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

A technology of nikkomycin and culture medium, applied in the field of production of nikkomycin Z, 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 reducing production costs and simplifying separation and purification

Active Publication Date: 2009-12-02
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

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Effect test

Embodiment 1

[0026] Example 1. Construction of Streptomyces ansochromogenes TH322 sanPD CGMCC No. 3086

[0027] The nikkomycin-producing strain used in this example is Streptomyces chromogenes TH322. Streptomyces chromogenes TH322 is a strain obtained by mutagenesis of Streptomyces chromogenes 7100 CGMCC 4.321 (purchased from the General Microbiology Center of China Microbial Culture Collection Management Committee). The spores of Streptomyces chromogenes 7100 CGMCC 4.321 (purchased from the General Microbiology Center of the China Microbial Species Collection and Management Committee) are filamentous hooks, or coils, rarely forming rings; the spores are spherical or oval. The initial yield of Nicomycin Z from Streptomyces chromogenes TH322 was 350 mg / L.

[0028] The genomic DNA of Streptomyces chromogenes 7100 CGMCC 4.321 was extracted, digested with Sau3AI, and separated by pulse electrophoresis to recover 40-50kb DNA fragments. At the same time, the cosmid plasmid supercos1 (Stratagene) was...

Embodiment 2

[0033] Example 2: HPLC analysis and biological activity detection of the fermentation product of Streptomyces ansochromogenes TH322 sanPD CGMCC No. 3086

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

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

[0036] The SP medium was prepared as follows: 30g glucose, 30g mannitol, 5g soy peptone, 8g yeast extract, dissolve...

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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 and its special culture medium and engineering bacteria. Background technique [0002] Nicomycin belongs to the nucleoside peptide antibiotic. It has a similar structure to UDP-N-acetylglucosamine, the natural substrate of chitin synthase, and is a strong competitive inhibitor of the enzyme. Since chitin is an important part of fungal cell walls, insects and other invertebrate exoskeletons, Nicomycin can effectively inhibit fungi, insects and mites, and is non-toxic or extremely toxic to mammals, bees and plants. Low, and nikkomycin is easily degraded in nature, so nikkomycin is an ideal agricultural antibiotic. Field trials have proved that Nicomycin has a good control effect on many plant diseases, such as wheat powdery mildew, tobacco red spot, cucumber sclerotinia, tomato leaf mold and apple rot. At the same time, studies have shown that nikkomycin can effectively inhibit the deep infection of...

Claims

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Application Information

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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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