4-Coumarate CoA Ligase Microplate Assay Kit

The enzyme known as EC 6.2.1.12 goes by several names, including 4-coumaroyl-CoA synthetase, p-coumaroyl CoA ligase, and p-coumaryl Coenzyme A synthetase. It is also sometimes referred to as p-coumaryl-CoA synthetase, p-coumaryl-CoA ligase, feruloyl CoA ligase, hydroxycinnamoyl CoA synthetase, or 4-coumarate:coenzyme A ligase.
Other names for this enzyme include caffeoyl Coenzyme A synthetase, p-hydroxycinnamoyl Coenzyme A synthetase, feruloyl Coenzyme A synthetase, sinapoyl Coenzyme A synthetase, and 4-coumaryl-CoA synthetase. You may also come across the terms hydroxycinnamate:CoA ligase, p-coumaryl-CoA ligase, and p-hydroxycinnamic acid:CoA ligase when reading about this enzyme.
To keep things simple, this enzyme is often abbreviated as 4CL. It plays a crucial role in various metabolic pathways and is involved in the biosynthesis of several essential compounds, including lignin, flavonoids, and phenylpropanoids. So, it is a vital enzyme in the field of biochemistry and biotechnology.

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If one wants to measure the activity of 4-coumarate CoA ligase, a crucial enzyme involved in plant phenolic metabolism, then detection and quantification methods must be employed. This enzyme catalyzes the formation of CoA thioesters from 4-hydroxycinnamic acids, a reaction that is essential for the biosynthesis of lignin, flavonoids, and other secondary metabolites.
Various assays have been developed for determining 4-coumarate CoA ligase activity, including spectrophotometric, radiometric, and HPLC-based methods. These assays rely on the detection and measurement of the product or the substrate of the enzyme reaction, usually by using specific substrates, coenzymes, or labeling techniques.
For example, a colorimetric assay can be used to assess the formation of CoA-SH from 4-coumaroyl-CoA and 4-methylumbelliferone in the presence of ATP and Mg2+. The reaction can be monitored by measuring the fluorescence of the generated compound, which is proportional to the level of enzyme activity. Alternatively, a radioisotope assay can be employed to measure the incorporation of radiolabeled 4-coumaric acid into CoA thioesters, which can be detected by scintillation counting.
Overall, the detection and quantification of 4-coumarate CoA ligase activity is essential for studying the regulation of plant phenolic metabolism and for identifying potential targets for metabolic engineering.

Shipped and store at 4°C for 6 months.