Production of specialties for food, aquaculture and non-food applications via multi-product biorefinery of microalgae
Highlights of the EU FP7 project MIRACLES
Multi-product Integrated bioRefinery of Algae: from Carbon dioxide and Light Energy to high-value Specialties (2013-2017)
Hans Reith, Leen Bastiaens, Hans Kleivdal, Dorinde Kleinegris, Lolke Sijtsma, Philippe Willems, Carlos Unamunzaga, Elke Breitmayer, Macarena Sanz, René Wijffels
AlgaeBiorefineries for Europe, Brussels, 16-17 October 2017
Successful scale-up of the algae industry requires reduction of production costs and enhancement of the (economic) output.
MIRACLES addresses these challenges by:
● improving cost-effectiveness of algae production andprocessing through technology development
● development of multiple-product biorefinery for specialtiesfrom algae
● development of new products for food, aquaculture and non-food
Aims
The MIRACLES consortium: 26 partners
• 6 Universities, 5 Research Organisations, • 12 SME’s, 3 MNI’s incl. 11 end user companies in target sectors• In 6 EU countries + Norway + Chile.
● Production of biomass for biorefinery and application research using established production strains:● Nannochloropsis ● Isochrysis● Phaeodactylum● Scenedesmus
● Development molecular monitoring tools / product optimization strategies
● Pilot scale production
Algae production
Closed photobioreactors. 100% automated and Scada controlled. Induced accumulation of bioactive compounds.
FITOPLANCTON MARINO FACILITIES (2015)
CO2 concentration from the atmosphere
Aim: to become independent of CO2 point source (e.g. power plant, flue gas) >>more remote locations Based on Supported amine adsorption
Activities√ Sorbent screening (method & sorbent)
√ Sorbent characterisation√ CO2 adsorption capacity√ H2O co-adsorption√ Kinetics√ Sorbent stability√ Regeneration (Pure CO2 vs. enriched
air)√ Process & Contactor design√ Construction of Prototype √ Proof of Concept
Screening sorbents
Adsorption kinetics (H2O >CO2)
Contactor design and testing
Proof of concept ongoing
Installation fully operational
Evaluation of (semi) continuous operation (>1% CO2; > 1kg/day) coupled to algae cultivation ongoing
Update in lecture Wim Brilman + poster
Molecular tools to monitor and control production of target products during cultivation
• Molecular tools available for monitoring production inNannochloropsis gaditana:
• TriAcylGlycerides (TAG)
• Eicosa Pentaenoic Acid (EPA) ω3-essential fatty acid
• Envisioned use of the tools to optimize production:
• Real-time monitoring for appropriate timing of biomassharvesting to reach optimal productivity of TAG / EPA
• Potential use in selection programs for clones with higher capacityand/or efficiency to accumulate TAG and/or EPA
Lecture Carlos Infante
• Based on establishing correlationsbetween relative gene expressionlevels and productivity of a selectedproduct.
Concept: growth of algae in liquid foams
Foam generation controlled by foaming agents and gas distributors
Development Liquid Foam Bed PhotoBioReactor
Feature ConsequenceVery short light pathLimited amount of water in reactor
Low weight, low pressure drop,
Large interface surface areaIncreased gas residence time
High biomass concentration Reduced harvesting costs
Reduced construction costs, energy costs
Enhanced mass transfer CO2, O2
• Validation on lab scale achieved • Substantial reduction of energy use and costs projected
Membrane technology for harvesting and growth medium recycling
Harvesting and dewatering: major impact on energy use and costs
Water & nutrients: significant cost factor in large scale cultivation.
Aim: develop novel membrane technology for combined harvesting and medium recycling capable of saving water, nutrients, energy and costs
Submerged Membrane Algae Filtration tests
AlgaeCentrifugation
160-200 g/L0,2-2 g/L
Submerged membranereactor
10-20 g/L
Nannochloropsis gaditana
MAF1 (lab scale – 0,3 m²)
10 tests performed
MAF2 (semi-pilot – 1 m²)
>15 tests performed (100-600L)
MAF1e – continuous(semi-pilot– 1,3 m²)
2-stages
Outlook
Submerged membrane filtration technology is a promising approach for integrated algae pre-concentration & medium recirculation
Medium recycling (up to > 90%) is possible; no negative impact on algae growth observed
Technology positively evaluated at lab and semi-pilot scale both in batch & continuous mode
• Substantial reduction of operational costs is expected
Lecture Leen Bastiaens, + posters
Bioprospecting, screening, selection of strains
Environmentalsamples collected
Enrichmentscultures in lab
Clonal culturesplated (FACS)
Clonal isolatesestablished for
cultivationScreenedstrains
Candidatestrains
Sub-tropics,Gran Canarias, Spain 213 639 - 70 24 9
Altiplanic lagoon Antofagasta, Chile 69 360 19200 66 20 13
Arctic/Nordic, Bergen, Norway 58 110 10560 149 43 5
Total 337 1 109 38 400 285 87 27
Demonstration
Downstream processing
Application testing
SelectionIndustrial screening criteria
SamplingIsolation Cultivation Screening
BEA CODE STRAIN % BEA 0024 Rhodomonas sp 0,1%
TYPE Phaeodactylum tricornutum 0,2% BEA 0650 Tetraselmis striata 0,3% BEA 0664 Pyrocystis lunula 0,4% BEA 0136 Chlorococcum sp. 0,4% BEA 0620 Erythotrichia cf carnea 0,4% BEA 0097 Tetraselmis striata 0,8% BEA 0304 Dunaliella sp 1,3% BEA 0078 Tetraselmis convolutae 1,4% BEA 0163 Dunaliella sp 1,4% BEA 0808 Closteriun sp. 1,7% BEA 0124 Rhodomonas sp 1,9% BEA 0398 Picochlorum oklahomense 2,0% BEA 0615 Graesiella vacuolata 2,0% BEA 0788 Coelastrella saipanensis 2,1%
TYPE Scenedesmus obliquus 2,2% BEA 0274 Crucigeniella sp 2,3% BEA 0317 Desmodesmus sp. 2,4% BEA 0544 Coccomyxa sp 2,4% BEA 0406 Cosmarium sp 2,8% BEA 0316 Desmodesmus sp. 3,0% BEA 0364 Protodesmus globulifer 3,4% BEA 0069 Halochlorella rubescens 4,5%
TYPE Isochrysis galbana 7,1% BEA 0313 Sarcinochrysis marina 8,4%
TYPE Nannochloropsis gaditana 10,5% BEA 1048 Euglena sp. 22,0% BEA 0937 Euglena cantabrica 57,5%CONTROL Yeast 46% ß-Glucan 44,6%
Enrichment
Bioprospecting in extreme climatic conditions to identify novel algal strains with suitable product profiles.
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Characterisation of selected strains
• Characterisation on lab scale
• Best strains evaluated for pilot scale outdoor productivity in identical photobioreactors
Outlook
• New, validated production strains available
• Process strategies to maximize yields of oils and EPA in Nannochloropsis (based on gene studies and metabolic modeling)
Lecture Dorinde Kleinegris + posters
Complete, mild cell disruption via bead milling or homogenization effective. Operation bead mill optimized..
Suitable enzymes for processing identified
Novel extraction routes using Supercritical fluids, Gas Expanded Liquids, Pressurized Liquids
Membrane filtration (UF/DF) for protein refinery
Results Integrated in Multiproduct flowsheets
Algae oil produced and extracted on pilot scale
Lectures Lolke Sijtsma, Elena Ibanez + several posters
Achievements, innovations in biorefinery development
Soluble, functional proteins as potentialsubstitute for dairy proteins in food products
● Protein characteristics differ from otherprotein sources>> processing challenging
● Promising emulsifying properties shownin product tests using crude soluble proteinfractions
Proteins for food applications
Aquaculture will be main contributor to global seafood supply: 140 MT by 2050 (more than double in the next 15 years)
Aquaculture sector is constantly seeking novel, sustainable raw materials of high nutritional value
In the framework of MIRACLES partners EWOS and Sparos assessed the potential of microalgae as ingredients in aquafeeds
Application of microalgae in aquafeeds
• Diets for trials evaluating digestibility for seawater salmon and growth of small freshwater salmon prepared containing Nannochloropsis (disrupted*) and Phaeodactylum produced at Fitoplancton Marino.
• Reduced nutrient digestibility observed for the algae diets (esp. disrupted Nannochloropsis) compared to reference diet needs to be considered regarding nutritional value of these algae in salmon feed formulations.
• Disrupted Nannochloropsis and Phaeodactylum added at 8% of diet fed to smallfreshwater salmon (2.5g initial weight) over 8 weeks (flowchart below). Algae feeds were non-inferior for growth compared to reference diet.
Salmon trials evaluating digestibility in seawaterand growth in freshwater
*Wet algae paste went through two passes of high pressure liquid homogeniser at ILVO-Technology & Food, BENatural antioxidant used to provide extra protection to algae in algae diets only so combined algae/natural antioxidant effect compared against reference diet. Note that oxidation parameters (TBARS on all diets and PV on small fish diets) were not lower in algae diets compared to reference diet.
Demonstration of functional role of microalgae in gilthead seabream(major farmed species in the Mediterranean)
Diet with residual biomass of
N. gaditana and algal oil allowed the
successful replacement of
fishmeal and fish oil by 80% and 30%,
respectively
Diet with 1% P. tricornutum resulted in a short-term benefical modulation of several immune-related pathways
A consumer panel characterised algae-fed seabream as having a more vivid and typical external pigmentation
Demonstration of functional role of microalgae in Senegalese sole(an emergent high value species in Europe)
Diet with 1% of fucoxanthin-rich algae (P. tricornutum) improved immune response to stressful events in sole juveniles
Diets with 5% of a broken cells extract of P. tricornutum
did not affect growth, but significantly enhanced the
survival of sole larvae
Microalgae show great potential as functional ingredients in aquaculture feeds
Demonstrated benefits on:
• Animal welfare (stress resistance, immunity)• Consumer quality traits (pigmentation)• Reduce reliance on finite marine-harvested resources (e.g.
replacement of fish meal and fish oil)
Poster #40 M.Sardinha et al.
Protein hydrolysates (prepared by cell disruption + enzymatic hydrolysis) tested for bioactive properties: antioxidant, antimicrobial, ....
Potent inhibitors of food spoilage-related micro-organisms identified (longer lag phase, lower growth rate)
Different concentrations of extracts were added to a bacterial culture and the growth was monitored at 30 C directly (left) or after pre-incubation at 4 C (right), to mimick food storage conditions.
Harry Laan, Poster #30
Antimicrobial peptides
Development of a cosmetic product: facial cream
MICROENCAPSULATION
IGE: Carotenoids fraction ofIsochrysis galbana extract(fucoxanthin)
ACTIVE INGREDIENT 1 FUNCTIONALITY TESTS 2Human dermal fibroblasts (ATCC® PCS-201-012™)
CYTOTOXICITY ASSAY
PROOXIDANT TOXICITY Neutralization of ROS overproductioninduced by external agents (oxidantsand UV radiation) in human fibroblasts.Antioxidant effect up to 100 timesgreater than that of pure fucoxanthin
Selection of the optimal dose
Phospholipid complex formation
3
Stabilized active ingredient
ANTIOXIDANT PROTECTION
COSMETIC FORMULATION4
Esther de la Fuente, Natac
Development of microalgae-based resins for the wood-based panel industry
Range of algae species, materials tested: a) Untreated biomass b) fractions incl. proteins, residues after extraction of lipids.
Development of effective PF type resins with 30% phenol replacement by microalgae-derived materials.
Produce plywood panels that meet the criteria of the European standards for exterior applications (EN314.1 and EN314.2)
Development of bio-based, formaldehyde free covering material suitable for lamination of wood-based panels.
Electra Papadopoulou c.s., Poster #37
Produced batches bioplastic compound suitable for:
● Injection moulding
● Sheet extrusion
Up to 15% in compound replaced by algae
Acceptable mechanical and processing properties
Product development ongoing
Contact: Philippe Willems, exploitation officer Miracles
Biopolymers
Use of whole algae or processing residues Application in flooring, table tops, light covers, ...
Contact: Philippe Willems, VFT/Orineo
Decorative biomaterials VFT
Specialty fertilisers/plant growth promotors
• Isochrysis extract has rejuvenating and stress reduction effects in bioassays using wheat, corn seedlings
• stimulated growth rate • delayed leaf senescence• higher leaf expansion rate
• Phaeodactylum extract shows positive effect on Nitrogen-use efficiency of plants. Wheat and corn seedlings show increased growth rate at low nitrogen nutrition level.
• Relevant for improving plant quality, durability in logistics phase, and customer satisfaction.
• Product development underway
Poster #36, Daan Kuiper, Philippe Willems
Conclusions
The MIRACLES consortium has successfully developed and demonstrated:
● technological innovations that contribute to improving cost-effectiveness of algae production, harvesting and processing
● new value-added applications and product leads for food, aquaculture and non-food
We are ready for further development and commercialisation
For publications, final report, contacts see Project Website: http://miraclesproject.eu/
• The project is supported by the European Commission through the 7th Framework Program under Grant Agreement No. 613588.
• Program Officer DG RTD: Andrea Grisorio [email protected]
• More information: Hans Reith, [email protected]
• www.miraclesproject.eu
Acknowledgements
Thanks!
For more information visit our website and follow us on social networks:
www.miraclesproject.eu@miraclesproject
Project Website: http://miraclesproject.eu/
Coordinator: Hans Reith, Bioprocess Engineering, WU Agrotechnology& Food Sciences. Tel. +31 (0)317 480 100 , e-mail [email protected]
Exploitation Officer: Philippe Willems, Value for Technology bvba. Tel.: +32 (0)2 759 33 10. e-mail [email protected]
Dissemination Officer: Macarena Sanz, IDAction and IDConsortium, Tel. +34 911 25 03 11. e-mail [email protected]
EC Project Officer: Andrea GRISORIO, European Commission, DG Research & Innovation. F2. Bio-based products and processing., Tel.: +32 229-90538. e-mail: [email protected]
Contacts