Abstract: Thermophilic lipases have great industrial application advantages on the stability of biological property over mesophilic enzyme and attracting a lot of attention. A predicted lipase from Thermomicrobium roseum DSM 5159 was expressed in E. coil BL21(DE3). The lipase enzymatic property was characterized after separation and purification. The Tm and ΔH were 97.53°C and 1 637 kJ/mol, respectively. The secondary structure of the lipase was 56.3% α-helix, 4.3% β-sheet, 19.9% β-turn and 20.3% random coil. The optimal temperature and pH of the lipase on p-nitrophenol palmitate were 85°C and 8.5, respectively. The enzyme activity could be maintained at 80% after an incubation at 80°C for 12 hours, suggesting a good thermal stability. Moreover, the lipase was verified with effective resistance ability to most of organic solvents. K+ and Ca2+ also had a promotion effect on the enzyme activity. Based on kinetic study, the lipase showed high affinity to p-nitrophenol palmitate. The Km, kcat and kcat/Km for this substrate were 0.01 mmol/L, 3.0 s-1 and 299.0 L/(mmol·s), respectively. The results could contribute to understanding the thermophilic lipases and its enzyme property, which providing the basis for its further application in biodiesel synthesis, detergents and industrial environment operated under high temperature.
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