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Research article2005Peer reviewedOpen access

Structural and biochemical studies of GH family 12 cellulases: improved thermal stability, and ligand complexes

Sandgren M, Stahlberg J, Mitchinson C

Abstract

In this review we will describe how we have gathered structural and biochemical information from several homologous cellulases from one class of glycoside hydrolases (GH family 12), and used this information within the framework of a protein-engineering program for the design of new variants of these enzymes. These variants have been characterized to identify some of the positions and the types of mutations in the enzymes that are responsible for some of the biochemical differences in thermal stability and activity between the homologous enzymes. In this process we have solved the three-dimensional structure of four of these homologous GH 12 cellulases: Three fungal enzymes, Humicola grisea Cell2A, Hypocrea jecorina Cell2A and Hypocrea schweinitzii Cell2A, and one bacterial, Streptomyces sp. 11AG8 Cell2A. We have also determined the three-dimensional structures of the two most stable H. jecorina Cell2A variants. In addition, four ligand-complex structures of the wild-type H. grisea Cell2A enzyme have been solved and have made it possible to characterize some of the interactions between substrate and enzyme. The structural and biochemical studies of these related GH 12 enzymes, and their variants, have provided insight on how specific residues contribute to protein thermal stability and enzyme activity. This knowledge can serve as a structural toolbox for the design of Cell2A enzymes with specific properties and features suited to existing or new applications. (C) 2004 Elsevier Ltd. All rights reserved

Keywords

glycoside hydrolase; endoglucanase; protein engineering; protein structure; thermal stability

Published in

Progress in Biophysics and Molecular Biology
2005, Volume: 89, number: 3, pages: 246-291
Publisher: PERGAMON-ELSEVIER SCIENCE LTD

      SLU Authors

    • Ståhlberg, Jerry

      • Department of Molecular Biology, Swedish University of Agricultural Sciences

      • Sandgren, Mats

        • Department of Molecular Biology, Swedish University of Agricultural Sciences

      UKÄ Subject classification

      Renewable Bioenergy Research
      Food Science

      Publication identifier

      DOI: https://doi.org/10.1016/j.pbiomolbio.2004.11.002

      Permanent link to this page (URI)

      https://res.slu.se/id/publ/8402