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