Polarimetry and 13C n.m.r. show that the hydrolyses of beta-D-glucopyranosyl fluoride by beta(1-->3)-glucanases from Phanerochaete chrysosporium and Sporotrichum dimorphosporum have opposite stereochemistries.
The time courses of optical rotation and fluoride ion release during hydrolysis of beta-D-glucopyranosyl fluoride by the beta(1-->3)-glucanase of Phanerochaete chrysosporium (J. L. Copa-Patiño and P. Broda, unpublished work) indicated that the initial sugar product was beta-D-glucopyranose. This was confirmed by monitoring the hydrolysis of 1-[13C]beta-D-glucopyranosyl fluoride by this enzyme with 13C n.m.r. (without proton decoupling). The same two techniques were used to confirm that hydrolysis of beta-D-glucopyranosyl fluoride by the exo beta(-->3)-glucanase of 'Basidiomycete QM 806' (identified as Sporotrichum dimorphosporum) yielded alpha-glucopyranose as first sugar product, in accordance with previous results using laminarin as substrate [Parrish and Reese (1963) Carbohydr. Res. 3, 424-429; Nelson (1970) J. Biol. Chem. 245, 869-872].
Copa-Patiño
J L
JL
Department of Biochemistry and Applied Molecular Biology, University of Manchester Institute of Science and Technology, U.K.
Zhang
Y
Y
Padmaperuma
B
B
Marsden
I
I
Broda
P
P
Sinnott
M L
ML
D001487:Basidiomycota / Q000201:enzymology*; D002247:Carbon Isotopes; D004693:Glucan Endo-1,3-beta-D-Glucosidase / Q000378:metabolism*; D005947:Glucose / Q000031:analogs & derivatives* / Q000378:metabolism; D006868:Hydrolysis; D009682:Magnetic Resonance Spectroscopy; D009904:Optical Rotation; D013057:Spectrum Analysis; D013175:Sporothrix / Q000201:enzymology*; D013237:Stereoisomerism
D002247:Carbon Isotopes; C030364:glucosyl fluoride; D004693:Glucan Endo-1,3-beta-D-Glucosidase; D005947:Glucose