Open AccessPurification, crystallization and preliminary crystallographic analysis of Est25: a ketoprofen‐specific hormone‐sensitive lipase
Author(s) -
Kim SeungBum,
Joo Sangbum,
Yoon Hyun C.,
Ryu Yeonwoo,
Kim Kyeong Kyu,
Kim T. Doohun
Publication year - 2007
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309107026152
Subject(s) - ketoprofen , escherichia coli , crystallization , recombinant dna , chemistry , lipase , crystallography , monoclinic crystal system , biochemistry , crystal structure , chromatography , enzyme , organic chemistry , gene
Ketoprofen, a nonsteroidal anti‐inflammatory drug, inhibits the synthesis of prostaglandin. A novel hydrolase (Est25) with high ketoprofen specificity has previously been identified using a metagenomic library from environmental samples. Recombinant Est25 protein with a histidine tag at the N‐terminus was expressed in Escherichia coli and purified in a homogenous form. Est25 was crystallized from 2.4 M sodium malonate pH 7.0 and X‐ray diffraction data were collected to 1.49 Å using synchrotron radiation. The crystals belong to the monoclinic space group C 2, with unit‐cell parameters a = 197.8, b = 95.2, c = 99.4 Å, β = 97.1°.