Rheological properties of guar galactomannan solutions during hydrolysis with galactomannanase and alpha-galactosidase enzyme mixtures.
Guar galactomannan, a naturally occurring polysaccharide, is susceptible to hydrolysis by three enzymes: beta-mannosidase, beta-mannanase, and alpha-galactosidase. The beta-mannosidase cleaves a single mannose unit from the nonreducing end of the guar molecule, the beta-mannanase cleaves interior glycosidic bonds between adjacent mannose units, and the alpha-galactosidase cleaves the galactose side branches off the guar. In this study, hydrolysis of guar solutions using hyperthermopilic versions of these enzymes together in different proportions and combinations are examined. The enzymatic reactions are carried out in situ in a rheometer, and the progress of the reaction is monitored through measuring the variation in zero shear viscosity. We find the presence of alpha-galactosidase to affect the action of both beta-mannanase and beta-mannosidase with respect to solution rheology. However, this effect is more pronounced when the alpha-galactosidase and beta-mannanase or beta-mannosidase enzymes were added sequentially rather than simultaneously. This likely is the result of debranching of the guar, which facilitates attack on beta-1,4-linkages by both the beta-mannanase and the beta-mannosidase enzymes and increases hydrolytic rates by the individual enzymes. A rheology-based kinetic model is developed to estimate the reaction rate constants and interpret synergistic effects of multiple enzyme contributions. The model fits the experimental data well and reveals that both the native and the debranched guar have the same activation energy for beta-mannanase action, although debranching considerably increases the frequency of enzyme-guar interactions.
Mahammad
Shamsheer
S
Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, North Carolina 27695-7905, USA.
Comfort
Donald A
DA
Kelly
Robert M
RM
Khan
Saad A
SA
D002236:Carbohydrate Conformation; D005685:Galactans / Q000378:metabolism; D005690:Galactose / Q000031:analogs & derivatives; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007700:Kinetics; D008351:Mannans / Q000737:chemistry* / Q000378:metabolism; D008358:Mannose / Q000737:chemistry*; D008361:Mannosidases / Q000737:chemistry*; D008956:Models, Chemical; D015233:Models, Statistical; D053149:Plant Gums / Q000378:metabolism; D011134:Polysaccharides / Q000737:chemistry; D012212:Rheology / Q000379:methods*; D013997:Time Factors; D000519:alpha-Galactosidase / Q000737:chemistry*
D005685:Galactans; D008351:Mannans; D053149:Plant Gums; D011134:Polysaccharides; C012990:galactomannan; C007894:guar gum; D008361:Mannosidases; C109435:galactomannanase; D000519:alpha-Galactosidase; D008358:Mannose; D005690:Galactose