| Full text | |
| Author(s): |
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]
Total Authors: 7
|
| Affiliation: | [1] Univ Sao Paulo, Fac Filosofia Ciencias & Letras Ribeirao Preto, Ribeirao Preto, SP - Brazil
[2] Fac Ciencias Farmaceut Ribeirao Preto, Ribeirao Preto, SP - Brazil
Total Affiliations: 2
|
| Document type: | Journal article |
| Source: | Protein Expression and Purification; v. 150, p. 1-11, OCT 2018. |
| Web of Science Citations: | 7 |
| Abstract | |
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) | |
| FAPESP's process: | 14/10466-0 - Transcriptome analysis of Aspergillus fumigatus grown on sugarcane bagasse |
| Grantee: | Taisa Magnani Dinamarco |
| Support Opportunities: | Program for Research on Bioenergy (BIOEN) - Regular Program Grants |
| FAPESP's process: | 16/19095-0 - Identification, characterization and functional expression of auxiliary activity enzymes (LPMOs) from Aspergillus fumigatus |
| Grantee: | Taisa Magnani Dinamarco |
| Support Opportunities: | Program for Research on Bioenergy (BIOEN) - Regular Program Grants |