The invention discloses recombinant escherichia coli with glucosamine accumulation and application thereof, and belongs to the technical field of genetic engineering. The escherichia coli is used as ahost, the recombinant escherichia coli is obtained by knocking out four genes of an acetylglucosamine transporter encoding gene nagE, a mannose phosphate transporter encoding gene manXYZ, a deacetylase gene nagA and a deaminase gene nagB gene in the glucosamine synthesis pathway of the escherichia coli, and the recombinant escherichia coli is applied to the production of the glucosamine to achieve the purpose of blocking the transport pathway of glucosamine from the extracellular to the intracellular. By means of overexpression of a glucosamine synthetase encoding gene and a glucosamine acetylase encoding gene, the synthesis pathway of the glucosamine is enhanced, and the yield of the glucosamine is increased accordingly.