title: Purification and Characterization of a Thermostable Lipase from Geobacillus thermodenitrificans IBRL-nra.
abstract: Thermostable lipase from Geobacillus thermodenitrificans IBRL-nra was purified and characterized. The production of thermostable lipase from Geobacillus thermodenitrificans IBRL-nra was carried out in a shake-flask system at 65°C in cultivation medium containing; glucose 1.0% (w/v); yeast extract 1.25% (w/v); NaCl 0.45% (w/v) olive oil 0.1% (v/v) with agitation of 200 rpm for 24 hours. The extracted extracellular crude thermostable lipase was purified to homogeneity by using ultrafiltration, Heparin-affinity chromatography, and Sephadex G-100 gel-filtration chromatography by 34 times with a final yield of 9%. The molecular weight of the purified enzyme was estimated to be 30 kDa after SDS-PAGE analysis. The optimal temperature for thermostable lipase was 65°C and it retained its initial activity for 3 hours. Thermostable lipase activity was highest at pH 7.0 and stable for 16 hours at this pH at 65°C. Thermostable lipase showed elevated activity when pretreated with BaCl(2), CaCl(2), and KCl with 112%, 108%, and 106%, respectively. Lipase hydrolyzed tripalmitin (C16) and olive oil with optimal activity (100%) compared to other substrates.
authors.affiliation: Industrial Biotechnology Research Laboratory, School of Biological Sciences, Universiti Sains Malaysia, 11800 Pulau Pinang, Malaysia.
authors.forename: Anuradha
authors.initials: A
authors.lastname: Balan
authors.forename: Darah
authors.initials: D
authors.lastname: Ibrahim
authors.forename: Rashidah
authors.initials: R
authors.lastname: Abdul Rahim
authors.forename: Fatimah Azzahra
authors.initials: FA
authors.lastname: Ahmad Rashid