title: Polyamine synthesis in plants: isolation and characterization of spermidine synthase from soybean (Glycine max) axes.
abstract: Spermidine synthase (EC 2.5.1.16) was purified to homogeneity for the cytosol of soybean (Glycine max) axes using ammonium sulfate fractionation and chromatography on DEAE-Sephacel, Sephacryl S-300, omega-aminooctyl-Sepharose and ATPA-Sepharose. The molecular mass of the enzyme estimated by gel filtration and SDS-PAGE is 74 kDa. Cadaverin and 1,6-diaminohexane could not replace putrescine as the aminopropyl acceptor. Kinetic behaviors of the substrate are consistent with a ping pong mechanism. The kinetic mechanism is further supported by direct evidence confirming the presence of an aminopropylated enzyme and identification of product, 5'-deoxy-5'-methylthioadenosine, prior to adding putrescine. The Km values for decarboxylated S-adenosylmethionine and putrescine are 0.43 microM and 32.45 microM, respectively. Optimum pH and temperature for the enzyme reaction are 8.5 and 37 degrees C, respectively. The enzyme activity is inhibited by N-ethylmaleimide and DTNB, but stimulated by Co2+, Cu2+ and Ca2+ significantly, suggesting that these metal ions could be the cellular regulators in polyamine biosynthesis.
authors.affiliation: Department of Biochemistry, College of Science, Yonsei University, Seoul, South Korea.
authors.forename: S O
authors.initials: SO
authors.lastname: Yoon
authors.forename: Y S
authors.initials: YS
authors.lastname: Lee
authors.forename: S H
authors.initials: SH
authors.lastname: Lee
authors.forename: Y D
authors.initials: YD
authors.lastname: Cho
mesh_terms: D002118:Calcium / Q000494:pharmacology; D002850:Chromatography, Gel; D003035:Cobalt / Q000494:pharmacology; D003300:Copper / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation / Q000187:drug effects; D007700:Kinetics; D010944:Plants / Q000378:metabolism*; D011073:Polyamines / Q000378:metabolism*; D013025:Glycine max / Q000201:enzymology*; D011436:Spermidine Synthase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D011073:Polyamines; D003035:Cobalt; D003300:Copper; D011436:Spermidine Synthase; D002118:Calcium