Cloning, Production and Characterization of a Glycoside Hydrolase Family 7 Enzyme from the Gut Microbiota of the Termite Coptotermes curvignathus

James Sy Keen Woon, Patricia Jie Hung King, Muhammad Mukram Mohamed Mackeen, Nor Muhammad Mahadi, Wan Mohd Khairulikhsan Wan Seman, William J. Broughton, Abdul Munir Abd. Murad, Farah Diba Abu Bakar

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Coptotermes curvignathus is a termite that, owing to its ability to digest living trees, serves as a gold mine for robust industrial enzymes. This unique characteristic reflects the presence of very efficient hydrolytic enzyme systems including cellulases. Transcriptomic analyses of the gut of C. curvignathus revealed that carbohydrate-active enzymes (CAZy) were encoded by 3254 transcripts and that included 69 transcripts encoding glycoside hydrolase family 7 (GHF7) enzymes. Since GHF7 enzymes are useful to the biomass conversion industry, a gene encoding for a GHF7 enzyme (Gh1254) was synthesized, sub-cloned and expressed in the methylotrophic yeast Pichia pastoris. Expressed GH1254 had an apparent molecular mass of 42 kDa, but purification was hampered by its low expression levels in shaken flasks. To obtain more of the enzyme, GH1254 was produced in a bioreactor that resulted in a fourfold increase in crude enzyme levels. The purified enzyme was active towards soluble synthetic substrates such as 4-methylumbelliferyl-β-d-cellobioside, 4-nitrophenyl-β-d-cellobioside and 4-nitrophenyl-β-d-lactoside but was non-hydrolytic towards Avicel or carboxymethyl cellulose. GH1254 catalyzed optimally at 35 °C and maintained 70% of its activity at 25 °C. This enzyme is thus potentially useful in food industries employing low-temperature conditions.

Original languageEnglish
Pages (from-to)1-13
Number of pages13
JournalMolecular Biotechnology
DOIs
Publication statusAccepted/In press - 1 Jun 2017

Fingerprint

Isoptera
Cloning
Glycoside Hydrolases
Organism Cloning
Enzymes
Gastrointestinal Microbiome
Cellulases
Carboxymethylcellulose Sodium
Pichia
Food Industry
Gold mines
Gene encoding
Bioreactors
Molecular mass
Cellulose
Gold
Biomass
Carbohydrates
Yeast
Industry

Keywords

  • Avicel
  • Biomass
  • Bioreactor
  • Cellulase
  • Saccharification
  • Symbiont

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology

Cite this

Cloning, Production and Characterization of a Glycoside Hydrolase Family 7 Enzyme from the Gut Microbiota of the Termite Coptotermes curvignathus. / Woon, James Sy Keen; King, Patricia Jie Hung; Mohamed Mackeen, Muhammad Mukram; Mahadi, Nor Muhammad; Wan Seman, Wan Mohd Khairulikhsan; Broughton, William J.; Abd. Murad, Abdul Munir; Abu Bakar, Farah Diba.

In: Molecular Biotechnology, 01.06.2017, p. 1-13.

Research output: Contribution to journalArticle

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AU - Mahadi, Nor Muhammad

AU - Wan Seman, Wan Mohd Khairulikhsan

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AU - Abu Bakar, Farah Diba

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