Fu_2013_Appl.Microbiol.Biotechnol_97_3965

Reference

Title : Functional and structural studies of a novel cold-adapted esterase from an Arctic intertidal metagenomic library - Fu_2013_Appl.Microbiol.Biotechnol_97_3965
Author(s) : Fu J , Leiros HK , de Pascale D , Johnson KA , Blencke HM , Landfald B
Ref : Applied Microbiology & Biotechnology , 97 :3965 , 2013
Abstract :

A novel cold-adapted lipolytic enzyme gene, est97, was identified from a high Arctic intertidal zone sediment metagenomic library. The deduced amino acid sequence of Est97 showed low similarity with other lipolytic enzymes, the maximum being 30 % identity with a putative lipase from Vibrio caribbenthicus. Common features of lipolytic enzymes, such as the GXSXG sequence motif, were detected. The gene product was over-expressed in Escherichia coli and purified. The recombinant Est97 (rEst97) hydrolysed various rho-nitrophenyl esters with the best substrate being rho-nitrophenyl hexanoate (K m and k cat of 39 muM and 25.8 s(-1), respectively). This esterase activity of rEst97 was optimal at 35 degrees C and pH 7.5 and the enzyme was unstable at temperatures above 25 degrees C. The apparent melting temperature, as determined by differential scanning calorimetry was 39 degrees C, substantiating Est97 as a cold-adapted esterase. The crystal structure of rEst97 was determined by the single wavelength anomalous dispersion method to 1.6 A resolution. The protein was found to have a typical alpha/beta-hydrolase fold with Ser144-His226-Asp197 as the catalytic triad. A suggested, relatively short lid domain of rEst97 is composed of residues 80-114, which form an alpha-helix and a disordered loop. The cold adaptation features seem primarily related to a high number of methionine and glycine residues and flexible loops in the high-resolution structures.

PubMedSearch : Fu_2013_Appl.Microbiol.Biotechnol_97_3965
PubMedID: 22832985
Gene_locus related to this paper: 9bact-Est97

Related information

Gene_locus 9bact-Est97
Family 9bact-Est97    Bacterial_Est97
Structure 9bact-Est97    Bacterial_Est97    4AO6    4AO7    4AO8

Citations formats

Fu J, Leiros HK, de Pascale D, Johnson KA, Blencke HM, Landfald B (2013)
Functional and structural studies of a novel cold-adapted esterase from an Arctic intertidal metagenomic library
Applied Microbiology & Biotechnology 97 :3965

Fu J, Leiros HK, de Pascale D, Johnson KA, Blencke HM, Landfald B (2013)
Applied Microbiology & Biotechnology 97 :3965