Aminosäurezusammensetzung kristallisierter Alkohol-Dehydrogenase aus Bäckerhefe

Abstract

Yeast alcohol dehydrogenase was purified and crystallized by the method of Racker as modified by Bucher and Christian. It was then refluxed for 12 hr. with 6 N HC1. The amino acids were separated by one-dimensional chromatography in the following systems: n-butanol:acetic acid:water, 80:20:20, for cystine + cysteine, lysTne, histidine, arginine, alanine, proline, tyrosine, and valine + methionine; phenol saturated with borate buffer pH 12 for aspartic acid, glutamic acid, serine, glycine, threonine, alanine, and tyrosine; and pyridine: n-amyl alcohol:water, 25: 50:25, for valine, tyrosine, isoleucine, leucine, and phenylalanine. The amino acids were rendered visible with ninhydrin, and the copper complexes were eluted from the paper with methanol and estimated colorimetrically. There were no unusual features in the amino acid composition of the protein.