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Identification of archaeon-producing hyperthermophilic α-amylase and characterization of the α-amylase

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Abstract

The extremely thermophilic anaerobic archaeon strain, HJ21, was isolated from a deep-sea hydrothermal vent, could produce hyperthermophilic α-amylase, and later was identified as Thermococcus from morphological, biochemical, and physiological characteristics and the 16S ribosomal RNA gene sequence. The extracellular thermostable α-amylase produced by strain HJ21 exhibited maximal activity at pH 5.0. The enzyme was stable in a broad pH range from pH 5.0 to 9.0. The optimal temperature of α-amylase was observed at 95°C. The half-life of the enzyme was 5 h at 90°C. Over 40% and 30% of the enzyme activity remained after incubation at 100°C for 2 and 3 h, respectively. The enzyme did not require Ca2+ for thermostability. This α-amylase gene was cloned, and its nucleotide sequence displayed an open reading frame of 1,374 bp, which encodes a protein of 457 amino acids. Analysis of the deduced amino acid sequence revealed that four homologous regions common in amylases were conserved in the HJ21 α-amylase. The molecular weight of the mature enzyme was calculated to be 51.4 kDa, which correlated well with the size of the purified enzyme as shown by the sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

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Acknowledgements

The authors would like to thank Mr. Fuchao Li, Ms. Li Chen, and Mr. Jinyu Yang for their technical assistance. This work was financially supported by National Natural Science Foundation of China (40746030), Natural Science Foundation of the Education Department of Jiangsu Province (06KJB550004, 06-A-017), Jiangsu Key Laboratory of Marine Biotechnology (2006HS008), and Science and Technology Department of Lianyungang City (KK06076).

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Correspondence to Zhaoxin Lu.

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Wang, S., Lu, Z., Lu, M. et al. Identification of archaeon-producing hyperthermophilic α-amylase and characterization of the α-amylase. Appl Microbiol Biotechnol 80, 605–614 (2008). https://doi.org/10.1007/s00253-008-1561-8

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Keywords

  • Hyperthermophilic α-amylase
  • Thermococcus sp. Thermostability
  • Archaeon
  • Identification
  • Characterization