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Genomic Mining for Novel FADH2-Dependent Halogenases in Marine Sponge-Associated Microbial Consortia

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Abstract

Many marine sponges (Porifera) are known to contain large amounts of phylogenetically diverse microorganisms. Sponges are also known for their large arsenal of natural products, many of which are halogenated. In this study, 36 different FADH2-dependent halogenase gene fragments were amplified from various Caribbean and Mediterranean sponges using newly designed degenerate PCR primers. Four unique halogenase-positive fosmid clones, all containing the highly conserved amino acid motif “GxGxxG”, were identified in the microbial metagenome of Aplysina aerophoba. Sequence analysis of one halogenase-bearing fosmid revealed notably two open reading frames with high homologies to efflux and multidrug resistance proteins. Single cell genomic analysis allowed for a taxonomic assignment of the halogenase genes to specific symbiotic lineages. Specifically, the halogenase cluster S1 is predicted to be produced by a deltaproteobacterial symbiont and halogenase cluster S2 by a poribacterial sponge symbiont. An additional halogenase gene is possibly produced by an actinobacterial symbiont of marine sponges. The identification of three novel, phylogenetically, and possibly also functionally distinct halogenase gene clusters indicates that the microbial consortia of sponges are a valuable resource for novel enzymes involved in halogenation reactions.

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Acknowledgments

We gratefully acknowledge the marine operations personnel at the Laboratoire Arago (Banyuls-sur-Mer, France) and at the Ruder Boskovic Institute (Rovinj/ Croatia) for the help during sponge collection and Prof. K.-H. van Pée (Dresden, Germany) for helpful advice. Prof. W. Wohlleben, S. Pelzer, and Claudia Kittel (Tübingen, Germany) are acknowledged for interesting discussions and activity testing of selected clones in an S. albus background. We thank Christine Gernert (Wuerzburg, Germany) for excellent technical assistance. Financial support was provided by the SFB630 (grant TPA5) to U.H.

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Authors and Affiliations

  1. Julius-von-Sachs Institute for Biological Sciences, University of Wuerzburg, Julius-von-Sachs Platz 3, D-97082, Wuerzburg, Germany

    Kristina Bayer & Ute Hentschel

  2. Center for Infectious Diseases, University of Wuerzburg, Josef-Schneider-Straße 2, D-97080, Wuerzburg, Germany

    Matthias Scheuermayer

  3. Institute for Food and Beverage Innovation, Zurich University of Applied Sciences, Campus Reidbach, P.O. Box, CH-8820, Wädenswil, Switzerland

    Lars Fieseler

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  1. Kristina Bayer
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  2. Matthias Scheuermayer
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  3. Lars Fieseler
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  4. Ute Hentschel
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Correspondence to Ute Hentschel.

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Kristina Bayer and Matthias Scheuermayer contributed equally to this work.

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Bayer, K., Scheuermayer, M., Fieseler, L. et al. Genomic Mining for Novel FADH2-Dependent Halogenases in Marine Sponge-Associated Microbial Consortia. Mar Biotechnol 15, 63–72 (2013). https://doi.org/10.1007/s10126-012-9455-2

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