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Method for reducing content of fusidic acid by-product by biotransformation method

A technology for fusidic acid and biotransformation, which is applied in the field of reducing the content of by-products of fusidic acid by biotransformation, can solve the problems of cumbersome chemical synthesis steps, no other bacterial species have been found, and chemical methods are difficult to apply, etc., and achieves good economic efficiency. Value and social benefit, mild cultivation conditions, simple operation effect

Pending Publication Date: 2020-03-06
YANCHENG INST OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Fusidic acid is a steroidal compound. Due to the existence of an asymmetric center in its molecule, the chemical synthesis steps are cumbersome, the cost is high, and the yield is low. It is difficult to apply chemical methods to actual industrial production. At present, microbial fermentation is mainly used at home and abroad. production method
[0005] Fusidic acid is isolated from the secondary metabolites of fungal fermentation. At present, except Fusidella sphaericus, no other strains have been found capable of synthesizing fusidic acid

Method used

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  • Method for reducing content of fusidic acid by-product by biotransformation method
  • Method for reducing content of fusidic acid by-product by biotransformation method
  • Method for reducing content of fusidic acid by-product by biotransformation method

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Experimental program
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Effect test

Embodiment 1

[0034] The preserved Rhizopus niger and Rhizopus oryzae glycerol tubes were respectively coated on the prepared PDA solid medium, incubated at 28 °C for 4-5 days, and then washed with sterile normal saline or 0.5% Tween-80 sterile aqueous solution plate, collect the spores, in 1 ml (10 8 cells / ml) were respectively inserted into the fermentation medium, and co-cultivated at 28 °C and 200 rpm for 24 h with constant temperature shaking and then set aside.

Embodiment 2

[0036] Prepare PDA solid medium, spread the preserved Fusidia sphaericus glycerol tube on the medium, culture the culture medium for 5-6 d fusidic acid-producing strains into primary seed medium, and cultivate at 26 °C After 72±5 hours, transfer 1% of the inoculum to the secondary expansion medium, culture at 26°C for 25±5 hours, then transfer to the fermentation medium at 10% of the inoculum, and culture at 26°C for 7-8 days , to prepare fusidic acid fermentation broth. After testing, the yield of fusidic acid produced by Fusidia sphaericus fermentation was 3205 μg / ml, and the yield of M impurity was 268 μg / ml (the content was 7.72%).

Embodiment 3

[0038] Collect the fermented fusidic acid fermentation broth, adjust the pH to 3.5-4.0 with dilute hydrochloric acid, let it stand for 30 minutes, and extract it with 4 times the volume of absolute ethanol for 30 minutes, during which the mixture was shaken continuously. After the end, appropriate amount of samples were taken and stored in a refrigerator at 4°C. The fermentation extract was centrifuged, and the supernatant was concentrated by low-temperature rotary evaporation.

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Abstract

The invention belongs to the technical fields of fermentation engineering and microbial steroid transformation, and particularly relates to a method for reducing content of a fusidic acid by-product through a biotransformation method. According to the method, rhizopus nigricans or rhizopus oryzae is used as transforming bacteria, and a M by-product in fusidic acid crude extract is transformed intofusidic acid. The method specifically comprises the following steps: inoculating a rhizopus nigricans spore suspension or a rhizopus oryzae spore suspension into a rhizopus nigricans or rhizopus oryzae fermentation culture medium, and carrying out fermentation culture for 24 hours; carrying out digestion and centrifugal treatment on fusidic acid fermentation liquor obtained by fermentation to obtain a supernatant, evaporating and concentrating the supernatant to obtain an M impurity ethanol concentrated solution, putting the M impurity ethanol concentrated solution into the rhizopus nigricansor rhizopus oryzae fermentation culture medium subjected to fermentation culture for 24 hours, adding an oxygen carrier, and continuously converting the M impurity. According to the method for reducing the content of M impurities in fusidic acid through biotransformation, the content of M impurities in fusidic acid is lower than 2%, and the quality of the fusidic acid product is remarkably improved.

Description

technical field [0001] The invention belongs to the technical field of fermentation engineering and microbial steroid transformation, and in particular relates to a method for reducing the content of fusidic acid by-products through biotransformation. Background technique [0002] Fusidic acid, the molecular formula is C 31 h 48 o 6 , molecular weight 516.69, chemical name: 16β-acetoxy [0003] Base-3α,11α-dihydroxy-4α,8α,14β-trimethyl-18-nor-5α,10β-cholesteryl-(17 Z ) -17(20) , 24-diene-21-acid, has a steroid-like structure, and its molecular structure has a high degree of similarity with fumagic acid and cephalosporin P, with only a small number of groups having differences , the presence of different groups endows it with a unique antibacterial effect. Studies have shown that fusidic acid has very little water solubility and good fat solubility, while sodium fusidate is easily soluble in water. [0004] Fusidic acid is an important antibiotic for G + bacteria have ...

Claims

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Application Information

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IPC IPC(8): C12P33/00C12R1/845
CPCC12P33/00
Inventor 高健黄巍巍张丽徐虹邓丽娜刘长青仇静刘伟
Owner YANCHENG INST OF TECH
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