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Phn and Nag-like dioxygenases metabolize polycyclic aromatic hydrocarbons in Burkholderia sp. C3

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Abstract

Burkholderia sp. C3 can transform polycyclic aromatic hydrocarbons (PAHs), a class of ubiquitous pollutants, through multiple pathways, indicating existence of multiple dioxygenases (Seo et al., in Biodegradation 18:123–131, 2006a). Both phn and nag-like genes in C3 were cloned and identified with the DNA sequence alignment and the gene organization in the clusters. When cloned and expressed in Escherichia coli, either the alpha- and beta-subunits of dioxygenase of the phn genes or the ferredoxin-, alpha- and beta-subunits of the nag-like genes transformed naphthalene, phenanthrene and dibenzothiophene but at different rates. The E. coli transformant containing the phn genes transformed phenanthrene faster than that containing the nag-like genes, which was consistent with higher transcription of the phnAc gene than the nagAc-like gene in C3 in response to phenanthrene. 1-Hydroxy-2-naphthanoic acid (1H2NA) and 2-hydroxy-1-naphthanoic acid (2H1NA) (3,4- and 1,2-dioxygenation metabolites of phenanthrene, respectively) were detected in the culture medium of the phn genes transformed E. coli. The concentration of 1H2NA was 262-fold higher than 2H1NA in the medium of the phn genes transformed E. coli. The results suggested that the phn genes play a major role in 1,2-/3,4-dioxygenation and 3,4-dioxygenation dominates. Twenty-eight PAH degradation-associated enzymes including those encoded by the nag-like and phn genes in phenanthrene-grown C3 cells were identified via alignment of amino acid sequences of the detected polypeptides with those in protein databases. The polypeptides were determined with nano liquid chromatography–ion trap mass spectrometry after tryptic in-gel digestion of the enzymes on 1D SDS-PAGE.

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Abbreviations

PAHs:

Polycyclic aromatic hydrocarbons

RHD:

Ring-hydroxylating dioxygenase

1H2NA:

1-Hydroxy-2-naphthanoic acid

2H1NA:

2-Hydroxy-1-naphthanoic acid

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Acknowledgments

This work was supported in part by grants from Hawaii State Civil Defense, Hawaii Department of Agriculture Pesticides Branch, the US-EPA award 989512-01-1, USDA TSTAR awards, US ONR NRL award N00173-05-2-C003, and the ONR HEET award N00014-09-1-0709.

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Authors and Affiliations

  1. Department of Molecular Biosciences and Bioengineering, University of Hawaii at Manoa, 1955 East-West Road, Honolulu, HI, 96822, USA

    Panlada Tittabutr, Il Kyu Cho & Qing X. Li

  2. School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima, 30000, Thailand

    Panlada Tittabutr

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  1. Panlada Tittabutr
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  2. Il Kyu Cho
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  3. Qing X. Li
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Correspondence to Qing X. Li.

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Tittabutr, P., Cho, I.K. & Li, Q.X. Phn and Nag-like dioxygenases metabolize polycyclic aromatic hydrocarbons in Burkholderia sp. C3. Biodegradation 22, 1119–1133 (2011). https://doi.org/10.1007/s10532-011-9468-y

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