Fermentation method of pneumocandin B0

A fermentation method, the technology of pneumocidine, which is applied in the field of microbial fermentation of pneumocidine B0, can solve the problems affecting the quality of caspofungin acetate, etc.

Active Publication Date: 2017-05-31
BRIGHTGENE PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

C0 impurity can participate in the follow-up reaction, seriously affecting the quality of caspofungin acetate

Method used

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  • Fermentation method of pneumocandin B0
  • Fermentation method of pneumocandin B0

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Fermentation strain: Glarea lozoyensis

[0018] Fermentation medium (wt): lactose 3.0%, threonine 1.0%, yeast powder 1.0%, proline 1.2%, KH 2 PO 4 0.15%, magnesium sulfate heptahydrate 0.05%, MES buffer salt 1.5%, pH 5.3.

[0019] 50L fermenter culture, 30L medium, sterilized at 121°C for 30 minutes. The seed volume is 1.5L, the culture temperature of the fermentation broth is 25°C, the initial ventilation volume is 0.9VVM, 200 rpm, and the tank pressure is 0.05Mpa. The maximum amount is 1.2VVM.

[0020] After 48 hours of fermentation, start to add vitamin b5, the added amount is 30mg / l. After 72 hours of fermentation (at this time, the ventilation rate and rotation speed have been adjusted to the upper limit), adjust the pH control range according to the change of dissolved oxygen until the end of fermentation: when the dissolved oxygen content is not lower than 45%, control the pH to 5.0-5.4, and the dissolved oxygen content in 96 hours 35%-45%, control pH 5.4-5....

Embodiment 2

[0021] Embodiment 2 (the comparative example that does not supplement vitamin b5)

[0022] Fermentation strain: Glarea lozoyensis

[0023] Fermentation medium (wt): lactose 3.0%, threonine 1.0%, yeast powder 1.0%, proline 1.2%, KH 2 PO 4 0.15%, magnesium sulfate heptahydrate 0.05%, MES buffer salt 1.5%, pH 5.3.

[0024] 50L fermenter culture, 30L medium, sterilized at 121°C for 30 minutes. The seed volume is 1.5L, the culture temperature of the fermentation broth is 25°C, the initial ventilation volume is 0.9VVM, 200 rpm, and the tank pressure is 0.05Mpa. The maximum amount is 1.2VVM.

[0025] After 72 hours of fermentation (at this time, the ventilation rate and rotation speed have been adjusted to the upper limit), adjust the pH control range according to the change of dissolved oxygen until the end of fermentation: when the dissolved oxygen content is not lower than 45%, control the pH to 5.0-5.4, and the dissolved oxygen content in 96 hours 35%-45%, control pH 5.4-5.8...

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Abstract

The invention provides a new fermentation method of pneumocandin B0, which can reduce the content of pneumocandin C0 in the fermentation process and remarkably lower the cost of downstream extraction technology. According to the method, the adopted strain is glarea lozoyensis, vitamin b5 is supplemented in the whole fermentation process, and pH is strictly controlled; and in the fermentation liquid obtained by the method, the content of pneumocandin C0 is reduced to 1.5% from 6%.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and relates to microbial fermentation of antifungal drugs, in particular to a microbial fermentation method of pneumocidine B0. Background technique [0002] Neomercontin B 0 It is a secondary metabolite synthesized by the fermentation of the fungus Glarea lozoyensis. In addition to the target product B0, the fermentation products of Glarealozoyensis include A0, B1, B2, C 0 , D0, E0 and other 12 analogues, among which the structure of C0 is the closest to B0, the structural difference between the two lies in the trans 3-hydroxyproline in B0, and the trans 4-hydroxyproline in C0. [0003] [0004] Neomercontin B0 Cas: 135575-42-7 [0005] [0006] Neomercontin C0 Cas: 144074-96-4 [0007] The existing technology mainly adopts the resin column process to purify Neomocontin B0. When column chromatography is applied to industrial production, it is time-consuming, uses a large amount ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/04C12R1/645
CPCC07K7/56
Inventor 袁建栋王其龙别一
Owner BRIGHTGENE PHARMA
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