title: A kinetic study of homocitrate synthetase activity in the yeast Saccharomycopsis lipolytica.
abstract: 1. A rapid method for estimating the activity of the first enzyme of lysine biosynthesis in yeasts (acetyl-coenzyme A: 2-ketoglutarate C-acetyl transferase, EC 4.1.3.21) is described. 2. In the wild type strain, the fixation of one substrate, S-acetyl coenzyme A, shows sigmoidal saturation kinetics. The initial rate experiments indicate that the reaction obeys an ordered mechanism, 2-ketoglutaric acid binding before S-acetyl coenzyme A. 3. The activity is completely inhibited in vitro by lysine and by some lysine analogs, which all show cooperative binding and have an heterotropic effect on 2-ketoglutaric binding sites. A second class of affectors is found, including 2-aminoadipic acid, pipecolic acid and dipicolinic acid, which all affect the cooperativity of S-acetyl coenzyme A binding sites. 4. Two types of mutations which modify these inhibition patterns without affecting the catalytic activity are described. One results in a desensitization towards lysine and lysine analogs only. The other entirely abolishes the susceptibility towards the second type of inhibitors, without affecting the susceptibility to lysine. 5. No variations of the specific activity could be detected in the wild type strain at all; mutants showing an increased or a reduced activity were isolated. 6. Our results do not support the existence of isoenzymes at the level of homocitrate synthetase in this yeast.
mesh_terms: D000105:Acetyl Coenzyme A / Q000378:metabolism; D001665:Binding Sites; D002951:Citrates; D007656:Ketoglutaric Acids / Q000378:metabolism; D007700:Kinetics; D008239:Lysine / Q000494:pharmacology; D007652:Oxo-Acid-Lyases / Q000378:metabolism*; D010848:Picolinic Acids / Q000494:pharmacology; D011485:Protein Binding; D012440:Saccharomyces / Q000201:enzymology*; D013329:Structure-Activity Relationship
chemical_list: D002951:Citrates; D007656:Ketoglutaric Acids; D010848:Picolinic Acids; D000105:Acetyl Coenzyme A; D007652:Oxo-Acid-Lyases; D008239:Lysine