Enzyme kinetics by mid-infrared spectroscopy: beta-fructosidase study by a one-step assay.
An alternate method for enzyme study is proposed. Multidimensional statistical analysis applied on mid-infrared attenuated total reflectance spectra (Cadet et al. (1991) Appl. Spectrosc. 42, 166-172) collected during a kinetic allows a direct and fast quantification of the remaining substrate, as well as a one step enzymatic assay. Furthermore, the combination of these techniques may be used as a structural tool. The method applied to the study of beta-fructosidase is developed in this paper as an example. With appropriate calibration, the method may be extend to any enzyme.
Cadet
F
F
Université de la Réunion, Laboratoire de Biochimie, Faculté des Sciences, Saint Denis, Reunion.
Pin
F W
FW
Rouch
C
C
Robert
C
C
Baret
P
P
D004187:Disaccharides / Q000737:chemistry*; D006026:Glycoside Hydrolases / Q000737:chemistry*; D007700:Kinetics; D013055:Spectrophotometry, Infrared; D013395:Sucrose / Q000737:chemistry; D043324:beta-Fructofuranosidase
D004187:Disaccharides; D013395:Sucrose; D006026:Glycoside Hydrolases; D043324:beta-Fructofuranosidase