title: Production of an oligosaccharide-specific cellobiohydrolase from the thermophilic fungus Thielavia terrestris.
abstract: OBJECTIVE
  To express and determine the hydrolytic activity of a cellobiohydrolase (TTCBH6B) from the thermophilic fungus Thielavia terrestris in Pichia pastoris.
  
  
  RESULTS
  Ttcbh6B encodes a protein of 507 amino acid residues with a predicted molecular mass of 54 kDa. TTCBH6B contains a familial 6-glycosyl hydrolase catalytic domain and a type I carbohydrate-binding module. TTCBH6B was expressed and purified to homogeneity but the purified enzyme was inactive against Avicel. It could, however, digest Celluclast-treated Avicel producing cellobiose (0.27 μmol min(-1) mg(-1)). To determine the substrate preferences of TTCBH6B, oligosaccharides of varying numbers of subunits were generated by acid hydrolysis of Avicel and fluorescently tagged. Peaks corresponding to oligosaccharides containing three to six glucose units were reduced to cellobiose after addition of TTCBH6B.
  
  
  CONCLUSIONS
  TTCBH6B is active against shorter oligosaccharides rather than polysaccharides.
authors.affiliation: School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM, Bangi, Selangor, Malaysia.
authors.forename: James Sy-Keen
authors.initials: JS
authors.lastname: Woon
authors.affiliation: School of Chemical Sciences and Food Technology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM, Bangi, Selangor, Malaysia.|Institute of Systems Biology (INBIOSIS), Universiti Kebangsaan Malaysia, 43600 UKM, Bangi, Selangor, Malaysia.
authors.forename: Mukram Mohamed
authors.initials: MM
authors.lastname: Mackeen
authors.affiliation: School of Chemical Sciences and Food Technology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM, Bangi, Selangor, Malaysia.
authors.forename: Amirul Husna Bin
authors.initials: AH
authors.lastname: Sudin
authors.affiliation: Institute of Systems Biology (INBIOSIS), Universiti Kebangsaan Malaysia, 43600 UKM, Bangi, Selangor, Malaysia.|Malaysia Genome Institute, Jalan Bangi Lama, 43000, Kajang, Selangor, Malaysia.
authors.forename: Nor Muhammad
authors.initials: NM
authors.lastname: Mahadi
authors.affiliation: Department of Bioprocess Engineering, Faculty of Chemical Engineering, Universiti Teknologi Malaysia, 81310, Skudai, Johor, Malaysia.
authors.forename: Rosli Md
authors.initials: RM
authors.lastname: Illias
authors.affiliation: School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM, Bangi, Selangor, Malaysia.
authors.forename: Abdul Munir Abdul
authors.initials: AM
authors.lastname: Murad
authors.affiliation: School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM, Bangi, Selangor, Malaysia. fabyff@ukm.edu.my.
authors.forename: Farah Diba Abu
authors.initials: FD
authors.lastname: Bakar
mesh_terms: D002475:Cellobiose / Q000378:metabolism; D043366:Cellulose 1,4-beta-Cellobiosidase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D008970:Molecular Weight; D009844:Oligosaccharides / Q000378:metabolism*; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D020037:Sordariales / Q000201:enzymology* / Q000235:genetics; D013379:Substrate Specificity
chemical_list: D009844:Oligosaccharides; D011994:Recombinant Proteins; D002475:Cellobiose; D043366:Cellulose 1,4-beta-Cellobiosidase
keywords: 2-Aminobenzamide; Avicel; Carbohydrate-binding module; Cellobiose; Celluclast; Endoglucanases; Glycosyl hydrolases