title: Cloning, expression and characterization of a halotolerant esterase from a marine bacterium Pelagibacterium halotolerans B2T.
abstract: An esterase PE10 (279 aa) from Pelagibacterium halotolerans B2(T) was cloned and overexpressed in Escherichia coli Rosetta in a soluble form. The deduced protein was 29.91 kDa and the phylogenetic analysis of the deduced amino acids sequence showed it represented a new family of lipolytic enzymes. The recombinant protein was purified by Ni-NTA affinity chromatography column and the characterization showed its optimal temperature and pH were 45 °C and pH 7.5, respectively. Substrate specificity study showed PE10 preferred short chain p-nitrophenyl esters and exhibited maximum activity toward p-nitrophenyl acetate. In addition, PE10 was a halotolerant esterase as it was still active under 4 M NaCl. Three-dimensional modeling of PE10 suggested that the high negative electrostatic potential on the surface may relevant to its tolerance to high salt environment. With this halotolerance property, PE10 could be a candidate for industrial use.
authors.affiliation: College of Life Sciences, Zhejiang University, 866 Yuhangtang Road, Hangzhou, 310058, Zhejiang, China.
authors.forename: Xiawei
authors.initials: X
authors.lastname: Jiang
authors.forename: Yingyi
authors.initials: Y
authors.lastname: Huo
authors.forename: Hong
authors.initials: H
authors.lastname: Cheng
authors.forename: Xinqi
authors.initials: X
authors.lastname: Zhang
authors.forename: Xufen
authors.initials: X
authors.lastname: Zhu
authors.forename: Min
authors.initials: M
authors.lastname: Wu
mesh_terms: D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D004950:Esterases / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D015870:Gene Expression; D041921:Hyphomicrobiaceae / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D012965:Sodium Chloride / Q000737:chemistry
chemical_list: D011994:Recombinant Proteins; D012965:Sodium Chloride; D004950:Esterases