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J EPPE L UND N IELSEN C ENTER FOR M ICROBIAL C OMMUNITIES D EPARTMENT OF B IOTECHNOLOGY, C HEMISTRY AND E NVIRONMENTAL E NGINEERING A ALBORG U NIVERSITY.

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Presentation on theme: "J EPPE L UND N IELSEN C ENTER FOR M ICROBIAL C OMMUNITIES D EPARTMENT OF B IOTECHNOLOGY, C HEMISTRY AND E NVIRONMENTAL E NGINEERING A ALBORG U NIVERSITY."— Presentation transcript:

1 J EPPE L UND N IELSEN C ENTER FOR M ICROBIAL C OMMUNITIES D EPARTMENT OF B IOTECHNOLOGY, C HEMISTRY AND E NVIRONMENTAL E NGINEERING A ALBORG U NIVERSITY S EARCHING FOR NEW FUNCTIONAL GENES INVOLVED IN DEGRADATION OF MICROPOLLUTANTS

2 Aalborg West WWTP Mio kWh 100 th anniversary… Future challenges: • High effluent water quality (micropollutants) • Reliable stable plants • Low energy and ressource consumption • Utilization of valuable materials C ENTER FOR M ICROBIAL C OMMUNITIES | A ALBORG U NIVERSITY First Pasveersloot ditch built 1953 (Dreisschor, NL)

3 Ecosystem Design Improve xenobiotic degradation Ecosystem Design Improve xenobiotic degradation ”Marker genes” Assess potential in pilot/full scale systems Improved degradation of micropollutants Identify degraders C ENTER FOR M ICROBIAL C OMMUNITIES | A ALBORG U NIVERSITY

4 Aalborg East Aabybro Hirtshals Aalborg West Triclosan removal hrs C ENTER FOR M ICROBIAL C OMMUNITIES | A ALBORG U NIVERSITY Plant design and operation predicts removal and number of cells involved…

5 C ENTER FOR M ICROBIAL C OMMUNITIES | A ALBORG U NIVERSITY Enumerations by FISH and relative abundance by amplicon sequences Ecophysiology by single cell Activity measures, i.e. MAR-FISH, SIP and various staining techniques Core groups found in 50 Danish treatment plants ?

6 Identifying the “tourists” Combined with FISH/Card-FISH Quantitative MAR MAR-positive EE2 utilising cells EUBmix Estimated biovolume fraction involved in EE2-degradation ~ 0,1 ‰ (~10 5 cells/ml) Uptake of of 3 H-EE2 Principle of MAR-FISH C ENTER FOR M ICROBIAL C OMMUNITIES | A ALBORG U NIVERSITY Uptake of of 3 H-EE2

7 Isolation DNA extraction Sequencing + Assembly Genome Gene sequence acquisition in the genomic era Total DNA extraction Sequencing + Assembly Assembly of individual genomes Identity Metabolic pathways (Functional potential / Blueprint) Genome C ENTER FOR M ICROBIAL C OMMUNITIES | A ALBORG U NIVERSITY Complex community Genomes The pre-genome era as the ‘dark ages’ (Ian Dunham, )

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

9 Abundance of mRNA and proteins in S. cerevisiea (Futcher et al., 1999) mRNA content in a cell ~42,000 (S. cerevisiea) and many are non-coding! Prokaryotes has less… (Aebersold et al., 2013) Proteins carry out the job… C ENTER FOR M ICROBIAL C OMMUNITIES | A ALBORG U NIVERSITY

10 Quantitative Proteomics pipeline C ENTER FOR M ICROBIAL C OMMUNITIES | A ALBORG U NIVERSITY Quantification of proteins combined with metabolomics (+/- micropollutant) provide insight into the proteins affected by the micropollutant! Quantification of proteins combined with metabolomics (+/- micropollutant) provide insight into the proteins affected by the micropollutant! Full or partial biochemical pathway established for degradation of: • p-cresol (effect on nitrifiers) • Gemfibrozil • Ibuprofen • Estradiol • Bisphenol A • Triclosan • Chrome Full or partial biochemical pathway established for degradation of: • p-cresol (effect on nitrifiers) • Gemfibrozil • Ibuprofen • Estradiol • Bisphenol A • Triclosan • Chrome

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

12 MAR-FISH to confirm Uptake of 14 C-Ibuprofen qPCR for quantification of Ibuprofen-degrading genes and their expression Better removal of micropollutants by introducing a polishing of effluent BIOSEP™ MBR C ENTER FOR M ICROBIAL C OMMUNITIES | A ALBORG U NIVERSITY Phylogeny to describe diversity Development of microflora in polishing step for complete removal of: Carbamazepine Diclofenac Ibuprofen Naproxen Triclosan,… Development of microflora in polishing step for complete removal of: Carbamazepine Diclofenac Ibuprofen Naproxen Triclosan,…

13 C ENTER FOR M ICROBIAL C OMMUNITIES | A ALBORG U NIVERSITY Thank you for your attention! and collaborations: K. Bester (AU) H. Gough (U. Washington) M. Von Bergen (Leipzig)


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