An Avicel-affinity site in an Avicel-digesting exocellulase from a Trichoderma viride mutant.
A single form of exo-type cellulase (Exo I; MW, 65,000), purified from a Trichoderma viride protease-depressed mutant, HK-75, digested Avicel to cellobiose exowise, and hydrolyzed cellotriose, cellotetraose, and cellopentaose in the strict manner of splitting off by cellobiose units. Exo I, however, hydrolyzed cellohexaose by both cellobiose and cellotriose units. Exo I was proteolyzed by papain into two fragments; GPExo (MW, 9,000) and Exo I' (MW, 56,000). The GPExo intensively adsorbed onto Avicel but did not hydrolyze it. Exo I' had nearly identical activity to that of intact Exo I toward cellooligosaccharides but was almost inert to Avicel in digestion and adsorption. Sequence analysis of N-terminal and C-terminal amino acids showed that GPExo was between Gly435 and Leu496 and Exo I' between Glu1 and Gly434 in Exo I. Exo I therefore consists of two domains, one for adsorption to Avicel, as demonstrated by the Avicel-affinity site, GPExo and the other for the cleavage of glycosidic linkages as demonstrated in Exo I'.
Department of Agricultural Chemistry, Kyushu University, Fukuoka, Japan.
M
M
Goto
K
K
Furukawa
S
S
Hayashida
D000327:Adsorption; D000595:Amino Acid Sequence; D001665:Binding Sites; D002241:Carbohydrates / Q000032:analysis; D002480:Cellulase; D002482:Cellulose / Q000378:metabolism* / Q000493:pharmacokinetics; D043366:Cellulose 1,4-beta-Cellobiosidase; D010450:Endopeptidases / Q000494:pharmacology; D005656:Fungal Proteins / Q000302:isolation & purification / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009154:Mutation; D010446:Peptide Fragments / Q000378:metabolism; D013696:Temperature; D014242:Trichoderma / Q000201:enzymology* / Q000235:genetics; D014783:Viscosity
D002241:Carbohydrates; D005656:Fungal Proteins; D010446:Peptide Fragments; D002482:Cellulose; D006026:Glycoside Hydrolases; D002480:Cellulase; C019826:carboxymethylcellulase; D043366:Cellulose 1,4-beta-Cellobiosidase; D010450:Endopeptidases