Identification and characterization of a novel (S)-ketoprofen-specific esterase.
A new (S)-ketoprofen specific esterase (EST-Y29) was identified from a metagenome library from environmental samples, which showed homologies with class C-beta lactamase, penicillin binding protein, and other lipases/esterases. In order to investigate the biochemical and biophysical properties, the recombinant protein was overexpressed, purified to homogeneity, and characterized. This EST-Y29 has high catalytic activity against p-nitrophenyl esters of short fatty acids (C(2) and C(4)) and alpha-naphthyl acetate with activation energy of 30.4 kJ/mol. We have further characterized EST-Y29 using high performance liquid chromatography (HPLC), circular dichroism (CD), dynamic light scattering (DLS) and size exclusion chromatography (SEC).
Yoon
Sangyoung
S
Department of Biological and Molecular Engineering, College of Engineering, Ajou University, Suwon 443-749, South Korea.
Kim
SeungBum
S
Ryu
Yeonwoo
Y
Kim
T Doohun
TD
D000595:Amino Acid Sequence; D020134:Catalytic Domain / Q000235:genetics; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D004950:Esterases / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D015698:Genomic Library; D006863:Hydrogen-Ion Concentration; D007660:Ketoprofen / Q000378:metabolism*; D008027:Light; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012542:Scattering, Radiation; D017386:Sequence Homology, Amino Acid; D012997:Solvents; D013379:Substrate Specificity; D013696:Temperature; D013816:Thermodynamics
D011994:Recombinant Proteins; D012997:Solvents; D007660:Ketoprofen; D004950:Esterases