Abstract: The invention discloses a method for improving the yield of Bacillus subtilis acetylglucosamine, which belongs to the technical field of genetic engineering. In the invention, the recombinant Bacillus subtilis S5 (S5-PxylA-glmS-P43-GNA1) is taken as a starting strain, and the glmS ribozyme is integrated into the mid of rbs and the promoter sequence of the glmM and pfkA gene, respectively. The ribozyme mutant has the advantage of prolonging the stability of the mRNA and integrated into the mid of rbs and the promoter sequence of the pgi gene. The yield of GlcNAc of the recombinant strain reaches 11.79-20.05 g/L. This laid the foundation for the further metabolic engineering of Bacillus subtilis to produce GlcNAc.

Abstract: The invention discloses a method for improving the yield of Bacillus subtilis acetylglucosamine, which belongs to the technical field of genetic engineering. In the invention, the recombinant Bacillus subtilis S5 (S5-PxylA-glmS-P43-GNA1) is taken as a starting strain, and the glmS ribozyme is integrated into the mid of rbs and the promoter sequence of the glmM and pfkA gene, respectively. The ribozyme mutant has the advantage of prolonging the stability of the mRNA and integrated into the mid of rbs and the promoter sequence of the pgi gene. The yield of GlcNAc of the recombinant strain reaches 11.79-20.05 g/L. This laid the foundation for the further metabolic engineering of Bacillus subtilis to produce GlcNAc.

Abstract: The present invention provides a recombinant Bacillus subtilis for producing acetylglucosamine and construction method thereof. The recombinant Bacillus subtilis is obtained by deletion of glmS ribozyme of Bacillus subtilis for regulating expression of glucosamine synthase, and insertion of a terminator and a constitutive promoter. The method comprises constructing a deleting cassette of a glmS ribozyme encoding gene, which includes an upstream homologous fragment, a resistance gene, a terminator sequence, a constitutive promoter sequence and a downstream homologous fragment in sequence; and transforming the deleting cassette into Bacillus subtilis, to obtain the recombinant Bacillus subtilis. In the invention, glucosamine synthase gene (glmS) ribozyme is deleted by homologous recombination, and in host cells, GlcN6P feedback inhibition of expression of glucosamine synthase gene glmS is blocked, and the accumulation of acetylglucosamine is improved.

Abstract: The present invention provides a recombinant Bacillus subtilis for producing acetylglucosamine and construction method thereof. The recombinant Bacillus subtilis is obtained by deletion of glmS ribozyme of Bacillus subtilis for regulating expression of glucosamine synthase, and insertion of a terminator and a constitutive promoter. The method comprises constructing a deleting cassette of a glmS ribozyme encoding gene, which includes an upstream homologous fragment, a resistance gene, a terminator sequence, a constitutive promoter sequence and a downstream homologous fragment in sequence; and transforming the deleting cassette into Bacillus subtilis, to obtain the recombinant Bacillus subtilis. In the invention, glucosamine synthase gene (glmS) ribozyme is deleted by homologous recombination, and in host cells, GlcN6P feedback inhibition of expression of glucosamine synthase gene glmS is blocked, and the accumulation of acetylglucosamine is improved.