In vitro and in silico characterization of a novel dextranase from Pochonia chlamydosporia

Detalhes bibliográficos
Autor(a) principal: Sufiate, Bruna
Data de Publicação: 2018
Outros Autores: Araújo, Jackson Victor de, Queiroz, José Humberto de, Gouveia, Angélica S, Cardoso, Evandro, Moreira, Samara Silveira, Soares, Filippe Elias de Freitas, Braga, Fabio Ribeiro
Tipo de documento: Artigo
Idioma: eng
Título da fonte: LOCUS Repositório Institucional da UFV
Texto Completo: https://doi.org/10.1007/s13205-018-1192-4
http://www.locus.ufv.br/handle/123456789/24107
Resumo: The objective of this study was to purify, characterize, and phylogenetically and structurally analyze the dextranase produced by the fungus Pochonia chlamydosporia. Dextranase produced by the fungus P. chlamydosporia was purified to homogeneity in two steps, with a yield of 152%, purification factor of 6.84 and specific activity of 358.63 U/mg. Its molecular weight was estimated by SDS-PAGE at 64 kDa. The enzyme presented higher activity at 50 °C and pH 5.0, using 100 mM citrate–phosphate buffer, was inhibited by Ag1+, Hg2+, Cu2+, Mg2+, and presented KM of 23.60 µM. Mature dextranase is composed of 585 amino acids residues, with a predicted molecular weight of 64.38 kDa and pI 5.96. This dextranase showed a strong phylogenetic similarity when compared to Trichoderma harzianum dextranase. Its structure consists of two domains: the first composed by 15 β strands, and the second composed by a right-handed parallel β-helix.
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spelling Sufiate, BrunaAraújo, Jackson Victor deQueiroz, José Humberto deGouveia, Angélica SCardoso, EvandroMoreira, Samara SilveiraSoares, Filippe Elias de FreitasBraga, Fabio Ribeiro2019-03-25T17:44:43Z2019-03-25T17:44:43Z2018-032190-5738https://doi.org/10.1007/s13205-018-1192-4http://www.locus.ufv.br/handle/123456789/24107The objective of this study was to purify, characterize, and phylogenetically and structurally analyze the dextranase produced by the fungus Pochonia chlamydosporia. Dextranase produced by the fungus P. chlamydosporia was purified to homogeneity in two steps, with a yield of 152%, purification factor of 6.84 and specific activity of 358.63 U/mg. Its molecular weight was estimated by SDS-PAGE at 64 kDa. The enzyme presented higher activity at 50 °C and pH 5.0, using 100 mM citrate–phosphate buffer, was inhibited by Ag1+, Hg2+, Cu2+, Mg2+, and presented KM of 23.60 µM. Mature dextranase is composed of 585 amino acids residues, with a predicted molecular weight of 64.38 kDa and pI 5.96. This dextranase showed a strong phylogenetic similarity when compared to Trichoderma harzianum dextranase. Its structure consists of two domains: the first composed by 15 β strands, and the second composed by a right-handed parallel β-helix.eng3 BiotechVolume 8, Issue 3, Pages 1-9, March 2018Springer-Verlag GmbH Germany, part of Springer Natureinfo:eu-repo/semantics/openAccessEnzymeVerticillium chlamydosporiumPurification3D structureIn vitro and in silico characterization of a novel dextranase from Pochonia chlamydosporiainfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVORIGINALartigo.pdfartigo.pdfTexto completoapplication/pdf1991774https://locus.ufv.br//bitstream/123456789/24107/1/artigo.pdf296d1149a7042edc44896742f62fbfc0MD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81748https://locus.ufv.br//bitstream/123456789/24107/2/license.txt8a4605be74aa9ea9d79846c1fba20a33MD52123456789/241072019-03-25 14:47:30.956oai:locus.ufv.br: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Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452019-03-25T17:47:30LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false
dc.title.en.fl_str_mv In vitro and in silico characterization of a novel dextranase from Pochonia chlamydosporia
title In vitro and in silico characterization of a novel dextranase from Pochonia chlamydosporia
spellingShingle In vitro and in silico characterization of a novel dextranase from Pochonia chlamydosporia
Sufiate, Bruna
Enzyme
Verticillium chlamydosporium
Purification
3D structure
title_short In vitro and in silico characterization of a novel dextranase from Pochonia chlamydosporia
title_full In vitro and in silico characterization of a novel dextranase from Pochonia chlamydosporia
title_fullStr In vitro and in silico characterization of a novel dextranase from Pochonia chlamydosporia
title_full_unstemmed In vitro and in silico characterization of a novel dextranase from Pochonia chlamydosporia
title_sort In vitro and in silico characterization of a novel dextranase from Pochonia chlamydosporia
author Sufiate, Bruna
author_facet Sufiate, Bruna
Araújo, Jackson Victor de
Queiroz, José Humberto de
Gouveia, Angélica S
Cardoso, Evandro
Moreira, Samara Silveira
Soares, Filippe Elias de Freitas
Braga, Fabio Ribeiro
author_role author
author2 Araújo, Jackson Victor de
Queiroz, José Humberto de
Gouveia, Angélica S
Cardoso, Evandro
Moreira, Samara Silveira
Soares, Filippe Elias de Freitas
Braga, Fabio Ribeiro
author2_role author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Sufiate, Bruna
Araújo, Jackson Victor de
Queiroz, José Humberto de
Gouveia, Angélica S
Cardoso, Evandro
Moreira, Samara Silveira
Soares, Filippe Elias de Freitas
Braga, Fabio Ribeiro
dc.subject.pt-BR.fl_str_mv Enzyme
Verticillium chlamydosporium
Purification
3D structure
topic Enzyme
Verticillium chlamydosporium
Purification
3D structure
description The objective of this study was to purify, characterize, and phylogenetically and structurally analyze the dextranase produced by the fungus Pochonia chlamydosporia. Dextranase produced by the fungus P. chlamydosporia was purified to homogeneity in two steps, with a yield of 152%, purification factor of 6.84 and specific activity of 358.63 U/mg. Its molecular weight was estimated by SDS-PAGE at 64 kDa. The enzyme presented higher activity at 50 °C and pH 5.0, using 100 mM citrate–phosphate buffer, was inhibited by Ag1+, Hg2+, Cu2+, Mg2+, and presented KM of 23.60 µM. Mature dextranase is composed of 585 amino acids residues, with a predicted molecular weight of 64.38 kDa and pI 5.96. This dextranase showed a strong phylogenetic similarity when compared to Trichoderma harzianum dextranase. Its structure consists of two domains: the first composed by 15 β strands, and the second composed by a right-handed parallel β-helix.
publishDate 2018
dc.date.issued.fl_str_mv 2018-03
dc.date.accessioned.fl_str_mv 2019-03-25T17:44:43Z
dc.date.available.fl_str_mv 2019-03-25T17:44:43Z
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 https://doi.org/10.1007/s13205-018-1192-4
http://www.locus.ufv.br/handle/123456789/24107
dc.identifier.issn.none.fl_str_mv 2190-5738
identifier_str_mv 2190-5738
url https://doi.org/10.1007/s13205-018-1192-4
http://www.locus.ufv.br/handle/123456789/24107
dc.language.iso.fl_str_mv eng
language eng
dc.relation.ispartofseries.pt-BR.fl_str_mv Volume 8, Issue 3, Pages 1-9, March 2018
dc.rights.driver.fl_str_mv Springer-Verlag GmbH Germany, part of Springer Nature
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Springer-Verlag GmbH Germany, part of Springer Nature
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv 3 Biotech
publisher.none.fl_str_mv 3 Biotech
dc.source.none.fl_str_mv reponame:LOCUS Repositório Institucional da UFV
instname:Universidade Federal de Viçosa (UFV)
instacron:UFV
instname_str Universidade Federal de Viçosa (UFV)
instacron_str UFV
institution UFV
reponame_str LOCUS Repositório Institucional da UFV
collection LOCUS Repositório Institucional da UFV
bitstream.url.fl_str_mv https://locus.ufv.br//bitstream/123456789/24107/1/artigo.pdf
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