wp no. 3: advanced biochemical conversion of biomass · project biosynergy ... from biotechnology...
TRANSCRIPT
Workshop ‘Development of multi-product lignocellulose biorefinery technology with focus on residues (pentoses, lignin) from cellulose ethanol production - Results of the Integrated Project BIOSYNERGY ’Biomass derived pentoses: from biotechnology to fine chemistry – Side eventNovember 17, 2010, Reims (France)
C. Aymard, C Boeriu, B. Clément, B. Houweling-Tan, T. Jing, A. Lopez Contreras, M.-L. Mattinen, R. Marchal, M. Penttilä, A. Rogut, J. Rogut, M. Ropars, T. Tamminen, A. Togtema, H. van der Wal, P. van Doeveren, M. Wiebe, F. Monot
WP No. 3: Advanced biochemical conversion of biomass
Background/Vision WP 3 activities (1)
Physicalchemical
pretreatment&
Fractionation
Ligno-cellulosicbiomass
Esp.STRAW
Lignin
Hemicellulose
Cellulose EnzymaticHydrolysis
Fermentation
ChemicalConversion
Ethanol
ABE
Xylonic acid
(HMF) > 2,5 FDA
Furfural
PentosideSurfactants
C5 sugars
SC Depolymeri-
sation
CatalyticPyrolysis
Fractionation &
(bio-)chemicalconversion
Phenolics
Poly-ethers
Resins / Thermosets
C6 sugars
CHP Heat & Power to process
B i o m a s s r e s i d u e s
Background/Vision WP 3 activities (2)
Hemicellulose
C6 sugarsC5 sugars
Cellulose
C6 sugars
Lignin
Aromatic alcohols
Acetone + butanol
Sugar derivatives
Ethanol
Dispersants AdhesivesFuelChemical Building block
Separation processes
Biological conversion
Functional lignin derivatives
Background/Vision WP 3 activities (4)
Main partners
• IFP Energies nouvelles: Scientific Research and industrialdevelopment center. Dvt of chemical (and biotechnological) processes.
• VTT - VTT Biotechnology: enzymology of lignocellulosic materials transformation, metabolic engineering and fermentation technology.
• WUR: Wageningen UR Food and Biobased Products. Separation, microbiology and fermentation.
• GIG: Glowny Instytut Gornictwa (Central Mining Institute). Separationprocesses.
WP3: Main results
3.1 Acetone-butanol fermentation (ABE). - ABE from wheat straw hemicellulose hydrolysates IFP EN
from DDGS WUR- Separation by pervaporation WUR
using Multiphase Rotating Contactors (MRCs) GIG
3.2 Sugar conversion to platform chemicals. VTT
3.3 Production of functional lignin derivatives. VTT
WP3: Acetone-butanol fermentation (ABE) (1)
Microbiology
• Clostridium acetobutylicum, beijerinckii, ...
• Anaerobic bacteria
• Solventogenesis coupled to sporulation
• Genome sequences available
WP3: Acetone-butanol fermentation (ABE) (2)
Fermentation
• Two-phases (acidogenesis-solventogenesis)
• Shift to solventogenesis related to pH
• Can use C5 and C6 sugars
• Toxicity of products limits solvent levels
ABE (3) – Screening of strains on glucose
12.00.234IFP T'110A
13.00.334IFP M410C
5.50.120DSMZ1739
3.00.416DSMZ6423
7.04.035DSMZ 1733
7.57.047DSMZ 792
9.07.026DSMZ 1731
8.57.548DSMZ 1732
Acids (g/L)Solvents (g/L)Glucose consumed(g/L)
Strain
6.015.553IFP903
6.017.559DSMZ 6422
4.510.540IFP904
5.514.557DSMZ 6228
2.51541DSMZ 923
5.515.560DSMZ 1738
5.01662DSMZ 4685
7.51863DSMZ 1737
Acids (g/L)Solvents (g/L)Glucose consumed(g/L)
Strain
Solvents
Acids
Solvents
& Acids
ABE (4) – Screening of strains on xylose
CaCO3 : 2 g/L
2.40.37DSMZ 6423
2.51.19IFP 9046.52.635DSMZ 4685
4,814,147DSMZ 17313.513.545IFP 903
3.715.751DSMZ 6422
8,52,730DSMZ 17337,63,826,4DSMZ 17378,23,824DSMZ 1738
3,614,551,9DSMZ 17322,714,749,1DSMZ 6228
Acids (g.L-1)Solvents (g.L-1)xylose consumed (g.L-1)Strain
ABE (5) – Production on wheat strawhemicellulose hydrolyzate 1
Lignocellulosic hydrolysate(IFP ref: 1019)
• Concentrated wash water from steam-exploded wheat straw
• Pretreatment conditions: 18.5 bars for 2 min (after soaking in 0.04 M H2SO4).
Composition :Cellobiose: 1.2 g/LGlucose : 6.5 g/LXylose : 43.5 g/LGalactose: 5.5 g/LArabinose: 5.5 g/LMannose : 0.7 g/LAcetate : 0.6 g/L5-HMF : 132 mg/LFurfural : 4 mg/L
• A preculture was performed in a fermentor (4-L of culture medium) on glucose with C. acetobutylicum NCIB 8052 (DSMZ 1732)
• The culture on hydrolysate rapidly stopped (probably because of the inhibitors in the hydrolysate).
ABE (6) – Effect of lignocellulose degradation products
0.04ferulic acid
Growth inhibitionCoumaric acid
0.10Syringic acid
0.20Vanilline
0.05p-hydroxybenzoic acid
0.48Catechol
0.18Furfural
0.045-hydroxymethylfurfural
Relative productionCompound added ( at 1 g/L)
Strain used: DSMZ 6228 ; substrate: xylose at 60 g/LABE production with no added compound : 14.7 g/L
ABE (7) – Production on hydrolyzate 2 (a)
� Fermentation using lime-pretreated H1121 added at variousconcentrations to a synthetic medium and using various strains
Solvent production
0,00
2,00
4,00
6,00
8,00
10,00
12,00
14,00
0% 20% 40% 60% 80% 100% 0% 20% 40% 60% 80% 100% 0% 20% 40% 60% 80% 100%
ATCC 39057 DSMZ 1732 NCIB 8052
So
lven
t (g
/l)
EthanolAcétoneButanolSolvants totaux
EtOHAcetoneBuOHtotal solvents
ABE (8) – Production on hydrolyzate 2 (b)
0
2
4
6
8
10
12
14
16
18
20
0 10 20 30 40 50 60 70
Time (h)
pH
or
gas
pro
du
ced
(L/L
)
5,70
5,80
5,90
6,00
6,10
6,20
6,30
6,40
6,50
6,60
Gas
flow
(mL
/min
)
Volume (L/L) Flow rate[mL/min] pH
05
10
1520253035
404550
0 10 20 30 40 50 60 70 80
Time (h)
Res
idua
l glu
cose
or x
ylo
se (
g/L)
0,002,004,00
6,008,0010,0012,0014,00
16,0018,0020,00
Aci
ds
or S
olve
nts
(g/L
)
Residual Glucose (g/L) Residual Xylose (g/L) Total solvents (g/L) Total acids (g/L)
C. beijerinckii NCIB 8052
Culture time (h)
Total acids(g/L)
Total solvents
(g/L)
Total sugarsconsumed
(g/L)
Yabe/sugars(g/g)
solvent productivityg/L.h
70.00 3.2 17.5 50 0.35 0.25
������������ ��������������������������������
� �����������������������������
� ��� ��� ��������� ��� ������������������������ ����
� ����������������������� ���!"#$%& '���! �"#$
� �(������������������ )��!%& *+��! ��� ����! ���������
� ��������� ������������������������������� �� ����������� �������������������������������
• ���(�����������,���������� -���������������• ����������,���.����������������%-�������% �
� "#$-�������/����� ������-�������/����,(���0��-�������0���� ��������0-�������������
� 1������� ���������
"#$,�� �������2234���
0 2����2���������3������4���.����2234����������--��� )&5�,������(������,2234��(��������.��
0 6���������������,2234���--��� '&5�,���(�����
�+%�!���
'6����7
+"��
*�%�4����-��� ���
)�%�$�������/��
2234
����&&��(�
1� -�������,2234
2234��������������
"#$,�� �������2234�*�
� 8���,2234����������--�� ��,��(��������(������������
0 2234��-�����(���������������,��,�� �������,-���(��������(������������
* 9�� �������,2234������������
0 "�:����0-��-�����������������,2234���,�� ���.��(������,��������������--�� ������
2234�����������
1��������;����-�����2234,�� ������
0 "#$0-���������������������������-��� �����������2234
0 2234������������-��-����������:���������������,�� ���.����"#$.����������������
0 "�������,2234��1�0����(��������(������������������,�� ������%����.���������������������� � �������������
0 2234���������������,�������,��"#$,�� �������,-�����.�������%,����� -���������������� ������������
ABE – Separation (1)
• Extraction of solvents from acetone-butanol fermentation brothwith membranes (WUR)� Suitability of PerVaporation membranes for Acetone-butanol extraction� Suitability of Reverse Osmosis membranes for Acetone-butanol extraction� Fermentation with on-line extraction
• Extraction of alcools using rotating disc contactors (GIG)
<� .�����-�������,"#$
������� ����������!�����"����������!����� �#�������
�����-�����/��� ���������=���,.����,�� ������-�������������������������,(����.����������������
������6������� ������ � .�����������.�������-����������/����������.������
$���������� ���!�����"����������!����� �#�������
�����0������������ �/���"#$������9�� �������.����-�������/���������� �/���,���.�������/���
>��������
4������=)?@1�&0�������������%"#$,���& *�� * �
9�� ������ =)?@1�&0�������������%"#$,���& &�0& ��� * �������,���A�
"#$9�� ���������&&0!��?�&0!�����
��&&0!�������%
(��:��/��� �?*�0!
"����.��%)?@1
9�� �������� ��'��
��&0!�������%
B��:��/��� �?�0!
9��0.����%�����/��� �%�&&0!
"����.��%)?@1
9�� �������� ��*�C�*&�
!�:����-��/�-������ �����
"#$9�� ��������?*�0!�����
��&&0!�������%(��:��/��� �?*�0!%"����.��%)?@1%9�� �������� ��'��
750 L
0
10
20
30
40
50
60
0 10 20 30 40 50
Time (h)
gluc
ose
(g/l)
0
4
8
12
16
20
24
OD
600
acid
s, s
olve
nts
(g/l)
glucose OD600 acetic acid butyric acidacetone ethanol butanol ABE
���,����,"#$,�� ������ �,�������.������ 71><#�&�*
��?*�0!�����
"#$�� �/�� ,�� ���,�� ������������"#$D��!�
Membrane separation of ABE
PV Screening
$%�������$� ���������&�����'((����)����������
*��!����(+(,��-�
) 1��������;-���-����/��
"�&&0!,�� ���������-�����-��/�-������ ��,,����,�� -���������.���� E�(�/��%��-��������,,��������(����.���/����������������-������%����������.��������/������,��,��,��������/���- ����� ������
���/�-������ ����0���-�������� �/�������F�������"#$,�� ��������-����.��.������,����������������
• �-�� �������,<� .���/� 9�� ���� /��� ���������������������������/� ��� � "#$-�������/����G�����������H����/��������������
• �-�� �������,9�� ������� � ����,"��9�� �����������������,�������/��� -����
• �-�� �������,1���������� � ���������������% *%�� -���������
Separation using Rotating disc contactors
• Rotating disc contactors (GIG)� Principle
� Mixtures of alcools� Mass transfer for water solutions of single and multiple components
Background Model - Based on Zanfir and Gavriilidis (2007) work
Rotating disc contactors: principle (1)
Chemical Reaction Engineering • Mass transfer • Heat transfer • Chemical conversions • Separation – reaction • Energy consumption
Biotechnology Issues
• Conversions efficiencies • Product purity • Adaptability to Bio-Processes • Ability for control • Flexibility to raw materials
Emission Capture
Processes addressed: • Removal of volatile products of fermentation ( ethanol, ABE – as model processes ) • Esterification of organic acids ( ethyl lactate production in pressurized RDC …? )
Rotating Disc Concept – Reaction – Separation Tool for Bioprocesses
Rotating disc contactors: principle (2)
Integration of two rotating disks
Experimental Units in use in GIG
Materials and geometry : polycarbonate discs ( D = 0,27 m ), volumes for liquid phase : 0,5 L ( small unit ) , 10 L ( large unit )
Rotating disc contactors (3)
• Demonstration of the suitability of the system for ABE separation
• Comparison with stripping favorable to RDC
• Elaboration of a model for broad applications underway
Separation using Rotating disc contactors -conclusions
Comparison of rates of direct ethanol recovery by natural stripping, forced stripping and in rotating disc systems
0 5000 10000-2
-1
0
1
2
3