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Article
Tertiary Structure and Characterization of a Glycoside Hydrolase Family 44 Endoglucanase from Clostridium acetobutylicum
Applied and Environmental Microbiology
  • Christopher D. Warner, Iowa State University
  • Julie A. Hoy, Iowa State University
  • Taran C. Shilling, Iowa State University
  • Michael J. Linnen, Iowa State University
  • Nathaniel D. Ginder, Iowa State University
  • Clark Ford, Iowa State University
  • Richard B. Honzatko, Iowa State University
  • Peter J. Reilly, Iowa State University
Document Type
Article
Publication Date
1-1-2010
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
Date Available
November 19, 2012
File Format
application/pdf
Citation Information
Christopher D. Warner, Julie A. Hoy, Taran C. Shilling, Michael J. Linnen, et al.. "Tertiary Structure and Characterization of a Glycoside Hydrolase Family 44 Endoglucanase from Clostridium acetobutylicum" Applied and Environmental Microbiology Vol. 76 Iss. 1 (2010) p. 338 - 346
Available at: http://works.bepress.com/peter_reilly/39/