PMID:7978236
| Citation |
Ramjee, MK, Coggins, JR and Thorneley, RN (1994) A continuous, anaerobic spectrophotometric assay for chorismate synthase activity that utilizes photoreduced flavin mononucleotide. Anal. Biochem. 220:137-41 |
|---|---|
| Abstract |
A sensitive, continuous, spectrophotometric assay for chorismate synthase has been developed utilizing photoreduced flavin mononucleotide (FMNH2) as a cofactor under anaerobic conditions. The assay monitors directly the formation of chorismate from 5-enolpyruvylshikimate-3-phosphate (EPSP) at 275 nm with a precision of +/- 2 microM product. The assay conditions have been optimized with respect to FMNH2 (cofactor), EPSP (substrate) and enzyme concentrations, buffer type, and pH. The potential of the assay for detailed steady-state kinetic studies to elucidate the mechanism of action of this commercially important enzyme is also demonstrated. |
| Links |
PubMed Online version:10.1006/abio.1994.1309 |
| Keywords |
Anaerobiosis; Escherichia coli/enzymology; Flavin Mononucleotide/chemistry; Kinetics; Lyases/analysis; Oxidation-Reduction; Phosphorus-Oxygen Lyases; Reproducibility of Results; Spectrophotometry, Ultraviolet |
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