title: Production, purification, and characterization of lipase from thermophilic and alkaliphilic Bacillus coagulans BTS-3.
abstract: A thermophilic isolate Bacillus coagulans BTS-3 produced an extracellular alkaline lipase, the production of which was substantially enhanced when the type of carbon source, nitrogen source, and the initial pH of culture medium were consecutively optimized. Lipase activity 1.16 U/ml of culture medium was obtained in 48 h at 55 degrees C and pH 8.5 with refined mustard oil as carbon source and a combination of peptone and yeast extract (1:1) as nitrogen sources. The enzyme was purified 40-fold to homogeneity by ammonium sulfate precipitation and DEAE-Sepharose column chromatography. Its molecular weight was 31 kDa on SDS-PAGE. The enzyme showed maximum activity at 55 degrees C and pH 8.5, and was stable between pH 8.0 and 10.5 and at temperatures up to 70 degrees C. The enzyme was found to be inhibited by Al3+, Co2+, Mn2+, and Zn2+ ions while K+, Fe3+, Hg2+, and Mg2+ ions enhanced the enzyme activity; Na+ ions have no effect on enzyme activity. The purified lipase showed a variable specificity/hydrolytic activity towards various 4-nitrophenyl esters.
authors.affiliation: Department of Biotechnology, Himachal Pradesh University, Summer Hill, Shimla-171 005, India.
authors.forename: Satyendra
authors.initials: S
authors.lastname: Kumar
authors.forename: Khyodano
authors.initials: K
authors.lastname: Kikon
authors.forename: Ashutosh
authors.initials: A
authors.lastname: Upadhyay
authors.forename: Shamsher S
authors.initials: SS
authors.lastname: Kanwar
authors.forename: Reena
authors.initials: R
authors.lastname: Gupta
mesh_terms: D001407:Bacillus / Q000201:enzymology*; D002852:Chromatography, Ion Exchange; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004795:Enzyme Stability; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000302:isolation & purification* / Q000378:metabolism; D008670:Metals / Q000494:pharmacology; D008970:Molecular Weight; D010168:Palmitates / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D004791:Enzyme Inhibitors; D008670:Metals; D010168:Palmitates; C050015:4-nitrophenyl palmitate; D008049:Lipase