Document Type

Article

Publication Date

1-2010

Journal or Book Title

Applied and Environmental Microbiology

Volume

76

Issue

1

First Page

338

Last Page

346

DOI

10.1128/AEM.02026-09

Abstract

A gene encoding a glycoside hydrolase family 44 (GH44) protein from Clostridium acetobutylicum ATCC 824 was synthesized and transformed into Escherichia coli.The previously uncharacterized protein was expressed with a C-terminal His tag and purified by nickel-nitrilotriacetic acid affinity chromatography. Crystallization and X-ray diffraction to a 2.2-Å resolution revealed a triose phosphate isomerase (TIM) barrel-like structure with additional Greek key and β-sandwich folds, similar to other GH44 crystal structures. The enzyme hydrolyzes cellotetraose and larger cellooligosaccharides, yielding an unbalanced product distribution, including some glucose. It attacks carboxymethylcellulose and xylan at approximately the same rates. Its activity on carboxymethylcellulose is much higher than that of the isolated C. acetobutylicum cellulosome. It also extensively converts lichenan to oligosaccharides of intermediate size and attacks Avicel to a limited extent. The enzyme has an optimal temperature in a 10-min assay of 55°C and an optimal pH of 5.0.

Comments

This is a post-print of an article from Applied and Environmental Microbiology, 76, no. 1 (January 2010): 338–346, doi: 10.1128/AEM.02026-09.

Copyright Owner

American Society for Microbiology

Language

en

File Format

application/pdf

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