Department of Chemical Engineering, Clarkson University, Potsdam, NY 13699-5705, USA.
Biotechnol Bioeng. 1992 Dec 5;40(10):1173-80. doi: 10.1002/bit.260401006.
In this article, the results from a theoretical and experimental investigation of enzyme immobilization in porous membranes are reported. A theoretical model of the immobilization process, which accounts for restricted diffusion of enzyme in the pores of the membrane, has been developed. The model predicts the effect of immobilization kinetics and time of immobilization on the enzyme distribution in the pores of the membrane. The immobilization of glucose oxidase and glucose oxidase-biotin conjugate on porous alumina membranes was experimentally investigated. Enzyme uptake data was correlated to the theory to determine the rate constant of immobilization and the distribution of the enzyme in the pore. Immobilization studies were carried out for enzyme adsorption and for enzyme attachment by covalent coupling. The distribution of enzyme was experimentally studied by assembling five membranes in the diffusion cell. Following immobilization, the membranes were separated and each was assayed for activity. The amount of active enzyme present in each membrane yielded a discrete distribution that compared well with that predicted by theory.
本文报道了多孔膜中酶固定化的理论和实验研究结果。已经开发了一种固定化过程的理论模型，该模型考虑了酶在膜孔中的受限扩散。该模型预测了固定化动力学和固定化时间对酶在膜孔中分布的影响。实验研究了葡萄糖氧化酶和葡萄糖氧化酶-生物素缀合物在多孔氧化铝膜上的固定化。将酶摄取数据与理论相关联，以确定固定化速率常数和酶在孔中的分布。通过吸附和共价偶联进行了酶固定化研究。通过在扩散池中组装五个膜来实验研究酶的分布。固定化后，将膜分离并对每个膜进行活性测定。每个膜中存在的活性酶的量呈现离散分布，与理论预测非常吻合。
