Abstract
A thermophilic esterase, SsoPEst, from Sulfolobus solfataricus P2 was cloned and expressed in E. coli AD494 (DE3). Gene sequencing indicated the encoded 353 amino acids had less than 32% identity with reported esterases. The recombinant enzyme hydrolyzed p-nitrophenyl esters but not tributyrin or tricaprylin, exhibiting the highest specific activity (1.1 U/mg) with p-nitrophenyl caprylate. The enzyme was optimally active at pH 5.5 and 80°C. It retained 50% activity after 1 h incubation at 80°C. Activity was significantly inhibited by PMSF. Five SsoPEst mutants were generated by site-directed mutagenesis. One mutant had a higher specific activity of 2.8 U/mg at 37°C and 14 U/mg at 80°C than the wild-type enzyme which exhibited 0.7 U/mg at 37°C and 3.8 U/mg at 80°C against p-nitrophenyl butyrate.
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Acknowledgment
This work was supported by the Chinese Academy of Sciences Foundation (No. KSCX2-YW-G-017) and National 863 Program (No. 2006AA020203).
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Shang, YS., Zhang, XE., Wang, XD. et al. Biochemical characterization and mutational improvement of a thermophilic esterase from Sulfolobus solfataricus P2. Biotechnol Lett 32, 1151–1157 (2010). https://doi.org/10.1007/s10529-010-0274-0
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DOI: https://doi.org/10.1007/s10529-010-0274-0