AlgaePARC biorefinery René Wijffels, Michel Eppink, Maria Barbosa @ReneWijffels

Contents Technical and economical feasibility Process Photobioreactors Lipids Biorefinery Projects AlgaePARC AlgaePARC biorefinery SPLASH FUEL4ME MIRACLES

Can algae replace commodities of today? European market: Fossil fuels: 400 Mtonnes/year Polymers and chemicals: 100 Mtonnes/year Palm oil: 7 Mtonnes/year Soy: 8 Mtonnes/year Global microalgae market today: < 10,000 tonnes/year

Develop this process is a sustainable and economical way within the next 10 -15 years To develop a more sustainable and economically feasible process, all biomass components should be used

From basic to applied research Resources Efficient use of sunlight Reduction of energy input Use of residual nutrients Strains Lipid accumulation Metabolic Engineering Implementation Scale-up Biorefinery Design scenarios/LCA’s

Photobioreactor design

Antenna size mutants Wild type cells Cells oversaturated: heat dissipation Antenna size mutants Light-saturation at higher light intensity Less energy wasted. Tim de Mooij Turbidostat

Behaviour during day/night cycles Lenneke de Winter The aim of this research is to obtain more knowledge in the circadian behaviour of the cell cycle of lipid producing microalgae grown in photobioreactors. This will provide ways to optimize the current production process under outdoor day/night cycles. Neochloris oleoabundans is grown in a lab-scale photobioreactor under simulated outdoor conditions. The circadian rhythm of the cell cycle is analysed using flow cytometry. In addition, cell composition (lipids, starch, proteins) during the cell cycle is investigated.

Lipids e e e e e

Lipid accumulation N- N+ Nauwelijks TAG onder groeicondities zonder stikstof limitatie Alle algen hopen TAG op maar sommige niet veel 6 algen hopen veel vet op, en 4 zelfs meer dan 35% TAG Hoeveelheid functionele vetten (vetzuren) neemt af onder stikstof limitatie Breuer G., Lamers P.P. , Martens D.E. , Draaisma R.B., Wijffels R.H. (2012) The impact of nitrogen starvation on the dynamics of triacylglycerol accumulation in nine microalgae strains. Bioresource Technology 124: 217-226

Genetic engineering Lipids Strain improvement towards a desired fatty acid profile Strain improvement towards a increased TAG production Hydrocarbons Botryococcus Insert Botryococcus genes for hydrocarbons in Chlamydomonas Mutagenesis and direct gene targeting approaches Transcriptomics to obtain insight in regulation Stefan Aanstoot Lenny de Jaeger

TAG production: process optimization Breuer et al. 2013

Continuous TAG production Different light:N ratios Anne Klok NO3 N- medium Klok et al. 2013

Richard Postma Proteins from Green Sources Development of a mild, continuous, scalable disruption and protein extraction technology with minimal energy requirements Soluble Insoluble Disruption Bead-milling (reference method) Enzymatic Pulsed Electronic Field Extraction Ionic liquid Polymer precipitation Ultrafiltration

AlgaePARC Basic research is not sufficient Gap between basic research and commercial applications Continuous interaction between basic research and pilots Follow up in demonstrations Products Scale Production chain

AlgaePARC International centre of applied research Intermediate between basic research and applications Development of competitive technology (economics, sustainability) Acquire information for full scale plants Algal biomass for food, feed, chemicals and fuels

Scenario: Lipids + Biogas energy Scenedesmus: €600/T Cost of production: €500/T

Scenario: Lipid & protein biorefinery Nannochloropsis: €8,000/T Cost of production: €500/T

AlgaePARC biorefinery Fractionate all components Maintain their functionality At low energy input

EU-FP7 projects INTESUSAL: demonstration project Portugal ALGADISK: biofilmreactor; pilot Spain FUEL4ME: biofuel project with pilots in Israël, Italy, Spain and Netherlands SPLASH: biomaterials project with pilots in Spain and Netherlands MIRACLES: biorefinery project for food and feed applications with pilots in Norway, Chile, Netherlands and Spain

Education Primary school High schools Design challenge and practical kit BSc Physiology and process design MSc Bioreactor design PhD Designing of microalgae-based production processes, for research and commercial purposes Next edition: June 2014

Algae for commodities Demonstrate that biobased products have the quality to replace existing products Convince we can produce economically in the future Imagine we can reach the right market volumes Create awareness: it is not going to be easy Join forces Create a long term commitment

www.AlgaePARC.com Rene.Wijffels@wur.nl @ReneWijffels