Method for improving yield of echimocandins B

A technology for echinocandin and yield is applied in the field of cultivation for increasing the yield of aniduckin precursor compound EchinocandinB, and can solve the problems of low fermentation yield and the like

Inactive Publication Date: 2016-07-06
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to provide a technology for improving the fermentation level of echinocandin B by adding metal ions to address the defects of low yield of echinocandin B fermentation

Method used

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  • Method for improving yield of echimocandins B
  • Method for improving yield of echimocandins B
  • Method for improving yield of echimocandins B

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028]Embodiment 1: the influence of adding calcium chloride on the output of echinocandin B in the fermentation medium;

[0029] (1) Slant culture: inoculate Aspergillus nidulans ZJB09223 into the slant medium, culture at 25-30°C for 28 hours, and obtain slant bacteria; the final concentration of the slant medium used is composed of: potatoes 200g / L (boiled supernatant), Sucrose 20g / L, agar powder 20g / L, pH natural, solvent is water, sterilized at 121°C for 20min and cooled before use;

[0030] (2) Seed culture: inoculate the slanted bacteria from step (1) into the seed medium, cultivate at 25-30°C for 48 hours, and obtain the seed liquid; the final concentration of the seed medium consists of: cottonseed powder 25g / L, glucose 10g / L, Glycerin 10g / L, pH 6.8-7.0, solvent is distilled water, sterilized at 115°C for 30 minutes and cooled before use.

[0031] (3) Fermentation culture: step (2) seed liquid is respectively inoculated to add 0g / L, 0.3g / L, 0.4g / L, 0.5g / L, 0.6g / L fina...

Embodiment 2

[0049] Example 2: Effects of different addition times of calcium chloride with a final concentration of 0.5g / L in the fermentation medium on the yield of echinocandin B

[0050] Slope culture and seed culture are the same as in Example 1.

[0051] Fermentation culture: inoculate the seed solution to the fermentation medium with an inoculation amount of 10% volume concentration, set up 1 control group (the control group does not add calcium chloride) and 4 experimental groups (group 1-group 4), group 1 - Group 4 added calcium chloride in 1, 2, 3, and 4 times respectively, starting from the initial time 0h, and adding once every 8h (group 1 (that is, the 1-time addition group) added calcium chloride at a concentration of 5g / L , Group 2 (i.e. 2 times of addition group) added calcium chloride concentration of 2.5g / L each time, group 3 (i.e. 3 times of addition group) added calcium chloride concentration of 1.67g / L each time, group 4 (i.e. 4 Second addition group) each addition of...

Embodiment 3

[0053] Example 3: Effects of different addition intervals of calcium chloride with a final concentration of 0.5g / L in the fermentation medium on the yield of echinocandin B

[0054] Slope culture and seed culture are the same as in Example 1.

[0055] Fermentation culture: inoculate the seed solution to the fermentation medium with an inoculation amount of 10% volume concentration, set up 1 control group (the control group does not add calcium chloride) and 4 experimental groups (group 1-group 4), group 1 - Group 4 added calcium chloride once at the beginning of fermentation 0h, and then added calcium chloride once at intervals of 6h, 8h, 10h, and 12h, with a concentration of 0.167g / L each time, until the final concentration of calcium chloride was 0.5g / L, cultured at 20-30°C, 220-280rpm for 12 days to obtain a fermentation broth, and no addition of metal ions was used as a control. The content of echinocandin B in the fermentation broth was detected by HPLC method, and the ...

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Abstract

The invention discloses a method for increasing the yield of echinocandin B. The method comprises: using Aspergillus nidulans CCTCC M 2012300 as a production strain, inoculating Aspergillus nidulans CCTCC M 2012300 into a fermentation medium added with metal ions , cultured at 20-30°C and 220-280rpm for 7-12 days to obtain a fermented liquid, which was separated and purified to obtain echinocandin B; The method of echinocandin B output, by adding manganese sulfate, calcium chloride, cupric sulfate, ferrous chloride, magnesium sulfate etc. in fermentation medium, improves anidifungin precursor compound echinocandin B output; Result Show that in the fermentation culture system, the highest yield reaches 1738.6mg / L, which is 64.7% higher than that of the control group without metal ions. important meaning.

Description

(1) Technical field [0001] The invention relates to a cultivation method for increasing the yield of anidulungin precursor compound EchinocandinB. (2) Background technology [0002] Echinocandin B (Echinocandin B) is a natural cyclic lipopeptide compound with a cyclic hexapeptide core and fatty acid side chain structure. The chemical name is and the molecular formula is C 52 h 81 N 7 o 16 , the molecular weight is 1060, and the structural formula is as follows: [0003] [0004] Echinocandin antibiotics are a group of natural products discovered in the 1970s, which have similar structural features of cyclic polypeptide cores and different fatty acid side chains, and can non-competitively inhibit fungal cell wall β-1, 3-glucan synthase activity, so as to achieve the purpose of antifungal. Anidulafungin is the third-generation systemic antifungal echinocandin derivative drug following Caspofungin and Micafungin. On February 21, 2006, it was released by Pfizer Anifung...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/04C12R1/66
CPCC07K7/56
Inventor 郑裕国邹树平熊严牛坤
Owner ZHEJIANG UNIV OF TECH
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