基于蛋白质结构知识解析活性多肽的酶解制备反应行为(Ⅱ)酶解反应过程的集总动力学模型

Analysis of enzymatic hydrolysis process for preparing active peptides based on protein structure(Ⅱ)Lumping kinetic model for enzymatic hydrolysis reaction

以酪蛋白胰蛋白酶反应体系为研究对象,建立了非均相酶解反应四集总动力学模型.研究中根据高效液相色谱分析结果及目标多肽所在区间,按分子量大小将酶解产物分段集总;依据反应机理与基本假设,构建集总反应网络;用包含酶失活、底物抑制和产物抑制常数的本征动力学方程描述各集总组分的反应行为,得到微分方程组形式的动力学模型;采用膜截分方法制得各集总组分,经实验测定和Marquardt法估算得到各集总组分动力学常数与反应网络各动力学参数;模型拟合与验证结果表明:所建多参数集总动力学机理模型具有良好的拟合性与外推性.

On the basis of chromatography analysis and reaction mechanism of enzymatic hydrolysis, the four-lumping kinetic model for the process of casein tryptic hydrolysis was established. According to the molecular mass, the hydrolysates were combined into four lumps and the simplified reaction network was described with intrinsic kinetics equation, which contained product inhibition, substrate inhibition and enzyme inactivation. The lumps were prepared by membrane separation, and the lumping kinetic constants were estimated by the Marquardt method. The ability of prediction and extrapolation of the lumping kinetic model was fitted and tested by comparing the computed values with the experimental ones, which demonstrated that the lumping method could characterize the kinetic behavior of complicated bio-reaction and predict the concentration distribution of multi-products.