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Effects of substrate branching characteristics on kinetics of enzymatic depolymerization of mixed linear and branched polysaccharides: II. Amylose/glycogen α‐amylolysis
Author(s) -
Park Jun T.,
Rollings James E.
Publication year - 1995
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.260460106
Subject(s) - depolymerization , chemistry , amylose , amylopectin , branching (polymer chemistry) , polysaccharide , glycogen branching enzyme , kinetics , hydrolysis , glycogen , enzymatic hydrolysis , substrate (aquarium) , organic chemistry , chromatography , stereochemistry , polymer chemistry , starch , biochemistry , glycogen phosphorylase , physics , oceanography , quantum mechanics , geology
Enzymatic depolymerization of polysaccharides with α‐amylase has been studied in mixed aqueous dimethylsulfoxide (DMSO)/water solvents. Polysaccharide substrate chemical compositions, configurational structures, and bonding pattersn are known to affect observed enzymatic reaction kinetics. The branching structures of polysaccharides and their effects on the kinetic mechanisms of depolymerization reactions via endo‐acting hydrolyzing enzyme was studied via size exclusion chromatography coupled to low angle laser light scattering (SEC/LALLS). The glycogen branching structure is a heterogeneously distributed “cluster” structure rather than a homogeneously distributed “treelike” structure. The action pattern of α‐amylase on glycogen, which is composed of highly branched clusters, as end‐products, has a “pseudo‐exo‐attack” in contrast to an expected “endoattack” as seen in the hydrolysis of amylose or amylopectin substrates. These effects of branched substrates for mixed amylose/glycogen α‐amylolysis have been predicted and demonstrated by both experimental and theoretical analysis using the kinetic model presented in this report. The “lumped” kinetic model employed, assumes that the enzyme simultaneously attacks both linear and branched substrates. In general, excellent agreement between the model predictions and the experimental observations, both qualitatively and quantitatively, was obtained. © 1995 John Wiley & Sons, Inc.