1984 Volume 30 Issue 1 Pages 27-33
A rapid and simple method for large-scale preparation of phosphoenolpyruvate carboxylase [EC 18.104.22.168] from Escherichia coli K-12 was established. A strain carrying the ColE1-ppc+ (the gene of the enzyme) hybrid plasmid was used as an enzyme source. The cell-free extract was fractionated by ammonium sulfate, and the enzyme was purified to homogeneity by hydrophobic chromatography on hexyl-Sepharose using specific elution with L-aspartate, one of the allosteric effectors. The pure enzyme (153mg) was obtained from 137g wet cells in 4 days with 41% yield.