Malmqvist T, Möllby R
Acta Pathol Microbiol Scand B. 1981 Oct;89(5):363-7. doi: 10.1111/j.1699-0463.1981.tb00201_89b.x.
An assay system for phospholipases C has been described with sphingomyelin immobilized to octyl-Sepharose CL-4B as substrate. The immobilization procedure was further developed and used with [14 C-choline]-sphingomyelin and [14C-choline] phosphatidylcholine (lecithin). These immobilized radioactive phospholipids made the enzymatic assays easier to perform and made it possible to increase the sensitivity. Furthermore, since release of the choline part instead of the phosphate part of the substrate molecule was measured, it was possible to use this assay for phospholipase D as well. The enzyme characteristics of phospholipase D from Corynebacterium ovis were compared in this test system with those of three phospholipases C (from Clostridium perfringens, Bacillus cereus and Staphylococcus aureus) with respect to hydrolysing capacities and optimal ion concentrations.
已经描述了一种以固定在辛基-琼脂糖CL-4B上的鞘磷脂为底物的磷脂酶C检测系统。固定化程序得到了进一步改进，并与[14C-胆碱]-鞘磷脂和[14C-胆碱]磷脂酰胆碱（卵磷脂）一起使用。这些固定化的放射性磷脂使酶促检测更容易进行，并提高了灵敏度。此外，由于测量的是底物分子胆碱部分而非磷酸部分的释放，因此该检测也可用于磷脂酶D。在该测试系统中，比较了来自羊棒状杆菌的磷脂酶D与三种磷脂酶C（来自产气荚膜梭菌、蜡状芽孢杆菌和金黄色葡萄球菌）在水解能力和最佳离子浓度方面的酶特性。
