A novel thermostable esterase from the thermoacidophilic archaeon Sulfolobus tokodaii strain 7.
We have characterized an esterase expressed from the putative esterase gene (ST0071) selected from the total genome analysis from the thermoacidophilic archaeon Sulfolobus tokodaii strain 7. The ORF was cloned and expressed as a fusion protein in Escherichia coli. The protein was purified with heat treatment, affinity column chromatography, and size exclusion filtration. The optimum activity for ester cleavage against p-nitrophenyl esters was observed at around 70 degrees C and pH 7.5-8.0. The enzyme exhibited high thermostability and also showed activity in a mixture of a buffer and water-miscible organic solvents, such as acetonitrile and dimethyl sulfoxide. From the kinetic analysis, p-nitrophenyl butyrate was found to be a better substrate than caproate and caprylate.
Suzuki
Yoichi
Y
Department of Biosciences and Informatics, Faculty of Science and Technology, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama 223-8522, Japan.
Miyamoto
Kenji
K
Ohta
Hiromichi
H
D003001:Cloning, Molecular; D004950:Esterases / Q000235:genetics* / Q000378:metabolism*; D019848:Gene Expression Regulation, Archaeal; D015971:Gene Expression Regulation, Enzymologic; D006358:Hot Temperature; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D016944:Sulfolobus / Q000201:enzymology* / Q000235:genetics*
D011993:Recombinant Fusion Proteins; D004950:Esterases