Beta-cyanoalanine synthase: Its purification and basic physico-chemical properties

• Published in: Biokhimiia (Moscow, Russia) Vol. 41; no. 5; p. 906
• Main Authors: Akopian, T N, Goriachenkova, E V
• Format: Journal Article
• Language: Russian
• Published: Russia (Federation) 01-05-1976
• Subjects: Amino Acids - analysis; Carbon-Sulfur Lyases - analysis; Carbon-Sulfur Lyases - isolation & purification; Chemical Fractionation; Chromatography, DEAE-Cellulose; Chromatography, Ion Exchange; Electrophoresis, Disc; Electrophoresis, Polyacrylamide Gel; Lyases - analysis; Mitochondria - enzymology; Molecular Weight; Plants - enzymology; Pyridoxal - analysis
• ISSN: 0320-9725

A method has been developed for the purification of beta-cyano-L-alanine synthase from etiolated 10-day-old seedlings of blue lupine. High purity preparations of the enzyme were obtained with specific activity exceeding 4000-fold that of the seedling homogenate. Preparations were homogeneous on electrophoresis in polyacrylamide gel. The yield of total activity after purification was approximately 20%. Glutamic acid is the enzyme's only N-terminal amino acid; the molecular weight of the enzyme (both native and treated with 6 M urea) is 52000. The synthase containes one mole of pyridoxal-P per mole of protein; its isoelectric point is situated at pH 4,8. The enzyme's absorption spectrum has a maximum at 410 nm i.e., in the characteristic range of many pyridoxal-U-containing enzymes. Data on the amino acid composition of the enzyme are presented.