title: Characterization of a Hyperthermostable Alkaline Lipase from Bacillus sonorensis 4R.
abstract: Hyperthermostable alkaline lipase from Bacillus sonorensis 4R was purified and characterized. The enzyme production was carried out at 80°C and 9.0 pH in glucose-tween inorganic salt broth under static conditions for 96 h. Lipase was purified by anion exchange chromatography by 12.15 fold with a yield of 1.98%. The molecular weight of lipase was found to be 21.87 KDa by SDS-PAGE. The enzyme activity was optimal at 80°C with t 1/2 of 150 min and at 90°C, 100°C, 110°C, and 120°C; the respective values were 121.59 min, 90.01 min, 70.01 min, and 50 min. The enzyme was highly activated by Mg and t 1/2 values at 80°C were increased from 150 min to 180 min when magnesium and mannitol were added in combination. The activation energy calculated from Arrhenius plot was 31.102 KJ/mol. At 80-120°C, values of ΔH and ΔG were in the range of 28.16-27.83 KJ/mol and 102.79 KJ/mol to 111.66 KJ/mol, respectively. Lipase activity was highest at 9.0 pH and stable for 2 hours at this pH at 80°C. Pretreatment of lipase with MgSO4 and CaSO4 stimulated enzyme activity by 249.94% and 30.2%, respectively. The enzyme activity was greatly reduced by CoCl2, CdCl2, HgCl2, CuCl2, Pb(NO3)2, PMSF, orlistat, oleic acid, iodine, EDTA, and urea.
authors.affiliation: DST-FIST Sponsored School of Life Sciences, Swami Ramanand Teerth Marathwada University, Nanded 431606, India.
authors.forename: Hemlata
authors.initials: H
authors.lastname: Bhosale
authors.affiliation: DST-FIST Sponsored School of Life Sciences, Swami Ramanand Teerth Marathwada University, Nanded 431606, India.
authors.forename: Uzma
authors.initials: U
authors.lastname: Shaheen
authors.affiliation: DST-FIST Sponsored School of Life Sciences, Swami Ramanand Teerth Marathwada University, Nanded 431606, India.
authors.forename: Tukaram
authors.identifier: 0000-0003-2952-0504
authors.initials: T
authors.lastname: Kadam