Functional characterization of GH7 endo-1,4-beta-glucanase from Aspergillus fumigatus and its potential industrial application

Autor(es)

Bernardi, Aline Vianna ^[1] ; de Gouvea, Paula Fagundes ^[1] ; Gerolamo, Luis Eduardo ^[1] ; Yonamine, Deborah Kimie ^[1] ; de Lima Balico, Lais de Lourdes ^[2] ; Uyemura, Sergio Akira ^[2] ; Dinamarco, Taisa Magnani ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Fac Filosofia Ciencias & Letras Ribeirao Preto, Ribeirao Preto, SP - Brazil

^[2] Fac Ciencias Farmaceut Ribeirao Preto, Ribeirao Preto, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Protein Expression and Purification; v. 150, p. 1-11, OCT 2018.
• Citações Web of Science: 7

Resumo

A gene encoding an endo-1,4-beta-glucanase (Afu6g01800) from A. fumigatus was cloned into the vector pET-28a (+) and expressed in the E. coli strain RosettaTM (DE3) pLysS. Sequence analysis indicated that the enzyme Af-EGL7 belonged to the GH7 family. The gene Af-egl7 encoded a protein comprising 460 amino acids, with a CBM1 domain at residues 424-460 and molecular mass of 52 kDa, as estimated by SDS-PAGE. This enzyme was optimally active at pH and temperatures ranging from 4.5 to 5.5 and from 40 to 60 degrees C, respectively. Mn2+ addition significantly enhanced the Af-EGL7 cellulase activity by 233%, whereas SDS addition fully inhibited this activity. Higher activity was observed toward beta-glucan than toward xyloglucan and CM-Cellulose, suggesting that the enzyme corresponds to a beta-1,3-1,4-glucanase. qRT-PCR in different culture media helped to establish the time course expression profile. Different polysaccharides induced the gene Af-egl7 in a time-dependent manner; in the particular case of the substrate sugarcane exploded bagasse (SEB), Af-egl7 was induced 2500-fold. Upon addition to a commercial cellulase cocktail, Af-EGL7 significantly improved SEB saccharification, which suggested that the enzyme Af-EGL7 had great potential to hydrolyze complex biomass. From a biotechnological point of view, A. fumigatus Af-EGL7 is a promising candidate to enhance enzyme cocktails used in biorefineries such as consolidated bioprocessing. (AU)

• Processo FAPESP: 14/10466-0 - Análise transcricional do fungo Aspergillus fumigatus quando crescido em bagaço de cana-de-açúcar
• Beneficiário: Taisa Magnani Dinamarco
• Modalidade de apoio: Auxílio à Pesquisa - Programa BIOEN - Regular
• Processo FAPESP: 16/19095-0 - Identificação, caracterização e expressão funcional de enzimas auxiliares hidrolíticas do tipo LPMOs do fungo Aspergillus fumigatus
• Beneficiário: Taisa Magnani Dinamarco
• Modalidade de apoio: Auxílio à Pesquisa - Programa BIOEN - Regular