Overexpression of the genes from thermophiles in Escherichia coli by high-temperature cultivation

Daisuke Koma, Toshiya Sawai, Shigeaki Harayama, Kuniki Kino

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15 Citations (Scopus)

Abstract

Twenty-nine aminotransferase genes from Pyrococcus horikoshii, Aeropyrum pernix, and Sulfolobus tokodaii were cloned and expressed in Escherichia coli. The expression of several of the genes at 15, 25, or 37°C resulted in the formation of insoluble protein aggregates. Therefore, we developed a simple method to express these genes into soluble proteins, by cultivating E. coli clones at a higher temperature. Thus, four genes could be expressed efficiently into soluble and active enzymes by cultivating the respective E. coli clones at 46°C. Subsequently, the method was applied to the expression into soluble proteins of other aminotransferase genes that were derived from nine species of thermophilic microorganisms.

Original languageEnglish
Pages (from-to)172-180
Number of pages9
JournalApplied Microbiology and Biotechnology
Volume73
Issue number1
DOIs
Publication statusPublished - 2006 Nov 1

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ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

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