低温果胶酶的纯化及酶学性质的研究  

Study on purification and characteristics of the cold adapted pectinase

作　　者：王伟[1] 李锋[2] 张磊[3] 茆军[4] 

机构地区：[1]新疆农业大学食品科学学院,新疆乌鲁木齐830052 [2]新疆农业大学化工学院,新疆乌鲁木齐830052 [3]新疆农业大学教务处,新疆乌鲁木齐830052 [4]新疆农业科学院微生物研究所,新疆乌鲁木齐830091

出　　处：《食品工业科技》2011年第4期162-165,共4页Science and Technology of Food Industry

摘　　要：进行了低温果胶酶产生菌的筛选,并对低温果胶酶的酶学性质、纯化及动力学常数进行了研究。结果表明:该酶最适酶反应温度25℃,最适酶反应pH为5.8;酶液的pH稳定范围在5～6.4;在25℃保温3h,酶活降至50.5%,在35℃保温2h,酶活降至55.1%,在45℃保温1.5h,酶活降至48.7%,在55℃保温1h,酶活降至43.1%;金属离子Mg2+、Ca2+、Mn2+、Zn2+、Fe2+、Na+对低温果胶酶有激活作用,而Ba2+、Hg2+、Cu2+、Fe3+、Pb2+对低温果胶酶有强烈的抑制作用;当以果胶为底物时,该酶的动力学常数Km为27.86mg/mL,Vmax为152.13μmol/(min·mL);采用不同的处理方法均可使低温果胶酶得到纯化,但40%～85%的硫酸铵分级沉淀纯化效果最好。The strain producing cold adapted pectinase was screened, and the characteristics of cold adapted pectinase, purification and kinetic constant were studied.The results showed that： the optimal temperature and pH for the enzyme reaction was 25℃ and 5.8 ,the enzyme was stable at pH5-6.4,after 3h at 25℃,the enzyme activity was 50.5 %, at 35 ℃ for 2h, the enzyme activity was 55. 1 %, at 45 ℃ for 1.5 h, the enzyme activity was 48.7/%, at 55 ℃ for 1h, the enzyme activity was 43.1%.The metal ion Mg2＋ , Ca2＋ , Mn2＋ , Zn2. , Fe2＋ , Na＋ enhanced the cold adapted pectinase activity, but the metal ion Ba2＋ , Hg2＋ , Cu2＋ , Fe3＋ , Pb2＋ inhibited the cold adapted pectinase activity strongly. The kinetic constant of the cold adapted pectinase was Km value 27.86mg/mL and Vmax 152.13μmol/（ min · mL）when pectin （sigma）as a substrate. The cold adapted pectinase was purified by ammonium sulfate salted out and the optimal range of ammonium sulfate saturation was 40%-85%.

关 键 词：低温果胶酶 最适酶反应温度 最适酶反应pH 分离纯化 

分 类 号：TS201.25[轻工技术与工程—食品科学]