Conversion of

cellulose,
hemicellulose and
lignin into platform
molecules:
biotechnological
approach
EuroBioRef
Summer school
Lecce, Italy
18-24 September 2011

Anders Frlander
Gudbrand Rdsrud
Borregaard Industries Ltd,
Norway

Outline
1. Introduction
2. History of second generation bioethanol production
3. Worlds most advanced biorefinery history and learning points
4. Lignocellulosic biomass
5. Biorefinery options
6. The biochemical route (sugar plattform)
7. Pretreatment processes
8. Hydrolysis of cellulose
9. Anaerobic and aerobic fermentation
10. Lignin options
11. Hemicellulose/pentose options
12. Process integration & closing remarks

Critical sources to replace fossile sources and

reduce CO2 footprint
Agricultural products

Food

Lignocellulose
Algae
Organic waste
Metals & minerals

Feed

Plastics (Materials)
Chemicals

Green electricity
Hydropower
Solar Power
Wind power

Transport
Building materials

Geo-thermal
Nuclear power

Gas and petroleum

Coal

Mechanical power
Heat

Outline
1. Introduction
2. History of second generation bioethanol production
3. Worlds most advanced biorefinery history and learning points
4. Lignocellulosic biomass
5. Biorefinery options
6. The biochemical route (sugar plattform)
7. Pretreatment processes
8. Hydrolysis of cellulose
9. Anaerobic and aerobic fermentation
10. Lignin options
11. Hemicellulose/pentose options
12. Process integration & closing remarks

Sulfite ethanol production all started in Sweden

The worlds first sulfite ethanol plant

The inventors of sulfite ethanol production

Gsta Ekstrm

Skutskr sulfite ethanol plant in

Sweden started operation 1909

Hugo Wallin

Experimentation with
fermentation of spent sulfite
liquor (SSL) started around 1903
They soon found out they had to
neutralize with lime

Source: Persson, Bertil. Sulfitsprit. Frhoppningar och besvikelser under 100 r. Bjsta : DAUS Tryck & Media, 2007. ISBN: 91
7542 258-1.

Sugar composition of spent sulfite liquor (SSL) from sulfite pulping

Sulfite
cooking

Filtration

Fibre

SSL

Monosaccharide

% of DM in
SSL from
Eucalyptus

% of DM in
SSL from
Spruce

Arabinose

(C5)

0,3

0,8

Xylose

(C5)

21,9

5,3

Galactose

(C6)

1,6

2,1

Rhamnose (C6)

0,6

0,2

Glucose

(C6)

1,6

3,7

Mannose

(C6)

1,0

14,6

Spruce SSL
20,6% of DM is C6 sugars
77% of sugars are C6 sugars
Eucalyptus SSL
22,1% of DM is C5 sugars
82% of sugars are C5 sugars

33 sulfite ethanol plants in Sweden from 1909 until today

First sulfite ethanol plant ever

opened 1909 in Skutskr, Sweden
33 plants have been in operation
in Sweden
Only one in operation after 1983:
Domsj, capacity of 15 000 m3/y

Source: Persson, Bertil. Sulfitsprit. Frhoppningar och besvikelser under 100

r. Bjsta : DAUS Tryck & Media, 2007. ISBN: 91 7542 258-1.

17 sulfite ethanol plants in Finland 1927 - 1977

Sulfite ethanol production
was stopped in 1977
The last sulfite mill in
Finland stopped production
in the early 1990ies

Source:
1. Biorefining in the pulp and paper industry.
Niemel, Klaus. Flensburg : s.n., 2008. 5th European
Biorefinery Symposium.
2. Kaukoranta, Antti. Sulfittispiriteollisuus Suomessa
vuosina 1918-1978 (Eng:"Sulphite alcohol industry in
Finland in 1918-1978"). s.l. : Paino Polar Oy, 1981.
ISBN 951-9479-25-2.
3. Niemel, Klaus. Private communication. s.l. : VTT
TECHNICAL RESEARCH CENTRE OF FINLAND , 2010.

Sulfite ethanol plants in Central Europe

Attizholts (later Borregaard) in Switzerland
Production from 1912 to 2008
Capacity 13 mill litres
Also produced yeast and yeast extracts
M-Real in Hallein in Austria
Sulfite ethanol production 1941 1988
Capacity 6 mill litres
Evaluating to restart production in 2016
Kirov only plant still in operation in Russia

Source:

1) Borregaard internal files

2) Conference Austria April 2011
3) IEA Report: Status of 2nd Generation Biofuels Demonstration Facilities in June 2010, A REPORT TO IEA
BIOENERGY TASK 39

Sulfite ethanol plants in USA

Georgia Pacific
Bellingham mill produced ethanol from 1976
2001
Capacity 24 million liters

Source:

1) Katzen customer reference list (http://www.katzen.com/projects.html)

2) Borregaard internal files
3) Graf and Koehler, June 2000, OREGON CELLULOSE-ETHANOL STUDY, An evaluation of the potential for
ethanol production in Oregon using cellulose-based feedstocks.

Hydrolysis of wood for ethanol, SCP and furfural

Initially developed in Germany around
1900. Yields up to 190 L/mt dry wood
Used in the USA during World War I and II
Converted further to butadien for
rubber during WW II
USSR 1935 1985: Construction of
18 Ethanol plants,
16 SCP yeast plants
15 furfural/xylitol plants
Feedstock hardwood:softwood 6:4
Technology: weak sulfuric acid (130
150C), 1 or 2 step hydrolysis
None are profitable without subsidies
Sources:
Wood hydrolysis industry in the Soviet Union and Russia: What can be learned from the history?
Rabinovich, M.L. Helsinki, September 2009. The 2nd Nordic Wood Biorefinery Conference (NWBC-2009),
111-120.
Wikipedia contributors. Cellulosic ethanol. Wikipedia, The Free Encyclopedia. March 2, 2011, 16:08 UTC.
Available at: http://en.wikipedia.org/w/index.php?title=Cellulosic_ethanol&oldid=416750931. Accessed
March 8, 2011.

USSR wood hydrolysis plants 1935 Production of ethanol, SCP and furfural

Borregaard
worlds largest producer of 2nd gen bioethanol
BRG capacity 20 mill litres of
bioethanol pr year
1/3 as 99,5% and
2/3 as 96%
From hemicellulose from spruce
in SSL (spent sulfite liquor)
Production started 1938
Yeast strain: Bakers yeast,
Saccharomyces cerevisiae
Adapted to industrial SSL
continuously since 1938

Comparison of CO2 footprint of ethanol produced in different ways

Source:
1. Brekke, A., Modahl, I.S. and Raadal, H.L. Konkurrentanalyser for cellulose, etanol, lignin og vanillin fra Borregaard (Eng: Competitive CO2 footprint analysis
for cellulose, ethanol, lignin and vanillin from Borregaard). Fredrikstad : Ostforld Research, Des. 2008. Confidential report. Will be published.
2. Sutter, J. Life cycle inventories of petrochemical solvents. [red.] H.-J., Chudacoff, M., Hischier, R. Jungbluth, N., Osses, M. and Primas, A. Althaus. Life cycle
inventories of chemicals. Final report ecoinvnet data v2.0. Duebendorf and St. Gallen : Swiss Centre for LCI, Empa - TSL, 2007, Vol. 8 / 22.
3. Jungbluth, N., Chudacoff, M., Dauriat, A., Dinkel, F., Doka, G., Faist Emmenegger, M., Gnansounou, E., Kljun, N., Speilmann, M., Stettler, C. and Sutter, J.
Life cycle inventories of bioenergy. Final report ecoinvnet v2.0. Volume 17. . Duebendorf and Uster : Swiss Centre for LCI, ESU, 2007.

Sulfite ethanol production 2011

Outline
1. Introduction
2. History of second generation bioethanol production
3. Worlds most advanced biorefinery history and learning points
4. Lignocellulosic biomass
5. Biorefinery options
6. The biochemical route (sugar plattform)
7. Pretreatment processes
8. Hydrolysis of cellulose
9. Anaerobic and aerobic fermentation
10. Lignin options
11. Hemicellulose/pentose options
12. Process integration & closing remarks

Borregaard
worlds most advanced biorefinery in operation

Leading supplier of specialty cellulose

Global leader in lignin performance chemicals, 50%+ market share
Only producer of vanillin from lignocellulosics
Production of lignocellulosic bioethanol since 1938
Worlds most advanced biorefinery in operation

Borregaard product tree

Prodction cont
Production stopped

Product tree from 2G bioethanol 1950 - 1980

Outline
1. Introduction
2. History of second generation bioethanol production
3. Worlds most advanced biorefinery history and learning points
4. Lignocellulosic biomass
5. Biorefinery options
6. The biochemical route (sugar plattform)
7. Pretreatment processes
8. Hydrolysis of cellulose
9. Anaerobic and aerobic fermentation
10. Lignin options
11. Hemicellulose/pentose options
12. Process integration & closing remarks

Composition of lignocellulosics

LIGNOCELLULOSICS
contain:
Lignin
Cellulose
Hemicellulose

CELLULOSE
Fiber
35 - 45%

LIGNIN
Binder
20- 30%

HEMICELLULOSE
Various sugars
25-30%

Lignocellulosic biomass structure

Cellulose fibres for chemicals

Width: m
Length: mm

Micro fibrillar cellulose

Width: nm
Length: m - mm

Plant cells
Width: m - mm
Length: mm

Planks
M and cm
Polymer chains

10 100

Glucose monomers
A few ngstrm

Logs
Meters, m

Cellulose

LIGNIN
CELLULOSE Binder
Fiber
30%
40%
HEMICELLULOSE
Various sugars
25%

Cellulose
Long chains of ONE type of
beads (polymer of glucose)
Forming crystals - crystalline
Same chemical structure in
every plant

Hemicellulose

CELLULOSE
Fiber
45%

LIGNIN
Binder
30%

HEMICELLULOSE
Various sugars
25%

Hemicellulose
Long branched sugar chains
(polymer, polysaccharide)
Amorphous
Composition varies largely
from species to species
C6 and/or C5 sugars

Lignin
CELLULOSE
Fiber
45%

LIGNIN
Binder
30%

HEMICELLULOSE
Various sugars
25%

HO
OH
HO
H3CO

OH

Carb.

HO

Carb.

OH

Lignin
Branched long-chain molecule
(polymer) made up of 3 types of
monomers
Amorphous (non-crystalline)
Composition varies from species to
species
Is the binder in all plants gluing the
cellulose fibres together

OH
O

HO

OH

H3CO

HO

OH
O

OCH 3

CH 3O

OH
H3CO
OCH 3

OH

O
O

HO

OH
HO

HO
OCH 3

OH
H3CO

OCH 3
OH
HO

OCH 3

OH

HO
H3CO
O

OH

H3CO
O

OCH 3

H3CO
OH

H3CO
O

HO

(Adler, 1977)

Composition of some lignocellulosic feedstocks

Outline
1. Introduction
2. History of second generation bioethanol production
3. Worlds most advanced biorefinery history and learning points
4. Lignocellulosic biomass
5. Biorefinery options
6. The biochemical route (sugar plattform)
7. Pretreatment processes
8. Hydrolysis of cellulose
9. Anaerob and aerob fermentation
10. Lignin options
11. Hemicellulose/pentose options
12. Process integration

Biomass to products conversion options

Pretreatment

(Partly degraded)
Natural polymers

Chemical and/or
mechanical
processing

Marketable
products

- Biocehmicals
Separation

Pyrolysis
BCD
Solvolysis

Liquefaction/
hydrolysis
Sugar
- Enzymatic in solution
- Weak acid
- Strong acid

Bio-monomers

Gasification

Synthesis gas,
CO + H2

Combustion

Heat, energy

Fermentation
CCS

- Biomaterials

Chemical
conversion

- Proteins

Extraction,
Chemical &
Catalytic conversion

- Biofuels

Purification
Catalytic synthesis
Refining, (CCS?)
CO2, CCS

- Energy

Outline
1. Introduction
2. History of second generation bioethanol production
3. Worlds most advanced biorefinery history and learning points
4. Lignocellulosic biomass
5. Biorefinery options
6. The biochemical route (sugar plattform)
7. Pretreatment processes
8. Hydrolysis of cellulose
9. Anaerobic and aerobic fermentation
10. Lignin options
11. Hemicellulose/pentose options
12. Process integration & closing remarks

Sugar plattform path ways

Hydrolysis processes
Dissolving celluose and hemicellulose
leaving hydrolysis lignins undissolved

Strong acid
Weak acid
Enzymatic
Microbial

Pulping processes
Dissolving lignin (and hemicellulose)
leaving cellulose undissolved

Hydrolysis
Lignin (S)

Cellulose (L)
LIQUID

Kraft
Soda
Sulfite
Solvent
Extrusion

Lignin quality depends strongly on process

and biomass source
Hemicellulose/xylan form and quality
depends on process and biomass

HemiCellulose (L)
SOLID

Lignin
(L)
Cellulose (S)
HemiCellulose (L)
LIQUID
SOLID

Outline
1. Introduction
2. History of second generation bioethanol production
3. Worlds most advanced biorefinery history and learning points
4. Lignocellulosic biomass
5. Biorefinery options
6. The biochemical route (sugar plattform)
7. Pretreatment processes
8. Hydrolysis of cellulose
9. Anaerobic and aerobic fermentation
10. Lignin options
11. Hemicellulose/pentose options
12. Process integration & closing remarks

Pretreatment processes

Hydrolysis processes
Strong acid pretreatment (low temp, large consumption of acids, need
regeneration of acids, low yields): Weyland, TNO, BlueFire
Weak acid pretreatment (high temp and pressure, creates large amounts of
inhibitiors): SEKAB, Iogen
Steam explosion (followed by enzymatic hydrolysis, also combined with acids
or SO2): Abengoa, Inbicon, BioGasol, University of Lund, Andritz

Microbial (microbes doing the whole job of hydrolysis and fermentation)

Mascoma, Arbor Fuel etc.
Solid state fermentation

Pulping processes
Kraft: evaluated by Innventia, most common commercial chemical pulping
process
Soda: evaluated by Innventia, old pulping process
Sulfite: Borregaard, Wisconsin Uni. (SPORL), modified sulfite pulping
processes
Solvent/Organosolv : Lignol, CIM-V
Extrusion: PureVision (autohydrolysis)

Formation of fermentation inhibitors

High temp, water, acidic conditions

Xylose

Glucose

Furfural

HMF Hydroxymethyl furfural

Acetic acid
Hemicellulose

Steam explosion pretreatment

http://www.youtube.com/watch?v=jpMAiyWoEFo

BALI the holistic pretreatment process

The pretreatment and separation process used in EuroBioRef for lignocellulosics

Supplying sugars in solution
A pretreatment process that enables production of valuable products out of all three
main lignocellulosic components
Cellulose
Hemicellulose
Lignin
A pretreatment process that facilitates low cost hydrolysis of cellulose
Low enzyme consumption (lignin inhibition avoided)
Resirculation of enzymes (no adsorption to lignin)

Lignocellulose

BALI
Pretreatment and
separation

C6

Ethanol
Chemicals

C5

Ethanol?
Chemicals
Yeast

Lignin

Performance
chemicals

BALI in a nutshell

BALI in a nutshell

pulp

hydrolysis

cellulases

fermentation

BALI
Step 1: pretreatment & separation

Pretreated and reactive pulp

Bagasse or other biomass

Water soluble lignin

Mass Balance of BALI pretreatment process

Flexibility from two optional processes

Lignin
(L)

Bagasse
Lignin
Hemicellulose
Cellulose

Cellulose (S)

BALI
Acidic

HemiCellulose (L)
LIQUID

SOLID
PULP

Lignin (L)

BALI
Alkaline

LIQUID

Cellulose (S)
SOLID
PULP

HemiCellulose (S)

BALI pilot plant

Location: Borregaard Sarpsborg, Norway

Flexible feedstock
1 metric ton dry matter/day
Start-up Q2 2012
Budget: 130 MNOK

Outline
1. Introduction
2. History of second generation bioethanol production
3. Worlds most advanced biorefinery history and learning points
4. Lignocellulosic biomass
5. Biorefinery options
6. The biochemical route (sugar plattform)
7. Pretreatment processes
8. Hydrolysis of cellulose
9. Anaerobic and aerobic fermentation
10. Lignin options
11. Hemicellulose/pentose options
12. Process integration & closing remarks

BALI
Step 2: Enzymatic hydrolysis to sugars in solution
Pretreated and reactive pulp is
hydrolyzed using cellulase enzymes

Hydrolysate = monosaccharides in solution

Cellulose hydrolysis

xylanases,
mannanases,
hemicellulases
Cellobiohydrolase
Endoglucanase

b- glycosidase

Enzymatic hydrolysis of BALI cellulose

Yield and Viscosity

46h

Enzyme hydrolysis of BALI pulp

better substrate than soda pulp

Enzymes not inhibited by residual lignins

BALI cooks

Soda cooks
140-160 C
120-180 min

Enzymatic hydrolysis - carbohydrate conversion - dose response

Accellerase DUET at 7% cellulose, 50C, pH 5.0, 72h
120,00%

100,00%

% total carbohydrate conversion

80,00%

60,00%
Reference
(hardwood pulp)
BALI bagasse A

40,00%

BALI bagasse B

20,00%

0,00%
-

0,10

0,20

0,30

0,40

ml Accelerase DUET / g glucan

0,50

0,60

Outline
1. Introduction
2. History of second generation bioethanol production
3. Worlds most advanced biorefinery history and learning points
4. Lignocellulosic biomass
5. Biorefinery options
6. The biochemical route (sugar plattform)
7. Pretreatment processes
8. Hydrolysis of cellulose
9. Anaerobic and aerobic fermentation
10. Lignin options
11. Hemicellulose/pentose options
12. Process integration & closing remarks

Fermentation
Established technology from 1G bioethanol
C6H12O6 > 2 CH3CH2OH + 2 CO2
glucose
ethanol
carbon dioxide

Mw (w%) 46 (51%) 44 (49%)

Saccharomyces cereviciae
(Bakers yeast)
Only fermenting hexoses, not
pentoses
Anaerobic fermentation for
production of ethanol
Aerobic fermentation for
production of yeast cells
GMOs for C5 fermentation

Aerob fermentation
Reproduction and production of yeast/bacteria/chemicals
Example of simple aerob fermentation to yeast from pentoses with
added nutrients:

Outline
1. Introduction
2. History of second generation bioethanol production
3. Worlds most advanced biorefinery history and learning points
4. Lignocellulosic biomass
5. Biorefinery options
6. The biochemical route (sugar plattform)
7. Pretreatment processes
8. Hydrolysis of cellulose
9. Anaerobic and aerobic fermentation
10. Lignin options
11. Hemicellulose/pentose options
12. Process integration & closing remarks

Technical challenges for 2nd generation bioethanol

Low % of feedstock useful for ethanol production

Only approx. 40%- 45% of biomass can be converted to product

Low yield in several process steps

Theoretically maximum 51% yield of ethanol from C6 sugars
No industrial solution for fermenting C5 sugars to ethanol
Several process steps with 80%-95% yield create loss and sidestreams
Lignocellulosic biomass is recalcitrant to degradation tough demands
on pre-treatment and liquefaction/hydrolysis steps
Sidestreams impure challenge to convert into valuable products

Properties of hydrolysis lignins

Low Mw high polydispersity

Strongly condensed (high temp)
Very few -O-4 bonds left mainly C-C bonds
Few OH groups left
Generally low O content relative to other lignins
Water insoluble
Low reactivity - hard to modify chemically at a reasonable cost
Impurity level will be high

hard to separate
impure products
many side streams

NOT A GOOD STARTING POINT FOR CHEMICALS

IAR Reims G Rdsrud

8.9.2010 Borregaard

BALI lignin is water soluble

BALI lignin is sulfonated and
therefore highly water
soluble at almost every pH
Major challenge is to make
high quality lignin specialty
chemicals
Extensive application tests
have been conducted
Possible uses:
dispersing agent, soil
conditioner, antioxidant,
emulsion stabilizer, crystal
modifier for batteries, dust
control, binding agent, etc.

Lignosulfonate structure

At least one SO3- for every four C9 units needed to be water soluble

Properties of Lignosulfonates
MW
Polydispersity
Sulfonate groups
Organic sulfur
Solubility
Color

5,000 80,000 Da
6-8
0.6-1.2 per monomer
4-8%
soluble in water at all pH
insoluble in most organic solvents
light to dark brown

Delivery

powder or
liquid form (40-50% DS)

Non-toxic:

LD50 > 5 g/kg

Quality

Softwood: good
Hardwood: medium
Annual plants: low

Intrinsic properties of lignosulfonates

In frequent use
Binder
Dispersing agent
Emulsifier
Complexing metal ions
Under exploration commercially
Corrosion reduction
Plant growth stimulation
Antioxidant
Not in commercial use
Flame retardant
Resins (old, not in use any more)
UV-absorption/UV-protectant
Protein precipitation (old, not in use any more)

BALI - Examples of possible product mixes

100%
90%

5
24

80%
70%

20

60%

50%

Energy

18

CO2
Yeast

Ethanol

16

15

Lignin

40%
30%
20%

41

46

10%
0%
Acidic

Neutral
% of incoming biomass + added chemicals

LS decreases viscosity in mortar and concrete

Flow table test

Lignosulfonate
- emulsifier and dispersing agent
stabilize emulsions

disperse color pigments

disperse pesticides

Future use:
disperse carotenoids and fat soluble
vitamins

Lignosulfonate in lead acid batteries

crystal growth modifier =>
better discharge/charge
performance

Soil conditioner

Oxidation of lignosulfonate to vanillin

Copper catalyst is
recycled due to strict
limitations on copper
in effluent

Outline
1. Introduction
2. History of second generation bioethanol production
3. Worlds most advanced biorefinery history and learning points
4. Lignocellulosic biomass
5. Biorefinery options
6. The biochemical route (sugar plattform)
7. Pretreatment processes
8. Hydrolysis of cellulose
9. Anaerobic and aerobic fermentation
10. Lignin options
11. Hemicellulose/pentose options
12. Process integration & closing remarks

Fermentation of C5 and C6 sugars from the BALI process

C6 sugars to ethanol (anaerobic)
Absence of fermentation inhibitors
High yields

C5 sugars to SCP single cell proteins (aerobic)

No inhibitors
No toxic compunds
Interesting yeast strains identified and tested
C5 sugars to ethanol (anaerobic)
Hydrolysates under testing with many GMO
microbes

Theoretical yield of ethanol from biomasses

How much do we gain from using GMO yeasts?

Outline
1. Introduction
2. History of second generation bioethanol production
3. Worlds most advanced biorefinery history and learning points
4. Lignocellulosic biomass
5. Biorefinery options
6. The biochemical route (sugar plattform)
7. Pretreatment processes
8. Hydrolysis of cellulose
9. Anaerobic and aerobic fermentation
10. Lignin options
11. Hemicellulose/pentose options
12. Process integration & closing remarks

Process flow for the BALI process

Integration into a 1st generation bioethanol plant

Economy of a biorefinery

Higher turnover
BUT

Also additional
manufacturing costs
and capital cost

Will it be more profitable ??????

Turnover for ethanol production and biorefineries

ROCE for a biorefinery compared to P&P

Sources:
1.CEPI. [Internett] http://www.cepi-sustainability.eu/uploads/graphs/CEPI_graph_18_3.eps.
2. [Internett] Poyry. http://www.poyry.com/linked/en/publications/FIC.pdf.
3. Orkla annual reports

Environmental Impact

Future limit for advanced

fuels in EU and US

BALI
+ CCS
BALI

Sources:
1. Directive 2009/28/EC of 23 April 2009 On the promotion of the use of energy from renewable sources and .
2. Modahl, I.S., 2011, Klimagasspotensialet ved komprimering, transport og lagring av biologisk CO2. Screening LCA.
Confidential report by Ostfoldforskning for Borregaard.

Funding

EuroBioRef
Borregaard granted EUR 3.0 mill
funding (2010 2013)
BALI pretreatment & hydrolysis

Suprabio
Borregaard granted EUR 1.1 mill
funding (2010 2013)
Microfibrillar cellulose

Acknowledgement
The research leading to these results has received funding from the European Union
Seventh Framework Programme (FP7/2007-2013) under grant agreement n
241718 EuroBioRef.

Acknowledgement
The research leading to these results has received funding from the European Union
Seventh Framework Programme (FP7/2007-2013) under grant agreement n
241640 SupraBio.

Biomass2Products B2P
Borregaard granted 2,3 mill EUR from
the Norwegian Research Council
(2009 2012)

BALIPILOT
Borregaard granted EUR 7,25 mill
from Innovation Norway
(2011-2012)

Acknowledgement
The research leading to these results has
received funding from the Norwegian Research
Council, the BIA programme, proj. no. 193217

BALI PILOT

Thank you