Sanchez J, Harwood J L
Biochem J. 1981 Oct 1;199(1):221-6. doi: 10.1042/bj1990221.
The synthesis of lipids and acyl thioesters was studied in microsomal preparations from germinating pea (Pisum sativum cv. Feltham First) seeds. Under conditions of maximal synthesis (in the presence of exogenous acyl-carrier protein) acyl-acyl-carrier proteins accounted for about half the total incorporation from [14C]malonyl-CoA. Decreasing the concentrations of exogenous acyl-carrier protein lowered the overall synthesis of fatty acids by decreasing, almost exclusively, the radioactivity associated with acyl-acyl-carrier proteins. A time-course experiment showed that acyl-acyl-carrier proteins accumulated most of the radioactive label at the beginning of the incubation but, eventually, the amount of radioactivity in that fraction decreased, while a simultaneous increase in the acyl-CoA and lipid fractions was noticed. Addition of exogenous CoA (1 mM) produced a decrease of total incorporation, but an increase in the radioactivity incorporated into acyl-CoA. The microsomal preparations synthesized saturated fatty acids up to C20, including significant proportions of pentadecanoic acid and heptadecanoic acid. Synthesis of these 'odd-chain' fatty acids only took place in the microsomal fraction. In contrast, when the 18,000g supernatant (containing the microsomal and soluble fractions) was incubated with [14C]malonyl-CoA, the radioactive fatty acid and acyl classes closely resembled the patterns produced by germinating in the presence of [14C]acetate in vivo. The results are discussed in relation to the role of acyl thioesters in the biosynthesis of plant lipids.
对萌发的豌豆(品种为Feltham First)种子的微粒体提取物中脂质和酰基硫酯的合成进行了研究。在最大合成条件下(存在外源酰基载体蛋白),酰基 - 酰基载体蛋白约占[14C]丙二酸单酰辅酶A总掺入量的一半。降低外源酰基载体蛋白的浓度几乎完全通过降低与酰基 - 酰基载体蛋白相关的放射性来降低脂肪酸的总体合成。一项时间进程实验表明,酰基 - 酰基载体蛋白在孵育开始时积累了大部分放射性标记,但最终该部分的放射性量减少,同时注意到酰基辅酶A和脂质部分的放射性增加。添加外源辅酶A(1 mM)导致总掺入量减少,但酰基辅酶A中掺入的放射性增加。微粒体提取物合成了直至C20的饱和脂肪酸,包括相当比例的十五烷酸和十七烷酸。这些“奇数链”脂肪酸的合成仅在微粒体部分发生。相比之下,当18,000g上清液(包含微粒体和可溶部分)与[14C]丙二酸单酰辅酶A一起孵育时,放射性脂肪酸和酰基类别与在体内[14C]乙酸盐存在下萌发产生的模式非常相似。结合酰基硫酯在植物脂质生物合成中的作用对结果进行了讨论。