Figure 1
Consumption of R- 3-hydroxyoctanoyl-CoA in crude cell extracts of P. putida U and P. putida U:: pha C1-. Panel A: Influence of free CoA on R- 3-hydroxyoctanoyl-CoA thioesterase activity in a crude cell extract of P. putida U::pha C1-. Assay conditions: 100 mM Tris-HCl, pH 8, 1 mg/ml BSA, 0.5 mM MgCl2, 0.5 mM R- 3-hydroxyoctanoyl-CoA, 0.1 mg/ml crude cell extract of P. putida U::pha C1- with no CoA (filled diamond) or 1 mM CoA (open diamond). Data represent the average of two measurements. Panel B: R- 3-hydroxyoctanoyl-CoA consumption in crude cell extracts of P. putida U::pha C1- and P. putida U in the presence of free CoA. Assay conditions: 100 mM Tris-HCl, pH 8, 1 mg/ml BSA, 0.5 mM MgCl2, 0.5 mM R- 3-hydroxyoctanoyl-CoA, 1 mM CoA, 4 mg/ml crude cell extract of either P. putida U::pha C1- (open diamond) or P. putida U (open square). R- 3-hydroxyoctanoyl-CoA depletion was measured by HPLC. Data represent the average of two measurements.