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Genetically engineered bacterium for producing natamycin as well as construction method and application of genetically engineered bacterium

A technology for producing natamycin and genetically engineered bacteria, which is applied in the field of genetic engineering to achieve huge social and economic benefits, reduce production costs, and increase production

Active Publication Date: 2014-07-16
SHANDONG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Cholesterol oxidase was first used to detect the cholesterol content in animal serum. In recent years, it has been found that it also has good application value in biochemical pharmaceuticals, food processing, and insect-resistant genetic engineering. The application of fungicides in terms of biological yield has not been reported in the literature

Method used

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  • Genetically engineered bacterium for producing natamycin as well as construction method and application of genetically engineered bacterium
  • Genetically engineered bacterium for producing natamycin as well as construction method and application of genetically engineered bacterium

Examples

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Embodiment 1

[0024] Construction of recombinant plasmid pSET152-pimE:

[0025] (1) According to the Emr gene promoter Pemr* sequence recorded in GenBank and the downstream fused gene fragments, respectively design upstream and downstream primers:

[0026] Upstream primer Pemr-F:

[0027] 5'-GC GAGGATGAAGAGGATGAGGAGGC-3' (the underlined solid line marks the XbaI restriction site);

[0028] Downstream primer Pemr-R:

[0029] 5'-CTGGTTCTCGAACAT -3' (the underlined dashed line indicates the partial sequence of the downstream fragment).

[0030] Using the plasmid pMG36e as a template, PCR amplification obtained the Emr gene promoter Pemr* sequence:

[0031] (2) According to the pimE gene sequence recorded in GenBank and the upstream fused gene fragment, design upstream and downstream primers respectively:

[0032] Upstream primer pimE-F:

[0033] 5'-CCAAATTAAAGAGGGTTATAATGTTCGAGAACCAG-3' (the underlined dashed line indicates the partial sequence of the upstream fragment);

[0034] Dow...

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Abstract

The invention discloses a genetically engineered bacterium for producing natamycin as well as a construction method and application of the genetically engineered bacterium. The genetically engineered bacterium is classified and named Streptomyces gilvosporeus and is collected with the serial number of CGMCC No.8901 in the China general microbiological culture collection center on march 7th, 2014. The construction method comprises the steps of constructing a recombinant plasmid by utilizing a cholesterol oxidase gene pimE; integrating the gene pimE into a Streptomyces gilvosporeus genome in an Escherichia coli and Streptomyces gilvosporeus conjugal transfer way; sieving to obtain a positive transconjugant. The genetically engineered bacterium can be used for increasing the density of pimE proteins in a fermentation liquid, and as signaling proteins, the pimE proteins can be used for improving the transcriptional level of a natamycin gene cluster and greatly increasing the fermentation yield of natamycin, so that the production cost is reduced, and huge social and economic benefits are brought.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a genetically engineered bacterium for producing natamycin and its construction method and application. Background technique [0002] Natamycin (Natamycin), also known as Pimaricin, is a 26-ring tetraene macrolide antifungal agent, which has the characteristics of natural, broad-spectrum, high efficiency and low toxicity. This antibiotic can not only effectively inhibit the growth of yeast and mold, but also effectively prevent the production of aflatoxin by filamentous fungi. In the field of medicine, it can be widely used in the treatment of diseases caused by fungi. In the food field, due to its broad-spectrum and low toxicity to mammals, natamycin was approved by the US FDA in 1982 as a food preservative and classified as a GRAS (generally regard as safe) product In 1997, the Ministry of Health of my country officially approved natamycin as a food pre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/63C12P19/62C12R1/465
Inventor 朱希强王淼王绍花宗工理刘飞侯重文于林艳
Owner SHANDONG UNIV
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