   2.7.8.28 | -999 |
- |
CofD activity is measured in a coupled assay with lactylphosphate guanylyltransferase (CofC, MJ1117), which produces lactyl-2-diphospho-5'-guanosine from 2-phospho-L-lactate and GTP. The reaction mixture contains 50 mM TES (pH 7.5), 0.1 mM GTP, 10 mM 2-phospho-L-lactate, 0.4 mM 7,8-didemethyl-8-hydroxy-5-deazariboflavin, 2 mM MnCl2, 0.01 mg of CofC, and 0.0004–0.0006 mg of CofD. The enzyme system is inhibited by high concentrations of GMP. Therefore, 2 mM phosphoenolpyruvate and 1 unit of pyruvate kinase are also included in the reaction mixture to regenerate GTP from GMP |
698744 |
