Functional expression of 8-hydroxy-5-deazaflavin-dependent DNA photolyase from Anacystis nidulans in Streptomyces coelicolor

F. Mayerl, J. Piret, A. Kiener, C. T. Walsh, A. Yasui

    Research output: Contribution to journalArticlepeer-review

    7 Citations (Scopus)

    Abstract

    The gene encoding Anacystis nidulans 5-deazaflavin-dependent photolyase (phr) was inserted into the Streptomyces vector pIJ385 to form a transcriptional fusion with the neomycin resistance (aph) gene. The resulting plasmid, pANPL, was introduced into Streptomyces coelicolor, a host which exhibits no detectable photolyase activity and provides 5-deazaflavins. Transformants expressed functional photolyase and could be cultured at much higher cell densities than A. nidulans. A two-step affinity protocol was used to purify photolyase to homogeneity. High-pressure liquid chromatographic analysis established the presence of 5-deazaflavin cofactors in the enzyme, showing that this expression system allows heterologous production of 5-deazaflavin-class photolyases.

    Original languageEnglish
    Pages (from-to)6061-6065
    Number of pages5
    JournalJournal of bacteriology
    Volume172
    Issue number10
    DOIs
    Publication statusPublished - 1990

    ASJC Scopus subject areas

    • Microbiology
    • Molecular Biology

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