Purification and physicochemical properties of lipase from thermophilic Bacillus aerius.
A thermophilic bacterial isolate producing lipase was isolated from soil of hot spring and identified as Bacillus aerius (MTCC 10978). Peak lipase activity was observed when 30 h old inoculum was used and incubated in shaking conditions for 48 h. The optimal temperature and pH for the bacterial growth and lipase production was found to be 55°C and 8.0 respectively with cottonseed oil as carbon source, yeast extract and beef extract as nitrogen source. The enzyme produced by thermophilic Bacillus aerius (MTCC 10978) was purified to 9-fold with 7.2% recovery by ammonium sulfate precipitation and DEAE-Cellulose Column Chromatography. The enzyme was found to be a protein having a molecular weight of 33 kDa on SDS-PAGE. The Km and Vmax value of lipase using p-nitrophenyl palmitate as calculated from Lineweaver-Burk plot was 2.13 mM and 0.66 µmol/ml/min respectively.
Department of Biotechnology, Himachal Pradesh University.
Nitin Kumar
NK
Saun
Poonam
P
Mehta
Reena
R
Gupta
D000645:Ammonium Sulfate; D001407:Bacillus / Q000201:enzymology*; D055598:Chemical Phenomena; D011232:Chemical Precipitation; D002848:Chromatography, DEAE-Cellulose; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D008970:Molecular Weight; D013696:Temperature
D008049:Lipase; D000645:Ammonium Sulfate