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Cylindrocarpon fungi strain and method for preparing dihydro ascochyta chlorine element through strain

A technology of dihydroascochlorin and Cylindrosporium, which is applied in the field of microorganisms, can solve the problems of complex metabolite components and low fermentation units of dihydroascochlorin, and achieve the effect of simple and easy preparation process

Active Publication Date: 2014-09-24
NCPC NEW DRUG RES & DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Dihydroascochlorin can be produced by Fusarium sp. ( Fusarium sp.) (Ralph Henry Evans, et al., US3546073), Congrum ( Nectria coccinea ) (Andridge, et al., J. Chem. Soc., Perkin Trans. 1, 2136-2141, 1972), Verticillium ( Verticillium sp.) (Satoshi T, et al., Chem. Pharm. Bull., 42(4):953-956, 1994), Cylindrosporium ( Cylindrocarpon sp.) (Mio K, et al., J. Antibiot, 66: 23–29, 2013) and other strains of fungal genera, but the metabolites of the above strains are complex and the content of dihydroascochlorin The fermentation units are very low, no more than 50mg / L

Method used

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  • Cylindrocarpon fungi strain and method for preparing dihydro ascochyta chlorine element through strain
  • Cylindrocarpon fungi strain and method for preparing dihydro ascochyta chlorine element through strain
  • Cylindrocarpon fungi strain and method for preparing dihydro ascochyta chlorine element through strain

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Embodiment 1

[0041] Embodiment 1, the separation and identification of Cylindrospora bacterial strain NCC3746

[0042] (1) Isolation of strain NCC3746

[0043] The medium used for the isolation of the fungal strain NCC3746 is PDA (Beijing Shuangxuan);

[0044] Take 1 gram of soil sample (collected from the original forest of Wawu Mountain, Hongya County, Meishan City, Sichuan Province, China), put it into a 50mL centrifuge tube filled with 9mL of sterile water, and shake fully. Take 1mL of the suspension and put it into a test tube filled with 9mL of sterile water, and mix well. Serial serial dilutions were carried out until 10 -10 . Take 0.2 mL of each gradient to plate. The coated plate was cultured at 26°C, and a fungus was obtained from it, which was named NCC3746.

[0045] (2) Identification of strain NCC3746

[0046] The spore liquid of bacterial strain NCC3746 was spread on a glass slide, and the micromorphological observation was carried out under a microscope at 400 times....

Embodiment 2、50

[0055] The preparation of embodiment 2, 50L tank fermentation dihydroascochlorin

[0056] (a) Preparation of Cylindrospora fungus NCC3746 seed solution

[0057] The Cylindrosporium fungus NCC3746 was inoculated in the seed medium, and cultured at 26 °C and 200 rpm for 72 h to obtain the seed liquid. The preparation method of the seed medium is: 20.0 grams of corn starch, 10.0 grams of glucose, 2.0 grams of hot-pressed soybean cake powder, 6.0 grams of malt powder, 2.5 grams of yeast powder, 2.0 grams of sodium chloride, 1.0 grams of magnesium sulfate heptahydrate, and tap water Dissolve and dilute to 1000 mL, pH 6.5-7.0, and sterilize at 121 °C for 30 min.

[0058] (b) Preparation of the fermentation broth of Cylindrospora fungus NCC3746

[0059] The seed liquid was inoculated into a fermenter with 30 L of fermentation medium at an inoculum volume of 5% by volume. The tank temperature was 26 °C, the tank pressure was 0.05±0.01 Mpa, the ventilation rate was 30 L / min, and th...

Embodiment 3

[0071] Embodiment 3, the preparation of shake bottle fermentation dihydroascochlorin

[0072] (a) Preparation of seed liquid of Cylindrospora fungus NCC3746

[0073] Cylindrospora ( Cylindrocarpon sp.) The fungus NCC3746 was inoculated on the seed medium, and cultured at 26 °C and 200 rpm for 72 h to obtain the seed liquid. The preparation method of the seed medium is: 20.0 grams of corn starch, 20.0 grams of glucose, 2.0 grams of hot-pressed soybean cake powder, 4.0 grams of malt powder, 3.0 grams of yeast powder, 2.0 grams of sodium chloride, 1.0 grams of magnesium sulfate heptahydrate, and tap water Dissolve and dilute to 1000 mL, pH 6.5-7.0, and sterilize at 121 °C for 30 min.

[0074] (b) Preparation of fermentation broth of Cylindrospora fungus NCC3746

[0075] The seed solution was inoculated into a 1000 mL shake flask containing 150 mL of fermentation medium at an inoculum size of 5% by volume, and fermented for 144 h at 28.5 °C with a rotation speed of 220 rpm....

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Abstract

The invention discloses a cylindrocarpon fungi strain and a method for preparing a dihydro ascochyta chlorine element through the strain. The fungi strain is the cylindrocarpon fungi NCC3746 (Cylindrocarponsp.), and the preservation number is CGMCCNo.7756. According to the method for preparing the dihydro ascochyta chlorine element through the strain, a fermentation unit of the dihydro ascochyta chlorine element reaches to above 1,300 mg / L which is higher than that of a currently-reported fermentation unit of the dihydro ascochyta chlorine element, a simple and practical crystal purifying technology is developed, the yield is more than 60%, the product purity reaches to above 97.0%, and the cylindrocarpon fungi strain can be used in industrial production of the dihydro ascochyta chlorine element.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a fungal strain of the genus Cylindrosporium and a method for preparing dihydroascochlorin by using the strain. Background technique [0002] Dihydroascochlorin is one of the members of the ascochlorin family and is a prenylphenolic antibiotic. Studies have proved that dihydroascochlorin has testosterone 5α reductase inhibitory effect (Satoshi T, et al., Chem. Pharm. Bull., 42(4):953-956, 1994), acetylcholinesterase inhibitory effect, β -Glucosidase inhibitory activity (Gutiérrez M, et al., J. Agric. Food Chem., 53: 7701-7708, 2005), retinoid X receptor agonist (Tamura Xuezao, et al., CN1334802A), etc. This compound has great application potential in the research and development of drugs for cardiovascular diseases, diabetes, tumors, and immunosuppression. [0003] Dihydroascochlorin can be produced by Fusarium sp. ( Fusarium sp.) (Ralph Henry Evans, et al., US3546073)...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P7/26C12R1/645
Inventor 路新华郑智慧任晓李业英徐岩可爱兵林洁朱京童崔晓兰石英范玉玲栗若兰霍培元沈文斌赵峰段宝玲
Owner NCPC NEW DRUG RES & DEV