Isolation and in silico characterization of novel esterase gene with β-lactamase fold isolated from metagenome of north western Himalayas

An esterase-producing clone Aph2 was isolated from the Apharwat soil metagenomic library, a mountain peak in NW Himalayas. ORF 2 (Est Ac) of clone Aph2 corresponds to 271 aa protein and showed 26 % sequence similarity to carboxylesterase gene of Synechococcus sp. JA-2-3B. Est Ac contains nucleophili...

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Autores principales: Sudan, Avneet Kour, Vakhlu, Jyoti
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2014
Materias:
Original Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4522730/
https://www.ncbi.nlm.nih.gov/pubmed/28324560
http://dx.doi.org/10.1007/s13205-014-0254-5
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author Sudan, Avneet Kour
Vakhlu, Jyoti
author_facet Sudan, Avneet Kour
Vakhlu, Jyoti
author_sort Sudan, Avneet Kour
collection PubMed
description An esterase-producing clone Aph2 was isolated from the Apharwat soil metagenomic library, a mountain peak in NW Himalayas. ORF 2 (Est Ac) of clone Aph2 corresponds to 271 aa protein and showed 26 % sequence similarity to carboxylesterase gene of Synechococcus sp. JA-2-3B. Est Ac contains nucleophilic Ser in S(68)-X-X-K(71) motif of β-lactamases with Tyr Y(103). The conserved sequences are common with family VIII carboxylesterase and class C β-lactamase sequences. Phylogenetic analysis revealed that Est Ac sequence is closely related to esterase than to β-lactamases. In silico 3D protein structure of Est Ac was generated using MODELLER software (9.10 version). Model was generated on the basis of carboxylesterase template (PDB:1CI8) of Est B (Burkholderia gladioli) and the stereochemical parameters of the model generated were satisfactory. Docking with diisopropyl-fluorophosphate confirmed catalytic activity of Ser(68) present in S-X-X-K motif.
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spelling pubmed-45227302015-08-05 Isolation and in silico characterization of novel esterase gene with β-lactamase fold isolated from metagenome of north western Himalayas Sudan, Avneet Kour Vakhlu, Jyoti 3 Biotech Original Article An esterase-producing clone Aph2 was isolated from the Apharwat soil metagenomic library, a mountain peak in NW Himalayas. ORF 2 (Est Ac) of clone Aph2 corresponds to 271 aa protein and showed 26 % sequence similarity to carboxylesterase gene of Synechococcus sp. JA-2-3B. Est Ac contains nucleophilic Ser in S(68)-X-X-K(71) motif of β-lactamases with Tyr Y(103). The conserved sequences are common with family VIII carboxylesterase and class C β-lactamase sequences. Phylogenetic analysis revealed that Est Ac sequence is closely related to esterase than to β-lactamases. In silico 3D protein structure of Est Ac was generated using MODELLER software (9.10 version). Model was generated on the basis of carboxylesterase template (PDB:1CI8) of Est B (Burkholderia gladioli) and the stereochemical parameters of the model generated were satisfactory. Docking with diisopropyl-fluorophosphate confirmed catalytic activity of Ser(68) present in S-X-X-K motif. Springer Berlin Heidelberg 2014-10-19 2015-08 /pmc/articles/PMC4522730/ /pubmed/28324560 http://dx.doi.org/10.1007/s13205-014-0254-5 Text en © The Author(s) 2014 https://creativecommons.org/licenses/by/4.0/ Open AccessThis article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited.
spellingShingle Original Article
Sudan, Avneet Kour
Vakhlu, Jyoti
Isolation and in silico characterization of novel esterase gene with β-lactamase fold isolated from metagenome of north western Himalayas
title Isolation and in silico characterization of novel esterase gene with β-lactamase fold isolated from metagenome of north western Himalayas
title_full Isolation and in silico characterization of novel esterase gene with β-lactamase fold isolated from metagenome of north western Himalayas
title_fullStr Isolation and in silico characterization of novel esterase gene with β-lactamase fold isolated from metagenome of north western Himalayas
title_full_unstemmed Isolation and in silico characterization of novel esterase gene with β-lactamase fold isolated from metagenome of north western Himalayas
title_short Isolation and in silico characterization of novel esterase gene with β-lactamase fold isolated from metagenome of north western Himalayas
title_sort isolation and in silico characterization of novel esterase gene with β-lactamase fold isolated from metagenome of north western himalayas
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4522730/
https://www.ncbi.nlm.nih.gov/pubmed/28324560
http://dx.doi.org/10.1007/s13205-014-0254-5
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