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Novel aromatic polymers for immobilizing B-D-glucosidase and their possible application to cellulolysis
Journal article   Peer reviewed

Novel aromatic polymers for immobilizing B-D-glucosidase and their possible application to cellulolysis

J M Sarkar and Richard G Burns
Applied Biochemistry and Biotechnology, Vol.9(4), p.413
1984
url
https://doi.org/10.1007/BF02799002View
Published Version

Abstract

biotechnology
We have synthesized, by enzymic and chemical means, a variety of novel polyaromatic-enzyme complexes that are extremely stable and show promise in the conversion of cellulose to glucose. Thus we have prepared a number of homo- and heteropolymeric supports (involving l-tyrosine, pyrogallol, resorcinol, phloroglucinol, orcinol, catechol, protocatechuic acid, and various hydroxybenzoic acids) and discovered that, for example, a resorcinol-?-d-glucosidase copolymer has high stability combined with low Km (10.5 m M vs commercial soluble (3-d-glucosidase 9.3 mM) and high Vmax values (104 ?mol ?NP mg-1H-1 vs 85 ?mol ?NP mg-1H-1). These properties are enhanced when the copolymer is complexed with bentonite clay. The kinetic constants of the resorcinol-?-d-glucosidase copolymer-bentonite complex were Km = 9.6 m M and Vmax = 73.5 ?mol ?NP mg-1H-1. Stability has been assessed against proteolysis, organic solvents, elevated temperatures, storage, and incorporation into fresh soil. A cellulase preparation from Trichoderma viride has also been copolymerized with a variety of phenolic macromolecules and displays varying degrees of stability and activity against carboxymethyl cellulose. The resorcinol ?-d-glucosidase-copolymer was immobilized on a PM10 ultrafiltration membrane (Km = 16.8 mM; Vmax = 42.4 (?mol ?NP mg-1H-1) and showed enhanced thermostability, a broader pH range for maximal activity, and could be reused without loss of activity. An ultrafiltration cell, containing the membrane-immobilized resorcinol-?-d-glucosida se copolymer, can be operated as a continuous reactor with substrate flow rates from 0.1 to 0.7 mL min-1 without decrease in product formation. © 1984 The Humana Press Inc.

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Biochemistry & Molecular Biology
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