Characterization of EST3: a metagenome-derived esterase with suitable properties for biotechnological applications
Autor(a) principal: | |
---|---|
Data de Publicação: | 2016 |
Outros Autores: | , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1007/s00253-016-7385-z http://hdl.handle.net/11449/168740 |
Resumo: | Metagenomic libraries from diverse environments have been extensive sources of many lipases and esterases; nevertheless, most of these enzymes remain biochemically uncharacterized. We previously built a metagenomic fosmid library from a microbial consortium specialized for diesel oil degradation and tested it for lipolytic activity. In the present study, we identified the PL14.H10 clone that was subcloned and sequenced, which enabled the identification of the EST3 protein. This enzyme exhibited 74 % amino acid identity with the uncharacterized alpha/beta hydrolase from Parvibaculum lavamentivorans [GenBank: WP012110575.1] and was classified into lipolytic enzyme family IV. Biochemical characterization revealed that EST3 presents high activity in a wide range of temperature with highest activity from 41 to 45 °C. Also, this thermostable esterase acts from mild acidic to alkaline conditions with an optimum pH of 6.0. The enzyme exhibited activity against p-nitrophenyl esters of different chain lengths and highest catalytic efficiency against p-nitrophenyl caprylate. The activity of the protein was increased in the presence of 0.5 mM of Mn+2, Li+, EDTA, and 1 % of CTAB and exhibited half of the activity in the presence of 10 % methanol and ethanol. Moreover, the homology model of EST3 was built and compared to other esterases, revealing a substrate channel that should fit a wide range of substrates. Taken together, the data presented in this work reveal the unique and interesting characteristics of EST3 that might be explored for further use in biotechnological applications. |
id |
UNSP_d03f21ec96c062cc3250045fadbbca78 |
---|---|
oai_identifier_str |
oai:repositorio.unesp.br:11449/168740 |
network_acronym_str |
UNSP |
network_name_str |
Repositório Institucional da UNESP |
repository_id_str |
2946 |
spelling |
Characterization of EST3: a metagenome-derived esterase with suitable properties for biotechnological applicationsEsteraseFamily IVLipaseMicrobial consortiumpNP estersMetagenomic libraries from diverse environments have been extensive sources of many lipases and esterases; nevertheless, most of these enzymes remain biochemically uncharacterized. We previously built a metagenomic fosmid library from a microbial consortium specialized for diesel oil degradation and tested it for lipolytic activity. In the present study, we identified the PL14.H10 clone that was subcloned and sequenced, which enabled the identification of the EST3 protein. This enzyme exhibited 74 % amino acid identity with the uncharacterized alpha/beta hydrolase from Parvibaculum lavamentivorans [GenBank: WP012110575.1] and was classified into lipolytic enzyme family IV. Biochemical characterization revealed that EST3 presents high activity in a wide range of temperature with highest activity from 41 to 45 °C. Also, this thermostable esterase acts from mild acidic to alkaline conditions with an optimum pH of 6.0. The enzyme exhibited activity against p-nitrophenyl esters of different chain lengths and highest catalytic efficiency against p-nitrophenyl caprylate. The activity of the protein was increased in the presence of 0.5 mM of Mn+2, Li+, EDTA, and 1 % of CTAB and exhibited half of the activity in the presence of 10 % methanol and ethanol. Moreover, the homology model of EST3 was built and compared to other esterases, revealing a substrate channel that should fit a wide range of substrates. Taken together, the data presented in this work reveal the unique and interesting characteristics of EST3 that might be explored for further use in biotechnological applications.Department of Technology São Paulo State University (UNESP), Prof. Paulo Donato Castellane highway, s/n.Department of Microbiology Institute of Biomedical Sciences II University of São PauloDepartment of Technology São Paulo State University (UNESP), Prof. Paulo Donato Castellane highway, s/n.Universidade Estadual Paulista (Unesp)Universidade de São Paulo (USP)Maester, Thaís Carvalho [UNESP]Pereira, Mariana Rangel [UNESP]Machado Sierra, E. G. [UNESP]Balan, Andreade Macedo Lemos, Eliana Gertrudes [UNESP]2018-12-11T16:42:48Z2018-12-11T16:42:48Z2016-07-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article5815-5827application/pdfhttp://dx.doi.org/10.1007/s00253-016-7385-zApplied Microbiology and Biotechnology, v. 100, n. 13, p. 5815-5827, 2016.1432-06140175-7598http://hdl.handle.net/11449/16874010.1007/s00253-016-7385-z2-s2.0-849752086962-s2.0-84975208696.pdf2-s2.0-84975208696.pdfScopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengApplied Microbiology and Biotechnology1,1821,182info:eu-repo/semantics/openAccess2023-11-07T06:14:58Zoai:repositorio.unesp.br:11449/168740Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462023-11-07T06:14:58Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Characterization of EST3: a metagenome-derived esterase with suitable properties for biotechnological applications |
title |
Characterization of EST3: a metagenome-derived esterase with suitable properties for biotechnological applications |
spellingShingle |
Characterization of EST3: a metagenome-derived esterase with suitable properties for biotechnological applications Maester, Thaís Carvalho [UNESP] Esterase Family IV Lipase Microbial consortium pNP esters |
title_short |
Characterization of EST3: a metagenome-derived esterase with suitable properties for biotechnological applications |
title_full |
Characterization of EST3: a metagenome-derived esterase with suitable properties for biotechnological applications |
title_fullStr |
Characterization of EST3: a metagenome-derived esterase with suitable properties for biotechnological applications |
title_full_unstemmed |
Characterization of EST3: a metagenome-derived esterase with suitable properties for biotechnological applications |
title_sort |
Characterization of EST3: a metagenome-derived esterase with suitable properties for biotechnological applications |
author |
Maester, Thaís Carvalho [UNESP] |
author_facet |
Maester, Thaís Carvalho [UNESP] Pereira, Mariana Rangel [UNESP] Machado Sierra, E. G. [UNESP] Balan, Andrea de Macedo Lemos, Eliana Gertrudes [UNESP] |
author_role |
author |
author2 |
Pereira, Mariana Rangel [UNESP] Machado Sierra, E. G. [UNESP] Balan, Andrea de Macedo Lemos, Eliana Gertrudes [UNESP] |
author2_role |
author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) Universidade de São Paulo (USP) |
dc.contributor.author.fl_str_mv |
Maester, Thaís Carvalho [UNESP] Pereira, Mariana Rangel [UNESP] Machado Sierra, E. G. [UNESP] Balan, Andrea de Macedo Lemos, Eliana Gertrudes [UNESP] |
dc.subject.por.fl_str_mv |
Esterase Family IV Lipase Microbial consortium pNP esters |
topic |
Esterase Family IV Lipase Microbial consortium pNP esters |
description |
Metagenomic libraries from diverse environments have been extensive sources of many lipases and esterases; nevertheless, most of these enzymes remain biochemically uncharacterized. We previously built a metagenomic fosmid library from a microbial consortium specialized for diesel oil degradation and tested it for lipolytic activity. In the present study, we identified the PL14.H10 clone that was subcloned and sequenced, which enabled the identification of the EST3 protein. This enzyme exhibited 74 % amino acid identity with the uncharacterized alpha/beta hydrolase from Parvibaculum lavamentivorans [GenBank: WP012110575.1] and was classified into lipolytic enzyme family IV. Biochemical characterization revealed that EST3 presents high activity in a wide range of temperature with highest activity from 41 to 45 °C. Also, this thermostable esterase acts from mild acidic to alkaline conditions with an optimum pH of 6.0. The enzyme exhibited activity against p-nitrophenyl esters of different chain lengths and highest catalytic efficiency against p-nitrophenyl caprylate. The activity of the protein was increased in the presence of 0.5 mM of Mn+2, Li+, EDTA, and 1 % of CTAB and exhibited half of the activity in the presence of 10 % methanol and ethanol. Moreover, the homology model of EST3 was built and compared to other esterases, revealing a substrate channel that should fit a wide range of substrates. Taken together, the data presented in this work reveal the unique and interesting characteristics of EST3 that might be explored for further use in biotechnological applications. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016-07-01 2018-12-11T16:42:48Z 2018-12-11T16:42:48Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1007/s00253-016-7385-z Applied Microbiology and Biotechnology, v. 100, n. 13, p. 5815-5827, 2016. 1432-0614 0175-7598 http://hdl.handle.net/11449/168740 10.1007/s00253-016-7385-z 2-s2.0-84975208696 2-s2.0-84975208696.pdf 2-s2.0-84975208696.pdf |
url |
http://dx.doi.org/10.1007/s00253-016-7385-z http://hdl.handle.net/11449/168740 |
identifier_str_mv |
Applied Microbiology and Biotechnology, v. 100, n. 13, p. 5815-5827, 2016. 1432-0614 0175-7598 10.1007/s00253-016-7385-z 2-s2.0-84975208696 2-s2.0-84975208696.pdf |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Applied Microbiology and Biotechnology 1,182 1,182 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
5815-5827 application/pdf |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
_version_ |
1799964853319761920 |