Production of an oligosaccharide-specific cellobiohydrolase from the thermophilic fungus Thielavia terrestris

James Sy Keen Woon, Muhammad Mukram Mohamed Mackeen, Amirul Husna Bin Sudin, Nor Muhammad Mahadi, Rosli Md Illias, Abdul Munir Abd. Murad, Farah Diba Abu Bakar

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Objectives: 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. Conclusion: TTCBH6B is active against shorter oligosaccharides rather than polysaccharides.

Original languageEnglish
Pages (from-to)1-8
Number of pages8
JournalBiotechnology Letters
DOIs
Publication statusAccepted/In press - 1 Feb 2016

Fingerprint

Cellulose 1,4-beta-Cellobiosidase
Oligosaccharides
Fungi
Cellulose
Cellobiose
Hydrolases
Pichia
Molecular mass
Polysaccharides
Carbohydrates
Glucose
Amino acids
Hydrolysis
Catalytic Domain
Enzymes
Proteins
Amino Acids
Acids
Substrates

Keywords

  • 2-Aminobenzamide
  • Avicel
  • Carbohydrate-binding module
  • Cellobiose
  • Celluclast
  • Endoglucanases
  • Glycosyl hydrolases

ASJC Scopus subject areas

  • Biotechnology

Cite this

Production of an oligosaccharide-specific cellobiohydrolase from the thermophilic fungus Thielavia terrestris. / Woon, James Sy Keen; Mohamed Mackeen, Muhammad Mukram; Sudin, Amirul Husna Bin; Mahadi, Nor Muhammad; Illias, Rosli Md; Abd. Murad, Abdul Munir; Abu Bakar, Farah Diba.

In: Biotechnology Letters, 01.02.2016, p. 1-8.

Research output: Contribution to journalArticle

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AU - Illias, Rosli Md

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