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Method for preparing oritavancin intermediate by fermentation

A technology for oritavancin and intermediates, which is applied in the preparation of oritavancin intermediate A82846B, the fermentation field of glycopeptide antibiotics, can solve the problem of increasing production cost, affecting extraction yield, low fermentation unit and component ratio, etc. question

Active Publication Date: 2021-08-06
JIANGSU SENRAN CHEM
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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, and the separation and purification work is quite difficult, which increases the production cost. The proportion of A82846B components 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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  • Method for preparing oritavancin intermediate by fermentation
  • Method for preparing oritavancin intermediate by fermentation

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Experimental program
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Effect test

Embodiment 1

[0031] The preparation of embodiment 1 seed solution

[0032] Seed medium: maltodextrin 20g / L, glucose 10g / L, soybean powder 15g / L, yeast extract 3g / L, calcium carbonate 1g / L, pH7.0.

[0033] Pack 150ml of seed culture medium into a 750ml Erlenmeyer bottle, and sterilize at 120°C for 30 minutes before use.

[0034] Inoculate the A82846B production strain Nocardia orientalis NRRL18099 preserved at low temperature into the prepared seed bottle, and culture the shake flask at a temperature of 30°C and a rotation speed of 250rpm for 40-50h to obtain mature shake flask seeds.

Embodiment 2

[0036] 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.

[0037] Pack 35L of fermentation medium in a 50L fermentation tank, and sterilize at 120°C for 30 minutes before use.

[0038] 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, and control dissolved oxygen above 30% by adjusting the speed. Cultivate for 5 days to obtain a fermented liquid, and the fermentation unit is 515 mg / L.

Embodiment 3

[0040] The fermented liquid obtained in Example 2 was adjusted to pH=10.3 with NaOH solution, stirred for 2 hours, and subjected to plate-and-frame pressure filtration 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 conduct nanofiltration on the desorption mixture, and concentrate to The unit is 35000 mg / L.

[0041] Introduce the concentrated solution into a well-balanced C18 column, and use 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 the extraction of the product. A82846B purity below 90% is impurity waste liquid.

[0042] Collect the impurity waste liquid, concentrate it by nanofiltration, and ...

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Abstract

The invention discloses a method for preparing A82846B by fermentation. The method is to extract the fermentation culture liquid obtained by fermenting and cultivating Amycolatopsis orientalis, and then carry out alkali degradation on the extracted impurity waste liquid to obtain the alkali degradation liquid. After the alkali degradation solution was added to the fermentation medium, the fermentation was carried out again. By adding different concentrations of impurity alkali degradation solution, the fermentation unit of A82846B was greatly increased, and the components were also 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 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 patients with...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P21/02C07K9/00C12R1/01
Inventor 袁建栋刘会明
Owner JIANGSU SENRAN CHEM
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