title: Enhanced activity and enantioselectivity of a hyperthermophilic esterase from archaeon Aeropyrum pernix K1 by acetone treatment.
abstract: 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.
authors.affiliation: College of Life Sciences, Jilin University, Changchun 130012, People's Republic of China.
authors.forename: Fangdi
authors.initials: F
authors.lastname: Cong
authors.forename: Kezhi
authors.initials: K
authors.lastname: Xing
authors.forename: Renjun
authors.initials: R
authors.lastname: Gao
authors.forename: Shugui
authors.initials: S
authors.lastname: Cao
authors.forename: Guirong
authors.initials: G
authors.lastname: Zhang
mesh_terms: D000096:Acetone / Q000737:chemistry*; D041522:Aeropyrum / Q000737:chemistry / Q000201:enzymology*; D019843:Archaeal Proteins / Q000235:genetics / Q000378:metabolism*; D055162:Biocatalysis; D002210:Caprylates / Q000378:metabolism; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D004950:Esterases / Q000235:genetics / Q000378:metabolism*; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007218:Industrial Microbiology / Q000379:methods*; D009154:Mutation; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013237:Stereoisomerism; D013379:Substrate Specificity
chemical_list: D019843:Archaeal Proteins; D002210:Caprylates; D011994:Recombinant Proteins; D000096:Acetone; C034106:4-nitrophenyloctanoate; D004950:Esterases