Alcalase 2.4L催化牛骨胶原蛋白水解反应及其动力学研究

Study on optimization and kinetic of Alcalase 2.4L catalyzed hydrolysis of collagen from ox bone

对碱性蛋白酶Alcalase 2.4L水解牛骨胶原蛋白的工艺以及动力学特征进行了研究。探讨底物浓度、酶量、温度、pH、时间等因素对胶原蛋白水解度的影响,研究确定了碱性蛋白酶Alcalase 2.4L水解牛骨胶原蛋白的最佳水解条件为:在pH8.5,60℃,底物浓度为80g/L,加酶量为40μL/g条件下水解3h,水解度可达13.5%,优于其他蛋白酶。根据所建立的pH-stat酶解体系确定了Km为1.6362mol/L,Vmax为0.3444mol/L·h,为更有效地利用胶原蛋白资源奠定了理论基础。

Enzymatic hydrolysis of collagen from ox bone,using Alcalase 2.4L as biocatalyst,was carried out in aqueous medium.The effect of key factors,including enzyme dosage,substrate concentration,reaction temperature,pH and reaction time,on the hydrolysis of collagen were investigated.Results showed that the optimal condition parameters were enzyme dosage of 40μL /g,substrate concentration of 80g/L,reaction temperature of 60℃,pH of 8.5,and reaction time of 3h,under which DH value reached 13.5%.Kinetic study showed that the maximum velocity and Km value were of 0.3444mol/L· h and 1.6362mol/L, respectively.