Article (Scientific journals)
Enzyme Activity Determination on Macromolecular Substrates by Isothermal Titration Calorimetry: Application to Mesophilic and Psychrophilic Chitinases
Lonhienne, T.; Baise, Etienne; Feller, Georges et al.
2001In Biochimica et Biophysica Acta, 1545 (1-2), p. 349-56
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Abstract :
[en] 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.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Lonhienne, T.
Baise, Etienne ;  Université de Liège - ULiège > Département des sciences de la vie > Macromolécules biologiques
Feller, Georges ;  Université de Liège - ULiège > Département des sciences de la vie > Labo de biochimie
Bouriotis, V.
Gerday, Charles ;  Université de Liège - ULiège > Services généraux (Faculté des sciences) > Relations académiques et scientifiques (Sciences)
Language :
English
Title :
Enzyme Activity Determination on Macromolecular Substrates by Isothermal Titration Calorimetry: Application to Mesophilic and Psychrophilic Chitinases
Publication date :
10 February 2001
Journal title :
Biochimica et Biophysica Acta
ISSN :
0006-3002
eISSN :
1878-2434
Publisher :
Elsevier, Amsterdam, Netherlands
Volume :
1545
Issue :
1-2
Pages :
349-56
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 26 January 2010

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