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Fermentation method of oritavancin intermediate

The technology of oritavancin and intermediates is applied in the field of microbial fermentation production, which can solve the problems of low fermentation unit and component ratio, high production cost, difficult separation and purification work, etc.

Active Publication Date: 2019-07-26
CHONGQING QIANTAI BIOLOGICAL MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] It can be seen from the above that the structures of the three substances are very close, the separation and purification work is quite difficult, and the production cost is thus increased. The proportion of the components of A82846B in the fermentation broth (the proportion of A82846B in the three substances) will greatly affect the extraction yield. Rate
At present, there are few studies on A82846B in my country, and the fermentation unit and component ratio are relatively low, resulting in high production costs

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] The preparation of embodiment 1 seed liquid

[0035] Seed medium: maltodextrin 20g / L, glucose 10g / L, soybean powder 15g / L, yeast extract 3g / L, calcium carbonate 1g / L, pH7.0.

[0036] Pack 150ml of seed culture medium into a 750ml Erlenmeyer bottle, and sterilize at 120°C for 30 minutes before use.

[0037] Inoculate the A82846B production strain Nocardia orientalis NRRL18098 preserved at low temperature into the prepared seed bottle, and shake the bottle for 40-50 hours at a temperature of 30°C and a rotation speed of 250 rpm to obtain mature shake bottle seeds.

Embodiment 2

[0039] Fermentation medium: maltodextrin 40g / L, molasses 20g / L, soybean powder 20g / L, yeast powder 10g / L, calcium carbonate 3g / L, calcium chloride 10g / L, pH7.0.

[0040] Pack 35L of fermentation medium in a 50L fermentation tank, and sterilize at 120°C for 30 minutes before use.

[0041] Inoculate 3.5L of mature shake flask seeds into a fermenter with 35L of feed liquid, culture conditions: temperature 30°C, ventilation volume 1VVM, tank pressure 0.05Mpa, initial speed 200rpm, control the dissolved oxygen by adjusting the speed to more than 30%. Cultivate for 5 days to obtain a fermented liquid, and the fermentation unit is 515 mg / L.

Embodiment 3

[0043] The fermented liquid obtained in Example 2 was adjusted to pH=10.3 with NaOH solution, stirred for 2 hours, and plate-and-frame press-filtered to obtain 35 L of press-filtered liquid, which was adjusted to pH=9.2, and then introduced into LX18 adsorption resin. Purify 9L of resin with water with a pH of 8, then use 8L of 1.0% acetic acid aqueous solution for desorption, and mix components with a concentration greater than 500mg / L to form a primary desorption mixture, then perform nanofiltration on the desorption mixture, and concentrate to The unit is 35000mg / L.

[0044] The concentrated solution is introduced into the C18 column that has been balanced, and the eluent (5% acetonitrile: 0.5% NH 4 h 2 PO 4 =3:97 (v / v)) for elution, and the components with a purity of more than 90% were collected for extracting the product. A82846B purity below 90% is impurity waste liquid.

[0045] Collect the impurity waste liquid, concentrate it by nanofiltration, and concentrate un...

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Abstract

The invention discloses a method for preparing A82846B by fermentation. The method includes extracting a fermentation broth obtained after fermentation and culture of amycolatopsis orientalis, and conducting alkali degradation on the obtained impurity waste liquid to obtain an alkali degrading solution; after the alkali degradation liquid is added to a fermentation medium, performing fermentationagain, culturing for 25-35h and adding calcium chloride to continue the fermentation; adding the alkali degrading solution in different concentrations of impurities and adding calcium chloride after the fermentation culture is performed for about 30h. The A82846B fermentation unit is greatly improved, and the composition is significantly improved.

Description

technical field [0001] The invention belongs to the technical field of microbial fermentation production, and relates to a fermentation method of glycopeptide antibiotics, in particular to a method for preparing oritavancin intermediate A82846B through fermentation. Background technique [0002] Glycopeptide antibiotics are a large class of substances produced by microorganisms, or produced and partially modified by microorganisms. Vancomycin and teicoplanin are commercially available antibacterial products. Among the glycopeptides discovered in the 1990s, there are those known as A82846A (also known as ereomomycin), A82846B (also known as chloroorienticinA), A82846C (also known as orienticinC) and orienticin A . A variety of modifications have been made to naturally occurring glycopeptides, one of which is the reductive alkylation of reactive amines in glycopeptides. [0003] On August 7, 2014, the FDA approved the antibiotic oritavancin for the treatment of adult patien...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/00
CPCC12P21/005
Inventor 袁建栋刘会明
Owner CHONGQING QIANTAI BIOLOGICAL MEDICINE
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