From Data Mining of Chitinophaga sp. Genome to Enzyme Discovery of a Hyperthermophilic Metallocarboxypeptidase

Autor(es)

Fernandes, Gabriela Cabral ^{[1, 2]} ; Sierra, Elwi Guillermo Machado ^{[1, 3]} ; Brear, Paul ^[4] ; Pereira, Mariana Rangel ^{[4, 5]} ; Lemos, Eliana G. M. ^[1]

Afiliação do(s) autor(es)

^[1] Sao Paulo State Univ UNESP, Dept Technol, BR-14884900 Jaboticabal, SP - Brazil

^[2] Sao Paulo State Univ UNESP, Sch Agr & Vet Sci, Grad Program Agr & Livestock Microbiol, BR-14884900 Jaboticabal, SP - Brazil

^[3] Univ Simon Bolivar, Fac Ciencias Basicas & Biomed, Lab Invest Microbiol, Barranquilla 080002 - Colombia

^[4] Univ Cambridge, Dept Biochem, Cambridge CB21GA - England

^[5] Minist Educ Brazil, CAPES Fdn, BR-7004002 Brasilia, DF - Brazil

• Tipo de documento: Artigo Científico
• Fonte: MICROORGANISMS; v. 9, n. 2 FEB 2021.
• Citações Web of Science: 0

Resumo

For 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 degrees C and 75 degrees C, and 62% activity was still displayed at 100 degrees C. We observed that Ca2+, Ba2+, Mn2+ and Mg2+ ions had a positive effect on the activity of ChtCP, and an increase of 30 degrees C in the melting temperature was observed in the presence of Co2+. These features together with the structure of ChtCP at 1.2 angstrom highlight the relevance of ChtCP for further biotechnological applications. (AU)

• Processo FAPESP: 16/23892-2 - Nova vertente para descoberta de biomoléculas, lacases, com aplicação biotecnológica (LBMPMetaDB)
• Beneficiário: Eliana Gertrudes de Macedo Lemos
• Modalidade de apoio: Auxílio à Pesquisa - Regular