The purification and properties of isocitrate lyase from Chlorella.
1. Isocitrate lyase (threo-d(s)-isocitrate glyoxylate-lyase, EC 4.1.3.1) has been purified from acetate-adapted cells of Chlorella pyrenoidosa. 2. The final preparation was homogeneous by the criteria of sedimentation, diffusion and polyacrylamide-gel electrophoresis. 3. The sedimentation coefficient (S(20,w)) was 9.04x10(-13)sec. and the diffusion coefficient (D(20,w)) 4.62x10(-7)cm.(2)/sec.; from these values the molecular weight of the enzyme was calculated to be 170000 and its Stokes radius to be 4.63x10(-7)cm. 4. The elution of the enzyme from Sephadex G-100 was studied and estimates of molecular weight and Stokes radius were obtained from the elution data. 5. The turnover number of the enzyme was 5950moles of glyoxylate formed/min./mole of enzyme at 30 degrees . 6. With threo-d(s)(+)-isocitrate as substrate, the K(m) of the enzyme was 0.023mm.
D002021:Buffers; D002482:Cellulose; D002501:Centrifugation, Zonal; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003956:Dialysis; D004589:Electrophoresis, Disc; D056890:Eukaryota / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008190:Lyases / Q000032:analysis*; D008970:Molecular Weight; D013053:Spectrophotometry
D002021:Buffers; D002482:Cellulose; D008190:Lyases