Purification and characterization of an alkaline lipase from Pseudomonas aeruginosa isolated from putrid mineral cutting oil as component of metalworking fluid.
Extracellular lipase was isolated and purified from the culture broth of Pseudomonas aeruginosa, an extremophile which naturally grows in water-soluble mineral cutting oil (pH 10) used as metalworking fluid (MWF) for cooling and lubrication in industrial metalworking processes. The molecular mass of the purified lipase was estimated by SDS-PAGE to be 54 kDa. The optimum pH and temperature were 11 and 70 degrees C, respectively. The enzyme is stabile over a broad pH range (pH 4-11.5). The lipase preferably acted on triacylglycerols with medium-chain fatty acids. The lipase was inhibited strongly by Zn(2+), Hg(2+), Cu(2+) and slightly by Ca(2+) and Mg(2+). Non-ionic detergents and sodiumdeoxycholate enhanced lipase activity. Alkaline lipase from P. aeruginosa, capable of growing in a water-restricted medium has excellent properties and good potential for biotechnological applications in the metal industry. Its marked stability and activity in organic solvents suggest that this lipase is highly suitable as a biotechnological tool in a water-restricted medium with a variety of applications including organosynthetic reactions and the control and prevention of MWF putrification in the metal industry.
Karadzic
Ivanka
I
School of Medicine, Department of Chemistry, Belgrade University, Visegradska 26, 11000 Belgrade, Serbia and Montenegro. Ivanka@eunet.yu
Masui
Akihiko
A
Zivkovic
Lidija Izrael
LI
Fujiwara
Nobuaki
N
D001709:Biotechnology / Q000379:methods; D002118:Calcium / Q000737:chemistry; D002625:Chemistry, Organic / Q000379:methods; D003300:Copper / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007221:Industry; D008049:Lipase / Q000737:chemistry*; D008274:Magnesium / Q000737:chemistry; D008628:Mercury / Q000737:chemistry; D008899:Mineral Oil / Q000737:chemistry*; D011550:Pseudomonas aeruginosa / Q000201:enzymology*; D013379:Substrate Specificity; D013696:Temperature; D014865:Waste Disposal, Fluid / Q000379:methods; D015032:Zinc / Q000737:chemistry
D003300:Copper; D008899:Mineral Oil; D008049:Lipase; D008628:Mercury; D008274:Magnesium; D015032:Zinc; D002118:Calcium