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Functional and structural analyses of a 1,4-β-endoglucanase from Ganoderma lucidum

Paper ID Volume ID Publish Year Pages File Format Full-Text
16800 42613 2016 8 PDF Available
Title
Functional and structural analyses of a 1,4-β-endoglucanase from Ganoderma lucidum
Abstract

•The first 1,4-β-endoglucanase from Ganoderma lucidum (GlCel5A) is characterized.•GlCel5A expressed in Pichia pastoris shows good activity and thermostability.•Crystal structure of GlCel5A in complex with cellobiose was solved.

Ganoderma lucidum is a saprotrophic white-rot fungus which contains a rich set of cellulolytic enzymes. Here, we screened an array of potential 1,4-β-endoglucanases from G. lucidum based on the gene annotation library and found that one candidate gene, GlCel5A, exhibits CMC-hydrolyzing activity. The recombinant GlCel5A protein expressed in Pichia pastoris is able to hydrolyze CMC and β-glucan but not xylan and mannan. The enzyme exhibits optimal activity at 60 °C and pH 3–4, and retained 50% activity at 80 and 90 °C for at least 15 and 10 min. The crystal structure of GlCel5A and its complex with cellobiose, solved at 2.7 and 2.86 Å resolution, shows a classical (β/α)8 TIM-barrel fold as seen in other members of glycoside hydrolase family 5. The complex structure contains a cellobiose molecule in the +1 and +2 subsites, and reveals the interactions with the positive sites of the enzyme. Collectively, the present work provides the first comprehensive characterization of an endoglucanase from G. lucidum that possesses properties for industrial applications, and strongly encourages further studying in the cellulolytic enzyme system of G. lucidum.

Keywords
Ganoderma lucidum; Endoglucanase; Crystal structure
First Page Preview
Functional and structural analyses of a 1,4-β-endoglucanase from Ganoderma lucidum
Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 86, May 2016, Pages 67–74
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering