Applied Microbiology and Biotechnology

, Volume 85, Issue 3, pp 597–604

Co-expression of the lipase and foldase of Pseudomonas aeruginosa to a functional lipase in Escherichia coli

Biotechnologically Relevant Enzymes and Proteins

DOI: 10.1007/s00253-009-2131-4

Cite this article as:
Madan, B. & Mishra, P. Appl Microbiol Biotechnol (2010) 85: 597. doi:10.1007/s00253-009-2131-4


The lipA gene, a structural gene encoding for protein of molecular mass 48 kDa, and lipB gene, encoding for a lipase-specific chaperone with molecular mass of 35 kDa, of Pseudomonas aeruginosa B2264 were co-expressed in heterologous host Escherichia coli BL21 (DE3) to obtain in vivo expression of functional lipase. The recombinant lipase was expressed with histidine tag at its N terminus and was purified to homogeneity using nickel affinity chromatography. The amino acid sequence of LipA and LipB of P. aeruginosa B2264 was 99–100% identical with the corresponding sequence of LipA and LipB of P. aeruginosa LST-03 and P. aeruginosa PA01, but it has less identity with Pseudomonas cepacia (Burkholderia cepacia) as it showed only 37.6% and 23.3% identity with the B. cepacia LipA and LipB sequence, respectively. The molecular mass of the recombinant lipase was found to be 48 kDa. The recombinant lipase exhibited optimal activity at pH 8.0 and 37°C, though it was active between pH 5.0 and pH 9.0 and up to 45°C. Km and Vmax values for recombinant P. aeruginosa lipase were found to be 151.5 ± 29 µM and 217 ± 22.5 µmol min−1 mg−1 protein, respectively.


LipaseCo-expressionFoldaseP. aeruginosaE. coli

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  1. 1.Department of Biochemical Engineering and BiotechnologyIndian Institute of Technology DelhiNew DelhiIndia