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A method for preparing ascochlorin by using Cylindrosporium fungal strains

A technology of ascochlorin and ascochlorin, which is applied in the field of microorganisms, can solve the problems of complex metabolite components and low fermentation unit of ascochlorin, and achieves a simple and easy preparation process and is suitable for industrial production. Effect

Active Publication Date: 2016-08-24
NCPC NEW DRUG RES & DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Ascochlorin can be produced by Fusarium sp. (Ralph Henry Evans, et al., US3546073), Nectria coccinea (Andridge, et al., J. Chem. Soc., PerkinTrans. 1, 2136-2141, 1972), Verticillium sp. (Satoshi T, et al., Chem. Pharm. Bull., 42(4):953-956, 1994), 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 fermentation units of ascochlorin are all Very low, no more than 50mg / L

Method used

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  • A method for preparing ascochlorin by using Cylindrosporium fungal strains
  • A method for preparing ascochlorin by using Cylindrosporium fungal strains
  • A method for preparing ascochlorin by using Cylindrosporium fungal strains

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Experimental program
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Effect test

Embodiment 1

[0040] Embodiment 1, the separation and identification of Cylindrospora bacterial strain NCC3746

[0041] (1) Isolation of strain NCC3746

[0042] The medium used for the isolation of the fungal strain NCC3746 is PDA (Beijing Shuangxuan);

[0043] 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.

[0044] (2) Identification of strain NCC3746

[0045] 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

[0054] The preparation of embodiment 2, 50L tank fermentation ascochlorin

[0055] (a) Preparation of Cylindrospora fungus NCC3746 seed solution

[0056] 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 to 1000mL, pH 6.5-7.0, and sterilize at 121°C for 30 min.

[0057] (b) Preparation of the fermentation broth of Cylindrospora fungus NCC3746

[0058] The seed solution 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 the stirring was perfor...

Embodiment 3

[0069]Embodiment 3, the preparation of shake bottle fermentation ascochlorin

[0070] (a) Preparation of seed liquid of Cylindrospora fungus NCC3746

[0071] The fungus NCC3746 of the genus Cylindrocarpon was inoculated in the seed medium, and cultured at 26 °C and 200 rpm for 72 h to obtain the seed solution. 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.

[0072] (b) Preparation of fermentation broth of Cylindrospora fungus NCC3746

[0073] 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. The preparation m...

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Abstract

The invention discloses a method for preparing ascochlorin by using a fungus strain of the genus Cylindrosporium. The bacterial strain utilized in the present invention is Cylindrospora fungus NCC3746 ( Cylindrocarpon sp.), the deposit number is CGMCC No.7756. In the method for preparing ascochlorin provided by the present invention, the fermentation unit of ascochlorin has reached more than 1600 mg / L, which is higher than the fermentation unit of ascochlorin reported so far, and the development is simple and easy The advanced crystallization purification process has a yield of more than 60% and a product purity of more than 97.0%, which can be used for the industrial production of ascochlorin.

Description

technical field [0001] The invention relates to the technical field of microbes, in particular to a method for preparing ascochlorin by using a fungal strain of the genus Cylindrosporium. Background technique [0002] Ascochlorin is a prenyl phenolic compound (Tamura G , etal. J. Antibiot, 21: 539-544, 1968), has antiviral, antitumor, antibacterial and sedative activities and has the effect of lowering cholesterol and triglyceride levels, lowering blood pressure, and improving type I and type II diabetes (Hong S, et al., J. Biol Chem,280(26): 25202-25209, 2005; Tsuruga M, et al., J. Antibiot, 60(1): 20-26,2007; Sakaguchi K, et al., Biochem Bioph Res Commun, 329(1): 46-50, 2005; Tsuruga M, et al., Apoptosis, 9(4): 429-435, 2004), has great application prospects. [0003] Ascochlorin can be produced by Fusarium sp. (Ralph Henry Evans, et al., US3546073), Nectria coccinea (Andridge, et al., J. Chem. Soc., PerkinTrans. 1, 2136-2141, 1972), Verticillium sp. (Satoshi T, et al.,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P7/26C12R1/645
Inventor 李业英徐岩陈书红路新华郑智慧任晓李丽红林洁崔晓兰石英范玉玲蔡喜田张雪霞段宝玲
Owner NCPC NEW DRUG RES & DEV