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PMID: 6796041 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Purification of porphobilinogen deaminase from Euglena gracilis and studies of its kinetics.

The Biochemical journal ·Vol. 193 ·No. 1 ·1981-01-01 ·Pages 301-10

Williams DC, Morgan GS, McDonald E, Battersby AR

Abstract

1. Porphobilinogen deaminase [porphobilinogen ammonia-lyase (polymerizing), EC 4.3.1.8] from Euglena gracilis was purified more than 200-fold. 2. The enzyme has a molecular weight of 41 000 +/- 2000, does not contain a chromophoric prosthetic group, and appears not to require metal ions for activity. 3. The stoicheiometry of the overall reaction at pH 7.4 was shown to be: 4 Porphobilinogen leads to uroporphyrinogen-I + 4 NH4+. This stoicheiometry for porphobilinogen and uroporphyrinogen was also observed over a wide range of pH values. 4. Initial-velocity studies showed a hyperbolic dependence of velocity on substrate concentration, demonstrating the existence of a displacement-type mechanism. 5. Vmax. varied with pH as a typical bell-shaped curve, indicating that two ionizable groups with pK values of 6.1 and 8.9 are important for catalysis. A plot of Vmax./Km against pH showed a single ionization (pK 8.2) to influence binding of substrate.

MeSH Terms
Ammonia-Lyases/metabolism Chromatography, DEAE-Cellulose Euglena gracilis/enzymology Hydrogen-Ion Concentration Hydroxymethylbilane Synthase/isolation & purification,metabolism Kinetics Molecular Weight Porphobilinogen/metabolism Temperature
Chemicals
Porphobilinogen Hydroxymethylbilane Synthase Ammonia-Lyases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Williams D C
Morgan G S
McDonald E
Battersby A R
References (22)
22 references, click to expand
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj1930301
Published
1981-01-01
Pages
301-10
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1162603
Subset
IM
External Links
PubMed source DOI: 10.1042/bj1930301 PMC Full Text View journal details
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