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

Daisuke Koma, Toshiya Sawai, Shigeaki Harayama, Kuniki Kino

    Research output: Contribution to journalArticle

    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

    Fingerprint

    Escherichia coli
    Genes
    Temperature
    Transaminases
    Aeropyrum
    Pyrococcus horikoshii
    Proteins
    Clone Cells
    Sulfolobus
    Escherichia coli Proteins
    Microorganisms
    Gene Expression
    Enzymes

    ASJC Scopus subject areas

    • Biotechnology
    • Microbiology
    • Bioengineering
    • Microbiology (medical)

    Cite this

    Overexpression of the genes from thermophiles in Escherichia coli by high-temperature cultivation. / Koma, Daisuke; Sawai, Toshiya; Harayama, Shigeaki; Kino, Kuniki.

    In: Applied Microbiology and Biotechnology, Vol. 73, No. 1, 11.2006, p. 172-180.

    Research output: Contribution to journalArticle

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