CN104099385A - Method for producing inonotus obliquus exopolysaccharides through submerged fermentation - Google Patents
Method for producing inonotus obliquus exopolysaccharides through submerged fermentation Download PDFInfo
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Abstract
The invention belongs to the technical field of microorganisms, and relates to a method for producing inonotus obliquus exopolysaccharides through submerged fermentation, particularly to a method for industrial-scale production of the inonotus obliquus exopolysaccharides. The method comprises the steps as follows: activated inonotus obliquus strains are inoculated into a seed tank for propagation, then propagated liquid seeds are inoculated into a specially-made polysaccharide-producing liquid culture medium for fermentation, and a fermentation liquid is separated to extract the exopolysaccharides. According to the method, research is conducted to overcome defects in the prior art, on the premises that a fermentation culture medium (especially a carbon source) is continuously optimized through small and medium tests and the biomass and the exopolysaccharides are increased, the problems of fermentation process parameter control, extracting equipment model selection, optimization and the like in the process of enlargement from the small tests to industrial production are solved, the strains are successfully inoculated into fermentation tanks of 10-20 t finally for industrial production, the high-quality inonotus obliquus polysaccharides are produced smoothly, and the continuous, automatic and industrial production is actually realized.
Description
Technical field
The invention belongs to microbial technology field, be specifically related to a kind of method of deep layer liquid state fermentation production Phaeopoms obliquus exocellular polysaccharide.
Background technology
Phaeopoms obliquus has another name called Chaga, belongs to Basidiomycotina, Hymenomycetes, non-brown Zoopagales, polyporaceae, brown transverse hole fungus genus, is a kind of large-scale medicinal fungi of parasitics of being born on birch.Its sporophore presents the black block-shape morphology that is similar to carbon, is mainly distributed in the rare medical fungus in the areas such as northern North america, Russia, Finland, China Dark Longjiang, Hokkaido, Japan
Phaeopoms obliquus main parasitic is under the bark of other birch classes such as white birch, elm, silvery birch, and sporophore is sclerotium surface black drastic crack, tube leading portion cracking, and bacterial context suberin and there is ring grain, spore ellipticity or ovum shape, be that a kind of life is the wood-decay fungi in frigid zone.Phaeopoms obliquus is widely used on Russia and other places, it is a kind of Russian common drug among the people, the existing at least applicating history of centuries, remarkable in aspect effects such as treatment gastroenteropathy, relieving cancer symptoms, preventing diseases because of it, be widely used in areas such as Russia, Central Europe, China northeast, be widely accepted, be locally known as " panacea ".
Along with the development of modern biotechnology, investigator finds that the polysaccharide of Phaeopoms obliquus may be its main pharmacodynamics composition, and that the Fuscoporia obliqua polysaccharide pharmacological action of having reported at present mainly comprises is antitumor, antiviral, hypoglycemic, reducing blood-fat, removing free radical, LPO inhibitor, anti-inflammatory, strengthening immunity, anticoagulation, anti-ageing, pain relieving etc.It is reported, the antitumor action of Phaeopoms obliquus mainly activates the immunity system of body by polysaccharose substances such as dextran, plays the effect of the antineoplastic immune of enhancing body.Aspect treatment tumour, in serum, the content of nitric oxide of higher concentration can affect the conduction of tumour cell signal and the formation of intratumoral vasculature, promotes tumor growth.Fuscoporia obliqua polysaccharide has obvious restraining effect to the growth of murine sarcoma.Fuscoporia obliqua polysaccharide is hypoglycemic, the research of reducing blood lipid shows, Fuscoporia obliqua polysaccharide can promote the reverse metabolism of cholesterol to be converted to cholesteryl ester, suppresses enteron aisle inner cholesterol and absorbs, and can improve function of vascular endothelium simultaneously, reduces blood fat.In Phaeopoms obliquus, polysaccharose substance has immune enhancing function, and the lymphocytic activity of Immune interrelation and function are had to promoter action, can remove interior free yl, extends cell fission, extends cell survival.
Increasing scholar effective component and the pharmacological action of Phaeopoms obliquus that begin one's study in the recent decade, Phaeopoms obliquus has become most one of bacterial classification of DEVELOPMENT PROSPECT in medicinal fungi.But at present domestic wild Phaeopoms obliquus resource scarcity, the low and poor repeatability of the artificial culture success ratio of sporophore, the solid state cultivation cycle is long, pollution rate is high, is subject to surrounding environment restriction, cannot meet the growing market requirement.And the advantage such as deep layer liquid state fermentation technology has that fermentation period is short, high yield, pollution rate are low, not limited by surrounding environment, resource apparatus utilization ratio advantages of higher, thereby be subject to increasing investigators' favor.Also there is the investigator of a small amount of universities and colleges to carry out research to the submerged fermentation of Phaeopoms obliquus both at home and abroad, but often its main special emphasis is how to obtain higher biomass, only have small part investigator to pay close attention to the research of the functional component polysaccharide of Phaeopoms obliquus, but be mainly to extract and obtain polysaccharide by mycelium, and the research that Phaeopoms obliquus exocellular polysaccharide is produced in liquid state fermentation has been carried out in the part Study life of Jin You Institutes Of Technology Of Zhejiang and University Of Science and Technology Of Tianjin, but its deficiency mainly contains: be 1. all only confined to the shake flask test of lab scale, belong to phase of basic research; 2. from their data, can find out: about 9 days of culture cycle; biomass is 1.0-1.1g/100ml; exopolysaccharides is 0.11-0.14g/100ml; the outer sugared yield of biomass and active result born of the same parents is too low; production cost is too expensive; its technique needs further to promote, and also needs to carry out a large amount of research and cut-and-try work if will reach the production Fuscoporia obliqua polysaccharide of mass-producing and industrialization.
Summary of the invention
The object of the invention is the technical bottleneck in order to solve commercial scale production Fuscoporia obliqua polysaccharide, adopt liquid state fermentation to produce Fuscoporia obliqua polysaccharide and overcome that the solid state fermentation cycle is long, investment is large, be subject to seasonal restrictions, be difficult to the drawbacks such as industrialization, adopt this invention can be continuously, the Fuscoporia obliqua polysaccharide of producing high-quality of automatization, industrialization.
To achieve these goals, the present invention mainly carries out according to following step:
The method that Phaeopoms obliquus exocellular polysaccharide is produced in industrialization deep layer liquid state fermentation of the present invention comprises the steps:
Prepare a method for Phaeopoms obliquus fermented liquid, by the fermention medium after Phaeopoms obliquus seed culture fluid access sterilizing, inoculum size is 10～20%, ventilating ratio be 1:0.3 ?1.0vvm, tank pressure 0.02～0.05Mpa, cultivates 3～4 days at 22～30 DEG C of bottom fermentations, obtains Phaeopoms obliquus fermented liquid; Wherein said fermentative medium formula is: the high temperature resistant α-amylase 0.001-0.007% (v/v) of glucose 1-3%, starch 1-3%, dried silkworm chrysalis meal 0.1-0.5%, peptone 0.1-0.5%, potassium primary phosphate 0.1-0.5%, magnesium sulfate heptahydrate 0.1-0.5%, defoamer 0.01-0.05%, 20000u/ml, regulates pH value 6.5-7.5." % " in above-mentioned substratum composition except amylase, all represents g/100ml.
Described defoamer can be the materials such as vegetables oil (soya-bean oil, rapeseed oil) class, organic ethers.
Wherein, described Phaeopoms obliquus seed culture fluid preferably prepares by the following method:
1) first order seed is cultivated: by the liquid seed culture medium after activated shaking flask bacterial classification access sterilizing, inoculum size is 0.1～2%, ventilating ratio be 1:0.3 ?1.0vvm, tank pressure 0.02～0.05Mpa, at 22～30 DEG C, cultivate 5～6 days, obtain Phaeopoms obliquus first order seed nutrient solution;
2) secondary seed is cultivated: by step 1) liquid seed culture medium after the first order seed nutrient solution of gained access sterilizing, inoculum size is 10～20%, ventilating ratio be 1:0.3 ?1.0vvm, tank pressure 0.02～0.05Mpa, at 22～30 DEG C, cultivate 3～4 days, obtain described Phaeopoms obliquus seed culture fluid.
Described liquid seed culture medium formula optimization is: glucose 2-4%, dried silkworm chrysalis meal 0.1-0.5%, soybean cake powder 1-3%, potassium primary phosphate 0.1-0.5%, magnesium sulfate heptahydrate 0.1-0.5%, defoamer 0.01-0.05%, regulate pH value 6.5-7.5." % " in above-mentioned substratum composition all represents g/100ml.
Described fermentation culture terminal is: bacterium ball becomes broken, filtrate is muddy; Reducing sugar is down to minimum and slightly fluctuation, below reducing sugar control 0.3% (g/100g); Microscopy mycelia is in small, broken bits, without living contaminants.
The method that Phaeopoms obliquus exocellular polysaccharide extracts, the method comprises the steps:
1) separate: the Phaeopoms obliquus fermentation culture of claim 1 gained, through separating, is obtained to Phaeopoms obliquus supernatant liquor;
2) concentrated: by step 1) filtrate of gained adopts secondary cryoconcentration, obtains the thickened pulp that concentrated proportion is 1.25～1.40;
3) alcohol precipitation, oven dry: 95% alcohol that adds thickened pulp weight 3-5 doubly to measure, with the Ebullioscope detection about 65-80% of mixed solution alcoholic strength (v/v), leave standstill 8-16h, throw out is carried out to vacuum drying, temperature 50-75 DEG C, vacuum tightness is not less than 0.085MPa; Drying to being dried thing moisture content is 3～8% (g/100g), obtains the outer sugar of born of the same parents.
Wherein, described step 1) described in separation adopt horizontal screw centrifuge to carry out, rotating speed be controlled at 2500 ?3500r/min.
Described step 2) in the secondary simmer down to that adopts: one-level adopt 1 ?3t double-effect evaporator concentrated, by fermented liquid be concentrated to proportion be 1.15 ?1.20 (50 DEG C heat survey), secondary adopt 1 ?3t haplo-effect concentrator concentrated, by proportion be 1.1 ?the concentrated solution of 1.20 (50 DEG C of heat are surveyed) continue to be concentrated to the thickened pulp that proportion is 1.25～1.40 (50 DEG C of heat are surveyed); Concentration process all requires to control temperature not higher than 75 DEG C, and vacuum tightness is not less than 0.085MPa.
In the present invention alcohol precipitation be adopt 1 ?3t Alcohol-settling tank, alcohol precipitation number of times is more, the content of the outer sugar of born of the same parents is higher.The vacuum drying oven that vacuum drying adopts, the outer sugared content of born of the same parents that the present invention extracts 20 ?50%.
The present invention has advantages of following outstanding:
The present invention is directed to above-mentioned the deficiencies in the prior art studies, continue to optimize fermention medium (especially carbon source), promote on the basis of biomass and exocellular polysaccharide at lab scale and pilot scale, solve by lab scale and be amplified to the difficult problems such as the Fermentation Process of Parameter control that runs in suitability for industrialized production, extraction equipment type selecting, optimization, final successful grafting to 10 ?20t fermentor tank carry out industrialization production, and produce smoothly the Fuscoporia obliqua polysaccharide of high-quality, really realize continuously, the production of automatization, industrialization.
In order to improve the output of object product exocellular polysaccharide, the present invention is optimized and combines in the carbon source of fermention medium: the compound prescription that adopts fugitive glucose and long-acting starch.Earlier fermentation is with growth mycelium, and it is main improving biological content, therefore in substratum, has added the glucose carbon source that thalline is preferentially selected, rapidly propagation; The fermentation later stage will be improved the content of object product, and therefore carbon source provides thalline relatively to utilize slower starch, and in substratum, adds appropriate amount of starch enzyme, controls thalline quantity in suitable scale, the outer sugar of the raw metabolite born of the same parents of not stopping pregnancy.
For born of the same parents are provided the output of outer sugar, the present invention optimizes the formula of fermention medium, has adopted compound prescription for carbon source part, has both ensured the biomass of thalline in early stage, ensures again the output of later stage meta-bolites.
Adopt the present invention can, according to the market requirement and product standard, make the polysaccharide of the high-quality of different content, polysaccharide content can 20 ?50%.
Embodiment
Further illustrate the present invention by specific embodiment below.But the detail of embodiment only, for explaining the present invention, should not be construed as limited overall technical solution.
The Phaeopoms obliquus that following examples are used is bought in China Forest microbial strains preservation administrative center, and bacterial classification coding cfcc60260, taking this strain bacterium as example, describes concrete scheme of the present invention in detail.But the inventive method is common to all Phaeopoms obliquus, be not limited in this bacterial strain.
Embodiment 1
Industrialization deep layer liquid state fermentation of the present invention is produced the method for Phaeopoms obliquus exocellular polysaccharide and is carried out in the steps below:
1, the preparation of Phaeopoms obliquus fermented liquid: the Phaeopoms obliquus shaking flask bacterial strain having activated is inoculated in to seed culture medium (300L) by inoculum size 1%, wherein seed culture medium consists of glucose 3%, dried silkworm chrysalis meal 0.3%, soybean cake powder 1.8%, potassium primary phosphate 0.3%, magnesium sulfate heptahydrate 0.15%, defoamer 0.02%, regulate pH value 7.0, ventilating ratio is 1:0.3vvm, tank pressure 0.03Mpa, 26～28 DEG C of shaking culture obtain Phaeopoms obliquus primary seed solution for 5 days; Primary seed solution is accessed to secondary seed medium (3000L) by inoculum size 15%, wherein secondary seed medium consists of glucose 3%, dried silkworm chrysalis meal 0.3%, soybean cake powder 1.8%, potassium primary phosphate 0.3%, magnesium sulfate heptahydrate 0.15%, defoamer 0.02%, regulate pH value 7.0, ventilating ratio is 1:0.3vvm, tank pressure 0.03Mpa, cultivates for 26～28 DEG C and within 3 days, obtains Phaeopoms obliquus secondary seed solution; Secondary seed solution is accessed to fermention medium (7500L) by inoculum size 18%, wherein fermention medium consists of glucose 2%, starch 2%, dried silkworm chrysalis meal 0.5%, peptone 0.3%, potassium primary phosphate 0.2%, magnesium sulfate heptahydrate 0.2%, defoamer soya-bean oil 0.03%, Nai Gaowen α ?amylase (Zaozhuang biological company limited of outstanding promise enzyme, 20000u/ml) 0.002% (v/v), regulates pH value 7.0; Ventilating ratio is 1:0.5vvm, tank pressure 0.03Mpa, 26～28 DEG C cultivate 3 days, ferment to bacterium ball become broken, filtrate is muddy; Reducing sugar is down to minimum 0.2% also slightly fluctuation; Microscopy mycelia is in small, broken bits, without living contaminants.Obtain Phaeopoms obliquus fermented liquid 8850Kg.
2, Phaeopoms obliquus separation of fermentative broth: above-mentioned Phaeopoms obliquus fermented liquid 8850Kg is separated by horizontal screw centrifuge, rotating speed is controlled at 3000r/min, disengaging time is at 2h, obtain the wet mycelium (water content is 80%) of Phaeopoms obliquus filtrate 8150Kg and 575Kg, wet mycelium directly cryodrying pulverizing makes fermentation Phaeopoms obliquus bacterium powder.
4, cryoconcentration: by the Phaeopoms obliquus filtrate 8150Kg vacuum transfer of step 2 gained to 3t double-effect evaporator, be 0.085MPa in vacuum tightness, thickening temperature is under 65 DEG C of conditions, to be concentrated into proportion (the 50 DEG C of heat are surveyed) concentrated solution that is 1.20, again by concentrated solution vacuum transfer to 1.5t single-action ball-type thickener, be 0.09MPa in vacuum tightness, thickening temperature is under 60 DEG C of conditions, to continue to be concentrated into the thickened pulp 310kg that proportion is 1.30 (50 DEG C of heat are surveyed).
5, alcohol precipitation, oven dry: to filling 95% alcohol that adds weight 1200kg in the Alcohol-settling tank of 310kg thickened pulp, with Ebullioscope detection mixed solution alcoholic strength 73% (v/v), leave standstill 12h, the separation of bottom settlings thing is dried with vacuum drying oven, temperature 60 C, vacuum tightness 0.09MPa; Dry 12h and obtain the outer sugared 85kg of born of the same parents.Detecting moisture content is 4%, and polysaccharide content is 30%.
Embodiment 2
1, the preparation of Phaeopoms obliquus fermented liquid: the Phaeopoms obliquus shaking flask bacterial strain having activated is inoculated in to seed culture medium (300L) by inoculum size 1.5%, wherein seed culture medium consists of glucose 3%, dried silkworm chrysalis meal 0.3%, soybean cake powder 1.8%, potassium primary phosphate 0.3%, magnesium sulfate heptahydrate 0.15%, defoamer 0.02%, regulate pH value 7.0, ventilating ratio is 1:0.3vvm, tank pressure 0.03Mpa, 25～27 DEG C of shaking culture obtain Phaeopoms obliquus primary seed solution for 6 days; Primary seed solution is accessed to secondary seed medium (3000L) by inoculum size 15%, wherein secondary seed medium consists of glucose 3%, dried silkworm chrysalis meal 0.3%, soybean cake powder 1.8%, potassium primary phosphate 0.3%, magnesium sulfate heptahydrate 0.15%, defoamer 0.02%, regulate pH value 7.0, ventilating ratio is 1:0.3vvm, tank pressure 0.03Mpa, cultivates for 25～27 DEG C and within 3 days, obtains Phaeopoms obliquus secondary seed solution; Secondary seed solution is accessed to fermention medium (15000L) by inoculum size 18%, wherein fermention medium consists of glucose 2%, starch 2%, dried silkworm chrysalis meal 0.5%, peptone 0.3%, potassium primary phosphate 0.2%, magnesium sulfate heptahydrate 0.2%, defoamer rapeseed oil 0.03%, Nai Gaowen α ?amylase (Zaozhuang biological company limited of outstanding promise enzyme, 20000u/ml) 0.002% (v/v), regulates pH value 7.0; Ventilating ratio is 1:0.5vvm, tank pressure 0.04Mpa, 25～27 DEG C cultivate 7 days, ferment to bacterium ball become broken, filtrate is muddy; Reducing sugar is down to minimum 0.2% also slightly fluctuation; Microscopy mycelia is in small, broken bits, without living contaminants, has both obtained Phaeopoms obliquus fermented liquid 17700Kg.
2, the separation of Phaeopoms obliquus fermented liquid: above-mentioned Phaeopoms obliquus fermented liquid 17700Kg is separated by horizontal screw centrifuge, obtain the wet mycelium (water content is 75%) of Phaeopoms obliquus filtrate 16550Kg and 950Kg, wet mycelium directly cryodrying pulverizing makes fermentation Phaeopoms obliquus bacterium powder.Concrete technology condition is: rotating speed is controlled at 3200r/min, and disengaging time is at 4.5h.
4, cryoconcentration: by the Phaeopoms obliquus filtrate 16750Kg vacuum transfer of step 2 gained to 3t double-effect evaporator, vacuum tightness Wei ?0.09MPa, thickening temperature is under 63 DEG C of conditions, to be concentrated into proportion (the 50 DEG C of heat are surveyed) concentrated solution that is 1.18, again by concentrated solution vacuum transfer to 1.5t single-action ball-type thickener, be 0.09MPa in vacuum tightness, thickening temperature is under 65 DEG C of conditions, to be concentrated into the thickened pulp 580kg that proportion is 1.35 (50 DEG C of heat are surveyed).
5, alcohol precipitation, oven dry: to filling 95% alcohol that adds weight 2200kg in the Alcohol-settling tank of 580kg thickened pulp, with Ebullioscope detection mixed solution alcoholic strength 70% (v/v), leave standstill 10h, the separation of bottom settlings thing is dried with vacuum drying oven, 65 DEG C of temperature, vacuum tightness 0.09MPa; Dry 18h and obtain the outer sugared 175kg of born of the same parents.Detecting moisture content is 4.5%, and polysaccharide content is 32%.
Embodiment 3
For the compound prescription of fugitive glucose and long-acting starch in better checking the present invention is produced Fuscoporia obliqua polysaccharide superiority in industrialization, several simultaneous tests are done again.In test, only adjusted carbon source glucose in liquid fermentation medium or (with) consumption of starch, in comparative example 1, do not contain starch, glucose concn is 1.5%; In comparative example 2, do not contain glucose, starch concentration is 2.5%, and in comparative example 3, glucose content is 2.5%, and starch content is 0.5%; All the other conditions are all with embodiment 1, and test-results is in table 1:
Table 1
Claims (7)
1. prepare the method for Phaeopoms obliquus fermented liquid for one kind, it is characterized in that the fermention medium after Phaeopoms obliquus seed culture fluid access sterilizing, inoculum size is 10～20% (v/v), ventilating ratio be 1:0.3 ?1.0vvm, tank pressure 0.02～0.05Mpa, cultivate 3～4 days at 22～30 DEG C of bottom fermentations, obtain Phaeopoms obliquus fermented liquid; Wherein said fermentative medium formula is: the high temperature resistant α-amylase 0.001-0.007% (v/v) of glucose 1-3%, starch 1-3%, dried silkworm chrysalis meal 0.1-0.5%, peptone 0.1-0.5%, potassium primary phosphate 0.1-0.5%, magnesium sulfate heptahydrate 0.1-0.5%, defoamer 0.01-0.05%, 20000 μ/ml, regulates pH value 6.5-7.5.
2. the method for preparing Phaeopoms obliquus fermented liquid according to claim 1, is characterized in that described Phaeopoms obliquus seed culture fluid prepares by the following method:
1) first order seed is cultivated: by the liquid seed culture medium after activated shaking flask bacterial classification access sterilizing, inoculum size is 0.1～2% (v/v), ventilating ratio be 1:0.3 ?1.0vvm, tank pressure 0.02～0.05Mpa, at 22～30 DEG C, cultivate 5～6 days, obtain Phaeopoms obliquus first order seed nutrient solution;
2) secondary seed is cultivated: by step 1) liquid seed culture medium after the first order seed nutrient solution of gained access sterilizing, inoculum size is 10～20% (v/v), ventilating ratio be 1:0.3 ?1.0vvm, tank pressure 0.02～0.05Mpa, at 22～30 DEG C, cultivate 3～4 days, obtain described Phaeopoms obliquus seed culture fluid.
3. one according to claim 2 is prepared Phaeopoms obliquus fermented liquid method, it is characterized in that described liquid seed culture medium formula is: glucose 2-4%, dried silkworm chrysalis meal 0.1-0.5%, soybean cake powder 1-3%, potassium primary phosphate 0.1-0.5%, magnesium sulfate heptahydrate 0.1-0.5%, defoamer 0.01-0.05%, regulate pH value 6.5-7.5.
4. one according to claim 1 is prepared Phaeopoms obliquus fermented liquid method, it is characterized in that described fermentation culture terminal is: bacterium ball becomes broken, filtrate is muddy; Reducing sugar is down to minimum and slightly fluctuation, and reducing sugar control is below 0.3%; Microscopy mycelia is in small, broken bits, without living contaminants.
5. the method that Phaeopoms obliquus exocellular polysaccharide extracts, is characterized in that the method comprises the steps:
1) separate: the Phaeopoms obliquus fermentation culture of claim 1 gained, through separating, is obtained to Phaeopoms obliquus supernatant liquor;
2) concentrated: by step 1) filtrate of gained adopts secondary cryoconcentration, obtains the thickened pulp that concentrated proportion is 1.25～1.40;
3) alcohol precipitation, oven dry: 95% alcohol that adds thickened pulp weight 3-5 doubly to measure, with the Ebullioscope detection about 65-80% of mixed solution alcoholic strength (v/v), leave standstill 8-16h, throw out is carried out to vacuum drying, temperature 50 ?75 DEG C, vacuum tightness is not less than 0.085MPa; Drying to being dried thing moisture content is 3～8% (g/100g), obtains the outer sugar of born of the same parents.
6. the method for the outer sugar of Phaeopoms obliquus born of the same parents according to claim 5, is characterized in that described step 1) described in separation adopt horizontal screw centrifuge to carry out, rotating speed be controlled at 2500 ?3500r/min.
7. the method for the outer sugar of Phaeopoms obliquus born of the same parents according to claim 1, it is characterized in that described step 2) in the secondary simmer down to that adopts: one-level adopts double-effect evaporator concentrated, by fermented liquid be concentrated to proportion be 1.15 ?1.20, secondary adopts haplo-effect concentrator concentrated, by proportion be 1.1 ?1.20 concentrated solution to continue to be concentrated to proportion be 1.25～1.40 thickened pulp; Concentration process all requires to control temperature not higher than 75 DEG C, and vacuum tightness is not less than 0.085MPa.
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| CN109965268A (en) * | 2019-04-12 | 2019-07-05 | 天津农学院 | A kind of edible fungus jelly and preparation method thereof |
| CN110527702A (en) * | 2019-07-05 | 2019-12-03 | 河南农业大学 | Utilize the method for mutagenic obtained Inonotus obliquus bacterial strain production Fuscoporia obliqua polysaccharide |
| CN114456943A (en) * | 2020-11-09 | 2022-05-10 | 浙江养生堂天然药物研究所有限公司 | Inonotus obliquus and extract and application thereof |
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| CN105535035A (en) * | 2015-12-31 | 2016-05-04 | 浙江省林业科学研究院 | Inonotus obliquus fermentation culture composition and preparation method thereof |
| CN105535035B (en) * | 2015-12-31 | 2019-05-28 | 浙江省林业科学研究院 | A kind of Inonotus obliquus fermented and cultured composition and preparation method thereof |
| CN105670944A (en) * | 2016-03-15 | 2016-06-15 | 江苏神华药业有限公司 | Method for producing inonotus obliquus powder through rapid and deep liquid state fermentation |
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| CN106978465B (en) * | 2017-04-06 | 2021-05-11 | 杭州娃哈哈科技有限公司 | Fermentation method for improving fermentation yield of total triterpenoids in inonotus obliquus |
| CN109965268A (en) * | 2019-04-12 | 2019-07-05 | 天津农学院 | A kind of edible fungus jelly and preparation method thereof |
| CN110527702A (en) * | 2019-07-05 | 2019-12-03 | 河南农业大学 | Utilize the method for mutagenic obtained Inonotus obliquus bacterial strain production Fuscoporia obliqua polysaccharide |
| CN110527702B (en) * | 2019-07-05 | 2022-12-06 | 河南农业大学 | Method for producing inonotus obliquus polysaccharide by using inonotus obliquus strain obtained by mutagenesis |
| CN114456943A (en) * | 2020-11-09 | 2022-05-10 | 浙江养生堂天然药物研究所有限公司 | Inonotus obliquus and extract and application thereof |
| CN114456943B (en) * | 2020-11-09 | 2023-12-22 | 浙江养生堂天然药物研究所有限公司 | Inonotus obliquus and extract and application thereof |
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