|
type |
Journal Article |
authors |
Volland, C.; Felix, F. |
title |
Isolation and properties of 5-aminolevulinate synthase from the yeast Saccharomyces cerevisiae |
journal |
Eur J Biochem |
Activity |
2.3.1.37 |
ui |
84285364 |
year |
(1984) |
volume |
142 |
number |
3 |
pages |
551-7. |
| |
keywords |
5-Aminolevulinate Synthetase/*isolation & purification/metabolism |
abstract |
5-Aminolevulinate synthase from yeast mitochondria has been purified to homogeneity for the first time. By using affinity chromatography on agarose-hexane-CoA, gel filtration and DEAE-Sepharose chromatography, the enzyme was purified about 7000-fold with an overall yield of 40%. The specific activity of the final preparation was 39000 nmol of 5- aminolevulinate h-1 mg-1 of protein at 30 degrees C. As judged by gel filtration, polyacrylamide gradient gel and sodium dodecyl sulfate/polyacrylamide gel electrophoresis, the enzyme appeared to be composed of two identical subunits of a relative molecular mass of 53000. Electrophoresis of sodium-dodecyl-sulfate-solubilized yeast homogenate followed by immune replica analysis showed that the value of 53000 is the Mr of a non-degraded form. The purified enzyme had an isoelectric point of 5.3 and a pH optimum of 7.4. Pyridoxal 5'- phosphate has been shown to be an essential cofactor. The enzyme activity was sensitive to thiol blocking reagents. Hemin, but not heme, inhibited the activity of the purified enzyme. |
last changed |
2002/11/04 17:41 |
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