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Immobilization of enzymes in porous supports: Effects of support–enzyme solution contacting
Author(s) -
Dennis K. E.,
Clark D. S.,
Bailey J. E.,
Cho Y. K.,
Park Y. H.
Publication year - 1984
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260260812
Subject(s) - carbodiimide , immobilized enzyme , chemistry , enzyme , diffusion , glucose oxidase , chromatography , porosity , kinetics , chemical engineering , sepharose , biophysics , biochemistry , organic chemistry , biology , physics , quantum mechanics , engineering , thermodynamics
Proteins have been immobilized in porous support particles held in a fixed‐bed reactor through which protein solution is continuously circulated. Changing the recirculation flow rate alters the observed immobilization kinetics and the maximum enzyme loading which can be achieved for glucose oxidase and glucoamylase on carbodiimide‐treated activated carbon and for glucoamylase immobilized on CNBr–Sepharose 4B. Direct microscopic examination of FITC‐labelled protein in sectioned Sepharose particles and indirect activity–loading studies with activated carbon–enzyme conjugates all indicate that immobilized enzyme is increasingly localized near the outer surface of the support particles at larger recirculation flow rates. Restricted diffusion of enzymes may be implicated in this phenomenon. These contacting effects may be significant considerations in the scaleup of processes for protein impregnation in porous supports, since apparent activity and stability of the final preparation depend on internal protein distribution.