Method for Improving Fermentation Level of Salinomycin

A technology of fermentation level and salinomycin, applied in the field of bioengineering, can solve problems such as unclear fermentation level of antibiotics, and achieve the effects of reducing fermentation cost, increasing fermentation yield, and increasing fermentation yield

Active Publication Date: 2017-10-10
SHANGHAI JIAO TONG UNIV
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Problems solved by technology

However, for Streptomyces albicans DSMZ41398, whether it is possible to change the flow and direction of precursor metabolic flow by inactivating the active type I polyketide synthase biosynthesis gene cluster, so that more of it flows to the biosynthesis of salinomycin pathways, thereby increasing the fermentation levels of antibiotics is not clear

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  • Method for Improving Fermentation Level of Salinomycin
  • Method for Improving Fermentation Level of Salinomycin
  • Method for Improving Fermentation Level of Salinomycin

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Experimental program
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Embodiment

[0027] Step 1: Construction of plasmid pLQ52

[0028] Using the genomic DNA of Streptomyces albicans DSMZ41398 as a template, the interruption region (1,200,120bp- 1,200,750bp) the homology arm of 1.41kb on the left and the homology arm of 1.35kb on the right, the correctness of the homology arm was confirmed by gene sequencing; the SpeI / KpnI site of plasmid pJTU1278 was inserted from the interrupted region (1,200,120bp-1,200,750 bp) The 1.41kb PCR fragment (SpeI / HindIII) on the left and the 1.35kb PCR fragment (HindIII / KpnI) on the right of the interruption region (1,200,120bp-1,200,750bp); the plasmid pLQ52 for gene cluster inactivation was obtained by the above method. Under the condition of water bath at 37 degrees Celsius, two target bands of 1.35 kb and 1.41 kb can be observed by using three restriction enzymes SpeI, HindIII and KpnI for enzyme digestion, indicating that the plasmid is constructed correctly.

[0029] Step 2: Introduce the plasmid pLQ52 for recombination...

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Abstract

A method for improving the fermentation level of salinomycin is to reduce its response to salinomycin biosynthesis by inactivating the type I PKS biosynthesis gene cluster located at SLNWT0868‑SLNWT0920 (1,173,724bp‑1,276,916bp) in Streptomyces albicans DSMZ41398. Body competition, to achieve the increase of salinomycin production. By adopting the method of the invention, the final fermentation yield of salinomycin can be increased by more than 8 times, and the laboratory shake flask level can reach 6.3g / L.

Description

technical field [0001] The invention relates to bioengineering technology, in particular to a method for improving the fermentation level of salinomycin. technical background [0002] Actinomycetes are a class of Gram-positive bacteria with high GC content, and more than 60% of known antibiotics are synthesized by actinomycetes. Streptomyces is a class of higher actinomycetes, which has strong antibiotic synthesis ability and complex morphological differentiation, so it has always been concerned by people. Polyketides are a class of important secondary metabolites produced by actinomycetes, which have extremely important applications in the fields of anti-tumor, anti-parasite, and antibiotics. Previous experiments have proved that the biosynthesis of polyketides mainly depends on type I, type II and type III polyketide biosynthesis gene clusters, and the main precursors used are acetyl-CoA, malonyl-CoA, methyl Malonyl-CoA, ethylmalonyl-CoA, etc. [0003] Salinomycin is a ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P17/18C12R1/47
Inventor 白林泉芦晨阳蒋明康前进
Owner SHANGHAI JIAO TONG UNIV
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