Synergistic interactions among beta-laminarinase, beta-1,4-glucanase, and beta-glucosidase from the hyperthermophilic archaeon Pyrococcus furiosus during hydrolysis of beta-1,4-, beta-1,3-, and mixed-linked polysaccharides.
The synergistic interaction among three beta-specific glycosidases from the hyperthermophilic archaeon Pyrococcus furiosus, namely two endoglucanases (EglA and LamA) and an exo-acting beta-glucosidase (Bgl), on barley-glucan and laminarin, was examined. In addition to following glucose release and the generation of reducing sugar ends, the distribution and amounts of oligomeric products from beta-1,3- and beta-1,4-linked substrates were determined as a function of extent of hydrolysis at 98 degrees C. Positive interactions were noted between endo/exo glucanase combinations, leading to enhanced and rapid degradation of the larger complex carbohydrates to oligosaccharides. The EglA/LamA endo-acting combination was also synergistic in degrading barley-glucan. However, hydrolysis was most efficient when a blend of all three hydrolases was used, possibly due to the relief of product inhibition by the exoglyosidase. Furthermore, by monitoring the distribution of oligosaccharides present during hydrolysis, patterns of enzymatic attack could be followed in addition to determining the specific contributions of each hydrolase to the overall process.
Driskill
L E
LE
Department of Chemical Engineering, North Carolina State University, Box 7905, Raleigh, North Carolina 27695, USA.
Bauer
M W
MW
Kelly
R M
RM
D001426:Bacterial Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002236:Carbohydrate Conformation; D004187:Disaccharides / Q000378:metabolism; D004357:Drug Synergism; D044643:Glucan 1,4-beta-Glucosidase; D004693:Glucan Endo-1,3-beta-D-Glucosidase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D005936:Glucans / Q000378:metabolism; D001467:Hordeum; D006868:Hydrolysis; D009844:Oligosaccharides / Q000494:pharmacology; D011134:Polysaccharides / Q000378:metabolism; D011135:Polysaccharides, Bacterial / Q000378:metabolism*; D020027:Pyrococcus furiosus / Q000201:enzymology*; D013379:Substrate Specificity; D013696:Temperature; D001617:beta-Glucosidase / Q000378:metabolism*
D001426:Bacterial Proteins; D004187:Disaccharides; D005936:Glucans; D009844:Oligosaccharides; D011134:Polysaccharides; D011135:Polysaccharides, Bacterial; C014637:laminaribiose; C008247:laminaran; D001617:beta-Glucosidase; D004693:Glucan Endo-1,3-beta-D-Glucosidase; D044643:Glucan 1,4-beta-Glucosidase