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Recombinant bacteria for expressing plectasin and application thereof

A technology of plectasin and antibacterial agents, applied to recombinant bacteria expressing plectasia and its application fields, can solve the problems of increasing the difficulty of curing diseases, patients and social and economic burdens, and achieve the effect of increasing expression

Active Publication Date: 2015-04-01
GUANGDONG HINAPHARM PHARMA CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] With the widespread use of antibiotics in the clinic, the problem of bacterial resistance is becoming more and more serious, increasing the difficulty of curing diseases and bringing serious economic burdens to patients and society

Method used

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  • Recombinant bacteria for expressing plectasin and application thereof
  • Recombinant bacteria for expressing plectasin and application thereof
  • Recombinant bacteria for expressing plectasin and application thereof

Examples

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Embodiment 1

[0021] Example 1: Construction method of recombinant Lactococcus lactis expressing plectasin

[0022] 1. Construction of recombinant strains

[0023] The nucleotide sequence of the Usp45 protein signal peptide was cloned from Lactococcus lactis MG1363, and the Usp45 gene fragment was combined with the shuttle plasmid pMG36e (the pMG36e map is shown in figure 1 shown) to construct plasmid pGMN1; connect the plasmid pGMN1 with the Pnis-MCS segment gene amplified from the lactic acid bacteria plasmid pNZ8408 to construct the plasmid pGMN2; artificially synthesized lactic acid bacteria codon-biased plectasin gene fragment, the optimized strain The base sequence of silk mycin is: GGT TTT GGT TGT AAT GGT CCA TGG GAT GAT GAT GAT ATG CAA TGT CAT AAT CAT TGT AAA TCA ATT AAA GGT TAT AAA GGT GGT TAT TGT GCT AAA GGT GGT TTT GTT TGT AAA TGT TAT (SEQ ID NO: 1), the optimized plectasin gene fragment was connected to plasmid pGMN2 to obtain plasmid pGMN3, in pGMN3 Usp45 was located upst...

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Abstract

The invention discloses recombinant bacteria for expressing plectasin and an application thereof. The classification name of the recombinant bacteria is lactococcus lactis NZ9700-pGMN3, the preservation serial number of the recombinant bacteria is CCTCC NO: M2014611, the recombinant bacteria can achieve upper-high expression of plectasin protein, the expression quantity of the recombinant bacteria far exceeds that of other anode recombinant bacteria, and the recombinant bacteria can be directly used for preparing anti-bacteria medicament, have no hidden safety risks and hardly have any side effects. The recombinant bacteria have the functions of inhibition of methicillin-resistant staphylococcus aureus, streptococcus agalactiae, streptococcus pyogenes, enterococcus faecalis and the like. Bacterial strains of the recombinant bacteria can achieve efficient and continuous expression under self-induction, the recombinant bacteria can express and secrete fusion protein containing plectasin in the fermentation process, the total yield of protein is more than 1.41g / L, plectasin accounts for more than 70%, and the recombinant bacteria have the potential of treating mammitis and infectious diseases of a genital tract.

Description

technical field [0001] The invention relates to a strain of new bacterial strain and its application in treating inflammation, in particular to a strain of recombinant bacteria expressing plectasin and its application. Background technique [0002] Lactic acid bacteria (lactis acid bacteria, LAB) is a class of Gram-positive bacteria that can ferment carbohydrates and produce lactic acid in large quantities. It includes more than 10 genera such as Lactococcus, Lactobacillus, and Bifidobacteria. Long-term application has proved its non-pathogenicity. Lactic acid bacteria are common bacteria in the intestines of humans and most animals, and are widely distributed in the digestive system, respiratory system, secretory system, oral system and feces. [0003] Defensin is a host defense peptide widely present in organisms, which constitutes the first line of defense against pathogenic microorganisms. Generally 2-6kDa, rich in positively charged amino acid residues, containing 6-8...

Claims

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Application Information

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IPC IPC(8): C12N1/21A61K35/744A61P31/00A61P31/04A61P15/00A61P17/00A61P1/00C12R1/46
CPCA61K35/744C07K14/37
Inventor 周玉岩孙丹丹逯佩凤杨虹坤
Owner GUANGDONG HINAPHARM PHARMA CO LTD
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