可溶性固定化胰蛋白酶的制备及性质研究

将胰蛋白酶偶联在亲水性高分子聚合物N-琥珀酰壳聚糖上,制备得到水溶性固定化胰蛋白酶,并对其部分酶学性质进行了研究。结果表明,可溶性固定化胰蛋白酶的最适pH为9·0,最适温度为60℃,Km值为0·28mg/mL;该酶在pH6·0以上能够完全溶解,在pH4·0左右基本不能溶解;可溶性固定化胰蛋白酶在60℃条件下,保温8h,仍有55%的活性;在4℃条件下,30d后活性保留达98%;在4℃下保存72h,活性没有明显降低。pH8·0~10·0稳定性最好(相对活性>90%)。

Preparation and Properties of Soluble-Immobilized Trypsin

To prepare a soluble-immobilized trypsin and determine its enzymatic properties. The soluble immobilized trypsin was obtained by conjugating the trypsin with the hydrophilic high molecular polymer of N-succinyl-chitosan.The results showed that the optimum pH of soluble- immobilized trypsin was pH 9.0. Their optimum temperature was 60℃. Its Km value was 0.28 mg/mL. Under 60℃, after 8 hours the relative activity of soluble-immobilized trypsin was 55%. Under 4℃, after 30 days the relative activity was still over 98%. Under the pH 6.0, after 72 hours the activity of immobilized trypsin did not decrease obviously, and its stability was high(relative activity was over 90% between pH 8.0 and 10.0).