A microbial enzymatic conversion method for echinocandin B

An echinocandin and its conversion technology can be used in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., and can solve problems such as negative effects on deacylase activity, cumbersome crude enzyme extraction process, and unfavorable scale-up production. To achieve the effect of less negative impact on enzyme activity, favorable for separation and purification, and good commercial value

Active Publication Date: 2016-06-08
CHONGQING QIANTAI BIOLOGICAL MEDICINE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of this method is that the crude enzyme extraction process is cumbersome, which is not conducive to industrial scale-up production; the extraction of crude enzyme by rotary steaming is easy to cause protein denaturation due to high temperature, and the activity of deacylase has a negative impact

Method used

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  • A microbial enzymatic conversion method for echinocandin B
  • A microbial enzymatic conversion method for echinocandin B
  • A microbial enzymatic conversion method for echinocandin B

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Fermentation and Crude Enzyme Production of Echinocandin B Deacylase Strain

[0040] Strain: Streptomycesalbus (Streptomycesalbus), preservation number: ATCC21725; -80°C cryopreservation;

[0041] Seed medium: hot fried soybean cake powder 0.5%, yeast powder 0.5%, peptone 0.5%, glucose 1%, pH about 6.8~7.2, culture at 30°C for 1~2 days;

[0042] Fermentation medium: hot fried soybean cake powder 1%, yeast powder 1%, peptone 1%, glucose 3%, sodium chloride 0.5%, magnesium sulfate heptahydrate 0.2%, dipotassium hydrogen phosphate 0.1%, pH about 6.8~7.2 , cultured at 30°C for 2-3 days;

[0043] Inoculate the genetically engineered strain of Streptomyces albicans into the seed medium, culture at 30°C for 1-2 days, then inoculate 5% of the fermentation volume in the fermentation medium, and culture at 30°C for 2-3 days;

[0044] After the fermentation is completed, the fermentation broth is filtered to obtain a supernatant, ammonium sulfate is added according to...

Embodiment 4

[0062] Example 4 Fermentation and Crude Enzyme Production of Echinocandin B Deacylase Strains

[0063] Strain: Streptomycesalbus (Streptomycesalbus), preservation number: ATCC21725; -80°C cryopreservation;

[0064] Seed medium: hot fried soybean cake powder 0.5%, yeast powder 0.5%, peptone 0.5%, glucose 1%, pH about 6.8~7.2, culture at 30°C for 1~2 days;

[0065] Fermentation medium: hot fried soybean cake powder 1%, yeast powder 1%, peptone 1%, glucose 3%, sodium chloride 0.5%, magnesium sulfate heptahydrate 0.2%, dipotassium hydrogen phosphate 0.1%, pH about 6.8~7.2 , cultured at 30°C for 2-3 days;

[0066] Inoculate the genetically engineered strain of Streptomyces albicans into the seed medium, culture at 30°C for 1-2 days, then inoculate 5% of the fermentation volume in the fermentation medium, and culture at 30°C for 2-3 days;

[0067] After the fermentation is completed, filter the fermentation broth to obtain a supernatant, add sodium chloride according to 40% (w / v...

Embodiment 6

[0078] Example 6 Fermentation and Crude Enzyme Production of Echinocandin B Deacylase Strain

[0079] Strain: Streptomycesalbus (Streptomycesalbus), preservation number: ATCC21725; -80°C cryopreservation;

[0080] Seed medium: hot fried soybean cake powder 0.5%, yeast powder 0.5%, peptone 0.5%, glucose 1%, pH about 6.8~7.2, culture at 30°C for 1~2 days;

[0081] Fermentation medium: hot fried soybean cake powder 1%, yeast powder 1%, peptone 1%, glucose 3%, sodium chloride 0.5%, magnesium sulfate heptahydrate 0.2%, dipotassium hydrogen phosphate 0.1%, pH about 6.8~7.2 , cultured at 30°C for 2-3 days;

[0082] Inoculate the genetically engineered strain of Streptomyces albicans into the seed medium, culture at 30°C for 1-2 days, then inoculate 5% of the fermentation volume in the fermentation medium, and culture at 30°C for 2-3 days;

[0083] After the fermentation is completed, filter the fermentation broth to obtain a supernatant, add sodium sulfate according to 80% (w / v) ...

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Abstract

The invention discloses a microbial enzymatic conversion method for echinocandin B, and particularly relates to a method of converting the echinocandin B into an echinocandin B nucleus. The method includes performing microbial fermentation to produce echinocandin B deacylase, extracting to obtain crude enzyme, and converting through adding the crude enzyme and the echinocandin B in a batch manner. The method is high in mole conversion ratio, high in product concentration, and beneficial to separation and purification. An echinocandin B substrate can be recovered and reutilized, thus reducing a production cost.

Description

technical field [0001] The invention relates to the field of biopharmaceuticals, in particular to an echinocandin B microbial enzyme conversion method. Background technique [0002] Echinocandins are a group of natural products discovered in the late 1970s. They 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, clinically used as an antifungal drug. [0003] Echinocandin B (ECB for short) is produced by the metabolism of Aspergilus nidulans and is one of the echinocandins. Its structure is as follows: [0004] [0005] Echinocandin B Nucleus (ECBN) is obtained by deacylase, and the cyclic peptide nucleus can be chemically modified to prepare the antifungal drug Anidulafungin. At present, for the preparation of ECB core, the cost of chemical synthesis is relatively high, and microbial transformation is mainly used. Microbial transformatio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/04C12R1/47
CPCY02P20/582
Inventor 别一郭明袁建栋
Owner CHONGQING QIANTAI BIOLOGICAL MEDICINE
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