Construction method and application of a genetically engineered bacterium for highly efficient synthesis of lipopolysaccharide

The genetically engineered bacteria of E.coli W3110△npr△yhbJ△fabF△PBAD (lpxD-fabZ-lpxA-lpxB) constructed through gene editing technology solved the problem of low yield of lipopolysaccharides in natural E. coli strains, and achieved the effect of efficient synthesis of lipopolysaccharides, which is suitable for large-scale production and application.

CN115820524BActive Publication Date: 2025-06-24JIANGNAN UNIV
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Patent Information

Application Number
CN202211538742.0
Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Filing Date
2022-12-01
Publication Date
2025-06-24
Estimated Expiration
2042-12-01

AI Technical Summary

Technical Problem

Natural E. coli strains have low yields in the synthesis of lipopolysaccharides, which are difficult to meet the needs of vaccine adjuvants and immune experimental reagents.

Method used

A non-resistant E. coli genetically engineered bacteria was constructed through gene editing technology. The specific methods included deleting npr, yhbJ and fabF genes in the E.coli W3110 strain, and inserting arabinose-induced promoter PBAD in front of the lpxD-fabZ-lpxA-lpxB gene cluster to form the genetically engineered bacteria WOZF01.

Benefits of technology

The synthesis of lipopolysaccharides was increased, with a yield of 13.0 mg/gDCW, which was 91.2% higher than that of wild-type E.coli W3110. The strain has no resistance marks and is in good growth condition, which is suitable for large-scale production.

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Abstract

The present invention discloses a method for constructing a genetically engineered bacterium capable of efficiently synthesizing lipopolysaccharide and its application, belonging to the technical field of genetic engineering. The genetically engineered bacterium of the present invention, E. coli W3110△npr△yhbJ△fabF△P BAD(lpxD ‑fabZ‑lpxA‑lpxB), has undergone deletion mutations of three genes, npr, yhbJ, and fabF, and at the same time, a promoter P has been inserted in front of the chromosomal gene cluster lpxD‑fabZ‑lpxA‑lpxB sequence BAD . The genetically engineered bacterium of the present invention has no resistance markers, has good growth conditions, can be adapted to large-scale production, and the yield of synthesized lipopolysaccharide is 13.0 mg / g DCW, which is 91.2% higher than that of the wild-type E. coli W3110.
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