Galliard T
Biochem J. 1971 Feb;121(3):379-90. doi: 10.1042/bj1210379.
An enzyme preparation that catalyses the deacylation of mono- and di-acyl phospholipids, galactosyl diglycerides, mono- and di-glycerides has been partially purified from potato tubers. The preparation also hydrolyses methyl and p-nitrophenyl esters and acts preferentially on esters of long-chain fatty acids. Triglycerides, wax esters and sterol esters are not hydrolysed. The same enzyme preparation catalyses acyl transfer reactions in the presence of alcohols and also catalyses the synthesis of wax esters from long-chain alcohols and free fatty acids. Gel filtration, DEAE-cellulose chromatography and free-flow electrophoresis failed to achieve any separation of the acyl-hydrolase activities towards different classes of acyl lipids (phosphatidylcholine, monogalactosyl diglyceride, mono-olein, methyl palmitate and p-nitrophenyl palmitate) or any separation of these activities from a major protein component. For each class of lipid the acyl-hydrolase activity was subject to substrate inhibition, was inhibited by relatively high concentrations of di-isopropyl phosphorofluoridate and the pH responses were changed by Triton X-100. The hydrolysis of phosphatidylcholine was stimulated 30-40-fold by Triton X-100. The specific activities of the potato enzyme with galactolipids were at least 70 times higher than those reported for a homogeneous galactolipase enzyme purified from runner bean leaves. The possibility that a single lipolytic acyl-hydrolase enzyme is responsible for the deacylation of several classes of acyl lipid is discussed.
一种能催化单酰基和二酰基磷脂、半乳糖基甘油二酯、单甘油酯和二甘油酯脱酰基反应的酶制剂已从马铃薯块茎中部分纯化出来。该制剂还能水解甲基酯和对硝基苯酯,且优先作用于长链脂肪酸的酯。甘油三酯、蜡酯和甾醇酯不被水解。同一酶制剂在醇存在的情况下催化酰基转移反应,还能催化长链醇和游离脂肪酸合成蜡酯。凝胶过滤、DEAE - 纤维素色谱法和自由流动电泳未能实现对不同类酰基脂质(磷脂酰胆碱、单半乳糖基甘油二酯、单油酸甘油酯、棕榈酸甲酯和对硝基苯棕榈酸酯)的酰基水解酶活性进行任何分离,也未能将这些活性与一种主要蛋白质成分分离。对于每一类脂质,酰基水解酶活性都受到底物抑制,被相对高浓度的二异丙基氟磷酸酯抑制,并且pH响应会因Triton X - 100而改变。Triton X - 100能使磷脂酰胆碱的水解活性提高30 - 40倍。马铃薯酶对半乳糖脂的比活性至少比从菜豆叶片中纯化出的一种纯半乳糖脂酶所报道的比活性高70倍。本文讨论了单一脂解酰基水解酶负责几类酰基脂质脱酰基反应的可能性。