SAXS Studies of the Endoglucanase Cel12A from Gloeophyllum trabeum Show Its Monomeric Structure and Reveal the Influence of Temperature on the Structural Stability of the Enzyme

Detalhes bibliográficos
Autor(a) principal: Miotto, Lis S.
Data de Publicação: 2014
Outros Autores: Reis, Caio V. dos, Neto, Mario de Oliveira [UNESP], Polikarpov, Igor
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://dx.doi.org/10.3390/ma7075202
http://hdl.handle.net/11449/117516
Resumo: Endoglucanases are key enzymes applied to the conversion of biomass aiming for second generation biofuel production. In the present study we obtained the small angle X-ray scattering (SAXS) structure of the G. trabeum endo-1,4-beta-glucanase Cel12A and investigated the influence of an important parameter, temperature, on both secondary and tertiary structure of the enzyme and its activity. The CD analysis for GtCel12A revealed that changes in the CD spectra starts at 55 degrees C and the T-m calculated from the experimental CD sigmoid curve using the Boltzmann function was 60.2 +/- 0.6 degrees C. SAXS data showed that GtCel12A forms monomers in solution and has an elongated form with a maximum diameter of 60 +/- 5 angstrom and a gyration radius of 19.4 +/- 0.1 angstrom as calculated from the distance distribution function. Kratky analysis revealed that 60 degrees C is the critical temperature above which we observed clear indications of denaturation. Our results showed the influence of temperature on the stability and activity of enzymes and revealed novel structural features of GtCel12A.
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spelling SAXS Studies of the Endoglucanase Cel12A from Gloeophyllum trabeum Show Its Monomeric Structure and Reveal the Influence of Temperature on the Structural Stability of the EnzymeendoglucanaseGloeophyllum trabeumbiophysicscircular dichroismsmall angle X-ray scattering (SAXS)Kratky analysisEndoglucanases are key enzymes applied to the conversion of biomass aiming for second generation biofuel production. In the present study we obtained the small angle X-ray scattering (SAXS) structure of the G. trabeum endo-1,4-beta-glucanase Cel12A and investigated the influence of an important parameter, temperature, on both secondary and tertiary structure of the enzyme and its activity. The CD analysis for GtCel12A revealed that changes in the CD spectra starts at 55 degrees C and the T-m calculated from the experimental CD sigmoid curve using the Boltzmann function was 60.2 +/- 0.6 degrees C. SAXS data showed that GtCel12A forms monomers in solution and has an elongated form with a maximum diameter of 60 +/- 5 angstrom and a gyration radius of 19.4 +/- 0.1 angstrom as calculated from the distance distribution function. Kratky analysis revealed that 60 degrees C is the critical temperature above which we observed clear indications of denaturation. Our results showed the influence of temperature on the stability and activity of enzymes and revealed novel structural features of GtCel12A.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Univ Sao Paulo, Inst Fis Sao Carlos, Grp Biotecnol Mol, BR-13566590 Sao Carlos, SP, BrazilUniv Estadual Paulista, Inst Biociencias Botucatu, Dept Fis & Biofis, BR-18618970 Botucatu, SP, BrazilUniv Estadual Paulista, Inst Biociencias Botucatu, Dept Fis & Biofis, BR-18618970 Botucatu, SP, BrazilFAPESP: 09/08233-9Mdpi AgUniversidade de São Paulo (USP)Universidade Estadual Paulista (Unesp)Miotto, Lis S.Reis, Caio V. dosNeto, Mario de Oliveira [UNESP]Polikarpov, Igor2015-03-18T15:56:21Z2015-03-18T15:56:21Z2014-07-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article5202-5211application/pdfhttp://dx.doi.org/10.3390/ma7075202Materials. Basel: Mdpi Ag, v. 7, n. 7, p. 5202-5211, 2014.1996-1944http://hdl.handle.net/11449/11751610.3390/ma7075202WOS:000339989700015WOS000339989700015.pdf8213371495151651Web of Sciencereponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengMaterials2.4670,732info:eu-repo/semantics/openAccess2023-10-06T06:08:53Zoai:repositorio.unesp.br:11449/117516Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T14:11:18.298579Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv SAXS Studies of the Endoglucanase Cel12A from Gloeophyllum trabeum Show Its Monomeric Structure and Reveal the Influence of Temperature on the Structural Stability of the Enzyme
title SAXS Studies of the Endoglucanase Cel12A from Gloeophyllum trabeum Show Its Monomeric Structure and Reveal the Influence of Temperature on the Structural Stability of the Enzyme
spellingShingle SAXS Studies of the Endoglucanase Cel12A from Gloeophyllum trabeum Show Its Monomeric Structure and Reveal the Influence of Temperature on the Structural Stability of the Enzyme
Miotto, Lis S.
endoglucanase
Gloeophyllum trabeum
biophysics
circular dichroism
small angle X-ray scattering (SAXS)
Kratky analysis
title_short SAXS Studies of the Endoglucanase Cel12A from Gloeophyllum trabeum Show Its Monomeric Structure and Reveal the Influence of Temperature on the Structural Stability of the Enzyme
title_full SAXS Studies of the Endoglucanase Cel12A from Gloeophyllum trabeum Show Its Monomeric Structure and Reveal the Influence of Temperature on the Structural Stability of the Enzyme
title_fullStr SAXS Studies of the Endoglucanase Cel12A from Gloeophyllum trabeum Show Its Monomeric Structure and Reveal the Influence of Temperature on the Structural Stability of the Enzyme
title_full_unstemmed SAXS Studies of the Endoglucanase Cel12A from Gloeophyllum trabeum Show Its Monomeric Structure and Reveal the Influence of Temperature on the Structural Stability of the Enzyme
title_sort SAXS Studies of the Endoglucanase Cel12A from Gloeophyllum trabeum Show Its Monomeric Structure and Reveal the Influence of Temperature on the Structural Stability of the Enzyme
author Miotto, Lis S.
author_facet Miotto, Lis S.
Reis, Caio V. dos
Neto, Mario de Oliveira [UNESP]
Polikarpov, Igor
author_role author
author2 Reis, Caio V. dos
Neto, Mario de Oliveira [UNESP]
Polikarpov, Igor
author2_role author
author
author
dc.contributor.none.fl_str_mv Universidade de São Paulo (USP)
Universidade Estadual Paulista (Unesp)
dc.contributor.author.fl_str_mv Miotto, Lis S.
Reis, Caio V. dos
Neto, Mario de Oliveira [UNESP]
Polikarpov, Igor
dc.subject.por.fl_str_mv endoglucanase
Gloeophyllum trabeum
biophysics
circular dichroism
small angle X-ray scattering (SAXS)
Kratky analysis
topic endoglucanase
Gloeophyllum trabeum
biophysics
circular dichroism
small angle X-ray scattering (SAXS)
Kratky analysis
description Endoglucanases are key enzymes applied to the conversion of biomass aiming for second generation biofuel production. In the present study we obtained the small angle X-ray scattering (SAXS) structure of the G. trabeum endo-1,4-beta-glucanase Cel12A and investigated the influence of an important parameter, temperature, on both secondary and tertiary structure of the enzyme and its activity. The CD analysis for GtCel12A revealed that changes in the CD spectra starts at 55 degrees C and the T-m calculated from the experimental CD sigmoid curve using the Boltzmann function was 60.2 +/- 0.6 degrees C. SAXS data showed that GtCel12A forms monomers in solution and has an elongated form with a maximum diameter of 60 +/- 5 angstrom and a gyration radius of 19.4 +/- 0.1 angstrom as calculated from the distance distribution function. Kratky analysis revealed that 60 degrees C is the critical temperature above which we observed clear indications of denaturation. Our results showed the influence of temperature on the stability and activity of enzymes and revealed novel structural features of GtCel12A.
publishDate 2014
dc.date.none.fl_str_mv 2014-07-01
2015-03-18T15:56:21Z
2015-03-18T15:56:21Z
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/10.3390/ma7075202
Materials. Basel: Mdpi Ag, v. 7, n. 7, p. 5202-5211, 2014.
1996-1944
http://hdl.handle.net/11449/117516
10.3390/ma7075202
WOS:000339989700015
WOS000339989700015.pdf
8213371495151651
url http://dx.doi.org/10.3390/ma7075202
http://hdl.handle.net/11449/117516
identifier_str_mv Materials. Basel: Mdpi Ag, v. 7, n. 7, p. 5202-5211, 2014.
1996-1944
10.3390/ma7075202
WOS:000339989700015
WOS000339989700015.pdf
8213371495151651
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Materials
2.467
0,732
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 5202-5211
application/pdf
dc.publisher.none.fl_str_mv Mdpi Ag
publisher.none.fl_str_mv Mdpi Ag
dc.source.none.fl_str_mv Web of Science
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
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