Characterization of a cold-active and salt tolerant esterase identified by functional screening of Arctic metagenomic libraries

BMC Biochemistry
Concetta De SantiNils-Peder Willassen

Abstract

The use of metagenomics in enzyme discovery constitutes a powerful approach to access to genomes of unculturable community of microorganisms and isolate novel valuable biocatalysts for use in a wide range of biotechnological and pharmaceutical fields. Here we present a novel esterase gene (lip3) identified by functional screening of three fosmid metagenomic libraries, constructed from three marine sediment samples. The sequenced positive fosmid revealed an enzyme of 281 amino acids with similarity to class 3 lipases. The 3D modeling of Lip3 was generated by homology modeling on the basis of four lipases templates [PDB ID: 3O0D, 3NGM, 3G7N, 2QUB] to unravel structural features of this novel enzyme. The catalytic triad of Lip3 was predicted to be Asp207, His267 and the catalytic nucleophile Ser150 in a conserved pentapeptide (GXSXG). The 3D model highlighted the presence of a one-helix lid able to regulate the access of the substrate to the active site when the enzyme binds a hydrophobic interface. Moreover an analysis of the external surface of Lip3 model showed that the majority of the surface regions were hydrophobic (59.6 %) compared with homologous lipases (around 35 %) used as templates. The recombinant Lip3 esterase, expre...Continue Reading

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Citations

Oct 14, 2017·FEMS Microbiology Letters·Francesca BeriniFlavia Marinelli
Sep 14, 2018·Frontiers in Microbiology·Premankur SukulLars I Leichert
Sep 25, 2019·Marine Drugs·Stefano BrunoCinzia Verde
Jan 8, 2021·Applied Biochemistry and Biotechnology·Noora BarzkarMahmoud Hafezieh
Jan 31, 2021·Extremophiles : Life Under Extreme Conditions·Xin ChangBaojuan Wang

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Datasets Mentioned

BETA
KJ538549

Methods Mentioned

BETA
PCR

Software Mentioned

BLASTp
HHpred
Procheck
APBS ( Adaptive Poisson - Boltzmann Solver )
SigmaPlot
VMD
PDB2PQR
GeneMark
FastCloning
Modeller

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