeling potatoes
3It is 100 μ m that fritter, corn, analysis for soybean powder are broken to granular size, and substratum consists of 10 ~ 20g/L starch, 15 ~ 25g/L potato, 10 ~ 20g/L sucrose, 10 ~ 30g/L corn, 1 ~ 10g/L soya bean, adds the water preparation; First potato, corn, soybean with water are boiled 25min, add again dissolving good starch and sucrose, sterilize after adding the water constant volume;
(5) preparation mixed fungus fermentation seed liquor: get needle mushroom, the single bacterium fermentation seed liquid of glossy ganoderma, be inoculated in the mixed fungus fermentation substratum, be total to ferment nutrient solution acidity-basicity ph 4.5 ~ 5.5, the inoculum size proportioning is 1:1 ~ 3:1, amounts to 5%, 22 ~ 26 ℃ of fermentation culture 144 ~ 240h;
(6) separating bio amount: filtering fermentation liquor is collected mycelia, dries to constant mass and claims quality for 60 ℃;
(7) crude extracellular polysaccharide extraction and determination: centrifuging and taking supernatant liquor behind the filtering fermentation liquor, 80 ~ 90 ℃ of water-baths are condensed into 0.2 ~ 0.25 times of original volume, add 4 ~ 5 times of volume 95% ethanol, 4 ℃ leave standstill, and concentrated solution is centrifugal, precipitate twice with absolute ethanol washing, dry to constant weight, get crude extracellular polysaccharide;
(8) extraction of triterpene compound in the born of the same parents: get and filter after 1 g mycelia adds 50 mL methyl alcohol standing over night, add 4 mL, 95% dissolve with ethanol behind the evaporate to dryness;
(9) extraction of the outer triterpene compound of born of the same parents: get fermented liquid 15mL, with the water-saturated n-butanol extraction of 3 times of volumes 3 times, merging n-butanol extracting liquid, evaporated under reduced pressure adds 4mL 95% dissolve with ethanol.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103156253A (en) * | 2013-04-02 | 2013-06-19 | 徐州工程学院 | Preparation method of needle mushroom and lucid ganoderma mixed fermentation functional beverage |
| CN105145115A (en) * | 2015-08-31 | 2015-12-16 | 兴安宸亿工贸有限公司 | Method for producing organic selenium-rich zinc-rich products |
| CN106852496A (en) * | 2015-12-09 | 2017-06-16 | 黄建钧 | The method of the triterpenes synthesis contained by induction mushroom gill fungus |
| CN108260808A (en) * | 2018-01-31 | 2018-07-10 | 万宁万维诺丽生物科技有限公司 | A kind of beautiful ferment of promise and preparation method thereof |
| CN109777847A (en) * | 2019-03-01 | 2019-05-21 | 宁德师范学院 | The method of body cell is affine ganoderma strain to common fermentation production strong anti-oxidative activity polysaccharide |
| CN110982868A (en) * | 2019-11-29 | 2020-04-10 | 贵州中医药大学 | Co-culture method for improving triterpene content of ganoderma lucidum and application |
| CN111635924A (en) * | 2020-06-03 | 2020-09-08 | 天津科技大学 | Liquid fermentation method for increasing yield of ganoderma lucidum extracellular polysaccharide |
-
2012
- 2012-11-14 CN CN201210455815XA patent/CN102925527A/en active Pending
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| JINGSONG ZHANG ET AL.: "Activation of B lymphocytes by GLIS, a bioactive proteoglycan from Ganoderma lucidum", 《LIFE SCIENCES》 * |
| 吴漾 等: "松茸、蛹虫草混菌共酵菌丝体多糖提取工艺的优化研究", 《食品研究与开发》 * |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103156253A (en) * | 2013-04-02 | 2013-06-19 | 徐州工程学院 | Preparation method of needle mushroom and lucid ganoderma mixed fermentation functional beverage |
| CN105145115A (en) * | 2015-08-31 | 2015-12-16 | 兴安宸亿工贸有限公司 | Method for producing organic selenium-rich zinc-rich products |
| CN105145115B (en) * | 2015-08-31 | 2018-02-06 | 唐健发 | A kind of production method of organic Se-rich zinc-rich product |
| CN106852496A (en) * | 2015-12-09 | 2017-06-16 | 黄建钧 | The method of the triterpenes synthesis contained by induction mushroom gill fungus |
| CN108260808A (en) * | 2018-01-31 | 2018-07-10 | 万宁万维诺丽生物科技有限公司 | A kind of beautiful ferment of promise and preparation method thereof |
| CN109777847A (en) * | 2019-03-01 | 2019-05-21 | 宁德师范学院 | The method of body cell is affine ganoderma strain to common fermentation production strong anti-oxidative activity polysaccharide |
| CN110982868A (en) * | 2019-11-29 | 2020-04-10 | 贵州中医药大学 | Co-culture method for improving triterpene content of ganoderma lucidum and application |
| CN110982868B (en) * | 2019-11-29 | 2023-09-19 | 贵州中医药大学 | Co-culture method for improving triterpene content of ganoderma lucidum and application thereof |
| CN111635924A (en) * | 2020-06-03 | 2020-09-08 | 天津科技大学 | Liquid fermentation method for increasing yield of ganoderma lucidum extracellular polysaccharide |
| CN111635924B (en) * | 2020-06-03 | 2023-04-07 | 天津科技大学 | Liquid fermentation method for increasing yield of ganoderma lucidum extracellular polysaccharide |
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Application publication date: 20130213 |