Gene cloning, expression, and biochemical characterization of a recombinant trehalose synthase from Picrophilus torridus in Escherichia coli

Yi Shan Chen, Guan Chiun Lee, Jei Fu Shaw

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A trehalose synthase (TSase) gene from a hyperacidophilic, thermophilic archaea, Picrophilus torridus, was synthesized using overlap extension PCR and transformed into Escherichia coli for expression. The purified recombinant P. torridus TSase (PTTS) showed an optimum pH and temperature of 6.0 and 45°C, respectively, and the enzyme maintained high activity at pH 5.0 and 60°C. Kinetic analysis showed that the enzyme has a 2.5-fold higher catalytic efficiency (kcat/KM) for maltose than for trehalose, indicating maltose as the preferred substrate. The maximum conversion rate of maltose into trehalose by the enzyme was independent of the substrate concentration, tended to increase at lower temperatures, and reached ≈71% at 20°C. Enzyme activity was inhibited by Hg2+, Al3+, and SDS. Five amino acid residues that are important for α-amylase family enzyme catalysis were shown to be conserved in PTTS (Asp203, Glu 245, Asp311, His106, and His310) and required for its activity, suggesting this enzyme might employ a similar hydrolysis mechanism.

Original languageEnglish
Pages (from-to)7098-7104
Number of pages7
JournalJournal of Agricultural and Food Chemistry
Issue number19
Publication statusPublished - 2006 Sep 20



  • High performance liquid chromatography
  • Maltose
  • Picrophilus torridus
  • Trehalose
  • Trehalose synthase

ASJC Scopus subject areas

  • Chemistry(all)
  • Agricultural and Biological Sciences(all)

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