The Use of Isothermal Titration Calorimetry for the Assay of Enzyme Activity: Application in Higher Education Practical Classes

Autor/inn/en	Siddiqui, Khawar S.; Poljak, Anne; Ertan, Haluk; Bridge, Wallace
Titel	The Use of Isothermal Titration Calorimetry for the Assay of Enzyme Activity: Application in Higher Education Practical Classes
Quelle	In: Biochemistry and Molecular Biology Education, 50 (2022) 5, S.519-526 (8 Seiten)Infoseite zur Zeitschrift PDF als Volltext Verfügbarkeit
Zusatzinformation	ORCID (Siddiqui, Khawar S.) ORCID (Poljak, Anne) ORCID (Ertan, Haluk) ORCID (Bridge, Wallace)
Sprache	englisch
Dokumenttyp	gedruckt; online; Zeitschriftenaufsatz
ISSN	1470-8175
DOI	10.1002/bmb.21657
Schlagwörter	Biochemistry; Biological Sciences; College Science; Heat; Science Activities; Laboratory Experiments; Undergraduate Students + Suchen Sie Ihr Suchwort? Biochemie; Abwasserbiologie; Hitze; Laboratory work; Laborarbeit
Abstract	Determination of enzyme activity is crucial for discovery, research, and development in life sciences. The activity of enzymes is routinely determined using spectrophotometric assays that measure rates of substrate consumption or product formation. Though colorimetric-based detection systems are simple, rapid, and economical to perform, the majority of enzymes are unsuitable for this technique as their substrates/products do not absorb in the UV or visible range. This limitation can be addressed by the use of coupled-enzyme assays or artificial chromogenic substrates; however these approaches have their own drawbacks. Here, we describe a method based on the use of an isothermal titration calorimeter (ITC) to measure the heat produced or absorbed during any enzyme-catalyzed reaction. The concept of calorimetric enzyme assays was demonstrated for the determination of enzyme hexokinase activity, which cannot be monitored colorimetrically without first coupling it to another enzymatic reaction. The assay is suitable for incorporation into undergraduate laboratory classes, providing students with an appreciation for; the versatility and ease of use of ITC assays; ITC as a flexible generic method for exploring the functional characteristics of uncharacterized enzymes; an activity detection parameter suitable for enzymes that either have no straightforward colorimetric methods available or require the use of nonartificial chromogenic substrates. (As Provided).
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Erfasst von	ERIC (Education Resources Information Center), Washington, DC
Update	2024/1/01