The family 21 carbohydrate-binding module of glucoamylase from Rhizopus oryzae consists of two sites playing distinct roles in ligand binding

The Biochemical Journal
Wei-I ChouMargaret D-T Chang

Abstract

The starch-hydrolysing enzyme GA (glucoamylase) from Rhizopus oryzae is a commonly used glycoside hydrolase in industry. It consists of a C-terminal catalytic domain and an N-terminal starch-binding domain, which belong to the CBM21 (carbohydrate-binding module, family 21). In the present study, a molecular model of CBM21 from R. oryzae GA (RoGACBM21) was constructed according to PSSC (progressive secondary structure correlation), modified structure-based sequence alignment, and site-directed mutagenesis was used to identify and characterize potential ligand-binding sites. Our model suggests that RoGACBM21 contains two ligand-binding sites, with Tyr32 and Tyr67 grouped into site I, and Trp47, Tyr83 and Tyr93 grouped into site II. The involvement of these aromatic residues has been validated using chemical modification, UV difference spectroscopy studies, and both qualitative and quantitative binding assays on a series of RoGACBM21 mutants. Our results further reveal that binding sites I and II play distinct roles in ligand binding, the former not only is involved in binding insoluble starch, but also facilitates the binding of RoGACBM21 to long-chain soluble polysaccharides, whereas the latter serves as the major binding site m...Continue Reading

References

Aug 1, 1997·Bioscience, Biotechnology, and Biochemistry·S Chiba
Nov 20, 1997·Proteins·P M Coutinho, P J Reilly
Mar 21, 1998·Journal of Molecular Medicine : Official Organ of the Gesellschaft Deutscher Naturforscher Und Ärzte·P BorkM Huynen
Feb 3, 1999·Bioinformatics·J A CuffG J Barton
Aug 19, 1999·FEBS Letters·S Janecek, J Sevcík
Mar 1, 2000·Trends in Biochemical Sciences·C CombetG Deléage
Jun 22, 2000·Journal of Molecular Biology·L A KelleyM J Sternberg
Jan 11, 2001·Biochimica Et Biophysica Acta·J SauerB Svensson
Mar 29, 2001·Bioscience, Biotechnology, and Biochemistry·M K AliK Ohmiya
Feb 13, 2003·Biochemistry·Lan GuanH Ronald Kaback
Jun 26, 2003·Nucleic Acids Research·Torsten SchwedeManuel C Peitsch
Jul 29, 2003·Applied Microbiology and Biotechnology·J Houghton-Larsen, P A Pedersen
Jun 25, 2004·The Biochemical Journal·Alisdair B BorastonGideon J Davies
Jun 9, 2005·Current Opinion in Microbiology·Romina Rodríguez-SanojaSergio Sánchez

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Citations

Feb 2, 2012·Chemical Communications : Chem Comm·Preyesh StephenPing-Chiang Lyu
May 23, 2013·Acta Crystallographica. Section D, Biological Crystallography·Jose A Cuesta-SeijoMonica M Palcic
Jun 27, 2007·BMC Biochemistry·Shu-Chuan LinMargaret Dah-Tsyr Chang
Oct 4, 2011·PloS One·Wei-Yao ChouMargaret Dah-Tsyr Chang
Nov 7, 2006·FEBS Letters·Martin Machovic, Stefan Janecek
Nov 7, 2012·Bioresource Technology·Jorge A FerreiraMohammad J Taherzadeh
Mar 20, 2009·Protein Expression and Purification·Shu-Chuan LinMargaret Dah-Tsyr Chang
Oct 28, 2008·Biochemical and Biophysical Research Communications·Wei-Ting LiuMargaret Dah-Tsyr Chang
Nov 11, 2006·Journal of Molecular Biology·Alicia Lammerts van Bueren, Alisdair B Boraston
May 4, 2016·Cellular and Molecular Life Sciences : CMLS·Matthew H FoleyNicole M Koropatkin
Aug 28, 2009·Critical Reviews in Biotechnology·Pardeep Kumar, T Satyanarayana
Nov 23, 2006·The Biochemical Journal·Yu-Nan LiuPing-Chiang Lyu
Sep 20, 2019·Biotechnology Advances·Štefan JanečekBirte Svensson

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