The stereochemical course of reactions catalysed by the cellobiohydrolases produced by Talaromyces emersonii.
Three forms of exocellobiohydrolase (EC 3.2.1.91), CBH IA, CBH IB and CBH II, were isolated to apparent homogeneity from culture filtrates of the aerobic fungus Talaromyces emersonii. CBH IA and CBH II appear to be native forms of these enzymes, while CBH IB may represent a proteolytic degradation product of the CBH IA enzyme. The hydrolysis of beta-cellobiosyl fluoride by each form was monitored by 1H-n.m.r. spectroscopy. The reactions catalysed by CBH IA and CBH IB proceed with retention of the anomeric configuration, whereas that catalysed by CBH II is one of inversion. Thus one may deduce that CBH IA (or CBH IB) and CBH II operate double and single displacement reactions respectively during catalysis of substrate. On the basis of these findings and the observed substrate specificities of the various forms, one may conclude that CBH IA (and CBH IB) is a family C enzyme, while CBH II belongs to family B [Henrissat, Claeyssens, Tomme, Lemesle & Mornon (1989) Gene 81, 83-95].
Brooks
M M
MM
Department of Chemistry, University College, Galway, Ireland.
Tuohy
M G
MG
Savage
A V
AV
Claeyssens
M
M
Coughlan
M P
MP
D001203:Ascomycota / Q000201:enzymology*; D050260:Carbohydrate Metabolism; D002475:Cellobiose / Q000031:analogs & derivatives / Q000378:metabolism; D043366:Cellulose 1,4-beta-Cellobiosidase; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005656:Fungal Proteins / Q000302:isolation & purification / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000302:isolation & purification / Q000378:metabolism*; D006868:Hydrolysis; D007525:Isoelectric Focusing; D009682:Magnetic Resonance Spectroscopy; D013237:Stereoisomerism
D005656:Fungal Proteins; C074099:cellobiosyl fluoride; D002475:Cellobiose; D006026:Glycoside Hydrolases; D043366:Cellulose 1,4-beta-Cellobiosidase