水生栖热菌FL-03海藻糖合酶的分离纯化及特性-吉林农业大学学报.PDFVIP

　28 1 　　　　　　　 Vol.28 No.1　 　2006 2 Journal　of　Jilin　Agricultural　University Feb.2006　 FL-03 胡耀辉, 于寒松, 王　欢 (吉林农业大学食品工程学院, 长春 130118) 　:将从水生栖热菌FL-03 中获得的海藻糖合酶粗酶液经硫酸铵分 沉淀、DEAE Sepharose CL-6B 离 子交换层析、Sephacryl S-200HR凝胶过滤层析后, 纯化68.48 倍, 产率为18.4%。 研究表明:该酶分子量约为 101 kD, 最适反应温度为 60℃, 最适pH 为7.0, 在40 ～ 80℃、pH 6.0 ～ 9.0 范围内具有较高的反应活性和稳定 3+ 2+ 2+ 性。Fe 对该酶具有 一定的激活作用, Cu 、 ris、Zn 对该酶具有较强的抑制作用。 该酶具有高度的底物特 异性, 只能专 一地将麦芽糖转化为海藻糖。 在40 ℃、48 h 条件下, 麦芽糖转化为海藻糖的转化率最大可达到 79%。 :水生栖热菌FL-03;海藻糖合酶;分离纯化;理化性质 :S201.25　　　:A　　　:1000-5684(2006)01-0027-05 Separation Purification and Characterization of Trehalose Sythase from Thermus aquaticus FL-03 HU Yao-hui, YU Han-song, WA NG Huan (College of Food E ngineering ,J ilin Agricultura l University , Changchun 130118, China ) Abstract:he trehalose sythase from Thermus aquaticus FL-03 was purifiled by chromatography meth- ods, and the purity of enzyme was identified by sodium dodecyl suphate polyacrylamide gel electophoresis (SDS).he enzyme was purified about 68.48 fold with a yield of 18.4%total activity by suc- cessive ammonium sulfate-fractionation, DEAE Sepharose CL-6B, Sephacryl S-200HR column chro- matography.When the purified enzyme sampleswere subjected to SDS, the molecular weight esti- mated for trehalose sythase was 101kD.he optimum temperature and pH toward trehalose sythase were 60 ℃and 7.0 respectively, and the enzyme was stable at temperature 40 ～ 80 ℃and pH 6.0 ～9.0 for 60min.he maximum yield of trehalose from maltose by the enzyme reached 79% at 40 ℃after 48 hours.he trehalose sythase acted specifically on maltose an