Lipase has important applications in many industrial fields,but its use for industriy applications is limited by its expensive nature.Pichia pastoris expression system has been proven to be an effective host for the production of both secreted and intracellular heterologous proteins.Glyceraldehyde-3-phosphate dehydrogenase promoter(pGAP),a constitutive promoter,is capable of providing a comparable expression level of methanol-induced alcohol oxidase promoter(pAOX1)in Pichia pastoris.In the present study,constitutive-inducible combined P.pastoris clones strains pPIC9K-pGAPZαA-RML/GS115 were generated on the base of strain pPIC9K-RML/GS115 and strain pGAPZαA-RML/GS115 in order to increase the expression of Rhizomucor miehei lipase(RML).PCR detection indicated that RML driven by GAP and AOX1 promoter was successfully integrated into the P.pastoris GS115 genome.SDS–PAGE analysis comfirmed the expression of RML and also showed that the secreted proteins consisted of an unglycosylated and glycosylated recombinant RML with different extent of glycosylation.