CN102108371A - Process for submerged fermentation and production of anti-oxidation active substances of white rot fungi by utilizing lignocellulosic agricultural and forestry wastes - Google Patents
Process for submerged fermentation and production of anti-oxidation active substances of white rot fungi by utilizing lignocellulosic agricultural and forestry wastes Download PDFInfo
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- CN102108371A CN102108371A CN2009101571644A CN200910157164A CN102108371A CN 102108371 A CN102108371 A CN 102108371A CN 2009101571644 A CN2009101571644 A CN 2009101571644A CN 200910157164 A CN200910157164 A CN 200910157164A CN 102108371 A CN102108371 A CN 102108371A
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Abstract
The invention relates to a method for improving the yield of submerged fermentation polysaccharides and polyphenols of white rot fungi and the anti-oxidation activity by adding lignocellulosic agricultural and forestry wastes. The fermentation is performed in a liquid fermentation tank by taking the submerged fermentation white rot fungi, such as inonotus obliquus polysaccharides and the polyphenols as objects and taking inonotus obliquus in a Netherlands CBS (centraalbureauvoor schimmelcultures) cell bank as a starting strain. On the basis of taking hydrolysis sugar of corn starch as a fermentation carbon source, crop straws (corn straws, paddy rice straws and wheat straws) and/or wood chips are added for decomposing and utilizing lignocellulose in added substances while enabling mycelia of the inonotus obliquus to utilize the hydrolysis sugar of the starch. The fermentation conditions are as follows: the temperature is 27-28 DEG C, the air flow rate is 0.5-1.0vvm, the stirring rotational speed is 70-150 turns/minute, the internal pressure is 0.1-0.25kg/cm3, and the fermentation period is 12-14 days. By adopting the method, the yield of the submerged fermentation polysaccharides and the polyphenols of the inonotus obliquus and activity can be improved, and the lignocellulose can be simultaneously and effectively decomposed.
Description
Technical field
The present invention relates to a kind of lignocellulose agriculture and forestry organic waste material such as agricultural crop straw and wood chip of utilizing and promote liquid submerged fermentation to produce the technology of white-rot fungi active substance, belong to biotechnology or bioengineering field.
Technical background
Fungi has been had very long history by mankind's discovery, understanding, utilization, and people are known no more than large-scale edible and medicinal fungi the most.They are not only nutritious, and the many compositions that contain have stronger physiology and pharmacologically active.Modern study shows that senior basidiomycetes has wide range of therapeutic and prophylactic function.The pore fungus fungi is the big monoid of one in the basidiomycetes, manyly in the mushroom medicine of China belongs to this class fungi, as famous traditional Chinese medicine glossy ganoderma, umbellate pore furgus, Poria cocos or the like.
Adopt the method for liquid submerged fermentation, compare with traditional fungi method of cultivation, have with short production cyclely, cost is low, and output and quality is stable, and product such as can control at superiority.The biological effective components that utilizes biotechnology the to produce fungi trend that is inevitable, the existing in the world at present commercial employing liquid submerged fermentation of many medicinal fungis obtains effective constituent.Successfully mushroom has been carried out since submerged fermentation and nineteen fifty-seven cultivate the deep liquid of morel with fermentor tank from the U.S. in 1948, the liquid submerged fermentation technology is extensive day by day in the application of aspects such as the edible medicinal fungus production of hybrid seeds, comprehensive utilization.At present, existing in the world more than 50 kind of edible medicinal fungus carried out the liquid fermenting experiment.The domestic edible medicinal fungus that has carried out the application of submerged fermentation and suitability for industrialized production has Cordyceps sinensis, rainbow conk, glossy ganoderma, white fungus, hedgehog hydnum, dictyophora phalloidea, umbellate pore furgus, honey mushroom, mushroom, ergot, cuvette goatsbeard, Hericium caput-medusae etc.
In the various activeconstituentss that edible and medicinal fungi contained, fungus polysaccharide is the most effective so far, also is that people are familiar with the most deep activeconstituents, and it has immunomodulatory, antitumor, anti-inflammatory, function such as anti-oxidant.Over nearly 40 years, along with deepening continuously that the edible and medicinal fungi polysaccharide is studied, multiple polysaccharide has been made into medicine, is applied to clinical treatment and also introduces to the market on a large scale.
When liquid state fermentation was cultivated, various chemical factors all can influence fungi growth speed, biomass and pathways metabolism, so exploitation suitable medium and fermentation condition are to improve edible and medicinal fungi active substance output, obtained the effective way of high reactivity product.
Class white-rot fungi in the edible and medicinal fungi can produce lignocellulose lytic enzyme system, and xylogen finally is decomposed into CO2, exposes Mierocrystalline cellulose and hemicellulose, and then is that corresponding monose is for the thalline utilization with Mierocrystalline cellulose and hydrolysis of hemicellulose.Because the zymoprotein molecule volume is too huge, can't be smoothly fine and close reticulated structure by the xylogen composition, so thalline oxygenolysis xylogen at first.White-rot fungi is a chain reaction process based on free radical to the biological degradation of xylogen.In order to improve the output and the activity of white-rot fungi biosynthetic polysaccharide and polyphenolic compound in liquid submerged fermentation; the present invention adds lignocellulose class agriculture and forestry organic waste material in the liquid submerged fermentation substratum; as agricultural crop straw (maize straw, rice straw, straw etc.) and wood chip; to induce white-rot fungi in decomposing lignocellulose, to produce hydroxyl radical free radical; thereby stimulate the self-protective mechanism of fermentation mycelium; just stimulate the more antioxidant of fermentation mycelium biosynthesizing, as polyphenols and polysaccharose substance.
It is example that the present invention has another name called Chaga with the rare medicinal fungi Phaeopoms obliquus (Inonotus obliquus) that belongs to white-rot fungi.Phaeopoms obliquus belongs to Basidiomycotina, Hymenomycetes, and non-brown Zoopagales, polyporaceae, brown transverse hole fungus belongs to.On the bark of being born in white birch, silvery birch, elm, alder etc. dried-up down or after the felling.Since 16th century, Phaeopoms obliquus is as a kind of medicinal fungi among the people, in Russia and siberian, the area, Eastern Europe is used to prevent and treat difficult and complicated cases, comprise diabetes, infective virus and multiple cancer, be considered to have significant pharmacologically active and a kind of medicinal fungi of human body without any side effect.Fuscoporia obliqua polysaccharide class and polyphenols are considered to one of activeconstituents of Phaeopoms obliquus, have very strong anti-oxidant activity.
The present invention utilizes the degraded of lignocellulose to promote Phaeopoms obliquus liquid fermenting polyphenol, the generation of polysaccharide and the raising of anti-oxidant activity, its meaning is the follow-up carbon source of lignocellulose as the Phaeopoms obliquus fermentation, in the fermentative production active substance, decompose the agricultural wastes that utilize lignocellulose, have important scientific meaning and application prospect.
Summary of the invention
The present invention is to be target to produce Phaeopoms obliquus liquid fermenting antioxidant polyphenol and polysaccharide, set up a kind of when the Phaeopoms obliquus liquid fermenting produces active substance, decompose the technology utilize the lignocellulose in the agricultural crop straw, improved the output of Phaeopoms obliquus fermentation activity polyphenol, polysaccharide greatly.
Technical scheme of the present invention: the Phaeopoms obliquus fungi is the bacterial classification that sets out, and adopts slant strains to carry out liquid seeds and cultivates.At the W-Gum hydrolysis sugar is on the basis of fermenting carbon source, adds a certain amount of agricultural crop straw or wood chip, carries out liquid submerged fermentation.Make the Phaeopoms obliquus mycelium when fermentation utilizes amylum hydrolysate of the sugar, decompose the lignocellulose that utilizes in stalk or the wood chip.Fermentation condition is: 27-28 ℃, air flow quantity 0.5-1.0vvm, mixing speed are 70-150 rev/min, internal pressure 0.1-0.25 kilogram/cubic centimetre, fermentation period 10-14 days.Add lignocellulose the output of Phaeopoms obliquus fermentation exocellular polysaccharide is improved more than 40%, the output of the outer polyphenol of born of the same parents improves more than 700%.
Realize that concrete steps of the present invention are as follows:
(1) take out the inclined-plane bacterial strain and insert in the slant medium of fresh configuration, the slant strains culture temperature is 25-30 ℃, cultivates 200-250 hour.
(2) slant strains in the step 1 is changed over to liquid seed culture medium is housed shake in the bottle liquid seeds culture condition: temperature is 27-28 ℃, shaking speed 70-150 rev/min.Cultivated 96-120 hour.
(3) liquid spawn in the step 2 is changed in the fermentor tank that the maize straw liquid fermentation medium is housed, carry out liquid submerged fermentation.Fermentation condition: temperature is 27-28 ℃, shaking speed 70-150 rev/min.Cultivated 288 hours.
The slant strains substratum is formed (g/100mL) among the present invention: malt extract 30, peptone 3, agar 15.The pH value is 5.4-5.6.
The shake-flask seed substratum is formed (g/100mL) among the present invention: glucose 2, peptone 0.3, yeast extract 0.2, KH
2PO
40.1, MgSO
40.15, CaCl
20.01.
Stalk liquid submerged fermentation substratum is formed (g/100mL) among the present invention: Semen Maydis powder 5, peptone 0.3, KH
2PO
40.1, ZnSO
42H
2O 0.001, K
2HPO
40.04, FeSO
47H
2O 0.005, MgSO
47H
2O 0.05, CuSO
45H
2O 0.002, CoCl
20.001, MnSO
4H
2O 0.008.PH value 6.0.
The standard that fermentation termination of the present invention is established is that reducing sugar content is lower than 2%, and microscopy finds that mycelium senesces.
Embodiment
Below in conjunction with specific embodiment the present invention is further described in detail
Embodiment 1:
The Phaeopoms obliquus mycelium in the Dutch CBS culture presevation of inoculation storehouse in slant medium.28 ℃ of culture temperature were cultivated 240 hours.Fresh slant strains is inserted the 500mL that the 200mL liquid seed culture medium is housed shake in the bottle, cultivated 150 rev/mins of shaking speed 96 hours for 28 ℃.Cultured liquid seeds is equipped with in the fermentor tank of maize straw fermention medium according to 10% (v/v) access.Cultivated about 288 hours for 28 ℃.Air flow quantity 1.0vvm, 0.2 kilogram/cubic centimetre of internal pressure, mixing speed is 140 rev/mins.
Slant strains is cultivated in the present embodiment 1, and it is (g/100mL) that substratum is formed: malt extract 30, and peptone 3, agar 15, the pH value is 5.4-5.6.
Shake-flask seed is cultivated in the present embodiment 1, and substratum is formed (g/100mL): glucose 2, peptone 0.3, yeast extract 0.2, KH
2PO
40.1, MgSO
40.15, CaCl
20.01.
The maize straw liquid submerged fermentation is cultivated in the present embodiment 1, and substratum is formed (g/100mL): Semen Maydis powder 3.5, maize straw powder 5%, peptone 0.3, KH
2PO
40.1, ZnSO
42H
2O 0.001, K
2HPO
40.04, FeSO
47H
2O 0.005, MgSO
47H
2O 0.05, CuSO
45H
2O 0.002, CoCl
20.001, MnSO
4H
2O 0.008.The pH value is 6.0.
Fermentation terminated standard is that reducing sugar content is lower than 2% in the present embodiment 1, and senile symptom appears in the microscopy mycelium.
The extraction of fermentation exocellular polysaccharide: isolate fermented liquid with the vacuum filtration method after the fermentation ends.Fermented liquid be concentrated into original volume 1/4 after, add the dehydrated alcohol of 4 times of volumes, high degree of agitation obtains polysaccharide precipitation, precipitates 4 ℃ and spends the night.Polysaccharide precipitation is through centrifugation, and neutral protease is handled, and the Sevage method is removed albumen, and the dialysis postlyophilization gets exocellular polysaccharide.
The ferment extraction of water-soluble intracellular polyse: the mycelia after the fermentation ends, behind distilled water wash, be suspended in the distilled water, behind High Temperature High Pressure heating or the supersonic wave wall breaking, extract the thermostability intracellular polyse, the filtrate concentrated solution adds 4 times 95% ethanol, fully vibration, put 4 ℃ of precipitations of spending the night, precipitation is removed albumen with 95% ethanol and washing with acetone, Sevage method, and lyophilize gets intracellular polyse.
The extraction of the outer polyphenol of fermentation born of the same parents: isolate fermented liquid with the vacuum filtration method after the fermentation ends, or the fermentation concentrated solution behind the ethanol sedimentation separating polyose, with 1: 1 ethyl acetate extraction of volume ratio, extraction liquid concentrating under reduced pressure, vacuum or lyophilize obtain total polyphenols.Methods such as employing chromatogram are further purified.
Fig. 1 is maize straw Mierocrystalline cellulose in the liquid submerged fermentation process, hemicellulose, the content of lignin Changing Pattern with fermentation time.At preceding 144 hours, stable the remaining on the higher level of residual rate of three kinds of major ingredient of lignocellulose, and change little.After 144 hours, Mierocrystalline cellulose, hemicellulose, xylogen are decomposed fast, and residual rate descends rapidly.During fermentation ends in the substratum residual rate of Mierocrystalline cellulose, hemicellulose, xylogen be respectively 79.1,82.1, and 80.2%.
Fig. 2 be in the liquid submerged fermentation process exocellular polysaccharide content with the Changing Pattern of fermentation time.The output of Phaeopoms obliquus cell free fermentation polysaccharide that produces in lignocellulose fermention medium and no lignocellulose fermention medium (contrast) improves with the prolongation of fermentation time.After 144 hours, the polysaccharide yield of two kinds of substratum presents than big-difference.During fermentation ends, in lignocellulose fermention medium and the no lignocellulose fermention medium, the output of Phaeopoms obliquus fermentation exocellular polysaccharide is respectively 1.4g/L and 1.0g/L.The adding of lignocellulose has significantly improved the output of Phaeopoms obliquus fermentation exocellular polysaccharide.The output of fermentation exocellular polysaccharide and three kinds of major ingredient content of lignocellulose have closely over time gets in touch, and is accompanied by the quickening of three kinds of major ingredient rate of decomposition of lignocellulose, and the cell free fermentation polysaccharide yield improves fast.
Fig. 3 be in the liquid submerged fermentation process the outer polyphenol content of born of the same parents with the Changing Pattern of fermentation time.Phaeopoms obliquus produces the cell free fermentation polyphenol at the lignocellulose fermention medium output improves with the prolongation of fermentation time, reaches the highest on the 8th day in fermentation.Output is 700% of the cell free fermentation polyphenol that produces in the no lignocellulose fermention medium (contrast).
Fig. 4 is the anti-oxidant activity of lignocellulose fermention medium and no lignocellulose fermention medium fermentation exocellular polysaccharide, and (a) hydroxyl radical free radical is removed active; (b) superoxide radical is removed active; (c) lipid peroxidation restraining effect.Compare with no lignocellulose fermention medium fermentation polysaccharide, the highest hydroxyl radical free radical clearance rate, the superoxide radical that the adding of lignocellulose has significantly improved the fermentation exocellular polysaccharide removed active and to the restraining effect of the spontaneous lipid peroxidation of mouse liver.
Embodiment 2:
The Phaeopoms obliquus mycelium in the Dutch CBS culture presevation of inoculation storehouse in slant medium.28 ℃ of culture temperature were cultivated 240 hours.Fresh slant strains is inserted the 500mL that the 200mL liquid seed culture medium is housed shake in the bottle, cultivated 140 rev/mins of shaking speed 96 hours for 28 ℃.Cultured liquid seeds is equipped with in the fermentor tank of wood chip fermention medium according to 10% (v/v) access.Cultivated about 302 hours for 28 ℃.Air flow quantity 0.5vvm, 0.1 kilogram/cubic centimetre of internal pressure, mixing speed is 140 rev/mins.
Slant strains is cultivated in the present embodiment 2, and it is (g/100mL) that substratum is formed: malt extract 30, and peptone 3, agar 15, the pH value is 5.4-5.6.
Shake-flask seed is cultivated in the present embodiment 2, and substratum is formed (g/100mL): glucose 2, peptone 0.3, yeast extract 0.2, KH
2PO
40.1, MgSO
40.15, CaCl
20.01.Shaking speed is 150 rev/mins.
The rice straw liquid submerged fermentation is cultivated in the present embodiment 2, and substratum is formed (g/100mL): Semen Maydis powder 3.5, wood chip 3.5%, peptone 0.3, KH
2PO
40.1, ZnSO
42H
2O 0.001, K
2HPO
40.04, FeSO
47H
2O 0.005, MgSO
47H
2O0.05, CuSO
45H
2O 0.002, CoCl
20.001, MnSO
4H
2O 0.008.PH value 6.0.
Fermentation terminated standard is that reducing sugar content is lower than 2% in the present embodiment 2, and senile symptom appears in the microscopy mycelium.
Description of drawings
Fig. 1 is the exploded view of three kinds of main components of lignocellulose in the Phaeopoms obliquus liquid fermenting process, (●) hemicellulose, (▲) xylogen, (■) Mierocrystalline cellulose;
Fig. 2 is cell free fermentation polysaccharide yield figure in the Phaeopoms obliquus liquid fermenting process, (●) no lignocellulose fermentation basic medium, (■) xylogen fermention medium;
Fig. 3 is the outer polyphenol output figure of born of the same parents in the Phaeopoms obliquus liquid submerged fermentation process, (●) lignocellulose fermention medium, (■) no xylogen fermention medium;
Fig. 4 is that Phaeopoms obliquus ferments exocellular polysaccharide to three kinds of oxidation activity free radical scavenging design sketchs, (●) xylogen fermention medium, (■) no lignocellulose fermentation basic medium.(a) hydroxyl radical free radical is removed active; (b) superoxide radical is removed active; (c) lipid peroxidation restraining effect.
Claims (4)
1. one kind is utilized the lignocellulose agriculture and forestry organic waste material, and as agricultural crop straw and wood chip, liquid submerged fermentation is produced the technology of Phaeopoms obliquus fermentation exocellular polysaccharide, polyphenols.It is characterized in that being the bacterial classification that sets out, adopt slant strains to carry out liquid shaking bottle seed culture, ferment tank with white-rot fungi.Remove adopting traditional W-Gum hydrolysis sugar in the fermenting process is fermenting carbon source, replenishes a certain amount of agricultural crop straw and/or wood chip in addition, makes bacterial classification during the fermentation except that utilizing the W-Gum hydrolysis sugar, decomposes and utilizes lignocellulose.This method has improved the output and the anti-oxidant activity of liquid submerged fermentation polysaccharide, polyphenols greatly, has also effectively decomposed lignocellulose simultaneously, has reached the purpose that agricultural wastes utilize again.
2. the technology of utilizing lignocellulose agriculture and forestry organic waste material liquid submerged fermentation to produce fermented by white rot fungus polysaccharide, polyphenols according to claim 1 is characterized in that being the bacterial classification that sets out with white-rot fungi as Phaeopoms obliquus.
3. utilize the maize straw liquid submerged fermentation to produce the ferment technology of polysaccharide, polyphenols of white-rot fungi such as Phaeopoms obliquus according to claim 1, it is characterized in that agriculture and forestry organic waste material is: lignocellulose wastes such as maize straw, rice straw, large and small wheat straw stalk, wood chip, waste paper.
4. utilize lignocellulose waste liquid submerged fermentation to produce the ferment technology of polysaccharide, polyphenols of white-rot fungi such as Phaeopoms obliquus according to claim 1, it is characterized in that adding lignocellulose in the liquid fermentation tank substratum, substratum is formed (g/100mL): Semen Maydis powder 3.5, agricultural crop straw powder or wood chip 2-10%, peptone 0.3, KH
2PO
40.1, ZnSO
42H
2O0.001, K
2HPO
40.04, FeSO
47H
2O 0.005, MgSO
47H
2O 0.05, CuSO
45H
2O 0.002, CoCl
20.001, MnSO
4H
2O 0.008.PH value 6.0.Liquid fermentation condition: 27-28 ℃, air flow quantity 0.5-1.0vvm, mixing speed 70-150 rev/min, internal pressure 0.1-0.25 kilogram/cubic centimetre, fermentation period 12-14 days.
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103146775A (en) * | 2011-12-07 | 2013-06-12 | 徐向群 | Method for preparing epigallocatechin-3-gallate and epigallocatechin gallate by liquid-submerged fermentation |
| CN103156052A (en) * | 2013-02-21 | 2013-06-19 | 徐向群 | Submerged fermentation technology of producing immunological enhancement active materials of inonotus obliquus |
| CN103283990A (en) * | 2013-05-07 | 2013-09-11 | 徐向群 | Application of inonotus obliquus polysaccharide used as feed additive for litopenaeus vannamei |
| CN104206776A (en) * | 2013-05-29 | 2014-12-17 | 徐向群 | Application of inonotus obliquus polysaccharide as fish feed additive |
| CN105624133A (en) * | 2016-01-20 | 2016-06-01 | 徐向群 | Technique for producing Inonotus obliquus lignocellulose catabolic enzyme by liquid submerged fermentation |
| CN105695529A (en) * | 2016-03-08 | 2016-06-22 | 中国人民解放军总后勤部军需装备研究所 | Method for preparing exopolysaccharides from ganoderma lucidum |
| CN107189996A (en) * | 2017-07-10 | 2017-09-22 | 浙江理工大学 | The technique that a kind of utilization Inonotus obliquus solid fermentation agriculture and forestry organic waste material produces acidic cellulase |
| CN108330072A (en) * | 2018-02-02 | 2018-07-27 | 山西康欣药业有限公司 | Inonotus obliquus liquid submerged fermentation culture composition and preparation method and application |
| CN112159789A (en) * | 2020-10-17 | 2021-01-01 | 江苏师范大学 | Application of eupolyphaga powder in promoting growth of inonotus obliquus and inducing accumulation of polyphenol |
| CN112899323A (en) * | 2019-12-04 | 2021-06-04 | 中国农业大学 | Method for promoting inonotus obliquus to produce hypoglycemic active polysaccharide by adding pretreated birch sawdust |
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Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103146775A (en) * | 2011-12-07 | 2013-06-12 | 徐向群 | Method for preparing epigallocatechin-3-gallate and epigallocatechin gallate by liquid-submerged fermentation |
| CN103156052A (en) * | 2013-02-21 | 2013-06-19 | 徐向群 | Submerged fermentation technology of producing immunological enhancement active materials of inonotus obliquus |
| CN103283990A (en) * | 2013-05-07 | 2013-09-11 | 徐向群 | Application of inonotus obliquus polysaccharide used as feed additive for litopenaeus vannamei |
| CN104206776B (en) * | 2013-05-29 | 2017-05-31 | 徐向群 | Fuscoporia obliqua polysaccharide as fish feed additive application |
| CN104206776A (en) * | 2013-05-29 | 2014-12-17 | 徐向群 | Application of inonotus obliquus polysaccharide as fish feed additive |
| CN105624133A (en) * | 2016-01-20 | 2016-06-01 | 徐向群 | Technique for producing Inonotus obliquus lignocellulose catabolic enzyme by liquid submerged fermentation |
| CN105695529A (en) * | 2016-03-08 | 2016-06-22 | 中国人民解放军总后勤部军需装备研究所 | Method for preparing exopolysaccharides from ganoderma lucidum |
| CN107189996A (en) * | 2017-07-10 | 2017-09-22 | 浙江理工大学 | The technique that a kind of utilization Inonotus obliquus solid fermentation agriculture and forestry organic waste material produces acidic cellulase |
| CN107189996B (en) * | 2017-07-10 | 2020-10-20 | 浙江理工大学 | Process for producing acid cellulase by utilizing inonotus obliquus solid fermentation agricultural and forestry waste |
| CN108330072A (en) * | 2018-02-02 | 2018-07-27 | 山西康欣药业有限公司 | Inonotus obliquus liquid submerged fermentation culture composition and preparation method and application |
| CN112899323A (en) * | 2019-12-04 | 2021-06-04 | 中国农业大学 | Method for promoting inonotus obliquus to produce hypoglycemic active polysaccharide by adding pretreated birch sawdust |
| CN112899323B (en) * | 2019-12-04 | 2022-03-22 | 中国农业大学 | Method for promoting inonotus obliquus to produce hypoglycemic active polysaccharide by adding pretreated birch sawdust |
| CN112159789A (en) * | 2020-10-17 | 2021-01-01 | 江苏师范大学 | Application of eupolyphaga powder in promoting growth of inonotus obliquus and inducing accumulation of polyphenol |
| CN112159789B (en) * | 2020-10-17 | 2022-01-14 | 江苏师范大学 | Application of eupolyphaga powder in promoting growth of inonotus obliquus and inducing accumulation of polyphenol |
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Application publication date: 20110629 |