Jiangnan University

Taichi Chen

Jian Chen

Guocheng Du

Jianghua Li

Long Liu

Yaokang Wu

The present invention provides a method for promoting N-acetylglucosamine synthesis by using the GlcN6P responsive element. In the present invention,BSGNY-P-glmS-P-GNA1 is used as a starting strain, in which a CRISPRi system regulated by GlcN6P responsive element is integrated into the genome to dynamically weaken the N-acetylglucosamine synthesis competitive pathway; a GlcN6P responsive promoter is used to regulate the expression of GNA1 on the plasmid to dynamically regulate the N-acetylglucosamine synthesis pathway; and the key gene alsSD involved in the acetoin synthesis pathway is knocked out. During fed-batch fermentation with this strain in a 15 L fermenter, the production of N-acetylglucosamine reaches 131.6 g/L and no by-product acetoin is accumulated, which lays a foundation for the production of GlcNAc by industrial fermentation.

Method for promoting n-acetylglucosamine synthesis by using GlcN6P responsive element

The present invention discloses a method for producing N-acetylglucosamine (GlcNAc) by co-utilizing glucose and xylose based on CRISPRi, and belongs to the field of genetic engineering. According to the method,BSGNY-P-glmS-P-GNA1 is used as an original strain, dCas9 induced by xylose and three sgRNA expression fragments targeting to genes zwf, pfkA and glmM respectively are integrated on the genome, and the strain is fermented in a shake flask, so that the titer of GlcNAc reaches 20.5 g/L, the yield of GlcNAc is 0.612 g/g glucose, at the same time, the efficient co-utilizing of glucose and xylose by the recombinantis achieved, and the foundation for further metabolic engineering transformation of theto produce GlcNAc and industrialization thereof is laid.

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Wuxi, China

Taichi Chen