From data mining of chitinophaga sp. Genome to enzyme discovery of a hyperthermophilic metallocarboxypeptidase

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MetadadosDescriçãoIdioma
Autor(es): dc.contributorUniversidade Estadual Paulista (Unesp)-
Autor(es): dc.contributorUniversidad Simón Bolívar-
Autor(es): dc.contributorUniversity of Cambridge-
Autor(es): dc.contributorMinistry of Education of Brazil-
Autor(es): dc.creatorFernandes, Gabriela Cabral [UNESP]-
Autor(es): dc.creatorSierra, Elwi Guillermo Machado [UNESP]-
Autor(es): dc.creatorBrear, Paul-
Autor(es): dc.creatorPereira, Mariana Rangel-
Autor(es): dc.creatorLemos, Eliana G. M. [UNESP]-
Data de aceite: dc.date.accessioned2022-02-22T00:45:32Z-
Data de disponibilização: dc.date.available2022-02-22T00:45:32Z-
Data de envio: dc.date.issued2021-06-25-
Data de envio: dc.date.issued2021-06-25-
Data de envio: dc.date.issued2021-01-31-
Fonte completa do material: dc.identifierhttp://dx.doi.org/10.3390/microorganisms9020393-
Fonte completa do material: dc.identifierhttp://hdl.handle.net/11449/205871-
Fonte: dc.identifier.urihttp://educapes.capes.gov.br/handle/11449/205871-
Descrição: dc.descriptionFor several centuries, microorganisms and enzymes have been used for many different applications. Although many enzymes with industrial applications have already been reported, different screening technologies, methods and approaches are constantly being developed in order to allow the identification of enzymes with even more interesting applications. In our work, we have performed data mining on the Chitinophaga sp. genome, a gram-negative bacterium isolated from a bacterial consortium of sugarcane bagasse isolated from an ethanol plant. The analysis of 8 Mb allowed the identification of the chtcp gene, previously annotated as putative Cht4039. The corresponding codified enzyme, denominated as ChtCP, showed the HEXXH conserved motif of family M32 from thermostable carboxypeptidases. After expression in E. coli, the recombinant enzyme was characterized biochemically. ChtCP showed the highest activity versus benziloxicarbonil Ala-Trp at pH 7.5, suggesting a preference for hydrophobic substrates. Surprisingly, the highest activity of ChtCP observed was between 55 °C and 75 °C, and 62% activity was still displayed at 100 °C. We observed that Ca2+, Ba2+, Mn2+ and Mg2+ ions had a positive effect on the activity of ChtCP, and an increase of 30 °C in the melting temperature was observed in the presence of Co2+. These features together with the structure of ChtCP at 1.2 Å highlight the relevance of ChtCP for further biotechnological applications.-
Descrição: dc.descriptionCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)-
Descrição: dc.descriptionFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)-
Descrição: dc.descriptionConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)-
Descrição: dc.descriptionDepartment of Technology São Paulo State University (UNESP)-
Descrição: dc.descriptionSão Paulo State University (UNESP) School of Agricultural and Veterinarian Sciences-
Descrição: dc.descriptionLaboratorio de Investigación en Microbiología Facultad de Ciencias Básicas y Biomédicas Universidad Simón Bolívar-
Descrição: dc.descriptionDepartment of Biochemistry University of Cambridge-
Descrição: dc.descriptionCAPES Foundation Ministry of Education of Brazil, DF-
Descrição: dc.descriptionDepartment of Technology São Paulo State University (UNESP)-
Descrição: dc.descriptionSão Paulo State University (UNESP) School of Agricultural and Veterinarian Sciences-
Descrição: dc.descriptionCAPES: 001-
Descrição: dc.descriptionFAPESP: 2016/23892-2-
Descrição: dc.descriptionCNPq: 401886/2016-1-
Formato: dc.format1-18-
Idioma: dc.languageen-
Relação: dc.relationMicroorganisms-
???dc.source???: dc.sourceScopus-
Palavras-chave: dc.subjectChitinophaga sp-
Palavras-chave: dc.subjectM32 family of peptidases-
Palavras-chave: dc.subjectMetallocarboxypeptidase-
Título: dc.titleFrom data mining of chitinophaga sp. Genome to enzyme discovery of a hyperthermophilic metallocarboxypeptidase-
Tipo de arquivo: dc.typelivro digital-
Aparece nas coleções:Repositório Institucional - Unesp

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