Cultivation of Dunaliella in closed photobioreactors in the
Cultivation of Dunaliella in closed photobioreactors in the scope of the D-Factory project Joana Galante 1, Pedro Nascimento 1, Celina Parreira 1, Rui Mota 1, Filipe Semião 1, Diana Fonseca 1, Diana Arroyo 2, Belen Perna 2, Laura Sanchez 2, Alberto Contreras 2, Javier Gomez 2, Luís Costa 1, Vítor Verdelho Vieira 1, Ami Ben-Amotz 3 1 A 4 F – Algae for future (Portugal); 2 Monzón Biotech (Spain); 3 Nature Beta Technologies Ltd. (Israel) Brussels, Belgium 18 October 2017
Cultivation of Dunaliella in closed photobioreactors In the D-Factory project, A 4 F with Monzon Biotech deployed closed PBR technology to cultivate Dunaliella salina in attempts to allow better control and achieve higher quality of the final product. Advantages: Challenges: • Prevent or minimize certain contaminants’ growth; • Higher Cap. Ex; • Better control of parameters (p. H, CO 2, temperature, light); • Cultivation challenges for carotenogenesis; • Lower CO 2 losses; • Formation of fungal & bacteria biofilms during summer term PBR cultivations; • Reduce water evaporation; • Allow cultivation in any climate; • Can use GMOs; • Higher Op. Ex; • Shear forces of pumping/mixing taking into account Dunaliella cells fragility. • Can use herbicides/other chemicals in controlled environment; • Can grow non-dominant local strains; • Produce high quality inoculum for cultivation in raceways. 17 -18/10/2017 Algae Biorefineries for Europe – Brussels, Belgium 2
Dunaliella salina – Cultivation challenges Halotolerant green microalgae; Fragile, biflagellated and cell wall-less → more susceptible to the shear forces of the pump of a tubular PBR; Severe conditions for carotenogenesis: • • • NO 3 depletion; High salinity; High light Michele Greque de Morais, Bruna da Silva Vaz, Etiele Greque de Morais, and Jorge Alberto Vieira Costa, “Biologically Active Metabolites Synthesized by Microalgae, ” Bio. Med Research International, vol. 2015, Article ID 835761, 15 pages, 2015. β-carotene Glycerol compounds of commercial value Dunaliella and salt crystal EM of Dunaliella bardawil (Avron & Ben-Amotz) 17 -18/10/2017 Algae Biorefineries for Europe – Brussels, Belgium 3
Dunaliella products It is possible to cultivate Dunaliella in 3 different growth stages (green, brown and orange) in closed PBRs. Get different compounds of commercial value 17 -18/10/2017 Algae Biorefineries for Europe – Brussels, Belgium Analyse the potential of the different products 4
Cultivation of Dunaliella in closed PBR in the past Most work performed on lab scale… → Innovative potential at pilot scale! 17 -18/10/2017 Algae Biorefineries for Europe – Brussels, Belgium 5
Cultivation of Dunaliella in closed PBRs in the past Industrial archaeology (failure) - Murcia, Spain, 1990 6
PBR Cultivation technologies tested for Dunaliella More than 50 PBR cultivation trials were performed so far in the D-Factory. Flat panel PBR (Green-walls) 7 17 -18/10/2017 Algae Biorefineries for Europe – Brussels, Belgium
PBR Cultivation technologies tested for Dunaliella More than 50 PBR cultivation trials were performed so far in the D-Factory. Flat panel PBR (Green-walls) 8 17 -18/10/2017 Algae Biorefineries for Europe – Brussels, Belgium
PBR Cultivation technologies tested for Dunaliella Multilayer horizontal tubular PBR 9 17 -18/10/2017 Algae Biorefineries for Europe – Brussels, Belgium
Multilayer horizontal tubular PBR for Dunaliella cultivation ü ü X X X Pilot scale system (30 m 2) tested with Dunaliella (A 4 F & MZN) Higher control of certain contaminants possible Carotenogenic cultivation for several months with possible cycles of harvest Productivity data are on process, with spring high cell density Productivity data not consistent yet 300 m 2 (industrial scale) MHT-PBR trials required Several years of cultivation trials required 17 -18/10/2017 Algae Biorefineries for Europe – Brussels, Belgium 10
Flat panel PBRs (Green-walls) for Dunaliella cultivation ü ü Commercial scale module (30 m 2) tested with Dunaliella (A 4 F & MZN) Higher control of certain contaminants possible Carotenogenic cultivation for one years with periodic non consistent cycles of harvest Productivity data are in process, with high spring cell density, consistently 11 17 -18/10/2017 Algae Biorefineries for Europe – Brussels, Belgium
Achievements and Conclusions • Dunaliella production requires careful control of many interconnected culture parameters: light, cell density, N concentration, salinity, temperature, shear stress, micronutrients, etc. ; • Control of certain contaminants with temperature modification was possible; • Culture medium optimization is still possible and greatly influences productivity results; • PBR is expected to require approximately 3 ton CO 2 per ton of AFDW biomass; • Full data sets will be analysed at the end of the project: productivity, energy requirements, Op. Ex, etc. ; 12 17 -18/10/2017 Algae Biorefineries for Europe – Brussels, Belgium
Achievements and Conclusions • Different cultivation conditions: • 3 Dunaliella strains; • 3 growth stages (green, brown and orange); • 2 regimes (batch and semi-continuous); • 2 types of closed PBRs; • 2 locations. • Cultivations in carotenogenesis endured for up to 6 months in the flat panel PBRs in semi-continuous regime; • Very high spring but not summer cell densities of carotenogenic Dunaliella achieved in closed PBRs; • Still a lot of challenges to pursue, namely regarding carotenogenesis, internal biofilm and data to collect about consistency of cultivation results throughout consecutive seasons and years. 13 17 -18/10/2017 Algae Biorefineries for Europe – Brussels, Belgium
Achievements and Conclusions • Different products for niches: green Dunaliella for enzymes, bioactive peptides, etc. ; potentially to be explored: GMO strains, cultivation under chemical induction of target compounds, control of the isomer ratio of beta-carotene; • An acknowledgement to the full consortium of the D-Factory and especially to the team work of the partners which enabled significant progress of Dunaliella cultivation in PBRs: • NBT: knowledge and experience; • A 4 F: audacity and ingenuity; • MZN: hardwork and unique conditions. 14 17 -18/10/2017 Algae Biorefineries for Europe – Brussels, Belgium
Thank you! Questions…? Joana Galante 1, Pedro Nascimento 1, Celina Parreira 1, Rui Mota 1, Filipe Semião 1, Diana Fonseca 1, Diana Arroyo 2, Belen Perna 2, Laura Sanchez 2, Alberto Contreras 2, Javier Gomez 2, Luís Costa 1, Vítor Verdelho Vieira 1, Ami Ben-Amotz 3 1 A 4 F – Algae for future (Portugal); 2 Monzón Biotech (Spain); 3 Nature Beta Technologies Ltd. (Israel) Brussels, Belgium 18 October 2017 15
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