CN1003454B - Culture process and utilization of puckery fungus - Google Patents
Culture process and utilization of puckery fungus Download PDFInfo
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- CN1003454B CN1003454B CN85103492.6A CN85103492A CN1003454B CN 1003454 B CN1003454 B CN 1003454B CN 85103492 A CN85103492 A CN 85103492A CN 1003454 B CN1003454 B CN 1003454B
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Abstract
The present invention belongs to the technical field of the culture and the application of basidiomycetes and aims to increase the yield of schizophyllum commune mycelia and schizophan. The present invention has the technical scheme that after Nan Da 843 schizophyllum commune strain on which tissue is isolated from rotten wood is purified, natural bean flour, glucose or sucrose, yeast extract, magnesium sulphate, potassium dihydrogen phosphate and thiamine hydrochloride are used as a culture medium for fermentation and culture, and schizophyllum commune fermentation liquor containing high mycelia and polysaccharide can be obtained; in addition, the anti-tumour, anti-radiogenic leukocytosis, antibiotic synergistic agent and anti-inflammatory schizophan and amino acid used as intensifying agents of condiments and food are extracted from the fermentation liquor.
Description
The invention belongs to basidiomycetous cultivation and utilize technical field.
A kind of Split-gill Latin formal name used at school Schizophyllum Commune Fr in the club fungi of the present invention.Nanjing University's 843 bacterial strains on May 16th, 1985 were preserved in Beijing China Committee for Culture Collection of Microorganisms common micro-organisms center, preserving number CGMCC NO.0012.
In prior art, Institute of Microorganism, Academia Sinica is used as the Split-gill bacterial strain of purebred preservation, and its female kind developing medium is potato, glucose, agar or wort agar, and 24 °-26 ℃ of optimum temperutures are not applied to industrial production as yet.External Japan " The Japanese Journal Antiobics " Vol.23(2): report in the 157-160 page or leaf.Split-gill is cultivated and is adopted synthetic developing medium, promptly with known organism of Chemical Composition or inorganics preparation developing medium, and its cost height, mycelia yield low (cultivate 5 days 1 liter of fermented liquids and contain the 1-5g mycelium), polysaccharide yield is also low.(cultivate 5 days 1 liter of fermented liquids and contain the 0.2-1g polysaccharide).
The objective of the invention is to do developing medium and cultivate Split-gill with analysis for soybean powder etc., improve the yield of mycelium and schizophan, and antitumor, anti-the putting of extraction rises bletilla adjustment body's immunity, the schizophan of producing the antibiotic synergistic agent and the amino acid of needed by human from Split-gill.
To be Split-gill from rotten wood go up through separate tissue technical scheme of the present invention obtains pure bacterial strain Nanjing University 843, it is seeded on PDA or the wort solid slant culture base, and 24 ℃ of temperature were grown 7-10 days, the tube purifying is put into 0 °-2 ℃ refrigerator and is preserved standby then.Liquid fermentation and culture is with preparation such as natural analysis for soybean powder developing medium, and its composition and proportioning are that the 1000ml developing medium contains natural analysis for soybean powder 5-10g, glucose or multitudinous sugared 30-40g, yeast extract paste 2g, sal epsom 0.5g, potassium primary phosphate 1g.The also available vitamin of developing medium is done additive, and every 1000ml developing medium can increase 0.01-10mg.250ml shakes bottled 75-150ml developing medium, and 500ml shakes bottled 150-250ml developing medium, adopts reciprocating type shaking table, hunting speed 80-130 time/minute, or rotary shaking table, hunting speed 100-200 rev/min, 24 ℃-26 ℃ of culture temperature.When the mycelia raised growth, bacterium liquid is become clearly gradually by muddiness from depth to shallow.Prolongation along with incubation time, reducing sugar, ammonia-state nitrogen, pH reduce gradually in the fermented liquid, mycelium weight constantly increases, the highest to the 6th day polysaccharide content, reducing sugar drops to below 1%, and ammonia-state nitrogen drops to 15%, pH drops to 4.6-4.8, the volume of bacterium ball accounts for fermented liquid cumulative volume 70%, and can stop fermentation this moment, and fermentation period is five days.
Solid fermentation is cultivated and to be used 70-78%(weight) sugarcane slag (or wood chip, sweet potato slag, cotton seed hull), 1%(weight) multitudinous sugar or glucose, 20-28%(weight) wheat bran or rice bran, 1%(weight) gypsum or lime carbonate, poach, pH nature, the mushroom bottle of packing into is at 1.5kg/m
2Down sterilization 1-2 hour of pressure, after the cooling, the female kind in test tube slant is inoculated in the mushroom bottle under aseptic technique, the female inoculation 1-3 bottle of planting of a pipe, culture temperature 22-26 ℃, 20-30 days mycelium are covered with full bottle and can gather.
Split-gill is grown under liquid fermentation and culture technology, contains 3-5g schizophan (Schizophyllan) (dry weight) in every liter of zymocyte liquid, contains 200-300g mycelium (weight in wet base); Every 100g mycelium (weight in wet base) contains the 0.8-1g polysaccharide, and aminoacids content is as follows in fermented liquid and the mycelium:
Ammonia in the 100g mycelium in the amino acid title 100ml fermented liquid
Aminoacids content base acid content (g)
（mg/ml） 2.33
Aspartic acid 8.99 1.22
Threonine 3.94 1.29
Serine 2.62 3.82
L-glutamic acid 11.0 1.13
Glycine 2.72 1.65
L-Ala 2.07 0.238
Gelucystine 1.26 1.37
Xie Ansuan 4.59 0.396
Methionine(Met) 0.22 2.35
Isoleucine 4.02 1.98
Leucine 6.01 0.886
Tyrosine 2.40 1.01
Phenylamino acid 2.50 1.70
Methionin 2.56 0.628
Histidine 1.19 2.08
Arginase 12 .08 2.45
Proline(Pro) 2.45
The schizophan that the Split-gill of turning out from above-mentioned technology extracts, can be used for clinical antitumor, anti-put rise white, anti-inflammatory, reduction antibiotic is paid effect, produce the antibiotic synergistic agent, improve antibiotic and tire, the amino acid of extraction can be used as seasonings and infant foods reinforcer.
Split-gill of the present invention is born on deciduous tree or the rotten wood of softwood tree, and scattered or all living creatures is imbricate, the wide 6-42mm matter of cap is tough, white to canescence, last villous or coarse wool, fan-shaped or kidney shape, the edge curls inward, the most rip-panels of tool, lamella is narrow, goes out from the base portion radiation, white or canescence, lavender is along the edge lobe and warp sometimes; Spore is colourless, rod shape, 5-5.5 * 2m.Go up through Nanjing University's 843 pure bacterial strains that separate tissue obtains from rotten wood, on artificial medium-wood chip skin, cultivate, after 25 days, grow sporophore, its shape is identical with natural sporophore, soft leather matter, and there is the rug hair on the cap surface, be canescence, the edge curls inward, lamella is to cap edge lobe and warp; Spore is colourless, and is cylindrical, 5-5.5 * 2m.Nanjing University's 843 bacterial strains are compared with the Split-gill bacterial strain of Beijing Institute of Micro-biology of Chinese Academy of Sciences preservation, its homomorphosis, but developing medium, range of application difference.The developing medium of Beijing Institute of Micro-biology is potato, glucose, agar, and 26 ℃ of optimum temperutures only are used as purebred preservation and use.
The cultivation proterties of Split-gill of the present invention Nanjing University 843 bacterial strains is: 1. is seeded on potato, glucose, the nutrient agar, and 24 °-26 ℃ of temperature, it is vigorous to grow, and bacterium colony pure white, velvet-like, smooth is inoculated ten days thalline diameters and is reached 15-20mm.2. be seeded on the sawdust medium, 26 ℃ of temperature were grown 15 days, and the high 12cm of mycelia post was respectively between fruit body primordium and sporophore differentiation phase 15 days, 20 days.3. when shaking bottle shaking culture, the reciprocating type shaking table 80-130 of hunting speed rev/min, form the bacterium ball, when the bacterium ball formed in a large number, bacterium liquid became clear by muddiness from depth to shallow, and viscosity increases gradually.4. in the analysis for soybean powder developing medium, when reducing sugar drops to 1%, amino nitrogen drops to 15%pH and drops to below 5, when the bacterium sphere volume accounts for fermentating liquid volume 70%, can stop fermentation, and fermentation period is 6 days.
The physiological property of Split-gill of the present invention Nanjing University 843 bacterial strains is: 1. the analysis for soybean powder developing medium is better than other developing medium growing way, and the mycelia yield height was cultivated mycelia yield 20-30g/1000ml 6 days.2. 24 °-26 ℃ of the suitableeest culture temperature.3. optimal pH 5-6.4. carbon, nitrogen consumption ratio are 100: 1, and be bigger to the carbon source demand amount.
The polysaccharide that extracts from Split-gill of the present invention Nanjing University 843 bacterial strain fermentation liquors is a schizophan.Its foundation is that 1. the infrared absorption spectrum peak position is similar to known schizophan infrared absorption spectrum peak position.The peak position of known Japanese schizophan is 885cm
-1, its fermented liquid schizophan of this schizophan peak position is 845.104cm
-1, mycelium schizophan peak position is 848.278cm
-12. all be neutral polysaccharide.3. all be white, fibrous, toughness.4. major ingredient all is a glucose.The characteristic of known schizophan is seen " Japanese chemurgy meeting will " Vol.45NO4.1971.P162～168.
The schizophan extraction process of Split-gill of the present invention Nanjing University 843 is: schizophyllary fermentating liquid is with sintered filter funnel suction filtration (factory with board and frame machine separate fermentation liquid and mycelium) separate fermentation liquid and mycelium, 1. fermented liquid is concentrated into 95% alcohol precipitation that thick (half of original volume) adds equivalent to 3 times or 5 times, then with 95% alcohol washing for several times to the reaction of effluent liquid sugar-free, drain Crude polysaccharides.2. mycelium is added 50 times of water loggings bubble and spends the night, boil with boiling water bath and propose filtrations in 24 hours, add equivalent to 3-5 95% alcohol precipitation doubly after filtrate is concentrated, wash several to effluent liquid with 95% alcohol then and do not have reducing sugar reaction, drain Crude polysaccharides.At last Crude polysaccharides is filtered with the hot water dissolving, remove insolubles, spraying drying gets polysaccharide dry powder.
Be embodiments of the invention below:
Gather schizophyllum sporophores through separate tissue from open-air rotten wood, obtain purebred, with the purebred bottle shaking culture of shaking, the composition of developing medium and prescription are: the 1000ml developing medium contains analysis for soybean powder 5g, glucose or multitudinous sugared 30g, yeast extract paste 2g, potassium primary phosphate 1g, sal epsom 0.5g, 26 ℃ of culture temperature, rotary shakers reciprocating type with 100 times/minute or 180 rev/mins vibrate, cultivate that to occur little bacterium concave-sphere inspection mycelial growth in second day in great numbers, multi-branched, the tool tabula, colourless, transparent, the tool clamp connexion, it is clear that 6 days bacterium liquid of fermentation to the look becomes, the bacterium sphere volume accounts for fermentating liquid volume 70%, polysaccharide content is the highest, and can stop fermentation this moment.
Claims (4)
1, the culture process of a kind of Split-gill Schizophyllum Commune Fr. Nanjing University's 843 bacterial strains (preserving number CGMCC NO.0012) in the club fungi, it is characterized in that the every 1000ml developing medium of liquid fermentation and culture is with natural analysis for soybean powder 5-10g, glucose or multitudinous sugared 30-40g, yeast extract paste 2g, sal epsom 0.5g, potassium primary phosphate 1g form substratum, culture temperature is 24-26 ℃, the hunting speed of developing medium is reciprocating type shaking table 80-130 time/minute, or rotary shaking table 100-200 rev/min.
2, culture process according to claim 1 is characterized in that can increasing additive-vitamin in the developing medium.
3, culture process according to claim 2 is characterized in that can increasing the 0.01-10mg vitamin in the 1000ml developing medium.
4, a kind of Split-gill Schizophyllum Commune Fr in the club fungi.The culture process of Nanjing University's 843 bacterial strains, it is characterized in that the solid fermentation cultivation, developing medium 70-80%(weight) sugarcane slag (or wood chip, sweet potato slag, cotton seed hull), 1%(weight) multitudinous sugar or glucose, 20-28%(weight) wheat bran or rice bran, 1%(weight) gypsum or lime carbonate form substratum, poach, the pH nature is at 1.5kg/m
2Down sterilization 1-2 hour of pressure, culture temperature 22-26 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN85103492.6A CN1003454B (en) | 1985-05-22 | 1985-05-22 | Culture process and utilization of puckery fungus |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN85103492.6A CN1003454B (en) | 1985-05-22 | 1985-05-22 | Culture process and utilization of puckery fungus |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN85103492A CN85103492A (en) | 1987-04-15 |
| CN1003454B true CN1003454B (en) | 1989-03-01 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN85103492.6A Expired CN1003454B (en) | 1985-05-22 | 1985-05-22 | Culture process and utilization of puckery fungus |
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| Country | Link |
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| CN (1) | CN1003454B (en) |
Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101444231B (en) * | 2008-12-30 | 2011-06-15 | 北京市农林科学院 | Schizophyllum commune protein extract, and preparation method and application thereof |
| CN102090266B (en) * | 2010-12-05 | 2012-10-31 | 余家贵 | High-efficient cultivation method of schizophyllum commune |
| CN104673674A (en) * | 2013-11-27 | 2015-06-03 | 瑞帝安有限公司 | Method for manufacturing schizophyllum commune mycelium pellets and method for producing beta-glucan by using same |
| CN104892253A (en) * | 2015-06-19 | 2015-09-09 | 桂林健成生物科技开发有限公司 | Application of siraitia grosvenorii fruit residues in cultivation of schizophyllumcommuneh |
| CN106613350B (en) * | 2016-12-21 | 2020-06-30 | 广东省微生物研究所（广东省微生物分析检测中心） | Schizophyllum commune and domestication and cultivation method and application thereof |
| CN109247189A (en) * | 2018-09-29 | 2019-01-22 | 云南菌视界生物科技有限公司 | A kind of white ginseng bacterial strain and its white ginseng low cost cultural method |
| CN110283595A (en) * | 2019-06-25 | 2019-09-27 | 西北师范大学 | A kind of preparation method of sand-fixation method and its sand-fixing liquid and sand-fixing liquid |
| CN114941019B (en) * | 2022-06-24 | 2024-03-05 | 广东丸美生物技术股份有限公司 | Method for reutilizing microbial fermentation fungus dreg, schizophyllum commune fungus dreg extract and application thereof |
-
1985
- 1985-05-22 CN CN85103492.6A patent/CN1003454B/en not_active Expired
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| CN85103492A (en) | 1987-04-15 |
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