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Jeppe Lund Nielsen Center for Microbial Communities Department of Biotechnology, Chemistry and Environmental Engin

Searching for new functional genes involved in degradation of micropollutants. Jeppe Lund Nielsen Center for Microbial Communities Department of Biotechnology, Chemistry and Environmental Engineering Aalborg University. Mio kWh. 100 th anniversary…. Aalborg West WWTP.

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Jeppe Lund Nielsen Center for Microbial Communities Department of Biotechnology, Chemistry and Environmental Engin

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  1. Searching for new functional genes involved in degradation of micropollutants Jeppe Lund Nielsen Center for Microbial Communities Department of Biotechnology, Chemistry and Environmental Engineering Aalborg University

  2. Mio kWh 100th anniversary… Aalborg West WWTP First Pasveerslootditchbuilt 1953 (Dreisschor, NL) • Future challenges: • High effluent water quality (micropollutants) • Reliable stable plants • Low energy and ressource consumption • Utilization of valuable materials Center for Microbial Communities | Aalborg University

  3. Improved degradation of micropollutants Ecosystem Design Improvexenobioticdegradation Identify degraders Center for Microbial Communities | Aalborg University

  4. Aalborg East Aabybro Hirtshals Aalborg West hrs Triclosan removal Plant design and operation predicts removal and number of cellsinvolved… Center for Microbial Communities | Aalborg University

  5. Ecophysiology by single cell Activity measures, i.e. MAR-FISH, SIP and variousstaining techniques Core groups found in 50 Danish treatment plants ? Enumerations by FISH and relative abundance by ampliconsequences Center for Microbial Communities | Aalborg University

  6. Identifying the “tourists” Uptake of of3H-EE2 Combined with FISH/Card-FISH Estimated biovolume fraction involved in EE2-degradation~ 0,1 ‰ (~105 cells/ml) Uptake of of3H-EE2 Quantitative MAR Principle of MAR-FISH MAR-positive EE2 utilising cells EUBmix Center for Microbial Communities | Aalborg University

  7. Gene sequence acquisition in the genomicera Complex community DNA extraction Sequencing + Assembly Isolation Genome Total DNA extraction Identity Metabolic pathways (Functional potential / Blueprint) Sequencing + Assembly Genome Assembly of individual genomes Genomes Center for Microbial Communities | Aalborg University The pre-genomeera as the ‘dark ages’ (Ian Dunham, )

  8. First 1000 microbialgenomescomprise ca. 3,3mio genes Transitive catastrophe When a mistake is inherited from gene to gene COMBREX (COMputationalBridges to EXperiments; http://combrex.bu.edu/)

  9. Proteins carry out the job… Abundance of mRNA and proteins in S. cerevisiea(Futcher et al., 1999) mRNAcontent in a cell~42,000 (S. cerevisiea) and manyare non-coding! Prokaryotes has less… (Aebersold et al., 2013) Center for Microbial Communities | Aalborg University

  10. QuantitativeProteomics pipeline Quantification of proteins combined with metabolomics (+/- micropollutant) provide insightintothe proteins affected by the micropollutant! • Full or partialbiochemicalpathwayestablishedfor degradation of: • p-cresol (effect on nitrifiers) • Gemfibrozil • Ibuprofen • Estradiol • Bisphenol A • Triclosan • Chrome Center for Microbial Communities | Aalborg University

  11. Established model of Ibuprofen degradation Extracellular solute binding protein family 1 Proposed pathway for ibuprofen degradation A - AMP-dependent synthase and ligase B – Dioxygenase containing Rieske (2Fe-2S) iron sulphur domain C - Enoyl-CoA hydratase/isomerase Ibuprofen Cell Membrane ABC 2 Type transporter Cytosol D E A B C D & E in same operon?

  12. Better removal of micropollutants by introducing a polishing of effluent qPCR for quantification of Ibuprofen-degrading genes and theirexpression BIOSEP™ MBR Development of microflora in polishing step for complete removal of: Carbamazepine Diclofenac Ibuprofen Naproxen Triclosan,… Phylogeny to describe diversity MAR-FISH to confirm Uptake of 14C-Ibuprofen Center for Microbial Communities | Aalborg University

  13. Thank you for your attention! and collaborations: K. Bester (AU) H. Gough (U. Washington) M. Von Bergen (Leipzig) Center for Microbial Communities | Aalborg University

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