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首页> 外文期刊>Journal of Lipid Research >Enhancement of hydrolytic activity of sphingolipid ceramide N-deacylase in the aqueous-organic biphasic system.
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Enhancement of hydrolytic activity of sphingolipid ceramide N-deacylase in the aqueous-organic biphasic system.

机译：在水-有机两相系统中鞘脂神经酰胺N-脱酰基酶的水解活性增强。

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摘要

Lysoglycosphingolipids were produced from glycosphingolipids by using sphingolipid ceramide N-deacylase, which cleaves the N-acyl linkage between fatty acids and sphingosine bases in various glycosphingolipids. The enzyme reaction was done in a biphasic media prepared with water;-immiscible organic solvent and aqueous buffer solution containing the enzyme. We investigated the effects of organic solvents and detergents on lysoglycosphingolipid production in the biphasic system. Among the organic solvents tested, n-butylbenzene, cumene, cyclodecane, cyclohexane, n-decane, diisopropylether, n-heptadecane, and methylcyclohexane promoted hydrolysis of GM1, whereas benzene, chloroform, ethyl acetate, and toluene inhibited GM1 hydrolysis. Hydrolysis of asialo GM1, GD1a, GalCer, and sulfatide was also enhanced by the addition of n-decane. The hydrolytic activity of the enzyme was enhanced by the addition of 0.8% sodium taurodeoxycholate or sodium cholate to the aqueous phase. The most effective hydrolysis of various glycosphingolipids by the enzyme was thus obtained in the aqueous-n-decane biphasic system containing 0.8% sodium taurodeoxycholate. Under this condition, the fatty acids released from GM1 by the action of the enzyme were trapped and diffused into the organic phase, while lysoGM1 remained in the aqueous phase.Thus the almost complete hydrolysis of GM1 was achieved using the biphasic system, while at most 70% of hydrolysis was obtained using normal aqueous media possibly due to the inhibition of hydrolysis reaction by accumulation of fatty acids in the reaction mixture.
机译：通过使用鞘脂神经酰胺N-脱酰基酶从糖鞘脂产生溶糖鞘糖脂，其裂解各种糖鞘脂中的脂肪酸和鞘氨醇碱之间的N-酰基键。酶反应是在两相介质中完成的，该介质是用水，不混溶的有机溶剂和含酶的缓冲水溶液制备的。我们研究了有机溶剂和去污剂对双相系统中溶血糖鞘脂生产的影响。在测试的有机溶剂中，正丁基苯，枯烯，环癸烷，环己烷，正癸烷，二异丙醚，正十七烷和甲基环己烷可促进GM1的水解，而苯，氯仿，乙酸乙酯和甲苯则可抑制GM1的水解。加入正癸烷也可增强无唾液酸GM1，GD1a，GalCer和硫化物的水解。通过向水相中添加0.8％的牛磺脱氧胆酸钠或胆酸钠来增强酶的水解活性。因此，在含有0.8％牛磺脱氧胆酸钠的正癸烷双相水溶液中，通过该酶最有效地水解了各种鞘糖脂。在这种条件下，通过酶的作用从GM1释放的脂肪酸被捕集并扩散到有机相中，而lysoGM1保留在水相中，因此使用双相体系几乎可以完全水解GM1，而最多使用正常的水性介质可获得70％的水解，这可能是由于脂肪酸在反应混合物中的积累而抑制了水解反应。

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