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(Referência obtida automaticamente do Web of Science, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

Biochemical characterization of an isolated 50 kDa beta-glucosidase from the thermophilic fungus Myceliophthora thermophila M.7.7

Texto completo
Autor(es):
Bonfa, Emily Colferai [1] ; de Souza Moretti, Marcia Maria [1] ; Gomes, Eleni [1] ; Bonilla-Rodriguez, Gustavo Orlando [1]
Número total de Autores: 4
Afiliação do(s) autor(es):
[1] Sao Paulo State Univ Unesp, Inst Biosci Humanities & Exact Sci Ibilce, Sao Jose do Rio Preto Campus, Sao Jose Do Rio Preto, SP - Brazil
Número total de Afiliações: 1
Tipo de documento: Artigo Científico
Fonte: BIOCATALYSIS AND AGRICULTURAL BIOTECHNOLOGY; v. 13, p. 311-318, JAN 2018.
Citações Web of Science: 3
Resumo

This study characterized a 50 kDa beta-glucosidase (BGL50) produced by the thermophilic fungus Myceliophthora thermophila M.7.7 in solid state cultivation using a mixture of (1:1) sugarcane bagasse and wheat bran. The crude extract zymogram showed two isoforms of beta-glucosidase with approximately 50 and 200 kDa, which were separated by gel filtration chromatography. The characterization of BGL50 showed optimum activity at 60 degrees C and pH 5.0 when 4-nitrophenyl beta-D-glucopyranoside (pNPG) was used as the substrate, whereas when using cellobiose, the highest activity was observed at 50 degrees C and pH 4.5. Several ions and reagents produced different effects on the enzyme activity depending on the substrate and there was complete inhibition with Cu2+ and Fe3+ for both substrates. In addition, nine phenolic compounds showed no inhibitory effects on the enzyme, a significant feature since beta-glucosidase is used for the saccharification of lignocellulosic biomass that generates several phenolic compounds. Kinetic studies revealed competitive inhibition by glucose when pNPG was used, with a K-i value of 1.5 mM and a significantly lower K-m (0.52 mM) than for cellobiose (8.50 mM). The thermodynamic parameters showed that BGL50 is very stable at 60 degrees C displaying a half-life of 855.6 min but it is easily denatured above this temperature. The results emphasize the importance of investigating potential beta-glucosidases based on cellobiose instead of using only pNPG since, in the industrial process, the enzyme will act on this natural substrate. In addition, understanding the thermostability of the enzyme is an important contribution to enzyme technology. (AU)

Processo FAPESP: 10/12624-0 - Aplicação de métodos físico-químicos e enzimáticos na sacarificação do bagaço de cana: estudos de microrganismos, processos fermentativos relacionados e métodos de hidrólise
Beneficiário:Eleni Gomes
Linha de fomento: Auxílio à Pesquisa - Temático
Processo FAPESP: 11/23991-7 - Análise funcional e estrutural de hidrolases microbianas e vegetais com uso potencial em processos biotecnológicos ou industriais
Beneficiário:Gustavo Orlando Bonilla Rodriguez
Linha de fomento: Auxílio à Pesquisa - Regular