Purification and Characterization of an Organic Solvent-Tolerant Lipase from Pseudomonas aeruginosa CS-2.

Ren Peng; Jinping Lin; Dongzhi Wei

An extracellular lipase secreted by Pseudomonas aeruginosa CS-2 was purified to homogeneity about 25.5-fold with an overall yield of 45.5%. The molecular mass of the lipase was estimated to be 33.9 kDa by SDS-PAGE and 36 kDa by gel filtration. The optimum temperature and pH were 50 °C and 8.0. The lipase was found to be stable at pH 4–10 and below 50 °C. Its hydrolytic activity was highest against p-nitrophenyl palmitate ( p-NPP) among p-nitrophenyl esters of fatty acids with various chain lengths. The lipase was activated in the presence of Ca2+, while it was inactivated by other metal ions more or less. EDTA significantly reduced the lipase activity, indicating the lipase was a metalloenzyme. Gum Arabic and polyvinyl alcohol 124 enhanced lipase activity but Tween-20, Tween-80, and hexadecyltrimethyl ammonium bromide strongly inhibited the lipase. It exhibited stability in some organic solvents. The lipase was activated in the presence of acetonitrile. Conversely, it was drastically inactivated by methanol and ethanol.

PSEUDOMONAS aeruginosa; LIPASES; ORGANIC solvents; ENZYMES; PSEUDOMONAS

Applied Biochemistry & Biotechnology, 2010, Vol 162, Issue 3, p733

0273-2289

Academic Journal

10.1007/s12010-009-8841-3