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KOORDINIERT VON

Improved use of carbon streams in wastewater through biotechnological processes –

new approaches within the project ZeroCarbFP

Daniel Klein, Dirk Bogaczyk, Linh-Con Phan; EmschergenossenschaftRenate Schulze, Guido Meurer; BRAIN AG

Re-Water Braunschweig 03.11.2015

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014Growth of world population: 7.3 Bill.

in 2015; estimated > 9 Bill. in 2050

Increase of food demand (60% by2050)

Increasing production of waste, sewage sludges, CO2, …

Growing demand of energy andother (fossil) resources

Innovation Alliance ZeroCarbFPBackground

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Key substance: Carbon

ZeroCarbFPApproach

Use of carbon-containing waste streams instead of fossil, limited resources(such as petrol)

Substitution of food- and agro-based raw materials and chemicals(fuel-food-discussion)

Implementation of bio-based processes instead of physical/chemical ones

Waste streams Functionalbiomass

Valuableproducts

Bulk and specialchemicals,

biopolymers…

Energy

SP 2

DeICE Plus

Waste glycerine of biodiesel production

De-icing and cooling liquids

SP 3

Green Mining

Recycling materials, landfills

Sustainable ore leaching processes

SP 4

Additives 1

Waste fats, oils, etc

High-performance additives

ZeroCarbFPZeroCarbFP

Conversion of waste streams to valuable products by functional biomass

ZeroCarbFPSubprograms (SP)

SP 1

Bioplastics

Waste streams; flue gas

Monomers for polymer chemistry

Was

te(s

)P

rodu

ct(s

)

SP 5

Additives 2

Municipal/industrial Wastewater

Single cell oils & fatty acids; energy

ZeroCarbFPSubprogram „Additives 2“

SP 5

Additives 2

Biotechnological production of singlecell oils (SCO) andlipids (FAME)

Energy productionby anaerobicdigestion andmicrobial fuel cells(MFC)

SCO / EnergyProduct line 1SCO / Energy SCO / Additives

Product line 2SCO / Additives MFC / Energy

Product line 3MFC / Energy

Municipal wastewater Municipal wastewaterIndustrial wastewater

Municipal wastewater

Functional biomass(microorganisms) able toproduce / accumulateSCO and lipids

Functional biomass (exo-electrogene organisms) on electrodes of microbialfuel cells

Increase of biogasproduction in digestertowers

SCO to be used in industry, e.g. as basis ofhigh-performance additives

Energy

Functional biomass(microorganisms) able toproduce / accumulateSCO and lipids

ZeroCarbFPCoordination and SP 5

Water courses 742 km

Sewers 1,189 km

Pumping stations 303

WWTP 60

Capacity of WWTP 7.4 Mio. PE

Quantity of waste water 0.9 Bill. m3/a

Area:4.200 km²

Inhabitants:4 Mio.

PL 1: Improving on-site biogas productionApproach

Substrates

(e.g. sludge) Organisms

Single Cell Oils

Energy

PL 1: Improving on-site biogas productionConsiderations

Carbon available for biogas production

Carbon available for SCO-production

Carbon available for both processes

Carbon not yet used

Assumptions

• A notable amount of organic C is not available for anaerobic digestion

• Likely, specifically adapted functional biomass is able to convert a part of this C into SCO

• SCO and lipids in general are easily degradable anaerobically

• Anabolism vs. catabolism of microorganisms

• Net-energy-balance should be positive

PL 1: Improving on-site biogas productionEvaluation of oleaginous organisms

Ramalingam Subramaniam, Stephen Dufreche, Mark Zappi, Rakesh Bajpai: Microbial lipids from renewable resources: production and characterization. J Ind Microbiol Biotechnol (2010) 37:1271–1287

PL 1: Improving on-site biogas productionSelection of organisms, screening and validation

• Pre-selection of microorganisms (MO) • Bioprospecting

• Water- and wastewater streams• Oil-containing environmental samples

• Collections of microorganisms • BRAIN BioArchive• DSMZ

• Enrichment and isolation of strains

• Screening and validation• Potential SCO-accumulation

(colour indicator)• Risk assessment• Composition of SCO (e.g. length of

fatty acids; saturated/unsaturated)

• Determination of most promising organisms and backu ps

Primary sludge

Digestedsludge

Dewateredsludge

Strain Type Origin

Mos

tpro

mis

ing

MO

1 Fungus Collection

2 Fungus WWTP

3 Bacterium Collection

4 Fungus Collection

… … … … … …

Bac

kup

1 Fungus WWTP

2 Fungus WWTP

3 Fungus WWTP

4 Fungus WWTP

… … … … … …

• Scale up:• Growth of promising

combinations (substrate/organism) in bioreactors

• Lab-scale digestion to assess biogas production (ongoing)

• Optimisation of growing media (e.g. pre-treatment) and organisms

PL 1: Improving on-site biogas production Growth of selected MO in wastewater

GrowthNo growth

ZeroCarbFPSubprogram „Additives 2“

SP 5

Additives 2

Biotechnological production of singlecell oils (SCO) andlipids (FAME)

Energy productionby anaerobicdigestion andmicrobial fuel cells

SCO / EnergyProduct line 1SCO / Energy SCO / Additives

Product line 2SCO / Additives MFC / Energy

Product line 3MFC / Energy

Municipal wastewater Municipal wastewaterIndustrial wastewater

Municipal wastewater

Functional biomass(microorganisms) able toproduce / accumulateSCO and lipids

Functional biomass (exo-electrogene organisms) on electrodes of microbialfuel cells

Increase of biogasproduction in digestertowers

SCO to be used in industry, e.g. as basis of high-performance additives

Energy

Functional biomass (microorganisms) able to produce / accumulate SCO and lipids

PL 2: SCO for industrial use Approach and overview

Substrates

(e.g. sludge) Organisms

Single Cell Oils

Lubricants / Additives

Lubricant industry

Organism No. 1 2 3

FAME (glucose) 33% 28% 25%

FAME (wastewater 1) 19% 18% 15%

FAME (wastewater 2) > 50% 47% 42%

Sum of unsaturatedFA (glucose)

66% 60% 89%

Sum of unsaturatedFA (ww 1)

68% 66% 88%

Sum of unsaturatedFA (ww 2)

91% 88% 82%

… … …

• Quantity and composition of accumulated/produced SCO (measured as FAME) depend on substrate

• Lab-scale results with artificial media (glucose) could be verified with real wastewaters

• Promising composition of SCO (e.g. high content of long-chain unsaturated fatty acids)

PL 2: SCO for industrial use Example: Composition of accumulated SCO

• Optimisation of growing media (e.g. separation of growth- and accumulation phase, using different types of wastewater)

• Growth phase: balanced growing medium• Accumulation phase: high C/N-ratio

• Upscaling (bioreactor)

• Separation of accumulated SCO from biomass• Extraction protocol

• Production of a specimen to be tested by industry

PL 2: SCO for industrial use Current work and next steps

Conclusion and outlook

• Microorganisms (functional biomass) able to accumulate SCO, using wastewater as substrate, have been identified

• Wastewater treatment was linked with biotechnological approaches

• Improvement of biogas production (PL 1) possible; depending on energy balance• Conversion of waste C to valuable products (SCO for industry) possible (PL 2) • Substrate and/or microorganisms will further be optimised and adapted

• Development of extraction protocol / Assessment of integration into systems

2013 2014 2015 2016 2017 2018 2019 2020 2021 20222012

Start Phase 1(Research)

Phase 2(Development)

Phase 3(Pilot production)

• Phase II of ZeroCarbFP (2016-2019): ‘Development’ (to be applied for)

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KOORDINIERT VON

Dr.-Ing. Daniel Klein

Dirk Bogaczyk

Dr.-Ing. Linh-Con Phan

klein.daniel@eglv.de

+49 (0)201 104- 2280

Emschergenossenschaft

Kronprinzenstraße 24

45128 Essen

www.eglv.de

Dr. Renate Schulze

Dr. Guido Meurer

BRAIN Aktiengesellschaft

Darmstädter Str. 34-36

64673 Zwingenberg

06251-9331-0

www.brain-biotech.de

Thank you for your attention