The Journal of General and Applied Microbiology
Online ISSN : 1349-8037
Print ISSN : 0022-1260
ISSN-L : 0022-1260
IMPROVED METHOD FOR LARGE SCALE PURIFICATION OF THE PHOSPHOENOLPYRUVATE CARBOXYLASE OF ESCHERICHIA COLI K-12
SUMIO ISHIJIMANOBUYUKI FUJITAHISATAKA SABEKATSURA IZUIHIROHIKO KATSUKI
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1984 Volume 30 Issue 1 Pages 27-33

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Abstract

A rapid and simple method for large-scale preparation of phosphoenolpyruvate carboxylase [EC 4.1.1.31] 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.

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