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FP7

D-FACTORY Report Summary

Project ID: 613870
Funded under: FP7-KBBE
Country: United Kingdom

Periodic Report Summary 3 - D-FACTORY (THE MICRO ALGAE BIOREFINERY)

Project Context and Objectives:
Context: The D-Factory is a biorefinery which produces nutraceuticals, chemicals, feed and fuels from halotolerant microalgae, such as Dunaliella. Dunaliella grows in saline, non-potable water, around the globe. It captures CO2 and harnesses sunlight energy, and naturally produces carotenoids, and other compounds of commercial value - bioactives, emulsifiers, polymers and glycerol. The D-Factory project will tune the algae to make these products in different proportions, then extract them using supercritical CO2, HPCCC and membrane or chromatography, and formulate them to meet market requirements.
The objectives of the project that will deliver the D-Factory biorefinery are as follows:
1. Establish a D-Factory reference library of at least 12 new monoclonal halotolerant microalgal strains isolated either from partner/stakeholder locations or developed by mutagenesis, cryopreserve where possible and make this available for D-Factory Partners to screen, characterise and use in the D-Factory demonstration of a microalgal biorefinery.
2. Establish production protocols by month 36 that enable suppliers of microalgal biomass to tailor their production of halophytic algae such as Dunaliella using techniques based on strain selection and/or cultivation, in order to meet biorefinery specifications.
3. Benchmark an open raceway facility producing kilogram quantities of Dunaliella biomass in terms of its technological, economic, environmental and social sustainability.
4. Develop and implement a pilot (10 m3) cultivation PBR by month 24, which, by using controlled conditions,
• Is capable of producing large quantities of high density (2.0 g/l DW) microalgal biomass tailored to meet D-Factory biorefinery demands,
• Offers sustainable scalability to industrial scale (several hectares),
• Provides data for assessment and modelling (by month 33 ),
• Informs the creation of the D-Factory business plan (month 39).
5. Develop and demonstrate cell harvesting technologies to process ~2500l saline water /h and recover at least 90% intact Dunaliella cells at lowest cost from open ponds or PBRs by month 12 for the D-Factory demonstration
6. Develop methods for microalgal cell storage, handling and disruption (by month 6) and for microalgal bioprocessing using technology schemes based on scCO2, HPCCC, and membrane technologies, to produce extracts/fractions for bio/chemical profiling, and bioactivity screening (month 24), and for D-Factory scale-up designs to industrial scale, taking into account technical, environmental and economic aspects using data obtained from mobile units installed at the D-Factory demonstration production site (month 48).
7. Confirm chemical structures on selected D-Factory biological materials, including by-products of microalgal processing, establish biological activity, measure product recoveries and purities, and identify any new compounds of interest (month 36), and develop product specifications for extracts, by month 48.
8. Draw up plans for adding value to Dunaliella extracts either through formulation, or through more rigorous purification for applications in pharmaceutical, nutraceutical, food, or cosmetic industries (for new business development), by month 46.
9. Establish a) conceptual designs for the construction and operation of a full-scale D-Factory demonstration facility, b) the basic functions of the high throughput platform and c) a thorough assessment of opportunities to improve materials and energy efficiency for the D-Factory demonstration facility, by month 36.
10. Provide a full sustainability assessment of the technological, environmental, economic and social sustainability including a SWOT analysis on strengths, weaknesses, opportunities and threats for establishing a Dunaliella production facility and D-Factory biorefinery for the production of named compounds from Dunaliella, including the sustainable management of by-products and wastes such as salt water by month 48.
11. Build an Innovation Platform to serve as an interactive forum able to capture the interests and activities of a wide range of stakeholders and integrate their local knowledge and experience with data from the D-Factory partners so that existing knowledge can be shared and new knowledge created.
12. Develop a comprehensive business case to show industrial investors the complete opportunity and scope for the creation of a Dunaliella microalgal bio-refinery and raise additional investment for the D-Factory demonstration.
13. Manage and disseminate information at the same time as protecting the continuing acquisition of IP.

Project Results:
Monzon Biotech has been incorporated as a beneficiary to develop the Demo: it has an onsite power plant emitting flue gas with ~6% CO2 from the combustion of natural gas, predator-free salt brine, water, energy and heat, available land, good communications and infrastructures, industrial environment, qualified staff and company commitment, as well as evidence for the growth of Dunaliella all year and no requirement for marine environment permits. A state-of the art raceway in Eilat supports harvesting trials using Evodos technology and provides data to inform development of the Monzon Demo plant. Both sites supply kg quantities of algal powder and paste for analysis and processing and both are well-placed to validate the concept of a microalgae biorefinery. The Eilat facility serves as a benchmark of algae cultivation in a raceway for sustainability assessment, but control of predators remains a problem.
22 monoclonal strains of D. salina have been isolated and preserved using cryogenics. Selected strains have been genotyped; CCAP19/30 was identified as a strain of D. tertiolecta and a new phylogenetic tree has been developed. The transcriptomes of two strains have been annotated through a series of Blast and KEGG metabolic pathway mapping protocols using expression profiles in log phase of growth and after transfer from low to high salt medium. The main factor influencing the amount of carotenoids produced was the algal strain. Increased salt in the extracellular medium increased glycerol yield but not for extended periods of growth.
In the green, non-carotenogenic stage, strains grew well in Green Wall Panels and tubular PBRs, but the orange carotenogenic stage was not as readily implemented. Pilot unit expansion construction has started and tests to cultivate orange cultures in tubular PBRs and inoculate a raceway in Monzon are underway.
Laboratory mutants were unable to compete with prevailing natural strains outdoors or to withstand environmental conditions. Use of herbicides continues under investigation.
Highly promising early results have been obtained for continuous harvesting using Evodos T50 spiral plate technology. Membrane ultrafiltration is under test to pre-concentrate carotenogenic biomass before centrifugation and trials are evaluating permeate reuse.
Extracts of carotenoids have been prepared from biomass using scCO2 and HPCCC methods and preparations with anti-diabetic, anti-inflammatory, anti-microbial and cytotoxic activity have been identified. Membrane resistance to solvents is being investigated. Positive acceptance results have been obtained for using residue after extraction in bread and fish sausage. An animal feed trial is now in planning. Carrier systems for carotenoid-rich extracts for incorporation in consumer products have been produced and tested. A powder with excellent free flowing properties has been produced.
Definitions, settings and system boundaries and scenarios include the demo facility. Three proposed scenarios and three conceptual processing flowsheets have been designed and integrated to provide basic mass, energy and water balances. Input blocks for screening and assessing value chains have been identified and the platform is now under development for the simultaneous evaluation of paths with respect to processing costs and product values. For the Demo, an optimization model for screening processing paths has been developed, but more reliable data are needed to provide realistic solutions.
Approximately 800 stakeholders have been mapped to assign levels of most/least importance and influence and develop the communication plan. The newly redesigned website includes a knowledge base aimed at integrating stakeholder partnerships and activities. Newsletters, press releases and oral presentations have disseminated outputs in Europe and internationally and IP has been monitored through 7 IP and steering committee meetings.

Potential Impact:
Expected Final Results:
A sustainable D-Factory demonstration in Europe that sets a world benchmark for a microalgae biorefinery based on biomass from halotolerant and halophilic microalgae.

Potential Impact and use
Global adoption of licenced microalgae biorefinery processes utilising non-potable saline waters for the production of nutraceuticals, pharmaceuticals, protein and green chemical feedstocks.

List of Websites:
www.d-factoryalgae.eu

Reported by

UNIVERSITY OF GREENWICH
United Kingdom
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