Xylanase and β-xylosidase from Penicillium janczewskii: Purification, characterization and hydrolysis of substrates
Autor(a) principal: | |
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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/10.1016/j.ejbt.2016.08.001 http://hdl.handle.net/11449/173465 |
Resumo: | Background: Xylanases and β-D-xylosidases are the most important enzymes responsible for the degradation of xylan, the second main constituent of plant cell walls. Results: In this study, the main extracellular xylanase (XYL I) and β-xylosidase (BXYL I) from the fungus Penicillium janczewskii were purified, characterized and applied for the hydrolysis of different substrates. Their molecular weights under denaturing and non-denaturing conditions were, respectively, 30.4 and 23.6 kDa for XYL I, and 100 and 200 kDa for BXYL I, indicating that the latter is homodimeric. XYL I is highly glycosylated (78%) with optimal activity in pH 6.0 at 65°C, while BXYL I presented lower sugar content (10.5%) and optimal activity in pH 5.0 at 75°C. The half-lives of XYL I at 55, 60 and 65°C were 125, 16 and 6 min, respectively. At 60°C, BXYL I retained almost 100% of the activity after 6 h. NH+ 4,Na+, DTT and β-mercaptoethanol stimulated XYL I, while activation of BXYL I was not observed. Interestingly, XYL I was only partially inhibited by Hg2+, while BXYL I was completely inhibited. Xylobiose, xylotriose and larger xylooligosaccharides were the main products from xylan hydrolysis by XYL I. BXYL I hydrolyzed xylobiose and larger xylooligosaccharides with no activity against xylans. Conclusion: The enzymes act synergistically in the degradation of xylans, and present industrial characteristics especially in relation to optimal activity at high temperatures, prolonged stability of BXYL I at 60°C, and stability of XYL I in wide pH range. |
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Xylanase and β-xylosidase from Penicillium janczewskii: Purification, characterization and hydrolysis of substratesEnzyme characterizationEnzyme purificationXylan hydrolysisXylanolytic enzymesXylooligosaccharides hydrolysisBackground: Xylanases and β-D-xylosidases are the most important enzymes responsible for the degradation of xylan, the second main constituent of plant cell walls. Results: In this study, the main extracellular xylanase (XYL I) and β-xylosidase (BXYL I) from the fungus Penicillium janczewskii were purified, characterized and applied for the hydrolysis of different substrates. Their molecular weights under denaturing and non-denaturing conditions were, respectively, 30.4 and 23.6 kDa for XYL I, and 100 and 200 kDa for BXYL I, indicating that the latter is homodimeric. XYL I is highly glycosylated (78%) with optimal activity in pH 6.0 at 65°C, while BXYL I presented lower sugar content (10.5%) and optimal activity in pH 5.0 at 75°C. The half-lives of XYL I at 55, 60 and 65°C were 125, 16 and 6 min, respectively. At 60°C, BXYL I retained almost 100% of the activity after 6 h. NH+ 4,Na+, DTT and β-mercaptoethanol stimulated XYL I, while activation of BXYL I was not observed. Interestingly, XYL I was only partially inhibited by Hg2+, while BXYL I was completely inhibited. Xylobiose, xylotriose and larger xylooligosaccharides were the main products from xylan hydrolysis by XYL I. BXYL I hydrolyzed xylobiose and larger xylooligosaccharides with no activity against xylans. Conclusion: The enzymes act synergistically in the degradation of xylans, and present industrial characteristics especially in relation to optimal activity at high temperatures, prolonged stability of BXYL I at 60°C, and stability of XYL I in wide pH range.Biochemistry and Microbiology Department Biosciences Institute Univ Estadual Paulista – UNESP, PO Box 199, Av. 24 A, no. 1515Departamento de Biocatálisis Instituto de Catálisis CSIC (Consejo Superior de Investigaciones Científicas), Campus Universidad Autónoma de Madrid, CantoblancoBiochemistry and Microbiology Department Biosciences Institute Univ Estadual Paulista – UNESP, PO Box 199, Av. 24 A, no. 1515Universidade Estadual Paulista (Unesp)CSIC (Consejo Superior de Investigaciones Científicas)Terrasan, César Rafael Fanchini [UNESP]Guisan, José ManuelCarmona, Eleonora Cano [UNESP]2018-12-11T17:05:40Z2018-12-11T17:05:40Z2016-09-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article54-62application/pdfhttp://dx.doi.org/10.1016/j.ejbt.2016.08.001Electronic Journal of Biotechnology, v. 23, p. 54-62.0717-3458http://hdl.handle.net/11449/17346510.1016/j.ejbt.2016.08.0012-s2.0-849876190332-s2.0-84987619033.pdfScopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengElectronic Journal of Biotechnology0,537info:eu-repo/semantics/openAccess2023-10-08T06:02:34Zoai:repositorio.unesp.br:11449/173465Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462023-10-08T06:02:34Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Xylanase and β-xylosidase from Penicillium janczewskii: Purification, characterization and hydrolysis of substrates |
title |
Xylanase and β-xylosidase from Penicillium janczewskii: Purification, characterization and hydrolysis of substrates |
spellingShingle |
Xylanase and β-xylosidase from Penicillium janczewskii: Purification, characterization and hydrolysis of substrates Terrasan, César Rafael Fanchini [UNESP] Enzyme characterization Enzyme purification Xylan hydrolysis Xylanolytic enzymes Xylooligosaccharides hydrolysis |
title_short |
Xylanase and β-xylosidase from Penicillium janczewskii: Purification, characterization and hydrolysis of substrates |
title_full |
Xylanase and β-xylosidase from Penicillium janczewskii: Purification, characterization and hydrolysis of substrates |
title_fullStr |
Xylanase and β-xylosidase from Penicillium janczewskii: Purification, characterization and hydrolysis of substrates |
title_full_unstemmed |
Xylanase and β-xylosidase from Penicillium janczewskii: Purification, characterization and hydrolysis of substrates |
title_sort |
Xylanase and β-xylosidase from Penicillium janczewskii: Purification, characterization and hydrolysis of substrates |
author |
Terrasan, César Rafael Fanchini [UNESP] |
author_facet |
Terrasan, César Rafael Fanchini [UNESP] Guisan, José Manuel Carmona, Eleonora Cano [UNESP] |
author_role |
author |
author2 |
Guisan, José Manuel Carmona, Eleonora Cano [UNESP] |
author2_role |
author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) CSIC (Consejo Superior de Investigaciones Científicas) |
dc.contributor.author.fl_str_mv |
Terrasan, César Rafael Fanchini [UNESP] Guisan, José Manuel Carmona, Eleonora Cano [UNESP] |
dc.subject.por.fl_str_mv |
Enzyme characterization Enzyme purification Xylan hydrolysis Xylanolytic enzymes Xylooligosaccharides hydrolysis |
topic |
Enzyme characterization Enzyme purification Xylan hydrolysis Xylanolytic enzymes Xylooligosaccharides hydrolysis |
description |
Background: Xylanases and β-D-xylosidases are the most important enzymes responsible for the degradation of xylan, the second main constituent of plant cell walls. Results: In this study, the main extracellular xylanase (XYL I) and β-xylosidase (BXYL I) from the fungus Penicillium janczewskii were purified, characterized and applied for the hydrolysis of different substrates. Their molecular weights under denaturing and non-denaturing conditions were, respectively, 30.4 and 23.6 kDa for XYL I, and 100 and 200 kDa for BXYL I, indicating that the latter is homodimeric. XYL I is highly glycosylated (78%) with optimal activity in pH 6.0 at 65°C, while BXYL I presented lower sugar content (10.5%) and optimal activity in pH 5.0 at 75°C. The half-lives of XYL I at 55, 60 and 65°C were 125, 16 and 6 min, respectively. At 60°C, BXYL I retained almost 100% of the activity after 6 h. NH+ 4,Na+, DTT and β-mercaptoethanol stimulated XYL I, while activation of BXYL I was not observed. Interestingly, XYL I was only partially inhibited by Hg2+, while BXYL I was completely inhibited. Xylobiose, xylotriose and larger xylooligosaccharides were the main products from xylan hydrolysis by XYL I. BXYL I hydrolyzed xylobiose and larger xylooligosaccharides with no activity against xylans. Conclusion: The enzymes act synergistically in the degradation of xylans, and present industrial characteristics especially in relation to optimal activity at high temperatures, prolonged stability of BXYL I at 60°C, and stability of XYL I in wide pH range. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016-09-01 2018-12-11T17:05:40Z 2018-12-11T17:05:40Z |
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/10.1016/j.ejbt.2016.08.001 Electronic Journal of Biotechnology, v. 23, p. 54-62. 0717-3458 http://hdl.handle.net/11449/173465 10.1016/j.ejbt.2016.08.001 2-s2.0-84987619033 2-s2.0-84987619033.pdf |
url |
http://dx.doi.org/10.1016/j.ejbt.2016.08.001 http://hdl.handle.net/11449/173465 |
identifier_str_mv |
Electronic Journal of Biotechnology, v. 23, p. 54-62. 0717-3458 10.1016/j.ejbt.2016.08.001 2-s2.0-84987619033 2-s2.0-84987619033.pdf |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Electronic Journal of Biotechnology 0,537 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
54-62 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 |
|
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1797789324750618624 |