Application of Liquid Chromatography to a Study on 4-Coumarate: Coenzyme A Ligase Activity

Wolfgang Knogge 1 , Gottfried Weissenböck 1  and Dieter Strack 1
  • 1 Botanisches Institut der Universität zu Köln, Gyrhofstr. 15, D-5000 Köln 41

This report describes the separation of components from a 4-coumarate: CoA ligase assay by means of liquid chromatography. With the aid of polyamide column chromatography it is possible to enrich and isolate chromatographically and UV spectroscopically pure p-coumaroyl- CoA using as a solvent 0.01% NH4OH in methanol subsequent to water and methanol alone.

High performance liquid chromatography on octadecylsilane-bonded silica stationary phase allows a discontinuous determination of ligase activity. All components - ATP, Coenzyme A, pcoumaric acid, and the products AMP and p-coumaroyl-CoA - can be separated and accurately quantified within 20 min using a water-acetonitrile gradient, containing 1% phosphoric acid. The presented HPLC method may be used to affirm the accuracy of optical tests

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A Journal of Biosciences: Zeitschrift für Naturforschung C (ZNC) is an international scientific journal for the emerging field of natural and natural-like products. ZNC publishes original research on the isolation, bio-chemical synthesis and bioactivities of natural products, their biochemistry, pharmacology, biotechnology, and biological activity and innovative developed computational methods for predicting their structure and/or function.