Isolation and characterization of enzymes involved in lysine catabolism from sorghum seeds.
Lysine is an essential amino acid synthesized in plants via the aspartic acid pathway. The catabolism of lysine is performed by the action of two consecutive enzymes, lysine 2-oxoglutarate reductase (LOR, EC 1.5.1.8) and saccharopine dehydrogenase (SDH, EC 1.5.1.9). The final soluble lysine concentration in cereal seeds is controlled by both synthesis and catabolism rates. The production and characterization of high-lysine plants species depends on knowledge of the regulatory aspects of lysine metabolism and manipulation of the key enzymes. We have for the first time isolated, partially purified, and characterized LOR and SDH from developing sorghum seeds, which exhibited low levels of activity. LOR and SDH were only located in the endosperm and were very unstable during the isolation and purification procedures. LOR and SDH exhibited some distinct properties when compared to the enzymes isolated from other plant species, including a low salt concentration required to elute the enzymes during anion-exchange chromatography and the presence of multimeric forms with distinct molecular masses.
Fornazier
Ricardo F
RF
Departamento de Genética e Evolução, Universidade Estadual de Campinas, Campinas, CEP 13083-970, Brazil.
Gaziola
Salete A
SA
Helm
Cristiane V
CV
Lea
Peter J
PJ
Azevedo
Ricardo A
RA
D000596:Amino Acids / Q000032:analysis; D006863:Hydrogen-Ion Concentration; D008239:Lysine / Q000378:metabolism*; D010940:Plant Proteins / Q000032:analysis; D012443:Saccharopine Dehydrogenases / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D012639:Seeds / Q000201:enzymology*; D045868:Sorghum / Q000201:enzymology*; D013379:Substrate Specificity
D000596:Amino Acids; D010940:Plant Proteins; D012443:Saccharopine Dehydrogenases; D008239:Lysine