Construction method and application of microorganism capable of realizing high production of lacto-N-tetrose

Abstract

Disclosed are a construction method and application of a microorganism capable of realizing high production of lacto-N-tetrose, belonging to the field of microbial genetic engineering. The present disclosure employs a strain which has been constructed in the early stage for efficiently producing a precursor lacto-N-triose II as an original strain to synthesize a key gene of the lacto-N-tetrose via over-expression, thus enabling the strain to have a synthesis capability of producing the lacto-N-tetrose. The present disclosure improves the synthesis of the lacto-N-tetrose by screening a high-efficiency β-1,3-galactosyl transferase gene, and reasonably designing the co-expression of the β-1,3-galactosyl transferase gene and a key UDP-glucose 4 epimerase gene (galE) for strengthening a UDP-galactose pathway on a vector pCDFDuet-1. In a shake flask experiment, the lacto-N-tetrose production capacity of Escherichia coli is 3.04 g/L. The lacto-N-tetrose yield in a 3 L fermentation tank reaches 25.49 g/L. Therefore, the microorganism has an industrial application prospect.

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