A kind of method utilizing Aspergillus fumigatus to produce fumagillin

A technology of fumagillin and Aspergillus fumigatus, which is applied in the field of microbial fermentation, can solve the problems of low yield of fumagillin, difficult industrial production or application, etc., and achieves the effect of increasing yield

Active Publication Date: 2019-10-11
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] There are only few reports on the optimized culture method of Aspergillus fumigatus producing secondary metabolite fumagillin and its derivatives (Wen Yang, Carbon and nitrogen source nutrition of fumagillin biosynthesis by Aspergillus fumigatus, Current Microbiology, 2003, 275– 279), but the yield of fumagillin in this report is low, it is difficult to be used for industrial production or application

Method used

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  • A kind of method utilizing Aspergillus fumigatus to produce fumagillin
  • A kind of method utilizing Aspergillus fumigatus to produce fumagillin
  • A kind of method utilizing Aspergillus fumigatus to produce fumagillin

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Embodiment 1

[0029]The strain activation culture medium used in the present invention is GMM culture medium, and the described solid culture medium of every 1L is prepared as follows: glucose 10g, nitrate mother liquor 50mL, trace element mother liquor 1mL, yeast extract 1g, agar 15-20g, Add the above ingredients in sequence, and finally add distilled water to make the final volume 1L, adjust the pH value to 6.5, and sterilize at 121°C for 20-30 minutes. Invert a 90mm plate, streak and inoculate, and culture at 37°C for 3 to 4 days. Collect spores in sterile water, collect spores from each plate, make a spore suspension with a final volume of 2 mL, and store at 4°C for culture inoculation.

[0030] Wherein, the nitrate mother liquor preparation (1L) method is as follows: sodium nitrate NaNO 3 120g, potassium chloride KCl 10.4g, magnesium sulfate MgSO 4 ·7H 2 O 10.4 g, dipotassium hydrogen phosphate K 2 HPO 4 ·3H 2 O 30.4g, add the above ingredients in order, and finally add distille...

Embodiment 2

[0033] The used fermentation medium is CYA medium, and the CYA solid medium per 1L is prepared as follows: sodium nitrate NaNO 3 2-5g, dipotassium hydrogen phosphate K 2 HPO 4 ·3H 2 O 0.5-2g, magnesium sulfate MgSO 4 ·7H 2 O 0.3-1g, potassium chloride KCl0.3-1g, ferrous sulfate FeSO 4 ·7H 2 O 0.01-0.03g, sucrose 20.0-40.0g, yeast extract 3-8g, agar 15-20g, add the above ingredients in order, and finally add distilled water to the final volume to 1L, and the pH value is natural. Autoclave at 121°C for 30 minutes, pour on a 90mm plate, and inoculate by streaking. The culture temperature is 25 to 37° C., the culture medium is solid culture medium, and the cultivation time is 4 days to 6 days.

[0034] Through the HPLC fingerprint analysis detection peak area calculation method combined with the actual separation calculation method, it is measured that the yield of fumagillin per liter of CYA medium is about 34-50 mg.

Embodiment 3

[0036] The fermentation medium used is CXMA medium, and the solid medium per 1L is prepared as follows: Magnesium sulfate MgSO 4 ·7H 2 O 0.3-0.5g, potassium chloride KCl 0.3-0.8g, ferrous sulfate FeSO 4 ·7H 2 O 0.01-0.02g, xylan 20-40g, mannose 30-80g, glutamic acid 5-12g, agar 15-20g. The above ingredients were added in sequence, and finally distilled water was added to the final volume to 1L, and the pH value was natural. Autoclave at 121°C for 30 minutes, pour on a 90mm plate, and inoculate by streaking. The culture temperature is 25° C. to 37° C., the medium is a solid medium, and the culture is static, and the culture time is 4 days to 6 days.

[0037] Through the HPLC fingerprint analysis detection peak area calculation method combined with the actual separation calculation method, it is measured that the yield of fumagillin per liter of CXMA medium is about 25-35 mg.

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Abstract

The invention relates to a method for producing fumagillin by aspergillus fumigatus. The method comprises the following steps: inoculating aspergillus fumigatus into a GMM solid medium to activate strains, wherein the pH value of the GMM medium is 5 to 7; preparing activated strain spores into spore suspension, and then inoculating the spore suspension into a CYA solid medium, a CXMA solid medium or an LMM liquid medium, wherein the pH values of the CYA solid medium and the CXMA solid medium are natural, and the pH value of the LMM liquid medium is adjusted to be 5 to 7, and carrying out culturing at the temperature of 25 to 37 DEG C; collecting cultured aspergillus fumigatus to extract fumagillin and fumagillin derivatives. According to the invention, carbon and nitrogen sources and trace elements of fumagillin which affect the growth of aspergillus fumigatus are aimed to be screened, and the concentration of corresponding trace elements is optimized to obtain higher yield of fumagillin.

Description

technical field [0001] The invention relates to the field of microbial fermentation, in particular to a method for producing fumagillin by using Aspergillus fumigatus. Background technique [0002] Aspergillus fumigatus is an important fungus widely distributed in nature, present in soil and animal rotting fur. Aspergillus fumigatus is an opportunistic pathogen and an important pathogen. The conidia produced are in the air and enter the human body through the human respiratory tract, causing infection in patients with special conditions such as low immune function or flora imbalance. On the other hand, various mycotoxins such as gliotoxin and fumagillin produced by Aspergillus fumigatus in feed seriously harm the health of animals and reduce production capacity, threatening human food safety and health. In recent years, the complete genome sequencing of Aspergillus fumigatus has provided a powerful tool for the study of mycotoxins derived from Aspergillus fumigatus. Studie...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P17/18C12P17/04A61K31/336A61K31/343A61P35/00A61P33/02A61P19/02A61P29/00A61P3/04C12R1/68
CPCY02A50/30
Inventor 尹文兵李伟
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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