Polar Organic Solvent Added to an Aqueous Solution Changes Hydrolytic Property of Lipase
Polar Organic Solvent Added to an Aqueous Solution Changes Hydrolytic Property of Lipase
复制标题
DOI：
10.1271/bbb.67.1660
复制
发表时间：
2003-01
期刊：
影响因子：
--
通讯作者：
W. Tsuzuki;Akemi Ue;A. Nagao
中科院分区：
文献类型：
--
作者：
W. Tsuzuki;Akemi Ue;A. Nagao
为了进一步开发脂肪酶作为生物催化剂的应用，研究了脂肪酶在缓冲液和极性有机溶剂组成的混合溶液中对某些酯类的水解活性。20%二甲基甲酰胺、35%二甲基亚砜、15%二氧六环、15%二甲氧基乙烷和2%二乙氧基乙烷促进了米黑根毛霉脂肪酶对4-甲基伞形酮油酸酯、4-甲基伞形酮棕榈酸酯和单油酸甘油酯的水解。而该脂肪酶对疏水性相对较弱的底物（4-甲基伞形酮庚酸酯和4-甲基伞形酮纳米酸酯）的水解则被这些溶剂抑制。荧光分析表明，缓冲液中的极性有机溶剂诱导了R.米黑脂肪酶除了溶剂对底物溶解度的影响外，溶剂对蛋白质构象的影响也会影响脂肪酶的反应特性。在水溶液中加入极性有机溶剂是改变脂肪酶水解性能的有效方法。
For developing further uses of lipase as a biocatalyst, its hydrolytic activity toward some esters was investigated in a miscible solution composed of a buffer and a polar organic solvent. Twenty percent dimethylformamide, 35% dimethylsulfoxide, 15% 1,4-dioxane, 15% dimethoxyethane, and 2% diethoxyethane promoted the hydrolysis by a lipase from Rhizomucor miehei toward some hydrophobic substrates, 4-methylumbelliferyl oleate, 4-methylumbelliferyl palmitate, and monoolein. While hydrolysis by this lipase toward the substrates with a relatively weak hydrophobicity (4-metylumbelliferyl heptanoate and 4-methylumbelliferyl nanoate) was suppressed by these solvents. A fluorometric analysis showed that the polar organic solvent in the buffer induced some conformational change around a tryptophan residue of R. miehei lipase. In addition to the influence of the miscible solvent on the solubility of the substrates, the conformational change of the protein induced by the miscible solvent would also affect the reactive properties of the lipase. Adding a polar organic solvent to an aqueous solution will be an efficient method for changing hydrolytic performance of lipases.