Improved method for large scale purification of the phosphoenolpyruvate carboxylase of Escherichia coli K-12.
- 1 January 1984
- journal article
- research article
- Published by Microbiology Research Foundation in The Journal of General and Applied Microbiology
- Vol. 30 (1) , 27-33
- https://doi.org/10.2323/jgam.30.27
Abstract
A rapid and simple method for large-scale preparation of phosphoenolpyruvate carboxylase [EC 4.1.1.31] from E. coli K-12 was established. A strain carrying the Col E1-ppc+ hybrid plasmid was used as an enzyme source. The cell-free extract was fractionated by (NH4)2 SO4, 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 (153 mg) was obtained from 137 g wet cells in 4 days with 41% yield.Keywords
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