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Method for producing nikkomycin Z, special culture medium of nikkomycin Z and special engineering bacteria of nikkomycin Z

A nikkomycin and culture medium technology, applied in the field of nikkomycin Z production, can solve the problems of expensive reagents, unsuitable for large-scale industrial production, inability to effectively separate X components and Z components, etc. The effect of simplifying separation and purification and reducing production costs

Active Publication Date: 2011-02-02
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Even with the method of HPLC, the use of trifluoroacetic acid as the mobile phase cannot effectively separate the X component and the Z component
Using HPLC, component X and component Z can be separated with ion-pairing reagents, but the reagents used are expensive and not suitable for large-scale industrial production

Method used

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  • Method for producing nikkomycin Z, special culture medium of nikkomycin Z and special engineering bacteria of nikkomycin Z
  • Method for producing nikkomycin Z, special culture medium of nikkomycin Z and special engineering bacteria of nikkomycin Z
  • Method for producing nikkomycin Z, special culture medium of nikkomycin Z and special engineering bacteria of nikkomycin Z

Examples

Experimental program
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Effect test

Embodiment 1

[0026] Example 1, Construction of Streptomyces ansochromogenes TH322 sanPD CGMCCNo.3086

[0027] The nikkomycin-producing bacterium used in this example is Streptomyces chromogenes strain TH322. Streptomyces cylindrochromogenes TH322 is a strain obtained by mutagenizing Streptomyces cylindrochromogenes 7100 CGMCC 4.321 (purchased from the General Microorganism Center of China Committee for Culture Collection of Microorganisms). The spores of Streptomyces chromogenes 7100 CGMCC 4.321 (purchased from the General Microorganism Center of China Microbial Culture Collection Management Committee) are in the shape of silk hooks, or coils, and rarely coils; the spores are spherical or elliptical. The initial nikkomycin Z production of Streptomyces circulogenes strain TH322 was 350 mg / L.

[0028] Genomic DNA was extracted from Streptomyces chromogenes 7100CGMCC 4.321, digested with Sau3AI, and separated by pulse electrophoresis to recover a 40-50kb DNA fragment. Phosphorylated and rec...

Embodiment 2

[0033] Example 2, HPLC analysis and biological activity detection of Streptomyces ansochromogenes TH322 sanPD CGMCCNo.3086 fermentation product

[0034] 1) Preparation of fermentation product of Streptomyces ansochromogenes TH322 sanPD CGMCC No.3086

[0035] Inoculate Streptomyces ansochromogenes (Streptomyces ansochromogenes) TH322 sanPD CGMCC No.3086 on YEME medium (Difco Yeast Extract 3g, Difco Tryptone 5g, OxoidMalt Extract 3g, Sucrose 200g, add distilled water to 1000ml, before use Add the following sterile solution: MgCl 2 ·6H 2 O (2.5M) 2ml, Glucose (50%) 20ml, Glycine (10%) 50ml), 28°C, speed 180rpm, after 48 hours of cultivation, transfer 1ml of bacterial liquid to 50ml fermentation medium SP medium, 28°C, speed 180rpm, cultivated for 5 days, and centrifuged to collect the fermentation broth. The fermentation experiment was repeated 3 times.

[0036] The SP medium is prepared as follows: 30g of glucose, 30g of mannitol, 5g of soybean peptone, 8g of yeast extract, ...

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Abstract

The invention discloses recombinant streptomyces ansochromogenes, which are streptomyces ansochromogenes having blocked sanP genes for biologically synthesizing nikkomycins. The recombinant streptomyces ansochromogenes are (Streptomyces ansochromogenes) TH322 sanPD CGMCC No.3086. The invention also provides a method for producing the nikkomycin Z and a culture medium of the nikkomycin Z. In the invention, the recombinant streptomyces ansochromogenes which only produce a single nikkomycin Z component are provided, the culture medium of the nikkomycin Z is optimized, and the recombinant streptomyces ansochromogenes can produce the single component nikkomycin Z in large scale with a yield which is 4.5 times that of the prior initial strain of streptomyces ansochromogenes TH322 and reduce production cost, and have a significant for the simplification of the separation and purification of products obtained after fermentation.

Description

technical field [0001] The invention relates to a method for producing nikkomycin Z, its special culture medium and engineering bacteria. Background technique [0002] Nikkomycin belongs to nucleoside peptide antibiotics, which is similar in structure to UDP-N-acetylglucosamine, the natural substrate of chitin synthase, and is a strong competitive inhibitor of this enzyme. Since chitin is an important part of the cell wall of fungi and the exoskeleton of insects and other invertebrates, nikkomycin can effectively inhibit fungi, insects and mites, and is non-toxic or extremely toxic to mammals, bees and plants Low, and nikkomycin is easy to degrade in nature, so nikkomycin is an ideal agricultural antibiotic. Field experiments have proved that nikkomycin has a good control effect on many plant diseases, such as wheat powdery mildew, tobacco red spot, cucumber sclerotinia, tomato leaf mold, and apple ring spot. At the same time, studies have shown that nikkomycin can effecti...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/21C12P21/02C12R1/465
Inventor 谭华荣廖国建
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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