Engineering bacterium producing gentamicin A and application thereof

Abstract

The invention belongs to the technical field of medicines, and relates to an engineering bacterium producing gentamicin A and an application thereof. The engineering bacterium is micromonospora purpurea (Micromonospora purpurea) KD1989 with a geneD1 inactivating function, which is registered and collected in China General Microbiological Culture Collection Center (CGMCC) with the collection No. CGMCC No. 8598. The genD1 function in gentamicin biosynthetic gene cluster is inactivated through a gene knockout method, and according to the gene knockout method, shuttle plasmids (such as pGD14 plasmid) are built and knocked out, the pGD14 plasmid is transformed into a gentamicin producing strain, double crossover engineering bacterium micromonospora purpurea GKD1989(delta genK+delta genD1) is screened, the engineering bacterium is fermented, and gentamicin A is determined by detection of metabolites. The engineering bacterium producing gentamicin A produces the single component gentamicin A, which has not been reported in China and foreign countries, so that the industrial production of gentamicin A can be realized; and the production process is simple and easily controlled, the extraction is convenient, the cost is low, and the engineering bacterium meets clean production levels, has stable product quality, and is the first innovative product in the world.

Description

technical field [0001] The invention belongs to the field of antibiotic pharmacy, and relates to a gentamicin A-producing engineering bacterium and its application. Background technique [0002] Micromonospora can produce abundant secondary metabolites, especially aminoglycoside antibiotics, such as gentamicin, sisomicin and fortimicin. The distribution of these micromonospora is peculiar, the growth is slow and the cell wall structure is special, so the research progress is slow. In addition, due to the lack of a general gene operating system, the genetic manipulation of Micromonospora is difficult, making the research on its molecular biology lag behind that of Streptomyces. [0003] Gentamicin (Gentamicin, GM) is produced by Micromonospora rubrum ( Micromonospora purpurea ) or Micromonospora aculeatus ( Micromonospora echinospora ) and other aminoglycoside broad-spectrum antibiotics produced by fermentation, have strong antibacterial effects on a variety of Gram-negat...

AI Technical Summary

Problems solved by technology

Streptomyces genetic engineering, which emerged in the 1980s, has made some progress in the cloning of antibiotic biosynthetic genes, yield improvement, component improvement, and hybrid antibiotic production. However, the application in Micromonospora Development is slow

Although gentamicin A is extremely important, it is an intermediate in the biosynthesis of amino oligosaccharides. It is very difficult to obtain this compound. It is rare to make a significant breakthrough at home and abroad. It has not been able to achieve industrialization in the world so far. This compound has been produced in large quantities. Therefore, it hinders the research and development of this class of antiprotozoal, antiviral and antitumor drugs

With the help of the operating skills of microbial molecular genetics, on the basis of establishing a systematic genome transformation of Micromonospora, the present invention successfully realized the genome improvement of Micromonospora crimson, and obtained the main production Qing for the first time in the world. The gentamycin A engineering bacteria has not been reported at home and abroad, which has laid a solid foundation for the industrialization of gentamicin A

Images