Rajgarhia V B, Priestley N D, Strohl W R
Department of Microbiology, College of Pharmacy, Ohio State University, Columbus 43210.
Anal Biochem. 1995 Jan 1;224(1):159-62. doi: 10.1006/abio.1995.1022.
The efficient microscale synthesis of [1-14C]propionyl-CoA from commercially available sodium [1-14C]-propionate using 1,1'-carbonyldiimidazole in yields of nearly 70% is reported for the first time. A substantial improvement in the process for making [1-14C]acetyl-CoA from sodium [1-14C]acetate was also achieved. Yields of greater than 90% were consistently obtained for the latter synthesis. The salt-free CoA-thioesters were obtained in homogenous form by reverse-phase HPLC. The products were judged to be pure by 1H NMR analysis: neither iso-CoA analogs nor contaminants frequently found in commercial samples could be detected. The samples of acetyl- and propionyl-CoA were shown to be radiochemically pure by HPLC and by analysis of the products of incubations with acetyl- and propionyl-CoA carboxylase. This highly efficient synthesis is a cost-effective method for the preparation of radiolabeled CoA thioesters and can easily be adapted to the production of other acyl-CoA analogs.
首次报道了使用1,1'-羰基二咪唑从市售的[1-¹⁴C]丙酸钠高效微尺度合成[1-¹⁴C]丙酰辅酶A，产率接近70%。由[1-¹⁴C]乙酸钠制备[1-¹⁴C]乙酰辅酶A的过程也有了显著改进。后者的合成产率始终大于90%。通过反相高效液相色谱法可得到均一形式的无盐辅酶A硫酯。通过¹H NMR分析判断产物为纯品：未检测到商业样品中常见的异辅酶A类似物或污染物。通过高效液相色谱法以及对乙酰辅酶A羧化酶和丙酰辅酶A羧化酶孵育产物的分析表明，乙酰辅酶A和丙酰辅酶A样品在放射化学上是纯的。这种高效合成方法是制备放射性标记辅酶A硫酯的一种经济有效的方法，并且可以很容易地适用于生产其他酰基辅酶A类似物。
