Expression of Recombinant Lipase from Serratia marcescens LII61 in Escherichia coli

Abstract

Recombinant lipase encoding gene of Serratia marcescens LII61 has been successfully expressed in Escherichia coli. The gene was amplified from genomic DNA of S. marcescens LII61 by PCR method using lipase specific primers. The amplified DNA was then subsequently inserted in pGEM ® -T easy vector and transformed into E. coli JM109. The inserted DNA fragment in the plasmid was analyzed for studying the lipase gene sequence. The recombinant gene was subcloned in pET-28b(+) vector/ E. coli BL21(DE3) system. The positive clones were selected by growing E. coli cells in antibiotic medium and lipase-specific medium. The analysis of recombinant DNA showed that the lipase gene of S. marcescens LII61 was 1845 bp in size. The inserted gene in the pGEM-T easy vector was composed of 1842 bp lipase gene and flanked by several restriction enzyme as stated on the map vector. The recombinant E. coli BL21 (DE3) showed a fluorescent orange color on LB-IPTG-rhodamine B-olive oil agar plate, indicated that the recombinant bacteria were able to express the lipase gene from S. marcescens LII61. This report is the first endeavor on cloning and expression of lipase from Indonesian isolate of S. marcescens.