Coenzyme B-12-dependent reactions. Part IV. Observations on the purification of ethanolamine ammonia-lyase.
Purification of ethanolamine ammonia-lyase (EC 4.3.1.7) from a Clostridium sp. grown at the University of Sussex, U.K. and the National Institutes of Health, U.S.A., has been compared and an improved isotopic assay for the enzyme has been developed. Successful purification of this enzyme from Sussex-grown cells requires modification of the published procedure (Kaplan and Stadtman (1968) J. Biol, Chem. 243, 1787-1793) principally a 70% decrease in volume during precipitation with 0.4 M NaCl. This modification also increases the yield from N.I.H.-grown cells. Purified enzyme, resolved of inactive cobalamins, has the same high specific activity from both sources and behaves in the same way on disc gel electrophoresis. Sussex enzyme, before resolution, has less than 20% of the specific activity of unresolved N.I.H. enzyme and contains over 50% more inactive cobalamin. The bound cobalamin from both preparations has been identified as a "base-on" Co11 psi-cobalamin.
D000642:Ammonia-Lyases / Q000302:isolation & purification*; D001665:Binding Sites; D003013:Clostridium / Q000201:enzymology; D004578:Electron Spin Resonance Spectroscopy; D004789:Enzyme Activation; D004981:Ethanolamine Ammonia-Lyase / Q000302:isolation & purification* / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D013045:Species Specificity; D013053:Spectrophotometry; D014805:Vitamin B 12 / Q000494:pharmacology*
D000642:Ammonia-Lyases; D004981:Ethanolamine Ammonia-Lyase; D014805:Vitamin B 12