1.1.1.B60 Escherichia coli(gosldh-lrenox) producing both GoSLDH for D-sorbitol oxidation and LreNOX (NAD(P)H oxidase from Lactobacillus reuteri) for NADP+ regeneration is generated and used for L-sorbose production. L-Sorbose production by Escherichi coli(gosldh-lrenox) reaches 4.1 g/l after 40 min, which was 20.5fold higher than that of Escherichia coli(gosldh). This system reduces the NADPH inhibition effect in the GoSLDH reaction and enables high production of L-sorbose from D-sorbitol 1.1.1.B60 expressed in Escherichia coli BL21(DE3)-CodonPlus RIL cells