Nagao S, Fukushima H, Nozawa Y
Biochim Biophys Acta. 1978 Aug 25;530(2):165-74.
[1-14-C]Palmitoyl-Co A was incubated with Tetrahymena microsomes containing the complete enzyme system for desaturation during various time periods. The level of [1-14C]palmitoleoyl-CoA increased to a maximum during the 1--3 min incubation time, while [1-14C]palmitoleic acid in the phospholipid reached a maximum level during 6--7 min incubation time. The radioactivity of [1-14C]palmitoleic acid in free fatty acid and the triglyceride fraction was not significantly observed upon 3 min incubation. Incubation of [1-14C]palmitoyl-CoA with microsomes in the absence of NADH produced [1-14C]palmitoyl lipid without desaturation. Radioactive palmitic acids in the microsomal lipids were not converted to palmitoleic acids after addition of NADH by the complete enzyme system. When microsomes prepared from cells labeled with [1-14C]palmitic acid or [1-14C]stearic acid were incubated alone in the presence of O2 and NADH, no significant increase in [1-14C]palmitoleic acid in the phospholipid was observed, wherease an increase in [1-14C]linoleic acid and gamma-[1-14C]linolenic acid did occur at the expense of [1-14C]oleic acid in the phospholipid. From these results it can be concluded that the enzyme involving desaturation of palmitic acid to palmitoleic acid requires palmitoyl-CoA as the substrate. However, the possibility of oleoyl and linoleoyl phospholipids being substrates in the desaturation of Tetrahymena microsomes was suggested.
[1-14-C]棕榈酰辅酶A与含有完整去饱和酶系统的四膜虫微粒体在不同时间段进行孵育。在孵育1至3分钟期间,[1-14C]棕榈油酰辅酶A的水平增加到最大值,而磷脂中的[1-14C]棕榈油酸在孵育6至7分钟期间达到最高水平。孵育3分钟后,游离脂肪酸和甘油三酯部分中[1-14C]棕榈油酸的放射性未被明显观察到。在没有NADH的情况下,[1-14C]棕榈酰辅酶A与微粒体孵育产生了未去饱和的[1-14C]棕榈酰脂质。在加入NADH后,完整的酶系统并未将微粒体脂质中的放射性棕榈酸转化为棕榈油酸。当用[1-14C]棕榈酸或[1-14C]硬脂酸标记的细胞制备的微粒体在O2和NADH存在下单独孵育时,磷脂中[1-14C]棕榈油酸没有明显增加,而磷脂中[1-14C]亚油酸和γ-[1-14C]亚麻酸却以[1-14C]油酸为代价增加了。从这些结果可以得出结论,涉及将棕榈酸去饱和为棕榈油酸的酶需要棕榈酰辅酶A作为底物。然而,有人提出油酰和亚油酰磷脂可能是四膜虫微粒体去饱和过程中的底物。