Functional characterization of starch-degrading enzymes using quartz crystal microbalance with dissipation monitoring (QCM-D)
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 te...
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| Published in | Sensors and actuators. B, Chemical Vol. 176; pp. 1038 - 1043 |
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| Main Authors | , , , , , |
| Format | Journal Article |
| Language | English |
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Elsevier B.V
01.01.2013
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| Abstract | 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. |
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| AbstractList | 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. |
| Author | Coche-Guérente, Liliane Armand, Sylvie Lenouvel, François Fort, Sébastien Bouchet-Spinelli, Aurélie Labbé, Pierre |
| Author_xml | – sequence: 1 givenname: Aurélie surname: Bouchet-Spinelli fullname: Bouchet-Spinelli, Aurélie email: aurelie.bouchet@cermav.cnrs.fr organization: CERMAV – CNRS, UPR5301, Affiliated with Université Joseph Fourier and ICMG (FR CNRS 2607), BP 53, 38041 Grenoble, France – sequence: 2 givenname: Liliane surname: Coche-Guérente fullname: Coche-Guérente, Liliane organization: Département de Chimie Moléculaire, UMR 5250, CNRS-Université Joseph Fourier, Affiliated with ICMG, BP 53, 38041 Grenoble, France – sequence: 3 givenname: Sylvie surname: Armand fullname: Armand, Sylvie organization: CERMAV – CNRS, UPR5301, Affiliated with Université Joseph Fourier and ICMG (FR CNRS 2607), BP 53, 38041 Grenoble, France – sequence: 4 givenname: François surname: Lenouvel fullname: Lenouvel, François organization: CERMAV – CNRS, UPR5301, Affiliated with Université Joseph Fourier and ICMG (FR CNRS 2607), BP 53, 38041 Grenoble, France – sequence: 5 givenname: Pierre surname: Labbé fullname: Labbé, Pierre organization: Département de Chimie Moléculaire, UMR 5250, CNRS-Université Joseph Fourier, Affiliated with ICMG, BP 53, 38041 Grenoble, France – sequence: 6 givenname: Sébastien surname: Fort fullname: Fort, Sébastien organization: CERMAV – CNRS, UPR5301, Affiliated with Université Joseph Fourier and ICMG (FR CNRS 2607), BP 53, 38041 Grenoble, France |
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| Keywords | Starch degradation Glycoside hydrolases Quartz crystal microbalance with dissipation monitoring Real-time label-free detection |
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| SubjectTerms | biomass colorimetry Glycoside hydrolases hydrolysis monitoring mutants quartz Quartz crystal microbalance with dissipation monitoring Real-time label-free detection Starch degradation transferases |
| Title | Functional characterization of starch-degrading enzymes using quartz crystal microbalance with dissipation monitoring (QCM-D) |
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