To implement heterologous expression of Burkholderia glumae PG1 lipase operon lipAB in Pseudomonas protegens Pf5 via Red/ET homologous recombineering. The vector p15AcmlipAB was obtained using Red/ET direct cloning technology. Then, two recombinant expression vectors pBBR1kmlipAB and pBBR1kmPapralipAB with different promoters were constructed by subcloning technology, and electrotransformated the resultant expression vectors into P.protegens Pf5. Transformants obtained by kanamycin or apramycin resistance screening. The tributyrin glyceryl trinitrate plate diffusion method and the pnitrophenol method were used for the assay of the activities of lipase，and the effect of promoter replacement on lipA expression was examined by qRTPCR. We successfully cloned the lipase operon lipAB (GenBank accession number:AJK49931.1 and AJK49932.1). After the achievement of engineering bacteria Pf5/pBBR1kmlipAB and Pf5/pBBR1kmPapralipAB, fermentation results indicated that the activity of extracellular lipase in Pf5 was accomplished. Moreover, it was found that the expression level of lipA gene was 2.1fold the original level after promoter optimization. When the flask in LB medium was fermented to 66 h, the lipase activity of Pf5/pBBR1kmlipAB supernatant was 13.51 U/mL, with that of Pf5/pBBR1kmPapralipAB supernatant was 46.85 U/mL resulting in 3.47fold variation after promoter optimization. PG1 lipase gene lipA can be successfully heterologously expressed in Pf5 via genetic engineering. Results reveal that the constitutive promoter Papra is more efficient than the original promoter PlipAB in Pf5 strain. Furthermore, the present study provides an important prerequisite for scale production and industrial application of the lipase.