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The hyperthermophilic α-amylase from Thermococcus sp. HJ21 does not require exogenous calcium for thermostability because of high-binding affinity to calcium

  • Microbial Physiology and Biochemistry
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Abstract

The hyperthermophilic α-amylase from Thermococcus sp. HJ21 does not require exogenous calcium ions for thermostability, and is a promising alternative to commercially available α-amylases to increase the efficiency of industrial processes like the liquefaction of starch. We analyzed the amino acid sequence of this α-amylase by sequence alignments and structural modeling, and found that this α-amylase closely resembles the α-amylase from Pyrococcus woesei. The gene of this α-amylase was cloned in Escherichia coli and the recombinant α-amylase was overexpressed and purified with a combined renaturation-purification procedure. We confirmed thermostability and exogenous calcium ion independency of the recombinant α-amylase and further investigated the mechanism of the independency using biochemical approaches. The results suggested that the α-amylase has a high calcium ion binding affinity that traps a calcium ion that would not dissociate at high temperatures, providing a direct explanation as to why the addition of calcium ions is not required for thermostability. Understanding of the mechanism offers a strong base on which to further engineer properties of this α-amylase for better potential applications in industrial processes.

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Authors and Affiliations

  1. Jiangsu Key Laboratory of Marine Pharmaceutical Compound Screening, Huaihai Institute of Technology, Lianyungang, 222005, P. R. China

    Huaixu Cheng, Zhidan Luo, Mingsheng Lu, Song Gao & Shujun Wang

  2. Jiangsu Marine Resources Development Research Institute, Lianyungang, 222005, P. R. China

    Huaixu Cheng, Mingsheng Lu, Song Gao & Shujun Wang

  3. Co-Innovation Center of Jiangsu Marine Bio-industry Technology, Huaihai Institute of Technology, Lianyungang, 222005, P. R. China

    Huaixu Cheng, Zhidan Luo, Mingsheng Lu, Song Gao & Shujun Wang

  4. Lianyungang City Academy of Agricultural Sciences, Lianyungang, 222001, P. R. China

    Zhidan Luo

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  1. Huaixu Cheng
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  2. Zhidan Luo
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  3. Mingsheng Lu
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Correspondence to Song Gao or Shujun Wang.

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Cheng, H., Luo, Z., Lu, M. et al. The hyperthermophilic α-amylase from Thermococcus sp. HJ21 does not require exogenous calcium for thermostability because of high-binding affinity to calcium. J Microbiol. 55, 379–387 (2017). https://doi.org/10.1007/s12275-017-6416-5

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  • DOI: https://doi.org/10.1007/s12275-017-6416-5

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