Determination of total and active immobilized enzyme distribution in porous solid supports
β Scribed by Rolf Scharer; Md. M. Hossain; D. D. Do
- Publisher
- John Wiley and Sons
- Year
- 1992
- Tongue
- English
- Weight
- 718 KB
- Volume
- 39
- Category
- Article
- ISSN
- 0006-3592
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β¦ Synopsis
The total and active immobilized enzyme (IME) distributions in porous supports are studied both theoretically and experimentally. In order to determine experimentally the enzyme distribution profiles within a single particle, we construct a diffusion cell containing controlled-pore glass particles such that t h e cell would mimic a large pellet support. Our purpose is to study the interplay between the diffusion process within the interparticle void space and irnrnobilization process in the controlled-pore glass particles onto the evolution of the (total and active) enzyme distributions. A mathematical model is developed to describe the interaction of various processes within the diffusion cell. The im- mobilized enzymes are determined for a system of trypsin and controlled-pore glass particles. The total amount of enzymes are determined by the amino acid analysis, and the active fraction is obtained by an active-site titration. The experimentally measured total IME profiles compare very well with that predicted by the model. The determined active enzyme profile is found to be nonuniform one, and it represents about 40% of the total enzyme immobilized in the support particles.
π SIMILAR VOLUMES
Clark et al. (1985) recently formulated a kinetic model for coupling restricted diffusion and immobilized reaction of enzyme molecules in a cylindrical pore using the concept of "pore central core restricted diffusion." This model takes into consideration the increase of diffusion resistance by t
Immobilization of native proteins, retaining their activity, on the solid support is often crucial for a variety of biochemical assays involving protein-protein interactions. In this study we describe a technique which allows binding of both complex (protein kinase CK2) and simple (calf intestine al