title: Kinetic parameters and mode of action of the cellobiohydrolases produced by Talaromyces emersonii.
abstract: Three forms of cellobiohydrolase (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. The three enzymes are single sub-unit glycoproteins, and unlike most other fungal cellobiohydrolases are characterised by noteworthy thermostability. The kinetic properties and mode of action of each enzyme against polymeric and small soluble oligomeric substrates were investigated in detail. CBH IA, CBH IB and CBH II catalyse the hydrolysis of microcrystalline cellulose, albeit to varying extents. Hydrolysis of a soluble cellulose derivative (CMC) and barley 1,3;1,4-beta-D-glucan was not observed. Cellobiose (G2) is the main reaction product released by CBH IA, CBH IB, and CBH II from microcrystalline cellulose. All three CBHs are competitively inhibited by G2; inhibition constant values (K(i)) of 2.5 and 0.18 mM were obtained for CBH IA and CBH IB, respectively (4-nitrophenyl-beta-cellobioside as substrate), while a K(i) of 0.16 mM was determined for CBH II (2-chloro-4-nitrophenyl-beta-cellotrioside as substrate). Bond cleavage patterns were determined for each CBH on 4-methylumbelliferyl derivatives of beta-cellobioside and beta-cellotrioside (MeUmbG(n)). While the Tal. emersonii CBHs share certain properties with their counterparts from Trichoderma reesei, Humicola insolens and other fungal sources, distinct differences were noted.
authors: Department of Biochemistry, National University of Ireland, Galway, Ireland. maria.tuohy@nuigalway.ie
authors: Maria G
authors: MG
authors: Tuohy
authors: Daniel J
authors: DJ
authors: Walsh
authors: Patrick G
authors: PG
authors: Murray
authors: Marc
authors: M
authors: Claeyssens
authors: Michelle M
authors: MM
authors: Cuffe
authors: Angela V
authors: AV
authors: Savage
authors: Michael P
authors: MP
authors: Coughlan
mesh_terms: D001667:Binding, Competitive; D002384:Catalysis; D002475:Cellobiose / Q000494:pharmacology; D002480:Cellulase / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000302:isolation & purification; D043366:Cellulose 1,4-beta-Cellobiosidase; D004795:Enzyme Stability; D005658:Fungi / Q000201:enzymology*; D006027:Glycosides / Q000737:chemistry; D007700:Kinetics; D013379:Substrate Specificity
chemical_list: D006027:Glycosides; D002475:Cellobiose; D002480:Cellulase; D043366:Cellulose 1,4-beta-Cellobiosidase