Structural characterization of carbohydrate attached to the glycoprotein cellulase enzymes of Trichoderma reesei QM 9414

Thesis/Dissertation · Sun Jan 01 00:00:00 EST 1984

OSTI ID:6195066

de Chasteigner du Mee, C.P.R.

Cellobiohydrolases I(D) and II were purified by ion exchange chromatography from an extracellular culture filtrate of Tricoderma reesei QM 9414. Neutral sugar composition of each was determined by gas-liquid chromatographic analysis of the peracetylated alditol and aldononitrile acetate derivatives of sugars released by either reductive US -elimination or acid hydrolysis. The alkaline borohydride-released carbohydrate was analyzed by high-pressure liquid chromatography (HPLC). The HPLC data demonstrated that each molecule of cellobiohydrolase I(D) contained 5.9% carbohydrate and that cellobiohydrolase II contained 18.9% carbohydrate. The purified oligosaccharides were shown by methylation analysis to contain (1-2) and (1-6) glycosidic linkages and the position of the 6-substituted residues was confirmed by acetolysis. The sequence and anomeric nature of the sugar residues in each oligosaccharide was determined by sequential glycosidase digestion, and all the residues were found to be -linked. Proton decoupled TC-NMR analysis suggested that for cellobiohydrolase I(D), each oligosaccharide was attached to a threonyl residue on the polypeptide and for cellobiohydrolase II, each oligosaccharide was attached to threonyl and seryl residues on the polypeptides. Coupled TC-NMR and H-NMR data to define the structures of the oligosaccharides attached to the enzyme were determined and are reported. Thus, work with the cellobiohydrolases and the endoglucanases indicated that the predominant cellulase enzymes secreted by T. reesei QM 9414 are each glycosylated with similar oligosaccharides.

• Research Organization: Florida Univ., Gainesville (USA)
• OSTI ID: 6195066
• Resource Relation: Other Information: Thesis (Ph. D.)
• Country of Publication: United States
• Language: English

Related Subjects

37 INORGANIC
ORGANIC
PHYSICAL AND ANALYTICAL CHEMISTRY
CELLULASE
MOLECULAR STRUCTURE
GLUCOPROTEINS
OLIGOSACCHARIDES
CARBON 13
ION EXCHANGE CHROMATOGRAPHY
NUCLEAR MAGNETIC RESONANCE
PROTONS
TRICHODERMA VIRIDE
BARYONS
CARBOHYDRATES
CARBON ISOTOPES
CHROMATOGRAPHY
ELEMENTARY PARTICLES
ENZYMES
EVEN-ODD NUCLEI
FERMIONS
FUNGI
GLYCOSYL HYDROLASES
HADRONS
HYDROLASES
ISOTOPES
LIGHT NUCLEI
MAGNETIC RESONANCE
NUCLEI
NUCLEONS
O-GLYCOSYL HYDROLASES
ORGANIC COMPOUNDS
PLANTS
PROTEINS
RESONANCE
SACCHARIDES
SEPARATION PROCESSES
STABLE ISOTOPES
TRICHODERMA
400301* - Organic Chemistry- Chemical & Physicochemical Properties- (-1987)
400105 - Separation Procedures