Primary structure of O-linked carbohydrate chains in the cellulosome of different Clostridium thermocellum strains.
The cell-free forms of the multiple cellulase-containing protein complex (cellulosome), isolated from the cellulolytic bacterium Clostridium thermocellum strains YS, ATCC 27405 and LQRI, have a total carbohydrate content of 5-7% (by mass), consisting of O-linked oligosaccharide chains. The carbohydrate chains were liberated by alkaline-borohydride treatment and fractionated as oligosaccharide alditols via gel-permeation chromatography and HPLC. The fractions were investigated by 500-MHz 1H-NMR spectroscopy in combination with monosaccharide and methylation analysis and with fast-atom-bombardment mass spectrometry (FAB-MS). In addition to the previously described major oligosaccharide, (formula; see text) [Gerwig, G. J., de Waard, P., Kamerling, J. P., Vliegenthart, J. F. G., Morgenstern, E., Lamed, R. & Bayer, E. A. (1989) J. Biol. Chem. 264, 1027-1035], the following partial structures of this compound could be established: (formula; see text). Cell-free and cell-associated forms of the cellulosome of C. thermocellum, as determined for strain YS, have the same oligosaccharide pattern. Based on the oligosaccharide structures, a biosynthetic pathway is suggested.
Gerwig
G J
GJ
Bijvoet Center, Department of Bio-Organic Chemistry, Utrecht University, The Netherlands.
Kamerling
J P
JP
Vliegenthart
J F
JF
Morag
E
E
Lamed
R
R
Bayer
E A
EA
D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry*; D002480:Cellulase / Q000737:chemistry*; D003013:Clostridium / Q000032:analysis*; D006023:Glycoproteins / Q000096:biosynthesis / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy
D001426:Bacterial Proteins; D006023:Glycoproteins; D002480:Cellulase