3 129-132 转148 基础研究 4+20 地衣芽孢杆菌2709蛋白酶分离纯化与性质.pdfVIP

现代食品科技 Modern Food Science and Technology 2010, Vol.26, No.2 地衣芽孢杆菌2709蛋白酶分离纯化与性质 张浩，张帅，张娜，马永强 （哈尔滨商业大学食品工程学院，黑龙江哈尔滨150076 ） 摘要：采用盐析、DEAE 阴离子交换层析、凝胶层析等步骤，从地衣芽孢杆菌(B·licheniformis)2709的发酵液中获得了电泳 纯的碱性蛋白酶。结果发现：提纯酶的活力达61069 U/mg ，纯化倍数：38.7 ，活性回收率：19.3%，去酰氨度：20.9%.并研究了该酶的基本酶学特性，碱性蛋白酶最适反应温度为50 ℃，最适作用pH为10.0。40 ℃保温2 h后该酶保持80%以上的活力，在pH 8~11之间有较高的pH稳定性。 关键词：碱性蛋白酶；提纯；去酰氨；性质 文章篇号:1673-9078(2010)2-129-132 Purification and properties of alkaline protease from B.licheniformis 2709 ZHANG Hao, ZHANG Shuai, ZHANG Na, MA Yong-qiang (College of Food Engineering of Harbin Commercial University, Harbin 150076, China) Abstract: The alkaline protease was purified from B·licheniformis 2709 by alcohol precipitation, ammonium sulphate precipitation, DEAE ion exchange chromatography, and gel layer chromatography and the purified alkaline was demonstrated to be electrophoretic by SDS. The results indicated that the enzyme activity of purified protease was 61069 U·mg-1 with final purification factor, activity recovery factor and deamidation reate of 38.7, 19.3%, and 20.9%, respectively. The optimal conditions for the highest activity of alkaline protease were as follows: pH 10.0, and temperature 50℃. After incubation at 40 ℃for 2h, the residual activity of the enzyme was above 80%, and the enzyme was relatively stable at pH 8-11. Key words: alkaline protease; purification; deamidation; properties 129 现代食品科技 Modern Food Science and Technology 2009, Vol.26, No.2 我国是大豆的故乡，品种资源极为丰富 1 材料与方法 ，产量极大，尤其北方种植甚为广泛，大豆 的加工历史十分悠久，早在2000多年前，在 1.1 材料与设备 中