Toida J, Kondoh K, Fukuzawa M, Ohnishi K, Sekiguchi J
Food Technology Research Institute of Nagano Prefecture, Japan.
Biosci Biotechnol Biochem. 1995 Jul;59(7):1199-203. doi: 10.1271/bbb.59.1199.
A lipase from Aspergillus oryzae was purified by ammonium sulfate fractionation, anion exchange chromatography, hydrophobic interaction chromatography, and anion exchange chromatography. The purified enzyme was a monomeric protein with a molecular mass of 41 kDa estimated by SDS-PAGE and 39 kDa by gel filtration. The optimum pH at 30 degrees C and optimum temperature at pH 7.0 were 7.0 and 30 degrees C, respectively. The enzyme was stable over a pH range of 6-9 at 25 degrees C for 18 h, and up to 30 degrees C at pH 7.0 for 3 h. Ag+, Fe3+, Hg2+, Cu2+, and Zn2+ inhibited the enzyme activity severely. The enzyme was a lipase that hydrolyzed monoacylglycerols and diacylglycerols, but did not hydrolyze triacylglycerols. The N-terminal amino acid sequence of the enzyme was highly homologous with that of the mono- and diacylglycerol lipase from Penicillium camembertii U-150.
通过硫酸铵分级沉淀、阴离子交换色谱、疏水相互作用色谱和阴离子交换色谱对米曲霉脂肪酶进行了纯化。纯化后的酶是一种单体蛋白，经SDS-PAGE估计分子量为41 kDa，经凝胶过滤法测定为39 kDa。在30℃时的最适pH值和pH 7.0时的最适温度分别为7.0和30℃。该酶在25℃下pH值6 - 9的范围内18小时保持稳定，在pH 7.0时30℃下3小时内保持稳定。Ag +、Fe3 +、Hg2 +、Cu2 +和Zn2 +严重抑制该酶的活性。该酶是一种能水解单酰甘油和二酰甘油，但不能水解三酰甘油的脂肪酶。该酶的N端氨基酸序列与来自卡门柏青霉U - 150的单酰甘油和二酰甘油脂肪酶高度同源。
