Detection of enzyme-catalyzed polysaccharide synthesis on….pdfVIP

Biocatalysis and Biotransformation, January–February 2010; 28(1): 64–71 ORIGINAL ARTICLE Detection of enzyme-catalyzed polysaccharide synthesis on surfaces 1 2 1 3 ˇ CARLA CLÉ , CATHIE MARTIN , ROBERT A. FIELD , PETR KUZMIC STEPHEN BORNEMANN1 1Department of Biological Chemistry, John Innes Centre, Norwich, UK, 2Department of Metabolic Biology, John Innes Centre, Norwich, UK, and 3BioKin Limited, Watertown, Massachusetts, USA Abstract Strategically important cellular components, such as the cell wall and the starch granule, present surfaces during their biosynthesis and degradation. The enzymology of such surfaces is experimentally challenging and goes well beyond clas- sical solution-state analyses. The kinetics of surface catalysis is complex but tractable. A number of approaches to monitor surface catalysis are reviewed and each is suited to a different biological problem. Particular attention is paid to a method we have recently developed for quantitatively monitoring polysaccharide synthesis on a surface in real time using surface plasmon resonance spectroscopy. This method has many attractive features with the potential to tackle both biological and industrial problems. Keywords: Enzyme, surface, catalysis, polysaccharide, surface plasmon resonance spectroscopy, kinetics Introduction Enzymic surface degradation of cellulose likewise has great potential (Lynd et al. 2002). Surface enzy- Many biological phenomena involve surfaces that mology is gaining importance in many other com- need to be biosynthesized, modifi ed an