A new NAD+-dependent opine dehydrogenase from Arthrobacter sp. strain 1C.
A new NAD+-dependent opine dehydrogenase was purified to homogeneity from Arthrobacter sp. strain 1C isolated from soil by an enrichment culture technique. The enzyme has a molecular weight of about 70,000 and consists of two identical subunits with molecular weights of about 36,000. The enzyme catalyzed a reversible oxidation-reduction reaction of opine-type secondary amine dicarboxylic acids. In the oxidative deamination reaction, the enzyme was active toward unusual opines, such as N-[1-R-(carboxyl)ethyl]-S-methionine and N-[1-R-(carboxyl)ethyl]-S-phenylalanine. In the reductive secondary amine-forming reaction with NADH as a cofactor, the enzyme utilized L-amino acids such as L-methionine, L-isoleucine, L-valine, L-phenylalanine, L-leucine, L-alanine, and L-threonine as amino donors and alpha-keto acids such as pyruvate, oxaloacetate, glyoxylate, and alpha-ketobutyrate as amino acceptors. The product enzymatically synthesized from L-phenylalanine and pyruvate in the presence of NADH was identified as N-[1-R-(carboxyl)ethyl]-S-phenylalanine.
Asano
Y
Y
Sagami Chemical Research Center, Kanagawa, Japan.
Yamaguchi
K
K
Kondo
K
K
D000600:Amino Acids, Dicarboxylic / Q000138:chemical synthesis; D001173:Arthrobacter / Q000201:enzymology*; D002412:Cations; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D007700:Kinetics; D009243:NAD / Q000378:metabolism; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000302:isolation & purification* / Q000378:metabolism; D013045:Species Specificity; D013379:Substrate Specificity
D000600:Amino Acids, Dicarboxylic; D002412:Cations; D009243:NAD; D000587:Oxidoreductases Acting on CH-NH Group Donors; C060511:opine dehydrogenase, NAD+