Diversity of nitrite reductase genes in "Candidatus accumulibacter phosphatis"-dominated cultures enriched by flow-cytometric sorting

Ryuki Miyauchi, Kazuma Oki, Yoshiteru Aoi, Satoshi Tsuneda

    Research output: Contribution to journalArticle

    25 Citations (Scopus)

    Abstract

    "Candidatus Accumulibacter phosphatis" is considered a polyphosphate-accumulating organism (PAO) though it has not been isolated yet. To reveal the denitrification ability of this organism, we first concentrated this organism by flow cytometric sorting following fluorescence in situ hybridization (FISH) using specific probes for this organism. The purity of the target cells was about 97% of total cell count in the sorted sample. The PCR amplification of the nitrite reductase genes (nirK and nirS) from unsorted and sorted cells was performed. Although nirK and nirS were amplified from unsorted cells, only nirS was detected from sorted cells, indicating that "Ca. Accumulibacter phosphatis" has nirS. Furthermore, nirS fragments were cloned from unsorted (Ba clone library) and sorted (Bd clone library) cells and classified by restriction fragment length polymorphism analysis. The most dominant clone in clone library Ba, which represented 62% of the total number of clones, was not found in clone library Bd. In contrast, the most dominant clone in clone library Bd, which represented 59% of the total number of clones, represented only 2% of the total number of clones in clone library Ba, indicating that this clone could be that of "Ca. Accumulibacter phosphatis." The sequence of this nirS clone exhibited less than 90% similarity to the sequences of known denitrifying bacteria in the database. The recovery of the nirS genes makes it likely that "Ca. Accumulibacter phosphatisbehaves as a denitrifying PAO capable of utilizing nitrite instead of oxygen as an electron acceptor for phosphorus uptake.

    Original languageEnglish
    Pages (from-to)5331-5337
    Number of pages7
    JournalApplied and Environmental Microbiology
    Volume73
    Issue number16
    DOIs
    Publication statusPublished - 2007 Aug

    Fingerprint

    Nitrite Reductases
    nitrite reductase
    sorting
    nitrite
    clone
    Clone Cells
    clones
    gene
    Genes
    genes
    organisms
    Polyphosphates
    polyphosphates
    cells
    denitrifying bacteria
    Denitrification
    Nitrites
    fluorescence in situ hybridization
    Fluorescence In Situ Hybridization
    denitrification

    ASJC Scopus subject areas

    • Environmental Science(all)
    • Biotechnology
    • Microbiology

    Cite this

    Diversity of nitrite reductase genes in "Candidatus accumulibacter phosphatis"-dominated cultures enriched by flow-cytometric sorting. / Miyauchi, Ryuki; Oki, Kazuma; Aoi, Yoshiteru; Tsuneda, Satoshi.

    In: Applied and Environmental Microbiology, Vol. 73, No. 16, 08.2007, p. 5331-5337.

    Research output: Contribution to journalArticle

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