Purification, characterization and thermostability of lipase from a thermophilic Bacillus sp. J33.
A thermostable lipase produced by a thermophilic Bacillus sp. J33 was purified to 175-fold with 15.6% recovery by ammonium sulphate and Phenyl Sepharose column chromatography. The enzyme is a monomeric protein having molecular weight of 45 kDa. It hydrolyzes triolein at all positions. The fatty acid specificity of lipase is broad with little preference for C12 and C4. The Km and Vmax for lipase with pNP-laurate as substrate was calculated to be 2.5 mM and 0.4 microM min(-1) ml(-1) respectively. The immobilized enzyme was stable for 12 h at 60 degrees C. Polyhydric alcohols such as ethylene glycol (2.5 M), sorbitol (2.5 M) and glycerol (2.5 M) were used as thermostabilizers. Lipase acquired a remarkable stability, since no deactivation occurred at 70 degrees C for 150 min in the presence of additives.
Nawani
N
N
Department of Biotechnology, Panjab University, Chandigarh, India.
Kaur
J
J
D001407:Bacillus / Q000201:enzymology*; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability / Q000502:physiology*; D019855:Ethylene Glycol; D005990:Glycerol; D008049:Lipase / Q000302:isolation & purification* / Q000378:metabolism; D013012:Sorbitol; D013379:Substrate Specificity
D013012:Sorbitol; D008049:Lipase; D019855:Ethylene Glycol; D005990:Glycerol