Application of relA gene in increase of moenomycin yield of streptomyces bambergiensis and strain

A technology of Streptomyces Bambergeri and moenomycin, applied in the field of application and bacterial strains of relA gene in improving the yield of Streptomyces Bambergeri moenomycin, can solve the problem of not being able to produce spores and improve the quality of Streptomyces Bambergeri problems such as plant yield and yield increase, to achieve the effect of increasing yield

Inactive Publication Date: 2013-11-27
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Gomez-Escribano et al. found that blocking the relA gene of Streptomyces clavuligerus could not form aerial hyphae and produce spores, but the activity of clavulanic acid and cephamycin C Yield significantly increased (Gomez-Escribano J P, Martín J F, Hesketh A, et al. Streptomyces clavuligerus relA-null mutants overproduce clavulanic acid and cephamycin C: negative regulation of secondary metabolism by (p)ppGpp. Microbiology, 2008, 154:744- 755.)
At present, there is no report on the use of relA gene to improve moenomycin production in Streptomyces bambergiensis

Method used

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  • Application of relA gene in increase of moenomycin yield of streptomyces bambergiensis and strain
  • Application of relA gene in increase of moenomycin yield of streptomyces bambergiensis and strain
  • Application of relA gene in increase of moenomycin yield of streptomyces bambergiensis and strain

Examples

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Effect test

Embodiment 1

[0020] Example 1: Construction of Streptomyces Bamburgh relA Gene Blocking Engineering Bacteria 1203

[0021] (1) Cloning and sequencing of the relA gene fragment of Streptomyces Bambergeri

[0022] The relA gene sequences in Streptomyces coelicolor A3(2) and Streptomyces clavulus ATCC27064 were analyzed by bioinformatics, and two pairs of primers P1 / P2 and P3 / P4 were designed according to the conserved region of the relA gene, and Streptomyces bamberger CGMCC NO.4.1439 (=ATCC13879) Genomic DNA was used as a template, and the two gene fragments of the 5' and 3' conserved regions of the relA gene were amplified by PCR respectively (hereinafter referred to as the left arm and the right arm of the relA gene, respectively), with a size of about 910bp (SEQ ID No. 1) and 705bp (SEQ ID No. 2). The sequences of the above two pairs of primers are as follows:

[0023] P1: 5′-GA GAATTC CTCTTGCCAGACGAGGCC-3′ (the underline is the EcoRI restriction site)

[0024] P2: 5′-AA GATATC GC...

Embodiment 2

[0046] Embodiment 2: Moenomycin fermentation test and HPLC analysis of Streptomyces Bamburgh

[0047] First, the original starting strain CGMCC NO.4.1439 of Streptomyces Bamberger and the relA gene blocking project 1203 (CGMCC No.7237) were respectively inoculated into moenomycin seed medium (prepared according to the following composition: 30 g of soluble starch, 5 g of yeast powder , soybean cake powder 10g, MgSO 4 ·7H 2 O0.5g, K 2 HPO 4 ·3H 2 O0.5g, 1L of distilled water, pH7.2~7.5, sterilized at 115°C for 30min), cultured on a shaker at 30°C at 200r / min for 48h. Then, 5 mL of seed culture was inoculated into 50 mL moenomycin liquid fermentation medium according to 10% inoculum size (prepared according to the following composition: 50 g of soluble starch, 5 g of yeast powder, 30 g of soybean cake powder, (NH 4 ) 2 SO 4 3g, MgSO 4 ·7H 2 O0.5g, K 2 HPO 4 ·3H 2 O0.5g, CaCO 3 5g, 1L of distilled water, pH7.2~7.5, sterilized at 115°C for 30min), fermented on a shaki...

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Abstract

The invention provides an application of a relA gene in increase of the moenomycin yield of streptomyces bambergiensis, as well as obtained relA gene blocking engineering bacteria, namely, streptomyces bambergiensis 1203 for producing moenomycin and an application of the streptomyces bambergiensis 1203, wherein the strain is preserved in China General Microbiological Culture Collection Centre, the preservation number is CGMCC No. 7237, and the collection date is January 31, 2013. According to the application and the strain provided by the invention, relA homologous genes on streptomyces bambergiensis chromosomes are blocked by virtue of a molecular biological technique, thus increasing the moenomycin yield, and overcoming the defects of blindness, high screening workload and the like of mutagenesis adopting traditional physical and chemical methods.

Description

(1) Technical field [0001] The present invention relates to the application of relA gene in increasing the yield of moenomycin of Streptomyces bambergiensis, and the obtained relA gene blocking engineering bacteria for producing moenomyces-Streptomyces bambergiensis ) 1203 and its applications. (2) Background technology [0002] Moenomycin is a class of phosphoglycolipid antibiotics, it has at least four active components: A, B1, B2 and C, of ​​which A is the main component. Moenomycin A can be produced by four species of Streptomyces: Streptomyces bambergiensis ATCC13879, Streptomyces ghanaensis ATCC14672, Streptomyces geysiriensis ATCC15303, Streptomyces ederensis ATCC15304 , of which Streptomyces bambergiensis is the main moenomycin-producing bacteria. Moenomycin can directly inhibit the peptidoglycan glycosyltransferase (PGT) in the biosynthetic pathway of bacterial cell wall, thereby effectively inhibiting the growth and reproduction of bacteria. Moenomycin is an ide...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P19/26C12R1/465
Inventor 赵春田陈艳萍余志良裘娟萍
Owner ZHEJIANG UNIV OF TECH
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