An organic solvent-tolerant alkaline lipase from cold-adapted Pseudomonas mandelii: cloning, expression, and characterization.

A gene encoding a novel organic solvent-tolerant alkaline lipase, lipS (GenBank ID JQ071496), was cloned from cold-adapted Pseudomonas mandelii. Recombinant LipS was expressed in Escherichia coli as a 32-kDa soluble protein and was purified by standard procedures. It maintained more than 80% of its activity under alkaline conditions, pH 8-10.5, with an apparent optimum temperature range of 40-50 °C. It maintained thermal stability from 4 to 50 °C. After 1 h of incubation at 60 °C, approximately 50% of its activity remained. It retained its activity in organic solvents, and activity increased in the presence of ethanol and of DMSO. Our data indicate that LipS is an alkaline lipase with relatively high thermal stability and notable tolerance of organic solvents.