The invention provides a recombinant Bacillus subtilis, construction method and use thereof, wherein the cell's own FMMs are used as a space scaffold, and a multi-enzyme complex is constructed from specific marker proteins FloA and FloT, such that an artificial substrate channel is formed, and the cell metabolic burden is effectively reduced. The recombinant Bacillus subtilis of the invention can efficiently synthesize GlcNAc without affecting cell life activity, and can also limit the toxic intermediate metabolite GlcN-6-P near the plasma membrane to reduce or eliminate its inhibition on cell activity. In the process of shaking flask fermentation using complex medium, the yield of acetyl glucosamine of the control strain BSG-C was only 0.45 g.Math.L.sup.−1, while that of BSG-AT, BSG-ATP, BSG-ATPB increased to 5.29 g.Math.L.sup.−1, 6.22 g.Math.L.sup.−1 and 8.48 g.Math.L.sup.−1 respectively. The construction method of recombinant Bacillus subtilis is simple, easy to use and has a good application prospect.

This Invention discloses a method for production of N-Acetyl-D-Glucosamine and/or D-Glucosamine Salt by microbial fermentation. The method is intended to manufacture N-Acetyl-D-Glucosamine and/or D-Glucosamine Salt in higher efficiency and higher yield, by expression of vitreoscilla hemoglobin in microorganism.

The present invention discloses a process for producing N-acetyl-D-glucosamine and D-glucosamine salts by microbial fermentation. The invention includes a method to produce N-acetyl-D-glucosamine and/or D-glucosamine salts with higher efficiency and higher yield by increasing the effect of N-acetyl-D-aminomannose-6-phosphate epimerase in microorganisms.

This Invention discloses a method for production of N-Acetyl-D-Glucosamine and/or D-Glucosamine Salt by microbial fermentation. The method is intended to manufacture N-Acetyl-D-Glucosamine and/or D-Glucosamine Salt in higher efficiency and higher yield, by increasing the effects of N-Acetyl-D-Mannosamine Kinase.

The present invention relates to a mutant microorganism in which a gene that encodes phosphofructokinase-2 is disrupted or deleted to reduce glycolytic flux to thereby improve the ability of the microorganism to produce N-acetylglucosamine, and to a method of producing N-acetylglucosamine using the mutant microorganism. The mutant microorganism according to the present invention has advantages in that it has high resistance to various chemical substances, grows rapidly, is easily cultured, and produces N-acetylglucosamine with high efficiency, indicating that it is useful for production of a large amount of N-acetylglucosamine.

The invention provides a recombinant bacillus subtilis, construction method and use thereof, wherein the cell's own FMMs are used as a space scaffold, and a multi-enzyme complex is constructed from specific marker proteins FloA and FloT, such that an artificial substrate channel is formed, and the cell metabolic burden is effectively reduced. The recombinant Bacillus subtilis of the invention can efficiently synthesize GlcNAc without affecting cell life activity, and can also limit the toxic intermediate metabolite GlcN-6-P near the plasma membrane to reduce or eliminate its inhibition on cell activity. In the process of shaking flask fermentation using complex medium, the yield of acetyl glucosamine of the control strain BSG-C was only 0.45 g.L.sup.1, while that of BSG-AT, BSG-ATP, BSG-ATPB increased to 5.29 g.L.sup.1, 6.22 g.L.sup.1 and 8.48 g.L.sup.1 respectively. The construction method of recombinant Bacillus subtilis is simple, easy to use and has a good application prospect.

The present invention discloses a process for producing N-acetyl-D-glucosamine and D-glucosamine salts by microbial fermentation. The invention includes a method to produce N-acetyl-D-glucosamine and/or D-glucosamine salts with higher efficiency and higher yield by increasing the effect of N-acetyl-D-aminomannose-6-phosphate epimerase in microorganisms.

This disclosure is in the technical field of synthetic biology and metabolic engineering. More particularly, this disclosure is in the technical field of cultivation of metabolically engineered cells. This disclosure describes a method for the production of a glycosylated product derived from UDP-GlcNAc and comprising a di- or oligosaccharide that is composed of at least two different monosaccharide subunits by a cell as well as the separation of the glycosylated product from the cultivation. Furthermore, this disclosure provides a metabolically engineered cell for production of a glycosylated product derived from UDP-GlcNAc and comprising a di- or oligosaccharide that is composed of at least two different monosaccharide subunits. This disclosure also provides a cell excreting a di- or oligosaccharide out of the cell.

This Invention discloses a method for production of N-Acetyl-D-Glucosamine and/or D-Glucosamine Salt by microbial fermentation. The method is intended to manufacture N-Acetyl-D-Glucosamine and/or D-Glucosamine Salt in higher efficiency and higher yield, by expression of vitreoscilla hemoglobin in microorganism.

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.