WO2012122931A1 - Souche de paecilomyces tenuipes et ses applications - Google Patents

Souche de paecilomyces tenuipes et ses applications Download PDF

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WO2012122931A1
WO2012122931A1 PCT/CN2012/072261 CN2012072261W WO2012122931A1 WO 2012122931 A1 WO2012122931 A1 WO 2012122931A1 CN 2012072261 W CN2012072261 W CN 2012072261W WO 2012122931 A1 WO2012122931 A1 WO 2012122931A1
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paecilomyces
cgmcc
strain
acid
tenuipes
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Chinese (zh)
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陈祝安
谭悠久
李成
魏宁
孙长胜
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SHANGHAI BIOASIA LIFE TECHNOLOGY Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/06—Fungi, e.g. yeasts
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/20—Hypnotics; Sedatives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A61P39/06—Free radical scavengers or antioxidants
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/145—Fungi isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/645—Fungi ; Processes using fungi
    • C12R2001/79—Paecilomyces

Definitions

  • Paecilomyces tenuipes is an asexual type of Coryceps takaomantana. The nickname is Spicaria heliothia, Paecilomyces heliothia, Isaria japonica. More than 10 species of the same name, such as Isaria dussii.
  • Paecilomyces clavularum can prevent the decrease of white blood cell count caused by cyclophosphamide, and has a repressive effect on the atrophy of immune organs such as spleen and thymus of immunosuppressed mice induced by cyclophosphamide, showing immunomodulatory activity to the body (Jin Liqin, Zhejiang Medical Journal, 2001, 23(4): 216-217
  • the liquid culture medium of Paecilomyces sinensis and its glycoprotein concentrate can improve the anemia caused by 5-fluorouracil in mice (Takata T, Biol. Pharm. Bull 2008, 31(8) 1565-1573).
  • the object of the present invention is to provide a strain of Paecilomyces sinensis which has application value in the fields of food, health care products, medicines and cosmetics.
  • the strain of the invention has the characteristics of easy cultivation, high yield and stable culture trait, and the culture product has multiple Efficacy, therefore has a wide range of application value.
  • the invention has been subjected to a large number of experimental studies to isolate and purify a strain of Paecilomyces tenuipes (Peck) Samson from the collected lepidopteran larvae susceptible body.
  • the strain was deposited on September 20, 2010 at the General Microbiology Center of the China Microbial Culture Collection Management Committee, with the accession number CGMCC No. 4176.
  • the colony On the potato sucrose agar medium, the colony was white, dense, and the initial back was light yellow. On the later stage, a large number of spore bundles were formed on the colony, and the back surface was light yellowish brown. Hyphae colorless, branched, smooth, with septum, 1.5-3.0 m wide. The conidiophores protrude from the mycelium, colorless, smooth, branched, up to llO.O m, and up to 4.0 m wide.
  • the spore stem is white to creamy yellow, 10-30 cm long, sporulated with cell-shaped, colorless, 2-6 clusters on the spore stalk, and the top is narrow and narrow, 5.0-8.0 ⁇ 2.5-3.2 m.
  • the conidia are mostly cylindrical, partially elliptical, also have sausage-shaped curvature, single spore, colorless, smooth, chain, 5.0-6.7 X 2.0-2.5 (3.0) ⁇ m.
  • sucrose sucrose, fructose, lactose, maltose, sorbitol, rhamnose, mannose, inositol, glycerol, sorbic acid, xylose, soluble starch, etc.
  • sucrose followed by glucose , maltose, fructose and soluble starch, can not use xylose and sorbic acid, rhamnose, sorbitol growth potential.
  • the optimal growth temperature of the bacteria is 24-26 ° C, and it can grow between pH 3-11, but the best growth is between pH 5.0-6.7. Fermented on solid grain medium, the lawn moss is grayish white, the fruit body (spore stem bundle) is bright yellow, and the fertility is very good. The harvest period is 30-40 days. The mycelium is obtained by culturing in a liquid medium containing sucrose, ammonium salt, dipotassium hydrogen phosphate or magnesium sulfate for 10-14 days.
  • the culture product of this bacterium showed that the chemical components contained aspartic acid, threonine, serine, glutamic acid, glycine, alanine, methionine, phenylalanine, tyrosine, etc.; trace elements containing potassium , calcium, magnesium, iron, zinc, copper, manganese, nickel, selenium, etc.; in addition, polysaccharides, adenosine, mannitol, alcohols, organic acids and alkaloids are substantially identical to or more than Cordyceps sinensis active ingredients.
  • the ITS sequence of Paecilomyces c recipes CGMCC No. 4176 of the present invention contains the polynucleoside as shown in SEQ ID NO: 1. Acid sequence.
  • Tenuipes CGMCC No. 4176 is widely used in the preparation of foods, health products, pharmaceuticals and cosmetics.
  • Paecilomyces sphaeroides The fermentation product of tenuipes CGMCC No. 4176 can be used as a main medicinal ingredient or a raw material for the preparation of foods, health products, medicines and cosmetics.
  • the addition and treatment of other raw materials in the preparation of foods, health products, medicines and cosmetics can be referred to conventional.
  • the fermentation product of the pseudomycin CGMCC No. 4176 of the present invention can be used for preparing a drug for immunomodulation, nutrition improvement, liver protection, anti-aging, anti-tumor, anti-virus, anti-hypoxia, sedation or analgesic function.
  • the method and dosage of the human body can be referred to the Cordyceps sinensis.
  • the specific dose should also consider the route of administration, the health status of the patient, etc., which are all within the skill of the skilled physician.
  • Rats were administrated with Penicillium chrysogenum CGMCC No. 4176 fruiting body, which significantly increased the activities of lactate dehydrogenase, acid phosphatase and arginase in rat peritoneal macrophages, and enhanced macrophage pairs. Neutral red ingestion ability also significantly increased hemoglobin levels in the blood of rats. Rats were intragastrically administered with Penicillium chrysogenum CGMCC No. 4176 fruit body, which significantly reduced the carbon tetrachloride-induced liver injury and the lipid peroxide levels in serum, brain, liver, spleen and kidney tissues of normal rats. Decreased, elevated levels of reduced glutathione.
  • the fermentation product of Penicillium sp. CGMCC No. 4176 has the characteristics of easy cultivation, high yield and stable culture characteristics.
  • the culture products have immune regulation, improve nutrition, protect liver, anti-aging, anti-tumor, anti-virus, and improve hypoxia tolerance.
  • Example 1 Plate culture of Mycobacterium tuberculosis strain [i3 ⁇ 4edtomyc tenuipes (Peck) Samson]: Pseudomonas sinensis CGMCC No. 4176 was placed on potato sucrose agar medium, cultured at 25 ° C for 14 days, colony diameter 30-60mm. Mycelium is villous, flocculent, grayish white, fluffy. The back is light yellow, and a large number of egg stalks are formed in the late stage of the culture. Spore-forming cells (bottle stems) consist of a group of spurs that are clustered on the short branches of the conidiophores.
  • the base of the bottle stem is spherically enlarged and narrowed upwards, 5-7.8 ( 3 ) X 2.3-3.5 m.
  • the conidia are mostly cylindrical, also have a sausage-like curvature, monospores, smooth walls, and chain, 5.0-6.7 X 2.0-2.5 (3) ⁇ m.
  • the mycelium is villous, dense, pale yellow, and the spore bundle is dense and developed.
  • Chad's medium the colonies were grayish white, arched, fluffy, and villous, and the spore stems grew poorly or not long. No colony growth was observed on Sa's medium.
  • Example 2 Example 2.
  • Paecilomyces ciliata strain Inoculation of Paecilomyces sphaeroides strain CGMCC No. 4176 into glucose, sucrose, fructose, lactose, maltose, sorbitol, rhamnose, mannose
  • the carbon source medium such as inositol, glycerol, sorbic acid, xylose, soluble starch, etc.
  • sucrose, followed by glucose, maltose, fructose and soluble starch can not use xylose and sorbic acid, rhamnose , sorbitol growth potential.
  • nitrogen sources such as potassium nitrate, ammonium chloride, ammonium sulfate, diammonium hydrogen phosphate, sodium nitrite, citric acid ammonia, thiourea, etc., except for the growth of thiourea and sodium nitrite, with citric acid ammonia and phosphoric acid Hydrogen diammonium is the best growth, and it is better to use amino nitrogen than nitro nitrogen.
  • Example 3 Culture of fermented product of Paecilomyces militaris strain [i3 ⁇ 4edtomyc tenuipes (Peck) Samson]: Cultured Paecilomyces c recipes CGMCC No. 4176 in cereal medium (wheat medium) at 23 ⁇ 26 °C , the lawn is luxuriant. Gray-white hairy mycelia began to appear and was covered with the culture medium. After that, the lawn is gradually thickened, and the bacteria are formed, and many needle-like protrusions are seen, and then golden or egg-yellow fruit bodies are grown.
  • the shape of the spore stems varies greatly, with a cylindrical shape and no branches at the top; there are also dendritic, staghorn, coral-like branches, 29-54 X 0.5-1.8 m.
  • the fruiting bodies were collected for 25 to 30 days, and pulverized after drying to obtain a fruit body powder.
  • Analysis shows that the fine sample contains aspartic acid, threonine, serine, glutamic acid, glycine, alanine, methionine, phenylalanine, tyrosine, etc.; trace elements containing potassium, calcium, magnesium, iron, zinc , copper, manganese, nickel, selenium; in addition, polysaccharides, adenosine, mannitol, alcohols, organic acids and alkaloids are basically the same as the active ingredients of Cordyceps sinensis.
  • test solution Weigh accurately 1.0118g of the powder of Pseudomonas sinensis obtained from Example 3, and add 10mL of petroleum ether (60-90 °C) in a lOOmL conical flask. Ultrasound for 30 min. Filter, drain the petroleum ether, and put it together with the filter paper. Medium. Add 20 mL of ultrapure water to the bottle, weigh the total weight (including the weight of the Erlenmeyer flask), and sonicate for 30 min. After rapid cooling, weighed, added to the total weight with ultrapure water, mixed and centrifuged, the supernatant was taken, and the sample extract was obtained through a syringe filter.
  • Standard curve drawing Refer to the reference 1 g/mL, 5 ⁇ g/mL, 10 ⁇ g/mL, 30 ⁇ g/mL, 50 ⁇ g/mL, 100 ⁇ g/mL solution, press 1.2.
  • the chromatographic conditions of 1 are determined.
  • the sample concentration (g/mL) is plotted on the abscissa, and the peak area (microvolt ⁇ second) is plotted on the ordinate.
  • the standard curve is drawn and the regression equation is obtained.
  • Nash reagent 150g ammonium acetate + 2mL glacial acetic acid + 2mL acetylacetone, diluted to 1L with distilled water (currently used).
  • L-rhamnose solution L-rhamnose lOOmg, make up to 100mL with distilled water.
  • test solution Accurately weigh 1.0034 g of the dried fruit body obtained from Example 3, placed in a dry 500 mL Erlenmeyer flask, record the total mass of the conical flask and the sample, and add to the conical flask. Boiling distilled water 100 mL, placed on an electric furnace for boiling for 15 min, then quickly placed in cold water to cool to room temperature, remove, dry the water droplets on the outer wall of the bottle, add distilled water to the final mass of ml + 100g, shake, filter, Take the filtrate, which is the sample to be tested, and store at -20 °C.
  • 5% phenol solution Accurately weigh 5g of phenol and dilute to 100ml with water.
  • Preparation of glucose standard solution Prepare a glucose standard solution with a concentration of 0.25 mg/mL, accurately weigh 0.2500 g of dried glucose, and dilute to 1000 mL with water.
  • test solution Accurately weigh 1.0034 g of the fruit body powder obtained from Example 3 and place it in a dry 500 mL Erlenmeyer flask. Record the total mass of the conical flask and the sample, and add it to the Erlenmeyer flask. Boiling distilled water 100 mL, placed on an electric furnace for boiling for 15 min, then quickly placed in cold water to cool to room temperature, remove, dry the water droplets on the outer wall of the bottle, add distilled water to the final mass of ml + 100g, shake, filter, Take the filtrate, which is the sample to be tested, and store at -20 °C.
  • 3.3.2 Standard curve drawing accurately weigh 0.2500g of anhydrous glucose, prepare 0.25mg/mL glucose standard solution with distilled water, and absorb Portuguese Glucose standard solution 0, 0.1, 0.3, 0.5, 0.7, 0.9, l.lmL, respectively, placed in a test tube, each with distilled water, the volume is 2.0mL, plus 5% phenol solution lmL, shake well, quickly 5 mL of 98% sulfuric acid was added dropwise, and the mixture was shaken for 5 minutes, and heated in a boiling water bath for 15 minutes.
  • the cold water was taken out and cooled to room temperature; 2 mL of distilled water was added, and the same operation was used as a blank control, and the absorbance (Abs) was measured at 490 nm. Taking the standard solution concentration as the abscissa and the absorbance as the ordinate, the standard curve is drawn and the regression equation is obtained.
  • the glucose standard curve uses the standard glucose concentration (mg/mL) as the abscissa and Abs as the ordinate.
  • Example 5 Penicillium sinensis artificial culture against carbon tetrachloride-induced acute liver injury test Weighing 10 g of Penicillium sinensis artificial culture sample (the fruit body powder obtained in Example 3), adding distilled water to 40 mL , mix well and use as a test sample.
  • Experimental animals Adult rats, single sex, 10 in each group, a total of 3 groups. One dose group was 100 mg/kg body weight and one blank control group and one model control group.
  • a model of liver injury was induced by carbon tetrachloride (analytical purity), which can be administered intragastrically or intraperitoneally.
  • the concentration of carbon tetrachloride in rats was 2%, and the amount of gastric perfusion was 5mIJkg.
  • the Paecilomyces militaris sample was given for 30 days.
  • the left lobe of the rat liver was fixed with 10% formalin, and the transverse section was taken from the middle left lobe of the liver.
  • the conventional pathological preparation (paraffin-embedded, HE staining) was performed. The pathological changes of the cells were recorded from the end of the liver, and the entire tissue section was continuously observed with a 40-fold objective lens.
  • Example 6 Penicillium sinensis artificial culture was used to resist hypoxia, sedation and analgesia in mice.
  • the fruiting body obtained in Example 3 was 50g, heated water 6 times and 4 times, and fried twice. cook. Each time the mixture was immersed for 10 minutes, it was boiled for 30 minutes and filtered. Then, the filtrate was combined and concentrated to obtain 150 ml, which was used as a test sample for refrigeration.
  • Experimental animals Mice, male and female, 10 in each group, a total of 11 groups. The mice were intraperitoneally injected with normal hypoxia tolerance, sedative and analgesic treatment components of 2.5 g/kg body weight and 5 g/kg body weight, and were treated with normal saline.
  • Atmospheric hypoxia tolerance test At 30 minutes after administration, the mice were placed in a 250 ml jar with a lid, and each mouse was used to record the survival time of each group of mice. Sedation test: 30 minutes after intraperitoneal injection of decoction, 30 mg/kg of pentobarbital sodium was injected, and the number of sleep heads of each group was observed within 15 minutes.
  • Analgesic test 30 minutes after the administration, each mouse was injected with 0.7% acetic acid 0.1 ml/10 g in the abdomen, and the number of mice writhing was recorded within 5 minutes of acetic acid, and morphine and physiological saline were used as a control at a dose of 10 mg/kg. Test results: Table 5 Effect of Paecilomyces ciliata on hypoxia tolerance in mice
  • Example 7 ITS sequence of the Penicillium spp. of the present invention (intermediate non-transcribed sequence)
  • the mycelium of Paecilomyces ciliata CGMCC No. 4176 obtained in Example 3 was ground with quartz sand, and the total DNA of the strain was extracted by the guanidinium isothiocyanate method.
  • the ITS region of Paecilomyces variabilis was amplified using total DNA as a template.
  • the target used was Foreword primer: GGAAGTAAAAGTCGTAACAAGG (SEQ ID NO: 2) and Reverse primer: TCCTCCGCTTATTGATATGC (SEQ ID NO: 3).
  • the PCR reaction system (25 L) was: 2.5 L10 X Taq enzyme buffer (TAKARA Ex Taq Buffer), dNTP 200 ⁇ mol/L, MgCl 2 1.5 mmol/L, primer 0.2 ng, Taq enzyme (TAKARA Ex Taq) 1.5 U, template DNA 20ng ⁇ l ⁇ g.
  • the reaction procedure of the amplification reaction on the PCR machine first 94 ° C for 3 minutes; then 94 ° C for 30 seconds, 54 ° C for 30 seconds, 72 ° C for 40 seconds, a total of 36 cycles; then 72 ° C extension for 5 minutes Finally, cool down to 4 °C.
  • the gel electrophoresis product was recovered by agarose gel electrophoresis using a DNA gel recovery kit (AxyPrepTM, Axygen Scientific, Inc. USA), and the sequence of the obtained sequence was as follows:

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Abstract

L'invention concerne un Paecilomyces tenuipes, CGMCC No. 4176, et ses applications dans la préparation d'aliments, de produits de santé, de produits pharmaceutiques et de produits cosmétiques. Le Paecilomyces tenuipes est caractérisé par sa facilité de culture, son haut rendement et la stabilité de ses caractères de culture. Un produit cultivé à partir de Paecilomyces tenuipes est largement applicable et présente les caractéristiques d'ajustement de l'immunité, de résistance aux tumeurs et aux virus, de protection du foie, d'amélioration de la nutrition, d'amélioration de la résistance à l'hypoxie, de soulagement de la douleur et est anti-vieillissement et tranquillisant.
PCT/CN2012/072261 2011-03-14 2012-03-13 Souche de paecilomyces tenuipes et ses applications Ceased WO2012122931A1 (fr)

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CN120737979A (zh) * 2025-08-04 2025-10-03 青岛海科生物技术有限公司 一种基于复合培养基的蝉拟青霉高效发酵工艺

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WO2016047638A1 (fr) * 2014-09-24 2016-03-31 国立大学法人岩手大学 Dérivé de peptide cyclique, procédé pour sa préparation et composition de celui-ci
CN106282033A (zh) * 2016-08-15 2017-01-04 郑毅男 一株新青霉菌及其代谢产物安他拟酸a
CN109266557A (zh) * 2018-10-17 2019-01-25 上海市农业科学院 一类具有保健作用的细脚棒束孢虫草复合种菌种的获得方法
CN111944698A (zh) * 2020-07-24 2020-11-17 江苏大学 一种麦角甾醇过氧化物的生物发酵制备方法

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EP1297746A1 (fr) * 2001-09-26 2003-04-02 Sumitomo Chemical Company, Limited Champignon entomopathogène destiné à être utilisé comme insecticide
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Publication number Priority date Publication date Assignee Title
EP1297746A1 (fr) * 2001-09-26 2003-04-02 Sumitomo Chemical Company, Limited Champignon entomopathogène destiné à être utilisé comme insecticide
CN101195805A (zh) * 2007-11-01 2008-06-11 华富生物科技(上海)有限公司 一种虫草真菌及其人工栽培方法

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WEN TING-CHI ET AL.: "Advances of Researches on the Anamorph of Cordyceps takaomontana-Paecilomyces tenuipes", JOURNAL OF FUNGAL RESEARCH, vol. 2, no. 3, 31 December 2004 (2004-12-31), pages 58 - 62 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN120737979A (zh) * 2025-08-04 2025-10-03 青岛海科生物技术有限公司 一种基于复合培养基的蝉拟青霉高效发酵工艺

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