Applied Microbiology and Biotechnology

, Volume 75, Issue 4, pp 813–820 | Cite as

Purification and characterization of a highly thermostable α-l-Arabinofuranosidase from Geobacillus caldoxylolyticus TK4

  • Sabriye Canakci
  • Ali Osman Belduz
  • Badal C. Saha
  • Ahmet Yasar
  • Faik Ahmet Ayaz
  • Nurettin Yayli
Biotechnologically Relevant Enzymes and Proteins


The gene encoding an α-l-arabinofuranosidase from Geobacillus caldoxylolyticus TK4, AbfATK4, was isolated, cloned, and sequenced. The deduced protein had a molecular mass of about 58 kDa, and analysis of its amino acid sequence revealed significant homology and conservation of different catalytic residues with α-l-arabinofuranosidases belonging to family 51 of the glycoside hydrolases. A histidine tag was introduced at the N-terminal end of AbfATK4, and the recombinant protein was expressed in Escherichia coli BL21, under control of isopropyl-β-D-thiogalactopyranoside-inducible T7 promoter. The enzyme was purified by nickel affinity chromatography. The molecular mass of the native protein, as determined by gel filtration, was about 236 kDa, suggesting a homotetrameric structure. AbfATK4 was active at a broad pH range (pH 5.0–10.0) and at a broad temperature range (40–85°C), and it had an optimum pH of 6.0 and an optimum temperature of 75–80°C. The enzyme was more thermostable than previously described arabinofuranosidases and did not lose any activity after 48 h incubation at 70°C. The protein exhibited a high level of activity with p-nitrophenyl-α-l-arabinofuranoside, with apparent K m and V max values of 0.17 mM and 588.2 U/mg, respectively. AbfATK4 also exhibited a low level of activity with p-nitrophenyl-β-d-xylopyranoside, with apparent K m and V max values of 1.57 mM and 151.5 U/mg, respectively. AbfATK4 released l-arabinose only from arabinan and arabinooligosaccharides. No endoarabinanase activity was detected. These findings suggest that AbfATK4 is an exo-acting enzyme.


α-l-Arabinofuranosidase Geobacillus caldoxylolyticus Exo-acting Thermostable Xylosidase 



We are grateful to the Karadeniz Technical University Research Foundation (grant no. for financial support.


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Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Sabriye Canakci
    • 1
  • Ali Osman Belduz
    • 1
  • Badal C. Saha
    • 2
  • Ahmet Yasar
    • 3
  • Faik Ahmet Ayaz
    • 1
  • Nurettin Yayli
    • 3
  1. 1.Department of Biology, Faculty of Arts and SciencesKaradeniz Technical UniversityTrabzonTurkey
  2. 2.Fermentation Biotechnology Research Unit, National Center for Agricultural Utilization ResearchUSDA-ARSPeoriaUSA
  3. 3.Department of Chemistry, Faculty of Arts and SciencesKaradeniz Technical UniversityTrabzonTurkey

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