Macroalgae biorefinery Production of bioethanol and protein for fish feed frombrown seaweed, Laminaria digitata

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1 Macroalgae biorefinery Production of bioethanol and protein for fish feed frombrown seaweed, Laminaria digitata Anne-Belinda Bjerre, Chem Eng PhD Senior Scientist, Danish Technological Institute Adjunct Professor, Ålborg University Co-authors Karin Svane Bech, Xiarou Hou, Lars Nikolaisen

2 Welcome to Danish Technological Institute

3 Division for Energy and Climate Center for Biomass & Biorefinery 4 locations in Denmark Several projects related to solid and liquid biofuels EU-projects / National 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 Background Processes on macroalgae Cultivation Harvest Conditioning Enzymatic hydrolysis Ethanol production 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 latissima into energy carriers, together with a protein enriched fish feed derived as a residual from the energy conversion processes Financed: the Danish Strategic Research Council Total budget of 24 million DKK Project period: March 1 st, March 1 st, Partners from Denmark, Germany, Ireland, Italy Education of 4 PhD and 2 postdoc Coordinator: Danish Technological Institute v/ Anne-Belinda Bjerre

6 MAB3 Partners Danish Technological Institute (Coordinator) (DK) Aarhus University (2 institutes) (DK) Technical University of Denmark (3 institutes) National 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 participating in the advisory board]

7 MacroAlgaeBiorefinery

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

9 For comparison: Land-based biomass production Grain and straw 20 yields, tons ha Grain and straw (total)

10 Marine biomass production Seaweed høstudbytte, tons ha korn og halm

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

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

13 Proces trin til dyrkning af makroalger Eksempel: Laminaria digitata Collectiion of fertile seaweed Preparation of L. digitata culture under controlled lab conditions Spraying on strings Development of baby seaweeds under controlled conditions Deployments of strings to strong ropes in sea waters Downstream processing - Bioraffinering Harvesting by boat Cultivation of seaweed at sea (6-7 months)

14 Formering af 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 til henholdsvis hanlige og hunlige gametofytter. 3. På de hanlige gametofytter dannes hanlige gameter med flageller og de svømmer hen til den hunlige ubevægelige ægcelle (og så kaldet gamet). 4. Hunlige feromoner tiltrækker de hanlige gameter til 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.

15 Produktion af Laminaria digitata i laboratoriet 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 til henholdsvis hanlige og hunlige gametofytter. 3. På de hanlige gametofytter dannes hanlige gameter med flageller og de svømmer hen til den hunlige ubevægelige ægcelle (og så kaldet gamet). 4. Hunlige feromoner tiltrækker de hanlige gameter til 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 timers mørke ved 6 grader C

16 Spray på dyrkningssnore 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 til henholdsvis hanlige og hunlige gametofytter. 3. På de hanlige gametofytter dannes hanlige gameter med flageller og de svømmer hen til den hunlige ubevægelige ægcelle (og så kaldet gamet). 4. Hunlige feromoner tiltrækker de hanlige gameter til 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 un strings under controlled lab conditions Development in open tanks in laboratory 1 (-2) måneder

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

19 Harvest Most harvesting nowadays is carried out by hand on a ship. Needs further development for automatic methods to be mature.

20 CULTIVATION MAB3 10 km of seeded lines Saccharina latissima Laminaria digitata Deployed in September 2012 Line mussel system Limfjorden, Denmark

21 HARVEST MAB3 Saccharina latissima Harvest time: May 2013 Growth period: 7-8 months Yield: 2 wet tons of S. latissima (2 km line) Harvest technology: line-mussel cultivation

22 HARVEST Laminaria digitata Natural population 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% 75% Laminaria: 78% 10% Saccharina: 89% 9% Saccharina 89% 88%

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

26 PRETREATMENT AND ENZYMATIC HYDROLYSIS MAB3 Wet Brown Seaweed Wet-milled by Disc mill Collect fibers by centrifugation Enzymatic Hydrolysis 1.0 mm 0.2 mm (Picture: Dirk Manns DTU) Enzymatic hydrolysis (Picture: Annette Bruhn) Laminaria digitata (approx. 2m, DM 27%) (Picture: Dirk Manns DTU) Sprout-Bauer 12 Lab disc mill (disc distances 1.0 and (Picture: Stinus 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] 0.2mm) Disc distance 0.2 mm 1.0 mm Glucose [% dry fibers] 22.8 ± ± 0.7

27 3. generations bioethanol Oprensning af Fucoidan Forbehandling Fucoidan extract Enzymer Gær Fucoidan (value-added product) Protein C6 sukker Protein Hydrolyse Hexose fermentering Destillering EtOH

28 H 2 0 CO 2 3G Bioethanol Fotosyntesen glucan enzymer glucose gær ethanol + C0 2

29 Fermentation to ethanol: SSF-proces Stenhus Gymnasium Holbæk 25. september 2013

30 Protein Concentration (g/100 g material) 12 ETHANOL PRODUCTION MAB3 Exp ID Lam 1 Lam 2 Lam 3 Lam 4 Pretreatment Wet-milled by disc mill (0.2 mm) Wet-milled by disc mill (1 mm) Dried, grinded by pin-mill and screened (ø 1 mm) Dried, grinded by pin-mill and screened (ø 1 mm) Substrate DM (%, w/v) Glucose (g/l) BF 3,8 11,5 AF 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 matter) Conditioning 1 kg Value added products (Fucoidan, etc.) 7,4 kg Ethanol + 7,1 kg CO 2 1 kg Protein Pretreatment C6-Fermentation Separation of liquid and solid Input: construction and building materials, enzymes, energy and electricity, water MAB3 MacroalgaeBiorefinery Output CO2eq Other output Emission in air, water and soil Enzymatic prehydrolysis Bioethanol Proteins 2 kg Amino acids Residual sugars fermentation 3 kg fertilizer (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 Fertilizers 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 communication.

33 ENVIRONMENTAL SUSTAINABILITY ASSESSMENT MAB3 Input: cultivation system (lines, buoys, water, nutrients) GHG savings on the climate mitigation bank account Macroalgae cultivation and harvesting Output: CO2eq Other outputs: Emissions in air and water Algae Amounts (g/kg) Macroalgae carbon content Macroalgae nitrogen content 3-5 CO 2 assimilation Avoided N 2 O emission from N assimilation 4-8 Total CO 2 eq According to IPCC guidelines

34 Conclusions Macroalgae cultivation has high potential for CO 2 saving providing water quality protection by assimilating excess nutrients Raw material price is essential for the overall feasibility Knowledge about biomass composition is important for the choice of biorefinery processes Ethanol and protein are realistic products from Laminaria digitata, however the algal biomass must be harvested at the right time 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 information: Thanks to co-authors: Xiaoru Hou, Karin Svane Bech, Jonas Høeg Hansen, Annette Bruhn, Bodo Saake, Anne Meyer, Dirk Manns, Michael Bo Rasmussen, Mette Nielsen, Jens Kjerulf Petersen, Ditte Tørring, Peter Daugbjerg Jensen, Michele Seghetta, Simone Bastanoni, Marianne Thomsen

36 DTI CONTACTS: Anne-Belinda Bjerre (Senior Scientist, Senior Consultant): Coordinator of MAB3 bioethanol design, pretreatment and enzymatic hydrolysis Biorefinery Lars Steenberg Nikolajsen (Senior Consultant): Macroalgae cultivation and harvesting Conditioning and Storage Karin Svane Bech (Consultant): Macroalgae cultivation Newsletter and dissemination Xiaoru Hou (Post Doc): Ethanol fermentation (C6 and C5)