PMID: 11342059May 9, 2001Paper

Enzyme activity determination on macromolecular substrates by isothermal titration calorimetry: application to mesophilic and psychrophilic chitinases

Biochimica Et Biophysica Acta
T LonhienneC Gerday

Abstract

Isothermal titration calorimetry has been applied to the determination of the kinetic parameters of chitinases (EC 3.2.1.14) by monitoring the heat released during the hydrolysis of chitin glycosidic bonds. Experiments were carried out using two different macromolecular substrates: a soluble polymer of N-acetylglucosamine and the insoluble chitin from crab shells. Different experimental temperatures were used in order to compare the thermodependence of the activity of two chitinases from the psychrophile Arthrobacter sp. TAD20 and of chitinase A from the mesophile Serratia marcescens. The method allowed to determine unequivocally the catalytic rate constant k(cat), the activation energy (E(a)) and the thermodynamic activation parameters (DeltaG(#), DeltaH(#), DeltaS(#)) of the chitinolytic reaction on the soluble substrate. The catalytic activity has also been determined on insoluble chitin, which displays an effect of substrate saturation by chitinases. On both substrates, the thermodependence of the activity of the psychrophilic chitinases was lower than that observed with the mesophilic counterpart.

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Citations

Dec 30, 2006·Marine Biotechnology·J-C MarxC Gerday
Aug 31, 2010·Applied Biochemistry and Biotechnology·Søren N OlsenPeter Westh
Aug 16, 2002·Current Opinion in Biotechnology·Ricardo CavicchioliKevin R Sowers
Jun 12, 2002·Current Opinion in Microbiology·Jody W Deming
Jan 22, 2008·Applied and Environmental Microbiology·Debora Frigi Rodrigues, James M Tiedje
Jul 1, 2008·Biochimica Et Biophysica Acta·Indalecio Quesada-SorianoLuis García-Fuentes
Jul 11, 2006·Biophysical Chemistry·M Lucia Bianconi
Nov 18, 2015·Scientific Reports·Kourosh Honarmand EbrahimiWilfred R Hagen
Nov 5, 2005·Analytical Biochemistry·Tina JeohWilliam S Adney
Jun 30, 2015·Analytical Biochemistry·Ksenia Maximova, Joanna Trylska
Sep 23, 2008·Chemistry & Biology·Bjarne VincentsUlrich von Pawel-Rammingen
Jan 3, 2007·Journal of Molecular Biology·Ellen WangPirkko Heikinheimo
Aug 31, 2013·Archives of Biochemistry and Biophysics·Priscila Lamb WinkDiogenes Santiago Santos
Jun 8, 2006·Annual Review of Biochemistry·Khawar Sohail Siddiqui, Ricardo Cavicchioli
Nov 3, 2009·Biochimie·Indalecio Quesada-SorianoLuis García-Fuentes
Jan 1, 2013·Biology·Charles Gerday
Apr 28, 2001·The Journal of Biological Chemistry·S D'AmicoG Feller
Sep 27, 2002·The Journal of Biological Chemistry·Salvino D'AmicoGeorges Feller
May 28, 2017·Journal of Computer-aided Molecular Design·I StawoskaM Szaleniec
Nov 17, 2020·Frontiers in Molecular Biosciences·Yun WangAnthony K Mittermaier
May 12, 2021·Journal of Molecular Recognition : JMR·Robert J FalconerAnthony K Mittermaier

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