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Streptomyces parvus and application thereof for preparing daptomycin

A technology of Streptomyces parvum and strains, applied in the field of Streptomyces parvum and its application in the preparation of daptomycin, which can solve the problems of many impurities, many fermentation by-products, and complicated extraction work

Active Publication Date: 2010-12-01
HANGZHOU HUADONG MEDICINE GRP PHARMA RES INST +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Generally, daptomycin-producing bacteria have unstable production capacity, low yield, more fermentation by-products, and more impurities, resulting in more complicated extraction work, which greatly increases the difficulty of subsequent purification, and it is difficult to obtain high-purity final product, so that it cannot be industrialized

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  • Streptomyces parvus and application thereof for preparing daptomycin
  • Streptomyces parvus and application thereof for preparing daptomycin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: Culture characteristics of Streptomyces parvum CCTCC NO: M2010136

[0042] The strain of Streptomyces parvum CCTCC NO: M2010136 was inoculated on Gaoshi No. 1, glucose asparagine and other medium for observing the morphological characteristics. Cultivate at 28°C for a certain period of time for 5, 8, and 10 days, observe and describe.

[0043] Table 1. Colony characteristics of strains of Streptomyces parvum CCTCC NO: M2010136 on various media (see Table 1)

[0044] culture medium

[0045] Glucose Yeast Extract Agar

Embodiment 2

[0046] Example 2: Physiological and biochemical characteristics and carbon source utilization

[0047] Physiological and biochemical properties used standard methods in the classification of Streptomyces minor.

[0048] The physiological and biochemical reactions of Streptomyces parvum CCTCC NO: M2010136 are as follows: the bacterium gelatin liquefies slowly and becomes a bright yellow pigment. The milk does not coagulate and is quickly peptonized. Starch hydrolysis. Cellulose does not grow. Nitrate reduction is weak and does not produce melanoids. Use the Biolog automatic microbial identification system to investigate the metabolism of Streptomyces parvum to 95 different carbon sources: Inoculate the strains on the BUG+B plate medium, culture at a constant temperature of 37°C for 16 hours, wash the bacteria on the plate with a sterile cotton swab, Mix with the inoculum (GN / GP-IF+T) to make a bacterial suspension, and adjust to 20% T with a turbidimeter. Use an 8-hole ele...

Embodiment 3

[0054] Embodiment 3: the production fermentation culture of daptomycin

[0055] 1. Seed strain cultivation and preservation

[0056] Solid medium: glucose 0.5g, yeast extract 0.3g, wort juice 0.3g, CaCO30.3g, agar 1.8g, water 100ml, pH7.0

[0057] Cultivation: The strain is inoculated on a solid medium slant and cultured at 30°C for 7-10 days.

[0058] After the solid culture is over, place it on an inclined plane at 4-10°C and refrigerate it for later use.

[0059] 2. Shake Flask Seed Culture

[0060] Medium: glycerin 20g, soluble starch 80g, yeast powder 6g, raw soybean powder 20g, trypsin soybean protein 20g, Fe(NH 4 ) 2 (SO 4 ) 2 .6H 2 O 10g, add water 2000ml, pH 7.0

[0061] Inoculum size: Dig out the slanted lawn block and insert it into the 2L shake flask seed medium

[0062] Culture: 30℃, 24h

[0063] Shaker speed: 250rpm

[0064] When the hyphae grow up, there is obvious wall-hanging phenomenon and the bacterial concentration is about 11%. Put the 2L shake ...

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Abstract

The invention discloses a streptomyces parvus and application thereof for preparing daptomycin. The strain is reserved in the CCTCC and has the reservation number of CCTCC NO: M2010136, and the reservation data is June 4, 2010. The method for preparing the daptomycin is as follows: a, taking streptomyces parvus CCTCC NO: M2010136 as a fermentation strain; b, strain cultivation: inoculating slope lawn into a shake flask seeding tank to cultivate to obtain shake flask seeding liquid; inoculating the shake flask seeding liquid into the seeding tank to be cultivated; inoculating the seeding tank culture solution into fermentation tank culture medium to be cultivated, and collecting fermentation liquor; carrying out feed supplement in the fermentation and cultivation process; and c, extracting a fermentation product. The fermentation technology provided by the invention has stable production capability, high fermentation unit and few fermentation byproducts by the pilot plant test and the experiment in a 10-ton fermentation tank, greatly lowers post-extracting difficulty and is suitable for industrial production.

Description

technical field [0001] The present invention relates to a novel microorganism and its use and application, in particular to a Streptomyces parvum and its application in preparing daptomycin. Background technique [0002] Daptomycin is the first product of a new family of antibiotics called cyclic lipopeptides. It is extracted from the fermentation broth of Streptomyces parvus. Not only does it have a novel chemical structure, but its mode of action is also different from any approved antibiotic. Daptomycin can disrupt bacterial cell membrane function in multiple ways, thereby rapidly killing Gram-positive bacteria. In addition to its ability to act on most clinically relevant Gram-positive bacteria, daptomycin is still potent in vitro against isolates that have been resistant to methicillin, vancomycin, and linezolid. active. [0003] Daptomycin is a fermentation product, and daptomycin can be finally obtained through a series of purification processes such as chromatogr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P21/02C12R1/465
Inventor 朱健陈晓霞许永锋王蓓李艳高兴蓉
Owner HANGZHOU HUADONG MEDICINE GRP PHARMA RES INST
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