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

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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

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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

Grenå – Algae Center Denmark

Aarhus – Biomass Laboratory and Offices

Sdr. Stenderup – Pilot plant and production facilities

Taastrup – Enzyme Laboratory and Offices

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Ø

 

Background

Ø

 

Processes on macroalgae

CulHvaHon

Harvest

CondiHoning

EnzymaHc hydrolysis

Ethanol producHon

Ø

 

Sustainability and economic feasibility

M

acro

A

lgae

B

ioreﬁnery for

3 G 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 1st, 2012 -‐-‐-‐-‐ March 1st, 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

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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]

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Why Algae? Growth rate of up to 0,35 day

-‐1

1 cm²

1 m² 9.000 3mes increase of the biomass

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For comparison: Land-‐based biomass

produc3on

Grain and straw

Biom ass yie ld , tons ha -‐1

Grain and straw (total)

20

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Marine biomass produc3on Seaweed høstudby_ e, tons ha -‐1

korn og halm

20 10 høstudby_ e, tons ha -‐1

korn og halm

20

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Marine biomass produc3on Seaweed To tal pr oduc 3on yie ld , tons ha -‐1

Grain and straw

20

10 20

10

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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)

a

c b

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CollecHion of

fer3le seaweed

Proces steps for culHvaHon of seaweeds:

Ex: Laminaria digitata

Spraying on strings Downstream processing -‐ Bioraﬃnering HarvesHng by boat CulHvaHon of seaweed at sea

(6-‐7 months)

Deployments of strings to strong ropes in deep sea

waters Development of baby seaweeds under controlled condiHons PreparaHon of L. digitata culture under controlled lab condi3ons

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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.

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1st step in culHvaHon of

Laminaria digitata

in

laboratory

velafgrænsede og mørke. 2.  Når sporerne frigives

(sporrolering) omdannes de Hl henholdsvis hanlige og hunlige

gametofyQer.

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.

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Spraying on culHvaHon-‐strings

velafgrænsede og mørke. 2.  Når sporerne frigives

(sporolering) omdannes de Hl henholdsvis hanlige og hunlige

gametofyQer.

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.

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Development of seaweed adsporbed to strings

under controlled lab condiHons

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Deployment of culHvaHon strings in Autumn at sea

DK potenHale: ~ 2-‐10 tons/ha (tørvægt)

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Harvest

Most harvests nowadays are carried out by hand on a ship.

Needs further development for

automaHc methods to be mature.

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• 10 km of seeded lines

–

 

Saccharina la)ssima

–

 

Laminaria digitata

• Deployed in September 2012

–

 

Line mussel system

–

 

Limmorden, Denmark

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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

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Laminaria digitata

• Natural populaHon

• 300 kg

• August 2012

HARVEST

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RAW MATERIAL CHARACTERISATON

Species Harvesting Time Protein (%) Sulphated fucoidan (%) Mannitol (%) Glucose (%) Minerals (%) Xylose +Mannose (%) Residues (%) SUM (%) Laminaria digitata 2012.04. 14.6 6.8 3.7 7.7 31.8 1.2 5,5 71.3 Laminaria digitata 2012.08. 3.9 3.5 6.4 56.9 8.1 0.5 2.7 82.1 Saccharina latissima 2013.05. 20.1 4.3 5.1 5.8 35.4 0.6 8.6 79.9

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• Screw Pressing

-‐Algae juice

• Drying

Laminaria 78% 75% Saccharina 89% 88% 50°C (4 days): Laminaria: 78% à 10% Saccharina: 89% à 9%

CONDITIONING MAB3

Water content Water content

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Wet Brown Seaweed Dried and Grinded Re-‐adjust DM by water Enzyma3c Hydrolysis

PRETREATMENT AND ENZYMATIC HYDROLYSIS

Wet Brown Seaweed Wet-‐ milled by Disc mill Collect

ﬁbers Hydrolysis Enzyma3c

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Laminaria digitata

(approx. 2m, DM 27%) (Picture: Anneie Bruhn)

Sprout-‐Bauer 12” Lab disc mill

(disc distances 1.0 and 0.2mm)

(Picture: Dirk Manns DTU)

PRETREATMENT AND ENZYMATIC HYDROLYSIS MAB3

Wet Brown

Seaweed by Disc mill Wet-‐milled

Collect ﬁbers by centrifug-‐ a3on Enzyma3c Hydrolysis Disc distance 0.2 mm 1.0 mm

Glucose [% dry ﬁbers] 22.8 ± 0.9 32.7 ± 0.7

1.0 mm

(Picture: SHnus Andersen DTU)

0.2 mm

EnzymaHc hydrolysis

(Picture: Dirk Manns DTU)

pH 5.1, T 40 ̊C, t 72h, 4% [S]/[V], CellicCTec2 (Novozymes): 5% [E]/[S] Alginate Lyase (EC 4.2.2.3 ): 0.25% [E]/[S]

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pretreatment Hydrolysis Hexose fermentation Destillation

EtOH

3. generaHon bioethanol

Fucoidan extract Protein C6 sugars Recovery of Fucoidan Enzymes yeast Fucoidan (value-‐added product) Protein

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Fotosyntesen H₂0 CO₂ glucan enzymer glucose gær ethanol + C0₂

3G Bioethanol

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FermentaHon

to ethanol:

SSF-‐proces

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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) 4 4 5 10 Glucose (g/L) _9.12 _13.1 _28.5 _56.9

Enzyme loading CellicCtec2:

5 % v/w [E]/[S] Alginate Lyase (EC 4.2.2.3): 0.25 % v/w [E]/[S]

CellicCtec2: 5 % v/w [E]/[S] Alginate Lyase (EC 4.2.2.3): 0.25 % v/w [E]/ [S]

Celluclast 1.5L: 40 U/g DM

Alginate lyase (EC 4.2.2.3):

2 U/g DM

Celluclast 1.5L: 40 U/g DM

Alginate lyase (EC 4.2.2.3):

2 U/g DM

Hydrolysis temperature (°C) 40 40 40 40

Yeast inoculation conc. (g/ L)

2 2 2 2

Fermentation temperature (°C)

32 32 32 32

Ethanol yield (% theoretical value), after 48 h fermentation 73 77 73 75 Ethanol concentration (g/L) _3.4 _5.2 _10.5 _21.8 0 2 4 6 8 10 12 BF AF 3,8 11,5 Pro tei n Co ncen tra 3o n (g /100 g m ate rial)

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100 kg wet algae biomass (Laminaria digitata)

CondiHoning

Pretreatment EnzymaHc pre-‐

hydrolysis C6-‐FermentaHon SeparaHon of liquid

and solid Residual sugars fermentaHon 1 kg Value added products (Fucoidan, etc.) 7,4 kg Ethanol + 7,1 kg CO₂ 1 kg Protein 2 kg Amino acids Residuals 3 kg ferHlizer

Input: construcHon and building materials, enzymes, energy and

electricity, water

Other output Emission in air,

water and soil

MAB3 MacroalgaeBioreﬁnery Output CO2eq Bio-‐ ethanol Proteins 2 kg Algae juice (dry maier)

ECONOMIC POTENTIAL

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Price (€/kg) €

Weight (kg) Scenario 1a Scenario ₂b Scenario ₁ Scenario ₂

Wet algae 100.0 1.12 0.08 112 8 Cost

Value added products (e.g.

Fucoidan) 1 2.9 2.9 2.9 2.9 Income Ethanol 7.4 1 1 7.4 7.4 Protein 1.0 1.5 1.5 1.5 1.5 Amino acids 2.0 1 1 2 2 Fer3lizers 3.0 0.35 0.35 1.05 1.05 -‐97.15 6.85 Margin

a _{Scenario 1: Price of macroalgae from}_{Watson, L. and Dring, M., 2011.} 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.

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Input: cul3va3on system (lines, buoys, water,

nutrients)

Other outputs: Emissions in air and water Macroalgae cul3va3on and harves3ng Output: CO2eq Algae Amounts (g/kg)

Macroalgae carbon content 43-‐50 Macroalgae nitrogen content 3-‐5

CO₂ assimilaHon 158-‐182

Avoided N₂O emission from N assimilaHon 4-‐8

Total CO₂ eq 1524-‐2519

GHG savings on the climate mi3ga3on bank account

According to IPCC guidelines

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• 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

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ACKNOWLEDGEMENT

Thanks to the Danish Council for Strategic Research for ﬁnancing

the MAB3 project

More informaHon: www.mab3.dk

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