Functional characterization of starch-degrading enzymes using quartz crystal microbalance with dissipation monitoring (QCM-D)

• Published in: Sensors and actuators. B, Chemical Vol. 176; pp. 1038 - 1043
• Main Authors: Bouchet-Spinelli, Aurélie, Coche-Guérente, Liliane, Armand, Sylvie, Lenouvel, François, Labbé, Pierre, Fort, Sébastien
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 01.01.2013; Elsevier
• Subjects: biomass; colorimetry; Glycoside hydrolases; hydrolysis; monitoring; mutants; quartz; Quartz crystal microbalance with dissipation monitoring; Real-time label-free detection; Starch degradation; transferases; Starch degradation; Glycoside hydrolases; Quartz crystal microbalance with dissipation monitoring; Real-time label-free detection
• ISSN: 0925-4005; 1873-3077
• DOI: 10.1016/j.snb.2012.10.013

During the last few decades, starch-degrading enzymes have emerged as promising tools for transforming biomass. Nevertheless, their functional characterization, based essentially on colorimetric tests, is still fastidious and time-consuming. In order to overcome these hurdles, we have developed a test for real-time and label-free detection of starch-degrading enzymes using quartz crystal microbalance with dissipation monitoring (QCM-D). By starch-functionalization of surfaces, we were able to discriminate clearly enzymes with different hydrolysis patterns (endo versus exo). The variation of the hydrolytic activity between the wild-type and a mutant of a cyclodextrin glucosyl transferases could also be monitored acoustically. The high sensitivity reached (down to 6.5nkat/L) opens the prospect of using fewer enzyme quantities for functional tests and is a further advantage for this easily used and efficient tool for enzyme characterization.