[Preparation and properties of isocitrate lyase isoforms from the cotyledons of Glycine max L].
A four-stage purification procedure including ammonium sulfate precipitation and ion exchange chromatography on DEAE cellulose has been elaborated for isolation of isocitrate lyase (EC 4.1.3.1) isoforms from the cotyledons of soybean Glycine max L. Electrophoretically homogeneous preparations of two forms of the enzyme with specific activity of 5.28 and 5.81 U/mg protein have been obtained. Comparison of physicochemical, kinetic, and regulation characteristics of the isoforms obtained revealed fundamental differences between them. Thus, the isoform that migrated quickly in PAAG had a much lower affinity to isocitrate (K(M) - 50 microM) than the slowly migrating form (K(M) - 16 microM). It has been shown that the conservation of activity of the isoforms obtained depends on the presence of divalent cations (Mn2+ and Mg2+) in the medium. It is suggested to use the isoforms of isocitrate lyase isolated from soybeans for the development of biosensors for biochemical and kinetic assays.
Eprintsev
A T
AT
Diachenko
E V
EV
Lykova
T V
TV
Kuen
C T H
CT
Popov
V N
VN
D000645:Ammonium Sulfate / Q000737:chemistry; D015374:Biosensing Techniques; D002848:Chromatography, DEAE-Cellulose; D002852:Chromatography, Ion Exchange; D018548:Cotyledon / Q000201:enzymology*; D007522:Isocitrate Lyase / Q000737:chemistry / Q000302:isolation & purification*; D007523:Isocitrates / Q000737:chemistry; D007527:Isoenzymes / Q000737:chemistry / Q000302:isolation & purification; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification*; D013025:Glycine max / Q000201:enzymology*; D013379:Substrate Specificity
D007523:Isocitrates; D007527:Isoenzymes; D010940:Plant Proteins; C034219:isocitric acid; D007522:Isocitrate Lyase; D000645:Ammonium Sulfate