Enhanced Activity and Enantioselectivity of a Hyperthermophilic Esterase from Archaeon Aeropyrum pernix K1 by Acetone Treatment.

Cong, Fangdi; Xing, Kezhi; Gao, Renjun; Cao, Shugui; Zhang, Guirong

To improve the activity and enantioselectivity of hyperthermophilic archaeon Aeropyrum pernix K1 esterase (APE1547) and its mutants, they were purified by acetone-treated method. It was found that the acetone treatment not only caused APE1547 and its mutants to display higher activity and enantioselectivity but also saved more than 90% of time spent in purifying them by Ni-chelating column. In hydrolysis of p-nitrophenyl caprylate, the acetone-treated APE1547 and mutant A containing the following substitutions R11G, L36P, V225A, I551L, and A564T showed 5.7- and 6.9-fold active increase, respectively. In the resolution of 2-octanol acetate, the acetone-treated mutant A had a 9-fold enantioselective increase relative to that purified by Ni-chelating column. In addition, the impact of pH, temperature, and chemical reagents on activity of APE1547 and mutant A was discussed in this paper.

ENANTIOSELECTIVE catalysis; ESTERASES; ACETONE; OCTYL alcohol; ACETATES

Applied Biochemistry & Biotechnology, 2011, Vol 165, Issue 3/4, p795

0273-2289

Academic Journal

10.1007/s12010-011-9297-9