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- Title
Production of tagatose by a recombinant thermostable l-arabinose isomerase from Thermus sp. IM6501.
- Authors
Jung-Woo Kim; Young-Wan Kim; Hoe-Jin Roh; Hae-Young Kim; Jae-Ho Cha; Kwan-Hwa Park; Cheon-Seok Park
- 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.
- Subjects
ISOMERASES; THERMOPHILIC bacteria; AMINO acid sequence; MOLECULAR weights; GENETIC recombination; POLYMERASE chain reaction; NUCLEOTIDES
- Publication
Biotechnology Letters, 2003, Vol 25, Issue 12, p963
- ISSN
0141-5492
- Publication type
Article
- DOI
10.1023/A:1024069813839