The endo-1,4-beta-glucanase I from Trichoderma reesei. Action on beta-1,4-oligomers and polymers derived from D-glucose and D-xylose.
The reaction mechanism of the non-specific endo-1,4-beta-glucanase from Trichoderma reesei QM 9414 (endoglucanase I) was investigated using both reducing-end3H-labelled and universally 14C-labelled cellooligosaccharides, as well as reducing-end3H-labelled xylooligosaccharides. The bond cleavage frequencies of cellooligosaccharides proved to be dependent upon the substrate concentration, especially in the case of cellotriose. In addition to simple hydrolytic cleavage, the enzyme catalyzes reactions along alternative pathways, including transglycosylations leading to products larger than the substrate. Some of these pathways were shown to be reversible. During cellotriose or cellopentaose degradation, substrate resynthesis was demonstrated by incorporation of added radioactive D-glucose or cellobiose. The endoglucanase I is active on xylan and xylooligosaccharides, but less than on soluble cellulose derivatives (e.g. hydroxyethylcellulose) and cellooligosaccharides. The fact that for these different types of substrates the same active site is operative is proven by the ability of the enzyme to utilize cellooligosaccharides and xylooligosaccharides as both glycosyl donors and acceptors. The mixed substrate reactions lead to products composed of D-glucosyl and D-xylosyl residues. The kinetic parameters for cellooligosaccharide degradation can be used for the description of an extended substrate binding site. Of the four putative glycosyl subsites, -II and +II show the highest affinities, 16.7 kJ.mol-1 and 7.1 kJ.mol-1, respectively.
Biely
P
P
Institute of Chemistry, Slovak Academy of Sciences, Bratislava, Czechoslovakia.
Vrsanská
M
M
Claeyssens
M
M
D002480:Cellulase / Q000737:chemistry* / Q000378:metabolism; D043364:Endo-1,4-beta Xylanases; D005947:Glucose / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000378:metabolism; D009844:Oligosaccharides / Q000378:metabolism*; D013379:Substrate Specificity; D014242:Trichoderma / Q000201:enzymology*; D014994:Xylose / Q000378:metabolism*
D009844:Oligosaccharides; D014994:Xylose; D006026:Glycoside Hydrolases; D002480:Cellulase; D043364:Endo-1,4-beta Xylanases; D005947:Glucose