Functional characterization of GH7 endo-1,4-ß-glucanase from Aspergillus fumigatus and its potential industrial application.
Protein Expr Purif
; 150: 1-11, 2018 10.
Article
em En
| MEDLINE
| ID: mdl-29715559
ABSTRACT
A gene encoding an endo-1,4-ß-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 52â¯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⯰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 ß-glucan than toward xyloglucan and CM-Cellulose, suggesting that the enzyme corresponds to a ß-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.
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MEDLINE
Assunto principal:
Polissacarídeos
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Aspergillus fumigatus
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Proteínas Fúngicas
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Celulase
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Saccharum
Idioma:
En
Ano de publicação:
2018
Tipo de documento:
Article