Macroalgae biorefinery Produc3on of bioethanol and protein for fish feed from brown seaweed, Laminaria digitata

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1 Macroalgae biorefinery Produc3on of bioethanol and protein for fish feed from brown seaweed, Laminaria digitata Anne- Belinda Bjerre, Chem Eng PhD Senior Scien)st, Danish Technological Ins)tute Adjunct Professor, Ålborg University Co- authors: Karin Svane Bech, Xiarou Hou, Lars Nikolaisen STAR*workshop Foggia 2014

2 Welcome to Danish Technological Ins3tute STAR*workshop Foggia 2014

3 Division for Energy and Climate Center for Biomass & Biorefinery 4 locahons in Denmark Several projects related to solid and liquid biofuels EU- projects / NaHonal projects / Industry Denmark Grenå Algae Center Denmark Aarhus Biomass Laboratory and Offices Taastrup Enzyme Laboratory and Offices Sdr. Stenderup Pilot plant and production facilities

4 Contents Ø Background Ø Processes on macroalgae CulHvaHon Harvest CondiHoning EnzymaHc hydrolysis Ethanol produchon Ø Sustainability and economic feasibility

5 BACKGROUND the MAB3 project MacroAlgaeBiorefinery for 3G bioenergy and fish feed An integrated biorefinery concept to be developed for conversion of two species of macro algae Laminaria digitata and Saccharina la)ssima into energy carriers, together with a protein enriched fish feed derived as a residual from the energy conversion processes Ø Financed: the Danish Strategic Research Council Ø Total budget of 24 million DKK Ø Project period: March 1 st, March 1 st, 2016 Ø 12 Partners from Denmark, Germany, Ireland, Italy Ø EducaHon of 4 PhD and 2 postdoc Ø Coordinator: Danish Technological InsHtute v/ Anne- Belinda Bjerre

6 MAB3 Partners Danish Technological InsHtute (Coordinator) (DK) Aarhus University (2 inshtutes) (DK) Technical University of Denmark (3 inshtutes) NaHonal University of Ireland, Galway (IRL) University of Hamburg (DE) University of Sienna (IT) Orbicon (DK) DONG Energy (DK) Aller Aqua (DK) VitaLys (DK) DanGrønt Products (DK) Affiliated partner: Novozymes (DK) [delivery of enzymes and parhcipahng in the advisory board]

7 MacroAlgaeBiorefinery

8 Why Algae? Growth rate of up to 0,35 day days mes increase of the biomass 1 cm² 1 m²

9 For comparison: Land- based biomass produc3on Grain and straw 20 Biomass yield, tons ha Grain and straw (total)

10 Marine biomass produc3on Seaweed høstudby_e, tons ha korn og halm

11 Marine biomass produc3on Seaweed Total produc3on yield, tons ha Grain and straw seaweed

12 Which species are of relevance? Experiences from the North AtlanHc Sea show promising results from Laminaria digitata Fingertang (a) Saccharina la)ssima Sukkertang (b) Palmaria palmata Søl (c) c a b

13 Proces steps for culhvahon of seaweeds: Ex: Laminaria digitata CollecHion of fer3le seaweed PreparaHon of L. digitata culture under controlled lab condi3ons Spraying on strings Development of baby seaweeds under controlled condihons Deployments of strings to strong ropes in deep sea waters Downstream processing - Bioraffinering HarvesHng by boat CulHvaHon of seaweed at sea (6-7 months)

14 Sexual breeding of Laminaria digitata Kønnet formering: 1. Der dannes sporer i specielle celler på både over- og underside på bladet. Disse områder kaldes sori, og som fremstår velafgrænsede og mørke. 2. Når sporerne frigives (sporolering) omdannes de Hl henholdsvis hanlige og hunlige gametofyqer. 3. På de hanlige gametofyier dannes hanlige gameter med flageller og de svømmer hen Hl den hunlige ubevægelige ægcelle (og så kaldet gamet). 4. Hunlige feromoner Hltrækker de hanlige gameter Hl ægcellen, og der sker en befrugtning. 5. Det befrugtede æg zygoten kan spire på fast materiale direkte på sten eller et f.eks. et reb. STAR*workshop Foggia 2014

15 1st step in culhvahon of Laminaria digitata in laboratory Kønnet formering: 1. Der dannes sporer i specielle celler på både over- og underside på bladet. Disse områder kaldes sori, og som fremstår velafgrænsede og mørke. 2. Når sporerne frigives (sporrolering) omdannes de Hl henholdsvis hanlige og hunlige gametofyqer. 3. På de hanlige gametofyier dannes hanlige gameter med flageller og de svømmer hen Hl den hunlige ubevægelige ægcelle (og så kaldet gamet). 4. Hunlige feromoner Hltrækker de hanlige gameter Hl ægcellen, og der sker en befrugtning. 5. Det befrugtede æg zygoten kan spire på fast materiale direkte på sten eller et f.eks. et reb. Udvikling i 3-5 måneder i laboratoriet, der kræves mindst 12 Hmers mørke ved 6 grader C

16 Spraying on culhvahon- strings Kønnet formering: 1. Der dannes sporer i specielle celler på både over- og underside på bladet. Disse områder kaldes sori, og som fremstår velafgrænsede og mørke. 2. Når sporerne frigives (sporolering) omdannes de Hl henholdsvis hanlige og hunlige gametofyqer. 3. På de hanlige gametofyier dannes hanlige gameter med flageller og de svømmer hen Hl den hunlige ubevægelige ægcelle (og så kaldet gamet). 4. Hunlige feromoner Hltrækker de hanlige gameter Hl ægcellen, og der sker en befrugtning. Det befrugtede æg zygoten kan spire på fast materiale direkte på sten eller et f.eks. et reb.

17 Development of seaweed adsporbed to strings under controlled lab condihons Development in open tanks in laboratory 1 (- 2) måneder

18 Deployment of culhvahon strings in Autumn at sea DK potenhale: ~ 2-10 tons/ha (tørvægt)

19 Harvest Most harvests nowadays are carried out by hand on a ship. Needs further development for automahc methods to be mature.

20 CULTIVATION MAB3 10 km of seeded lines Saccharina la)ssima Laminaria digitata Deployed in September 2012 Line mussel system Limmorden, Denmark

21 HARVEST MAB3 Saccharina la)ssima Harvest Hme: May 2013 Growth period: 7-8 months Yield: 2 wet tons of S. la)ssima (2 km line) Harvest technology: line- mussel culhvahon

22 HARVEST Laminaria digitata Natural populahon 300 kg August 2012 Adams et al., Bioresource Technology 102 (2011)

23 RAW MATERIAL CHARACTERISATON Species Harvesting Time Protein (%) Sulphated fucoidan (%) Mannitol (%) Glucose (%) Minerals (%) Xylose +Mannose (%) Residues (%) SUM (%) Laminaria digitata Laminaria digitata Saccharina latissima ,

24 CONDITIONING MAB3 Screw Pressing - Algae juice Drying Water content Water content 50 C (4 days): Laminaria 78% Saccharina 89% 75% 88% Laminaria: 78% à 10% Saccharina: 89% à 9%

25 PRETREATMENT AND ENZYMATIC HYDROLYSIS Laminaria digitata (harvested in August 2012) Wet Brown Seaweed Dried and Grinded Re- adjust DM by water Enzyma3c Hydrolysis Wet Brown Seaweed Wet- milled by Disc mill Collect fibers Enzyma3c Hydrolysis

26 PRETREATMENT AND ENZYMATIC HYDROLYSIS MAB3 Wet Brown Seaweed Wet- milled by Disc mill Collect fibers by centrifug- a3on Enzyma3c Hydrolysis 1.0 mm 0.2 mm (Picture: Dirk Manns DTU) EnzymaHc hydrolysis (Picture: Anneie Bruhn) Laminaria digitata (approx. 2m, DM 27%) (Picture: Dirk Manns DTU) Sprout- Bauer 12 Lab disc mill (disc distances 1.0 and 0.2mm) (Picture: SHnus Andersen DTU) ph 5.1, T 40 C, t 72h, 4% [S]/[V], CellicCTec2 (Novozymes): 5% [E]/[S] Alginate Lyase (EC ): 0.25% [E]/[S] Disc distance 0.2 mm 1.0 mm Glucose [% dry fibers] 22.8 ± ± 0.7

27 3. generahon bioethanol Recovery of Fucoidan pretreatment Fucoidan (value- added product) Fucoidan extract Enzymes yeast Protein C6 sugars Protein Hydrolysis Hexose fermentation Destillation EtOH

28 H20 3G Bioethanol CO2 Fotosyntesen glucan glucose enzymer gær ethanol + C02 STAR*workshop Foggia 2014

29 FermentaHon to ethanol: SSF- proces

30 ETHANOL PRODUCTION MAB3 Exp ID Lam 1 Lam 2 BF AF Lam 3 Lam 4 Pretreatment Wet-milled by disc Wet-milled by disc mill (1 mill (0.2 mm) mm) Dried, grinded by pin-mill and screened (ø 1 mm) Substrate DM (%, w/v) Glucose (g/l) Protein Concentra3on (g/100 g material) ,8 11,5 Dried, grinded by pin-mill and screened (ø 1 mm) Enzyme loading CellicCtec2: 5 % v/w [E]/[S] Alginate Lyase (EC ): 0.25 % v/w [E]/[S] CellicCtec2: 5 % v/w [E]/[S] Alginate Lyase (EC ): 0.25 % v/w [E]/ [S] Celluclast 1.5L: 40 U/g DM Alginate lyase (EC ): 2 U/g DM Celluclast 1.5L: 40 U/g DM Alginate lyase (EC ): 2 U/g DM Hydrolysis temperature ( C) Yeast inoculation conc. (g/ L) Fermentation temperature ( C) Ethanol yield (% theoretical value), after 48 h fermentation Ethanol concentration (g/l)

31 ECONOMIC POTENTIAL 100 kg wet algae biomass (Laminaria digitata) 2 kg Algae juice (dry maier) CondiHoning 1 kg Value added products (Fucoidan, etc.) 7,4 kg Ethanol + 7,1 kg CO 2 1 kg Protein Pretreatment EnzymaHc pre- hydrolysis C6- FermentaHon SeparaHon of liquid and solid Input: construchon and building materials, enzymes, energy and electricity, water MAB3 MacroalgaeBiorefinery Output CO2eq Other output Emission in air, water and soil Bio- ethanol Proteins 2 kg Amino acids Residual sugars fermentahon 3 kg ferhlizer (inorganic salts and silicium) Residuals

32 ECONOMIC POTENTIAL MAB3 Price ( /kg) Weight Scenario Scenario Scenario Scenario (kg) 1 a 2 b 1 2 Wet algae Cost Value added products (e.g. Fucoidan) Ethanol Protein Income Amino acids Fer3lizers Margin a Scenario 1: Price of macroalgae from Watson, L. and Dring, M., Business plan for the establishment of a seaweed hatchery & grow- out farm. Irish sea Fisheries Board, pp 41. B Scenario 2: Price of macroalgae from Michael Bo Rasmussen personal communicahon.

33 ENVIRONMENTAL SUSTAINABILITY ASSESSMENT MAB3 Input: cul3va3on system (lines, buoys, water, nutrients) GHG savings on the climate mi3ga3on bank account Amounts (g/kg) Macroalgae cul3va3on and harves3ng Algae Macroalgae carbon content Macroalgae nitrogen content CO 2 assimilahon Output: CO2eq Other outputs: Emissions in air and water Avoided N 2 O emission from N assimilahon Total CO 2 eq According to IPCC guidelines

34 Conclusions Macroalgae culhvahon has high potenhal for CO 2 saving providing water quality protec3on by assimilahng excess nutrients Raw material price is essen3al for the overall feasibility Knowledge about biomass composi3on is important for the choice of biorefinery processes Ethanol and protein are realishc products from Laminaria digitata, however the algal biomass must be harvested at the right 3me in August/September due to the need of high C6- sugar contents

35 ACKNOWLEDGEMENT Thanks to the Danish Council for Strategic Research for financing the MAB3 project More informahon:

36 DTI CONTACTS: Anne- Belinda Bjerre (Professor, Chem. Eng. PhD): Coordinator of MAB3 bioethanol design, pretreatment and enzymahc hydrolysis Biorefinery Lars Steenberg Nikolajsen (Senior Consultant): Macroalgae culhvahon and harveshng CondiHoning and Storage Karin Svane Bech (Consultant): Macroalgae culhvahon Newsleier and disseminahon Xiaoru Hou (Post Doc): Ethanol fermentahon (C6 and C5)

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