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Identification and characterization of a novel l-arabinose isomerase from Anoxybacillus flavithermus useful in d-tagatose production

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Abstract

d-Tagatose is a highly functional rare ketohexose and many attempts have been made to convert d-galactose into the valuable d-tagatose using l-arabinose isomerase (l-AI). In this study, a thermophilic strain possessing l-AI gene was isolated from hot spring sludge and identified as Anoxybacillus flavithermus based on its physio-biochemical characterization and phylogenetic analysis of its 16s rRNA gene. Furthermore, the gene encoding l-AI from A. flavithermus (AFAI) was cloned and expressed at a high level in E. coli BL21(DE3). l-AI had a molecular weight of 55,876 Da, an optimum pH of 10.5 and temperature of 95°C. The results showed that the conversion equilibrium shifted to more d-tagatose from d-galactose by raising the reaction temperatures and adding borate. A 60% conversion of d-galactose to d-tagatose was observed at an isomerization temperature of 95°C with borate. The catalytic efficiency (kcat/Km) for d-galactose with borate was 9.47 mM−1 min−1, twice as much as that without borate. Our results indicate that AFAI is a novel hyperthermophilic and alkaliphilic isomerase with a higher catalytic efficiency for d-galactose, suggesting its great potential for producing d-tagatose.

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Acknowledgments

This work was supported by Science and Technology Projects of Tianjin (10YFYBJC00100), the grant from Chinese Academy of Sciences (KSCX2EWG5) and Visiting Professorships for Senior International Scientists (2010T1S4).

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Authors and Affiliations

  1. College of Food Engineering and Biotechnology, Tianjin University of Science and Technology, Tianjin, 300457, China

    Yanjun Li & Anjun Liu

  2. Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, China

    Yueming Zhu & Yuanxia Sun

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  1. Yanjun Li
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  2. Yueming Zhu
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  3. Anjun Liu
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  4. Yuanxia Sun
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Corresponding author

Correspondence to Yuanxia Sun.

Additional information

Communicated by T. Matsunaga.

The authors Y. Li and Y. Zhu contributed equally to this work.

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Li, Y., Zhu, Y., Liu, A. et al. Identification and characterization of a novel l-arabinose isomerase from Anoxybacillus flavithermus useful in d-tagatose production. Extremophiles 15, 441–450 (2011). https://doi.org/10.1007/s00792-011-0375-2

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  • DOI: https://doi.org/10.1007/s00792-011-0375-2

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