The invention discloses a recombinant lipase mutant, engineering bacteria and application of the recombinant lipase mutant in the preparation of (2S,3R)-N-acetyl-piperidine-2,3-dicarboxylic acid dimethyl ester. The mutant is prepared by the step of carrying out single mutation or combined mutation on a site 140, a site 141, a site144, a site 189 and a site 190 of an amino acid sequence representedby SEQ ID NO.2. Compared with wild enzyme, the mutant has the advantages that the catalytic activity and the substrate tolerance are greatly improved, and the time consumed in the reaction progress is obviously shortened. Compared with (S,S)-2,8-diazabicyclo[4,3,0]decane prepared by virtue of a chemical method, the produced prepared by virtue of the technique provided by the invention has the advantages of high stereoselectivity, relatively mild reaction condition, low facility request and environmental friendliness, and the reaction cost is lowered.