嗜盐性蛋白酶产生菌VirgibacillusspP-4的筛选鉴定及其特性分析.PDF

102 2017, Vol.38, No.22 食品科学 ※生物工程 嗜盐性蛋白酶产生菌Virgibacillus sp. P-4 的 筛选鉴定及其特性分析 * 张延杰，王静雪 ，牟海津 （中国海洋大学食品科学与工程学院，山东青岛 266003 ） 摘  要：利用酪蛋白琼脂平板从传统虾酱中分离嗜盐性蛋白酶产生菌，经形态学、生理生化和分子生物学鉴定， 确定该菌隶属于枝芽孢杆菌属Virgibacillus sp. P-4 ，并用此菌为后续研究菌株。结果表明，该菌最适生长温度为 30～37 ℃，最适生长NaCl质量分数为5%～15%；所产胞外蛋白酶在20～50 ℃条件下酶活力较高且保持稳定，最适 反应温度为40 ℃，当NaCl质量分数为15%时酶活力达到最高，并且所产胞外蛋白酶种类可能不只一种，但不包含 巯基蛋白酶；通过分析酶解液中游离氨基酸释放速率，预测其酶切位点为Phe- 、Tyr- 、Lys- 、His- 、Pro-及Leu- 。 关键词：蛋白酶产生菌；嗜盐菌；虾酱；性质 Virgibacillus Isolation, Identification and Properties of Protease-Producing Halophilic Bacterium sp. P-4 from Traditional Shrimp Sauce * ZHANG Yanjie, WANG Jingxue , MOU Haijin (College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China) Abstract: Protease-producing bacteria were isolated from traditional shrimp sauce by using casein agar medium containing 10% NaCl. Based on its morphological, biochemical and molecular properties, Virgibacillus sp. P-4 was determined as target strain for further study. The strain grew best at 30–37 ℃ with a salinity of 5%–15%. The protease produced by Virgibacillus sp. P-4 showed the optimum activity at 40 ℃, and was stable in the range of 20–50 ℃. The enzymatic activity reached its peak when NaCl concentration was 15%. Besides, Virgibacillus sp. P-4 might produce more than one extracellular protease except sulfhydryl protease. According to the rate of release of free a