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

被引：0
作者：
Jung-Woo Kim
Young-Wan Kim
Hoe-Jin Roh
Hae-Young Kim
Jae-Ho Cha
Kwan-Hwa Park
Cheon-Seok Park
机构：
[1] Seoul National University,National Laboratory for Functional Food Carbohydrate, and Center for Agricultural Biomaterials, Department of Food Science and Technology
[2] Tong Yang Confectionery Co.,R & D Center
[3] Kyunghee University,Department of Food Science & Technology
[4] Pusan National University,Department of Microbiology
来源：
Biotechnology Letters | 2003年 / 25卷
关键词：
arabinose isomerase; galactose; isomerization; tagatose;
D O I：
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摘要：
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.
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页码：963 / 967
页数：4
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