Hyperthermophilic aldolases as biocatalyst for C–C bond formation: rhamnulose 1-phosphate aldolase from Thermotoga maritima

The TM1072 gene from Thermotoga maritima codifies for a putative form of a rhamnulose-1-phosphate aldolase (Rha-1PATm). To investigate this enzyme further, its gene was cloned and expressed in Escherichia coli. The purified enzyme was activated by Co2+ as a divalentmetal ion cofactor, instead of Zn2...

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Detalhes bibliográficos
Autores: Oroz-Guinea, Isabel, Sánchez-Moreno, Israel, Mena, Montaña, García-Junceda, Eduardo
Tipo de documento: artigo
Data de publicação:2015
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositório:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/113297
Acesso em linha:http://hdl.handle.net/10261/113297
Access Level:Acceso aberto
Palavra-chave:Aldolases
Biocatalysis
Enzyme catalysis
Hyperthermophilic enzymes
Thermozyme
Thermostability
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spelling Hyperthermophilic aldolases as biocatalyst for C–C bond formation: rhamnulose 1-phosphate aldolase from Thermotoga maritimaOroz-Guinea, IsabelSánchez-Moreno, IsraelMena, MontañaGarcía-Junceda, EduardoAldolasesBiocatalysisEnzyme catalysisHyperthermophilic enzymesThermozymeThermostabilityThe TM1072 gene from Thermotoga maritima codifies for a putative form of a rhamnulose-1-phosphate aldolase (Rha-1PATm). To investigate this enzyme further, its gene was cloned and expressed in Escherichia coli. The purified enzyme was activated by Co2+ as a divalentmetal ion cofactor, instead of Zn2+ as its E. coli homologue, and exhibited a maximum of activity at 95 °C. Furthermore, the enzyme displayed a high stability against extreme reaction conditions, retaining 90 % of its activity in the presence of 40 % of acetonitrile and showing a half-life greater than 3 h at 115 °C. The kinetic parameters at room temperature (R/T) were also studied; the KM was calculated to be 3.6±0.33 mM, while kcat/KM was found to be 0.7× 103 s−1 M−1. Given these characteristics, Rha-1PA Tm is an attractive enzyme for use as a biocatalyst for industrial applications, offering intriguing possibilities for practical biocatalysis.We thank the Spanish Ministerio de Economía y Competitividad (Grant PI11/01436) and Comunidad de Madrid (Grant S2009/PPQ-1752) for financial support. I. O.-G. is a JAEPredoc fellow from CSIC.Peer reviewedSpringer NatureConsejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]201520152015info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501http://hdl.handle.net/10261/113297reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Ingléshttp://dx.doi.org/10.1007/s00253-014-6123-7Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/1132972026-05-22T06:33:51Z
dc.title.none.fl_str_mv Hyperthermophilic aldolases as biocatalyst for C–C bond formation: rhamnulose 1-phosphate aldolase from Thermotoga maritima
title Hyperthermophilic aldolases as biocatalyst for C–C bond formation: rhamnulose 1-phosphate aldolase from Thermotoga maritima
spellingShingle Hyperthermophilic aldolases as biocatalyst for C–C bond formation: rhamnulose 1-phosphate aldolase from Thermotoga maritima
Oroz-Guinea, Isabel
Aldolases
Biocatalysis
Enzyme catalysis
Hyperthermophilic enzymes
Thermozyme
Thermostability
title_short Hyperthermophilic aldolases as biocatalyst for C–C bond formation: rhamnulose 1-phosphate aldolase from Thermotoga maritima
title_full Hyperthermophilic aldolases as biocatalyst for C–C bond formation: rhamnulose 1-phosphate aldolase from Thermotoga maritima
title_fullStr Hyperthermophilic aldolases as biocatalyst for C–C bond formation: rhamnulose 1-phosphate aldolase from Thermotoga maritima
title_full_unstemmed Hyperthermophilic aldolases as biocatalyst for C–C bond formation: rhamnulose 1-phosphate aldolase from Thermotoga maritima
title_sort Hyperthermophilic aldolases as biocatalyst for C–C bond formation: rhamnulose 1-phosphate aldolase from Thermotoga maritima
dc.creator.none.fl_str_mv Oroz-Guinea, Isabel
Sánchez-Moreno, Israel
Mena, Montaña
García-Junceda, Eduardo
author Oroz-Guinea, Isabel
author_facet Oroz-Guinea, Isabel
Sánchez-Moreno, Israel
Mena, Montaña
García-Junceda, Eduardo
author_role author
author2 Sánchez-Moreno, Israel
Mena, Montaña
García-Junceda, Eduardo
author2_role author
author
author
dc.contributor.none.fl_str_mv Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Aldolases
Biocatalysis
Enzyme catalysis
Hyperthermophilic enzymes
Thermozyme
Thermostability
topic Aldolases
Biocatalysis
Enzyme catalysis
Hyperthermophilic enzymes
Thermozyme
Thermostability
description The TM1072 gene from Thermotoga maritima codifies for a putative form of a rhamnulose-1-phosphate aldolase (Rha-1PATm). To investigate this enzyme further, its gene was cloned and expressed in Escherichia coli. The purified enzyme was activated by Co2+ as a divalentmetal ion cofactor, instead of Zn2+ as its E. coli homologue, and exhibited a maximum of activity at 95 °C. Furthermore, the enzyme displayed a high stability against extreme reaction conditions, retaining 90 % of its activity in the presence of 40 % of acetonitrile and showing a half-life greater than 3 h at 115 °C. The kinetic parameters at room temperature (R/T) were also studied; the KM was calculated to be 3.6±0.33 mM, while kcat/KM was found to be 0.7× 103 s−1 M−1. Given these characteristics, Rha-1PA Tm is an attractive enzyme for use as a biocatalyst for industrial applications, offering intriguing possibilities for practical biocatalysis.
publishDate 2015
dc.date.none.fl_str_mv 2015
2015
2015
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/113297
url http://hdl.handle.net/10261/113297
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv http://dx.doi.org/10.1007/s00253-014-6123-7
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Springer Nature
publisher.none.fl_str_mv Springer Nature
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15.812429

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