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Biotechnology Letters

, Volume 25, Issue 12, pp 963–967 | Cite as

Production of tagatose by a recombinant thermostable l-arabinose isomerase from Thermus sp. IM6501

  • Jung-Woo Kim
  • Young-Wan Kim
  • Hoe-Jin Roh
  • Hae-Young Kim
  • Jae-Ho Cha
  • Kwan-Hwa Park
  • Cheon-Seok ParkEmail author
Article

Abstract

A gene (thaI) corresponding to l-arabinose isomerase from Thermus strain IM6501 was cloned by PCR. It comprised 1488 nucleotides and encoded a polypeptide of 496 residues with a predicted molecular weight of 56019 Da. The deduced amino acid sequence had 96.8% identity with the l-arabinose isomerase of Geobacillus stearothermophilus. Recombinant ThaI with N-terminal hexa-tistidine tags was over-expressed in Escherichia coli and purified by affinity chromatography using Ni-NTA resin. The purified ThaI was thermostable with maximal activity at 60 °C at pH 8 for 30 min of reaction. Zn2+ and Ni2+ inactivated the catalytic activity of ThaI, 5 mM Mn2+ enhanced the bioconversion yield by 90%. The bioconversion yield of 54% from d-galactose to d-tagatose was obtained by recombinant ThaI at 60 °C over 3 d.

arabinose isomerase galactose isomerization tagatose Thermus 

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Copyright information

© Kluwer Academic Publishers 2003

Authors and Affiliations

  • Jung-Woo Kim
    • 1
  • Young-Wan Kim
    • 1
  • Hoe-Jin Roh
    • 2
  • Hae-Young Kim
    • 3
  • Jae-Ho Cha
    • 4
  • Kwan-Hwa Park
    • 1
  • Cheon-Seok Park
    • 3
    Email author
  1. 1.National Laboratory for Functional Food Carbohydrate, and Center for Agricultural Biomaterials, Department of Food Science and TechnologySeoul National UniversitySuwonKorea
  2. 2.R & D CenterTong Yang Confectionery Co.SeoulKorea
  3. 3.Department of Food Science & TechnologyKyunghee UniversityYounginKorea
  4. 4.Department of MicrobiologyPusan National UniversityPusanKorea

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