Rock C O, Garwin J L
J Biol Chem. 1979 Aug 10;254(15):7123-8.
We have used purified preparations of acyl-acyl carrier protein synthetase to prepare pure, native acyl-acyl carrier proteins (acyl-ACP) ranging in chain lengths from C10:0 to C delta 9 18:1. Factors affecting yield are explored and reaction conditions are presented that yield 0.8 to 0.9 mg of C16:0-ACP/ml of reaction mix. Ohter acyl groups, such as C10:0 and C delta 9 18:1 are poorer substrates and gave correspondingly lower yields. Acyl-Acp synthetase may be recovered from the reaction mixture using blue-Sepharose CL-6B and recycled. ACP and acyl-ACP are separated by hydrophobic chromatography on octyl-Sepharose CL-4B. Mixtures of acyl-ACPs could be resolved according to acyl chain length using octyl-Sepharose CL-4B columns eluted with a 2-propanol gradient. The high resolution obtained using 2-propanol gradients to separate acyl-ACP species suggests that similar techniques would be applicable to the chromatography of protein mixtures on hydrophobic supports.
我们使用纯化的酰基 - 酰基载体蛋白合成酶制剂来制备链长范围从C10:0到CΔ9 18:1的纯的天然酰基 - 酰基载体蛋白（酰基 - ACP）。探索了影响产量的因素，并给出了反应条件，该条件下每毫升反应混合物可产生0.8至0.9毫克的C16:0 - ACP。其他酰基，如C10:0和CΔ9 18:1是较差的底物，产量相应较低。酰基 - Acp合成酶可使用蓝色琼脂糖凝胶CL - 6B从反应混合物中回收并循环使用。ACP和酰基 - ACP通过在辛基琼脂糖凝胶CL - 4B上的疏水色谱法分离。使用辛基琼脂糖凝胶CL - 4B柱，用2 - 丙醇梯度洗脱，可以根据酰基链长度分离酰基 - ACP混合物。使用2 - 丙醇梯度分离酰基 - ACP种类所获得的高分辨率表明，类似技术将适用于在疏水支持物上对蛋白质混合物进行色谱分析。
