Stenotrophomonas maltophilia DHHJ分泌角蛋白酶理化性质研究.doc

  Stenotrophomonas maltophilia DHHJ 分泌 角蛋白酶理化性质研究# 龚润1,3，蒋晶君1,3，曹张军1,3，周美华2** 5 10 15 20 25 30 （1. 东华大学生态纺织教育部重点实验室， 上海 201620； 2. 东华大学环境科学与环境工程学院，上海 201620； 3. 东华大学化学化工与生物工程学院，上海 201620） 摘要：本文利用嗜麦芽窄食单胞菌 DHHJ 发酵并通过盐析、层析纯化角蛋白酶，进而研究其 理化性质。角蛋白酶对温度敏感，易于保存在低温环境（4）下，4 天后，纯酶液的酶活 损失率为 22.2%。10 mmol/L Ca2+，Ba2+，有机溶剂甘油和乙醇（浓度范围在 0-20%之间） 对角蛋白酶活有显著的促进作用，10mmol/LNa+对酶活有一定促进作用，而 10mmol/L Zn2+， Cd2+，表面活性剂 Tween 80 和 SDS 则可显著的抑制酶活，其中，ZnCl2 抑制角蛋白酶的类 型主要是与底物竞争性地抑制角蛋白酶活性。比起羽毛底物，角蛋白酶与酪蛋白的亲和能力 更好。本研究为工业化应用提供一定的理论基础。 关键词：嗜麦芽窄食单胞菌 DHHJ; 角蛋白酶; 米氏方程 Characterization of keratinase from keratin-degrading bacterial Stenotrophomonas maltophilia DHHJ Gong Rui1, Jiang Jingjun1,3, CAO Zhangjun1, ZHOU Meihua2 (1. Key Laboratory of Science Technology of Eco-Textile (Donghua University), Ministry of Education，Shanghai 201620; 2. College of Environmental Science and Engineering，Donghua University，Shanghai 201620; 3. College of Chemistry, Chemical Engineering and Biotechnology, Shanghai 201620) Abstract: In this study, Stenotrophomonas maltophilia DHHJ was used to produce keratinase by fermentation. The keratinase was purified to illustrate its physical and chemical characteristics. Keratinase is sensitive to temperature, and it is stable to conserve in the refrigerator at 4℃. The loss rate of enzyme activity is 22.2% at 4℃, while the keratinase shows no activity at 25℃ after 4 days. The keratinase activity was stimulated obviously by 10mmol/L Ca2+, Ba2+, organic solvents glycerol and ethanol (concentration between 0-20%) and enhanced slightly by 10mmol/LNa+. However, the keratinase activity was inhibited by 10mmol/L Zn2+, Cd2+, surfactant Tween 80 and SDS. In addition, the inhibition of pattern of ZnCl2 on keratinase activity belongs to competitive one. Compared to feathers, keratinase has a better affinity for casein. These physical and chemical characteristics provide a theoretical foundation for industrial application. Key words: Stenotrophomonas maltophilia DHHJ; keratinase; Michaelis const