Toida J, Arikawa Y, Kondou K, Fukuzawa M, Sekiguchi J
Food Technology Research Institute of Nagano Prefecture, Japan.
Biosci Biotechnol Biochem. 1998 Apr;62(4):759-63. doi: 10.1271/bbb.62.759.
Triacylglycerol lipase (L3) was purified from Aspergillus oryzae RIB128 by ammonium sulfate fractionation, acetone precipitation, anion-exchange chromatography, and gel filtration. The purified enzyme was formed from a glycoprotein and a monomeric protein with molecular masses of 25 and 29 kDa, by SDS-PAGE and gel filtration, respectively. The optimum pH at 40 degrees C was 5.5 and the optimum temperature at pH 5.5 was 40 degrees C. The enzyme was stable between a pH range of 4.0-7.5 at 30 degrees C for 24 h, and at up to 30 degrees C at pH 5.5 for 1 h. Heavy metal ions, detergents, DFP, and DEP strongly inhibited the enzyme activity. The lipase hydrolyzed not only triacylglycerols but also monoacylglycerols and diacylglycerols. The enzyme had higher specificity toward triacylglycerols of middle-chain saturated fatty acids than short-chain or long-chain fatty acids. The enzyme had 1,3-positional specificity. The N-terminal amino acid sequence of the enzyme was not significantly similar to that of other lipases with published sequences.
通过硫酸铵分级沉淀、丙酮沉淀、阴离子交换色谱和凝胶过滤从米曲霉RIB128中纯化了三酰甘油脂肪酶（L3）。通过SDS-PAGE和凝胶过滤分别测定，纯化后的酶由一种糖蛋白和一种分子量分别为25 kDa和29 kDa的单体蛋白组成。40℃时的最适pH为5.5，pH 5.5时的最适温度为40℃。该酶在30℃下pH 4.0 - 7.5的范围内24小时稳定，在pH 5.5时30℃下1小时内稳定。重金属离子、去污剂、DFP和DEP强烈抑制酶活性。该脂肪酶不仅能水解三酰甘油，还能水解单酰甘油和二酰甘油。该酶对中链饱和脂肪酸的三酰甘油的特异性高于短链或长链脂肪酸。该酶具有1,3-位特异性。该酶的N端氨基酸序列与已发表序列的其他脂肪酶没有明显相似性。
