Pohl B, Raichle T, Ghisla S
Eur J Biochem. 1986 Oct 1;160(1):109-15. doi: 10.1111/j.1432-1033.1986.tb09946.x.
The kinetic properties of general acyl-CoA dehydrogenase from pig kidney have been investigated using normal butyryl-CoA as well as an alpha-deutero, beta-deutero- and perdeutero-butyryl-CoA. In turnover catalysis, isotope effects of 2, 3.6, and 9 were found respectively. In the reductive half reaction the isotope effects were 2.5, 14, and 28 for the same substrates, and 21 for (2R,3R)-(2,3-D2)butyryl-CoA. No intermediates are apparent during the reduction of oxidized enzyme to the presumed complex of reduced enzyme and crotonyl-CoA. The results are interpreted as indicating a high degree of concertedness during the rupture of the alpha and beta C-H bonds. They are compatible with a mechanism in which simultaneously the alpha-hydrogen is abstracted as a proton, while the beta-hydrogen is transferred to the oxidized flavin as a hydride.
利用正常的丁酰辅酶A以及α-氘代、β-氘代和全氘代丁酰辅酶A，对猪肾中的通用酰基辅酶A脱氢酶的动力学性质进行了研究。在周转催化中，分别发现了2、3.6和9的同位素效应。在还原半反应中，相同底物的同位素效应分别为2.5、14和28，而(2R,3R)-(2,3-D2)丁酰辅酶A的同位素效应为21。在将氧化型酶还原为假定的还原型酶与巴豆酰辅酶A复合物的过程中，没有明显的中间体。结果表明，α和β碳氢键断裂过程具有高度协同性。这与一种机制相符，即α-氢同时作为质子被提取，而β-氢作为氢化物转移到氧化型黄素上。
