Purification and characterization of polyamine aminotransferase of Arthrobacter sp. TMP-1.
Polyamine aminotransferase of Arthrobacter sp. TMP-1 was induced by 1,3-diaminopropane (DAP), N-3-aminopropyl-1,3-diaminopropane (norspermidine), spermidine, and spermine, but not by putrescine. The enzyme was purified to homogeneity. Its molecular weight and subunit size were 129,000 and 64,000, respectively. Its absorption spectrum had maxima at 280 and 420 nm and a shoulder at about 350 nm, and changes were observed upon the addition of DAP, putrescine, and sodium borohydride. The spectrum and its changes indicated that the enzyme contained pyridoxal-5'-phosphate as the coenzyme. The coenzyme content was found to be 1 mol per mol of subunit. DAP, putrescine, norspermidine, spermidine, and spermine were active amino donors and gave relative rates of 100, 73, 24, 30, and 23%, respectively. Pyruvate was the most active amino acceptor, while 2-ketoglutarate and oxaloacetate were inert. The equilibrium constant of the DAP-pyruvate transamination was 0.34. DAP was suggested to be a minor product of the norspermidine-pyruvate reaction.
Yorifuji
T
T
Department of Bioscience and Biotechnology, Shinshu University, Minamiminowa, Nagano. yoriftk@gipmc.shinshu-u.ac.jp
Ishihara
T
T
Naka
T
T
Kondo
S
S
Shimizu
E
E
D001173:Arthrobacter / Q000201:enzymology*; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000302:isolation & purification* / Q000378:metabolism*; D015317:Biogenic Polyamines / Q000378:metabolism*; D001894:Borohydrides / Q000494:pharmacology; D002855:Chromatography, Thin Layer; D003959:Diamines / Q000494:pharmacology; D004790:Enzyme Induction / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D011700:Putrescine / Q000494:pharmacology; D019289:Pyruvic Acid / Q000378:metabolism; D013095:Spermidine / Q000031:analogs & derivatives / Q000494:pharmacology; D013379:Substrate Specificity; D000637:Transaminases / Q000096:biosynthesis / Q000302:isolation & purification* / Q000378:metabolism*
D001426:Bacterial Proteins; D015317:Biogenic Polyamines; D001894:Borohydrides; D003959:Diamines; D004791:Enzyme Inhibitors; C006760:norspermidine; D019289:Pyruvic Acid; C025364:sodium borohydride; C009475:trimethylenediamine; D000637:Transaminases; D013095:Spermidine; D011700:Putrescine