A broad pH range and processive chitinase from a metagenome library
| Autor(a) principal: | |
|---|---|
| Data de Publicação: | 2017 |
| Outros Autores: | , , , , , |
| Tipo de documento: | Artigo |
| Idioma: | por |
| Título da fonte: | Repositório Institucional da FIOCRUZ (ARCA) |
| Texto Completo: | https://www.arca.fiocruz.br/handle/icict/20956 |
Resumo: | Universidade Federal do Paraná. Departamento de Bioquímica e Biologia Molecular. Curitiba, PR, Brasil. |
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Thimoteo, S. S.Glogauer, A.Faoro, HelissonSouza, E. M. deHuergo, L. F.Moerschbacher, B. M.Pedrosa, Fábio Oliveira2017-09-12T13:30:31Z2017-09-12T13:30:31Z2017THIMOTEO, S. S. et al. A broad pH range and processive chitinase from a metagenome library. Brazilian Journal of Medical and Biological Research, v. 50, n. 1, p. 1-13, 2017.1414-431Xhttps://www.arca.fiocruz.br/handle/icict/2095610.1590/1414-431X20165658porAssociação Brasileira de Divulgação CientíficaAeromonasChitinasesMetagenomicsKineticsQuitinasasMetagenómicaCinéticaQuitinasesMetagenômicaCinéticaA broad pH range and processive chitinase from a metagenome libraryinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleUniversidade Federal do Paraná. Departamento de Bioquímica e Biologia Molecular. Curitiba, PR, Brasil.Universidade Federal do Paraná. Departamento de Bioquímica e Biologia Molecular. Curitiba, PR, Brasil / Instituto de Tecnologia do Paraná. Agência de Inovação. Curitiba, PR, Brasil.Universidade Federal do Paraná. Departamento de Bioquímica e Biologia Molecular. Curitiba, PR, Brasil / Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Universidade Federal do Paraná. Departamento de Bioquímica e Biologia Molecular. Curitiba, PR, BrasilUniversidade Federal do Paraná. Departamento de Bioquímica e Biologia Molecular. Curitiba, PR, Brasil.Institute for Biology and Biotechnology of Plants, WWU Münster University, Münster, Germany.Universidade Federal do Paraná. Departamento de Bioquímica e Biologia Molecular. Curitiba, PR, Brasil.Chitinases are hydrolases that degrade chitin, a polymer of N-acetylglucosamine linked β(1-4) present in the exoskeleton of crustaceans, insects, nematodes and fungal cell walls. A metagenome fosmid library from a wastewater-contaminated soil was functionally screened for chitinase activity leading to the isolation and identification of a chitinase gene named metachi18A. The metachi18A gene was subcloned and overexpressed in Escherichia coli BL21 and the MetaChi18A chitinase was purified by affinity chromatography as a 6xHis-tagged fusion protein. The MetaChi18A enzyme is a 92-kDa protein with a conserved active site domain of glycosyl hydrolases family 18. It hydrolyses colloidal chitin with an optimum pH of 5 and temperature of 50°C. Moreover, the enzyme retained at least 80% of its activity in the pH range from 4 to 9 and 98% at 600 mM NaCl. Thin layer chromatography analyses identified chitobiose as the main product of MetaChi18A on chitin polymers as substrate. Kinetic analysis showed inhibition of MetaChi18A activity at high concentrations of colloidal chitin and 4-methylumbelliferyl N,N'-diacetylchitobiose and sigmoid kinetics at low concentrations of colloidal chitin, indicating a possible conformational change to lead the chitin chain from the chitin-binding to the catalytic domain. The observed stability and activity of MetaChi18A over a wide range of conditions suggest that this chitinase, now characterized, may be suitable for application in the industrial processing of chitin.info:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-83084https://www.arca.fiocruz.br/bitstream/icict/20956/1/license.txt783568c2893d2e25a99990b126be1772MD51ORIGINALThimoteoabroadph.pdfThimoteoabroadph.pdfapplication/pdf944551https://www.arca.fiocruz.br/bitstream/icict/20956/2/Thimoteoabroadph.pdf6cd4f2bd464074f8f440fe163802860cMD52TEXTThimoteoabroadph.pdf.txtThimoteoabroadph.pdf.txtExtracted texttext/plain56042https://www.arca.fiocruz.br/bitstream/icict/20956/3/Thimoteoabroadph.pdf.txt9e9bcdd4f5d9c7d14134a35a6a7c8fd1MD53icict/209562019-11-23 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| dc.title.pt_BR.fl_str_mv |
A broad pH range and processive chitinase from a metagenome library |
| title |
A broad pH range and processive chitinase from a metagenome library |
| spellingShingle |
A broad pH range and processive chitinase from a metagenome library Thimoteo, S. S. Aeromonas Chitinases Metagenomics Kinetics Quitinasas Metagenómica Cinética Quitinases Metagenômica Cinética |
| title_short |
A broad pH range and processive chitinase from a metagenome library |
| title_full |
A broad pH range and processive chitinase from a metagenome library |
| title_fullStr |
A broad pH range and processive chitinase from a metagenome library |
| title_full_unstemmed |
A broad pH range and processive chitinase from a metagenome library |
| title_sort |
A broad pH range and processive chitinase from a metagenome library |
| author |
Thimoteo, S. S. |
| author_facet |
Thimoteo, S. S. Glogauer, A. Faoro, Helisson Souza, E. M. de Huergo, L. F. Moerschbacher, B. M. Pedrosa, Fábio Oliveira |
| author_role |
author |
| author2 |
Glogauer, A. Faoro, Helisson Souza, E. M. de Huergo, L. F. Moerschbacher, B. M. Pedrosa, Fábio Oliveira |
| author2_role |
author author author author author author |
| dc.contributor.author.fl_str_mv |
Thimoteo, S. S. Glogauer, A. Faoro, Helisson Souza, E. M. de Huergo, L. F. Moerschbacher, B. M. Pedrosa, Fábio Oliveira |
| dc.subject.other.pt_BR.fl_str_mv |
Aeromonas |
| topic |
Aeromonas Chitinases Metagenomics Kinetics Quitinasas Metagenómica Cinética Quitinases Metagenômica Cinética |
| dc.subject.en.pt_BR.fl_str_mv |
Chitinases Metagenomics Kinetics |
| dc.subject.es.pt_BR.fl_str_mv |
Quitinasas Metagenómica Cinética |
| dc.subject.decs.pt_BR.fl_str_mv |
Quitinases Metagenômica Cinética |
| description |
Universidade Federal do Paraná. Departamento de Bioquímica e Biologia Molecular. Curitiba, PR, Brasil. |
| publishDate |
2017 |
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2017-09-12T13:30:31Z |
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2017-09-12T13:30:31Z |
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2017 |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/article |
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article |
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THIMOTEO, S. S. et al. A broad pH range and processive chitinase from a metagenome library. Brazilian Journal of Medical and Biological Research, v. 50, n. 1, p. 1-13, 2017. |
| dc.identifier.uri.fl_str_mv |
https://www.arca.fiocruz.br/handle/icict/20956 |
| dc.identifier.issn.pt_BR.fl_str_mv |
1414-431X |
| dc.identifier.doi.none.fl_str_mv |
10.1590/1414-431X20165658 |
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THIMOTEO, S. S. et al. A broad pH range and processive chitinase from a metagenome library. Brazilian Journal of Medical and Biological Research, v. 50, n. 1, p. 1-13, 2017. 1414-431X 10.1590/1414-431X20165658 |
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openAccess |
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Associação Brasileira de Divulgação Científica |
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Associação Brasileira de Divulgação Científica |
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