Mode of action of exoglucanases from the cellulolytic fungus Trichoderma reesei: activity on reducing end-labeled cellooligosaccharides and topography of active sites of. beta. -glucosidase, cellobiohydrolase I(D), and cellobiohydrolase II
Thesis/Dissertation
·
OSTI ID:5941959
The activity of purified ..beta..-glucosidase, cellobiohydrolase I(D) and cellobiohydrolase II from Trichoderma reesei was studied using (1-/sup 3/H)cellooligosaccharides. Reducing end-labeled cellooligosaccharides of high specific radioactivity were synthesized by reducing lactones of cellooligosaccharide aldonic acids with sodium boro(/sup 3/He)hydride. A thin-layer chromatographic method for separating (1-/sup 3/H)cellooligosaccharides with high resolution and quantitatively extracting them from silica gel was developed. The ..beta..-glucosidase was shown to bind (1-/sup 3/H)cellooligosaccharides in one productive mode and sequentially remove glucosyl residues from the nonreducing end. Cellobiohydrolase I(D) was shown to bind (1-/sup 3/H)cellooligosaccharides in more than one productive mode. Cellobiohydrolase II was also shown to bind (1-3H) cellooligosaccharides in more than one productive model; however, glycosidic bonds at the nonreducing end are preferentially hydrolyzed. The results are discussed in terms of the individual roles of exoglucanases during the saccharification of cellulose. A mechanism by which cellobiohydrolase I(D) and cellobiohydrolase II may synergistically degrade crystalline cellulose is proposed.
- Research Organization:
- Florida Univ., Gainesville (USA)
- OSTI ID:
- 5941959
- Country of Publication:
- United States
- Language:
- English
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OSTI ID:6910310
Related Subjects
550201* -- Biochemistry-- Tracer Techniques
550701 -- Microbiology-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
BIOCHEMICAL REACTION KINETICS
BIOCHEMISTRY
CARBOHYDRATES
CARBON 14 COMPOUNDS
CELLOBIOSE
CELLULOLYTIC ACTIVITY
CELLULOSE
CHEMICAL REACTIONS
CHEMISTRY
DECOMPOSITION
DISACCHARIDES
ENZYME ACTIVITY
ENZYMES
FUNGI
GLUCOSIDASE
GLYCOSYL HYDROLASES
HYDROLASES
HYDROLYSIS
KINETICS
LABELLED COMPOUNDS
LYSIS
O-GLYCOSYL HYDROLASES
OLIGOSACCHARIDES
ORGANIC COMPOUNDS
PLANTS
POLYSACCHARIDES
REACTION KINETICS
SACCHARIDES
SACCHARIFICATION
SOLVOLYSIS
TRICHODERMA
TRITIUM COMPOUNDS
550701 -- Microbiology-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
BIOCHEMICAL REACTION KINETICS
BIOCHEMISTRY
CARBOHYDRATES
CARBON 14 COMPOUNDS
CELLOBIOSE
CELLULOLYTIC ACTIVITY
CELLULOSE
CHEMICAL REACTIONS
CHEMISTRY
DECOMPOSITION
DISACCHARIDES
ENZYME ACTIVITY
ENZYMES
FUNGI
GLUCOSIDASE
GLYCOSYL HYDROLASES
HYDROLASES
HYDROLYSIS
KINETICS
LABELLED COMPOUNDS
LYSIS
O-GLYCOSYL HYDROLASES
OLIGOSACCHARIDES
ORGANIC COMPOUNDS
PLANTS
POLYSACCHARIDES
REACTION KINETICS
SACCHARIDES
SACCHARIFICATION
SOLVOLYSIS
TRICHODERMA
TRITIUM COMPOUNDS