Optimized production method of abamectin

A technology of abamectin and its production method, applied in the field of biomedicine, can solve problems such as toxicity, no relevant research reports on genes, complex chemical components, etc.

Inactive Publication Date: 2012-06-27
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Second, the environment in which Streptomyces directly contacts is rich in compounds, the chemical composition is extremely complex, and many compounds are toxic to it
[0009] At present, there is no relevant research report on the genes related to the efflux of abamectin

Method used

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  • Optimized production method of abamectin
  • Optimized production method of abamectin
  • Optimized production method of abamectin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1 The improvement of Streptomyces avermitilis target antibiotic output

[0039] With the application of molecular biology and bioengineering technology in the field of antibiotic breeding, genetic engineering breeding technology has become the main means of improving industrial production strains. The invention improves the output of antibiotics by improving the active efflux efficiency of antibiotics. The specific operation is to connect the target gene responsible for antibiotic efflux to the carrier in vitro through genetic engineering technology, and then transfer it into recipient cells, so that the antibiotic efflux gene can be efficiently expressed and inherited in the recipient.

[0040] (1) Construction of plasmids (used for high expression of target genes transferred into hosts)

[0041] Plasmid construction for overexpression of sav933 and sav934 genes:

[0042] The F cosmid Fosmid 20B4 of Streptomyces avermitilis NRRL 8165 genomic library was di...

Embodiment 2

[0078] Embodiment 2 prepares the industrial high-yield bacterial strain of Abamectin

[0079] Step 1. Transfer of plasmids into Streptomyces hosts and screening and verification of conjugative transfers

[0080] The high-copy plasmid pFAveT of sav933 and sav934 was transferred into Streptomyces avermitilis industrial strain 3-115 cells by conjugative transfer, through the verification of thiostrepton resistance, the plasmid was extracted from Streptomyces and reversed Escherichia coli DH10B, and screened A correct genetically engineered strain was named 3-115 / pFAveT. Then the empty vector pJTU1278 was transformed into Streptomyces avermitilis 3-115 cells by conjugative transfer, and the control strain 3-115 / pJTU1278 was screened by the same method.

[0081] The above 3-115 / pFAveT, that is, Streptomyces avermitilis 3-115 / pFAveT was submitted on December 15, 2011. ), the General Microorganism Center CGMCC of the China Committee for Culture Collection of Microorganisms, and its...

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Abstract

An optimized production method of abamectin belongs to the technical field of biomedicine, and includes of constructing over-expression plasmid pFAveT containing gene sav933 and gene sav934, transferring the expression plasmid into streptomyces avermitilis 3-115, and finally realizing yield increase. The yield of specific antibiotics is increased by means of over expression of efflux gene in strains for industrial production of antibiotics.

Description

technical field [0001] The invention relates to a method in the technical field of biomedicine, in particular to an optimized production method of abamectin. Background technique [0002] Abamectin has high-efficiency and broad-spectrum activity against various nematodes and human parasites, and is widely used in the prevention and control of diseases and insect pests in agriculture and animal husbandry. A total of eight abamectin components can be isolated from Streptomyces avermitilis, and the only one with anti-insect activity is component B1a. In addition, Streptomyces avermitilis also produces oligomycin, a toxic by-product during the fermentation process. The whole genome sequencing of Streptomyces avermitilis has been completed, and the abamectin biosynthetic gene cluster has also been cloned and mapped (Ikeda H, Ishikawa J, Hanamoto A, Shinose M, Kikuchi H, Shiba T, Sakaki Y, Hattori M, Omura S .Complete genome sequence and comparative analysis of the industrial mi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/76C12N1/21C12P19/62C12R1/465
Inventor 白林泉邱竞帆朱冬青邓子新
Owner SHANGHAI JIAO TONG UNIV
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