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BIOPROCESS SYSTEMS ENGINEERING (BSE) APPLIED TO THE PRODUCTION OF BIOETHANOL FROM SUGARCANE
BAGASSE
Coord.: Roberto C. Giordano
roberto@ufscar.br (DEQ/UFSCar)
Proc. 2008/56246-0
WORKSHOP BIOEN – November 2012
Software Engineering:
Biorefinery simulation, optimization
Enzyme Engineering +
Bioreactor Engineering (“industrial” yeast strains)
Validation: Process
Non-conventional
bioreactors
Where we contributing (for the best of our knowledge) Biorefinery multiobjective optimization in EO simulator Application of immobilized enzymes Production of cellulases (in situ) in air lift reactors Simultaneous isomerization and fermentation (SIF) of C5 3
Subproject 1: Development, implementation and validation of a user-friendly computational environment
Senior researchers (UFSCar):
Antonio J. G. Cruz Caliane B. B. Costa Jose A. S. Gonçalves Luiz F. Moura Roberto C. Giordano Ruy Sousa Jr
Graduate students:
Cassia M. Oliveira (MSc) Fabio H. P. B. Pinto (PhD) Felipe F. Furlan (PhD) Gabriel C. Fonseca (MSc) Herbert A. S. P. M. Jardim (MSc) José I. S. Silva (MSc) Karina Matugi (MSc) Renato Tonon Filho (MSc)
Undergraduate students:
Anderson R. A. Lino Fernando Foramiglio Jorge A. R. Fanton Leonardo C. G. Gonçalves Lucas Passos
Collaboration (EMSO developers):
Argimiro R. Secchi (PEQ-COPPE/UFRJ) Rafael P. Soares (DEQ/UFRGS)
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Simulation platform: EMSO (www.enq.ufrgs.br/alsoc)
Novelty: global simulator, open access models; efficient solver for static & dynamic models; our group has access to the source code.
Research horizon: Optimization: static dynamic (natural for this process) Super-structural optimization, coupling economics,
sustainability, energetic integration, water balance, etc (tool: Pareto + PSO, Particle Swarm Optimization)
Dynamic real time optimization
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Biorefinery flowsheet: an example (autonomous distillery, integrated 1G + 2G, organosolv/ethanol pretreatment; other pretreatments are implemented)
Dimensionless cash flow: burning sugarcane trash (50%); C6 + C5 fermentation
Infeasible region
Dimensionless cash flow: burning sugarcane trash (50%); only C6 fermentation
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Furlan et al. Assessing the production of first and second generation bioethanol from sugarcane through the integration of global optimization and process detailed modeling. Computers & Chemical Engineering, 2012.
Bagasse partition in the biorefinery.
Pareto + PSO in EMSO – an example: Cellulignin hydrolysis (hydrotherm pretreatment) + Vapor demand in evaporator
Pareto + PSO in EMSO – an example: Cellulignin hydrolysis (hydrotherm pretreatment) + Vapor demand in evaporator
Ethanol mol fraction in outflow side stream
Mass balances: (a) ethanol; (b) global.
Distillation: Transients due to variations in the feed tank
Subproject 2: Cultivation of microorganisms for the production of cellulases and xylanases in non-conventional triphase reactors
Senior researchers: UFSCar:
Alberto C. Badino Jr. Teresa C. Zangirolami Paulo W. Tardioli Rosineide G. Silva
EMBRAPA: Cristiane S. Farinas
Graduate students: Fernanda M. Cunha (PhD) Camila Florencio (PhD) Gabriel D. Torresam (MSc) Mateus N. Esperança (MSc)
Undergraduate students: Daniel Botta RafaelRodrigues Esteves
Project Goals: Cellulase production by Aspergillus sp. and Streptomyces sp. in conventional and pneumatic bioreactors Enzymes purification and characterization Methodology: Inoculum development (with/without bagasse) Effect of the mass transfer and shear conditions Chromatographic purification
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Collaboration: Michael Ladisch and Eduardo Ximenes Purdue University, USA
13 Cunha et al. Biotechnology and Bioprocess Engineering 17:100-108, 2012.
24h under SSF
Methodology for inoculum cultivation
in a triphase medium established.
Hydrodymanics and
transport parameters in
pneumatic bioreactor in the
presence of sugarcane
bagasse determined.
14 Cunha et al. Bioresource Technology 112:270-274, 2012.
BIN <0,5
BIN 0,5-1
,0
BIN 1,0-2
,0
BEX <0,5
BEX 0,5-1,0
BEX 1,0-2,0
0
200
400
600
800
1000
1200
1400
1600
1800
En
do
glu
ca
nas
e a
cti
vit
y (
IU.L
-1)
Submerged Fermentation
Sequencial Fermentation
Effects of sugar
cane bagasse
particle size and
pretreatment
Almost 2-fold
improvement of
cellulase
biosynthesis
Sub-Project 3: Physical-chemical pretreatment of SCB bagasse: removal of hemicellulose, delignification. Biomass characterization.
Senior Researchers: Antonio J. G. Cruz (UFSCar) Cristiane S. Farinas (EMBRAPA) Raquel L. C. Giordano (UFSCar)
Pos-Doc Fellow: Úrsula F. Rodrigues-Zuñiga
Graduate Students: Gislene M. Silva (PhD) Luciano J. Corrêa (PhD) Renata B. A. Souza (PhD)
Specially acknowledged collaboration: Adilson R. Gonçalves (EEL/USP)
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Collaboration: Celina L. Duarte (IPEN, Brazil)
Claus Felby (University of Copenhagen, Denmark)
Doris Schieder and Martin Faulstich (TUM, Germany)
George J. M. Rocha (CTBE, Brazil)
Mark R. Wilkins (Oklahoma State University, USA)
Renato L. Carneiro (Chemistry Department, UFSCar, Brazil)
Biomass characterization (Methodology)
Biomass pretreatment: bagasse deconstruction
Cellulose loss;
Hemicellulose: oligos or monomers?
Lignin removal: cost-effectiveness
Enhancement of enzymatic hydrolysis
Data for simulations of the biorefinary
(Sub-project 1)
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4000 4500 5000 5500 6000 6500 7000 7500 0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
Setting the conventional methodology for an accurate compositional analysis
Based in: Laboratory Analytical Procedure: provided by the National Renewable Energy Laboratory (NREL) (Sluiter et al., 2006) with modifications proposed by Gouveia et al. (2009).
Bagasse components In-natura (w/w %) Steam exploded (w/w %)
Cellulose 39,0% 41,8%
Hemicellulose 29,7% 20,7%
Soluble lignin 2,5% 4,3%
Insoluble lignin 19,2% 30,0%
Extractives 5,0% -
Ashes 4,9% 2,2%
Mass balance (total) 100,3% 99,0%
Rapid alternative methodology based in Near Infrared Spectroscopy (NIR) Database generated in the subproject 3 + Chemiometrics management
4000 4500 5000 5500 6000 6500 7000 7500-0.06
-0.04
-0.02
0
0.02
0.04
0.06
0.08
0.1
0.12
0.14
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Biomass pretreatment
Organosolv-ethanol 1, 2, 3
Experimental conditions:
Temperature: 150, 170 and 190 oC.
Reaction time: 10, 30, 60 and 90 minutes.
Degree of severity: 3.2 to 4.7
Agitation speed: 300 rpm.
Ethanol solution: 30, 50 and 70% (w/w)
Solid liquid ratio: 1:10 (w/v)
1 – Autohydrolysis only;
2 – Autohydrolysis in presence of 1% w/w of sulfuric acid;
3 – Autohydrolysis followed by alkaline delignification (1% w/v of sodium hydroxide).
Parr reactor
Best condition: 190 oC, 10 min., 50% ethanol
• Loss of cellulose (%): < 2.0
• Hemicellulose removal (%): 86.7
• Lignin removal (%): 78.3
• Enzymatic Cellulose Conversion *, ECC (%): 61.2
• Specific produtivity (mgglucoseFPU-1h-1): 0.303
* Enzyme load: 20 FPU/gcellulignin (Accellerase 1500,
Genencor)
Solid load: 10% w/w
100x(%)(g)m
0.9(g)mECC(%)
initial
glucose
ECC: Enzymatic cellulose
conversion
x: cellulose content in the
lignocellulosic material
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Subproject 4: C6 processing
Senior researchers: Antonio J. G. Cruz Paulo W. Tardioli Ruy Sousa Jr Teresa C. Zangirolami Raquel L. C. Giordano Roberto C. Giordano
Graduate students: Carlos E. Galeano-Suárez (PhD) Inti D. Cavalcante-Montaño (PhD) Gislene M. Silva (PhD) Luciano J. Corrêa (PhD) Pedro L. M. Aquino (PhD) Renata B. A. Souza (PhD) William Kopp (PhD)
Pos-doc fellow:
Ursula F. Rodríguez-Zúñiga
Collaboration
Jose M. Guisan (ICP/CSIC, Spain) Claus Felby (Univ. of Copenhagen, Denmark)
Challenge: Immobilized cellullolitic enzymes •Which in the pool? •Mechanism of action? •Half-life vs immobilization cost? •Biorreactor operation?
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Sequential batches with immobilized pool
0 10 20 30 40
0
5
10
15
20
25
30
35
40
45
50T
ota
l R
ed
ucin
g S
ug
ar
Co
nce
ntr
atio
n (
g.L
-1)
Time (hours)
1st batch
0 10 20 30 40
Time (hours)
2nd
batch
0 10 20 30 40
Time (hours)
3rd batch
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Silica Magnetic Micro-Particles SMMP
Superparamagnetic Enzyme Aggregates,
SEAs)
Subproject 5: C5 processing
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Senior researchers:
Raquel L. C. Giordano Roberto C. Giordano Ruy Sousa Jr. Teresa C. Zangirolami
Graduate students:
Anny Manrich (PhD) Carlos E. Galeano-Suarez (PhD) Claudia R. Silva (PhD) Inti D. Cavalcante-Montano (PhD)
Guilherme S. Moraes (MSc) Patrícia M. Aquino (MSc)
Challenges: Xylulose metabolism in S. cerevisiae Directed evolution of strains Stabilizing ex-vivo immobilized enzyme (XI) Controlled hydrolysis of hemicellulose for production of xylooligosaccharides (XOS)
Collaboration: Eugénio. Ferreira, Isabel Rocha and Sónia Carneiro (Pos Doc) UMinho, Portugal
Andreas K Gombert EPUSP, Brazil
GI comercial livre
IGI-Ch
IGI- comercial
30 oC; pH 5,0;
60 g/L de xilose ISOMERIZATION UNDER FERMENTATION
CONDITIONS MADE FEASIBLE BY
IMMOBILIZATION
Silva CR, Zangirolami TC, Rodrigues JP, Matugi K, Giordano RC, Giordano RLC. . An innovative biocatalyst for production of ethanol from xylose in a continuous bioreactor. Enzyme and Microbial Technology 50:35-42, 2012.
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SCREENING FOR SUITABLE S. cerevisiae STRAINS
Experimental conditions: batches conducted at T= 35 oC; biocatalyst containing 20 % enzyme, 10 % yeast and 1 % CaCO3;
microaerophile. Fleischmann and Itaiquara: bakery yeasts; BG-1, CAT-1 and PE-2: industrial strains; CEN.PK117-7D: lab strain.
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ASSESSING PROCESS PERFORMANCE
EtOH – Ethanol; XOH – Xylitol; X – Conversion; PrEtOH – Productivity in ethanol; Se – Selectivity, YEtOH/S0 – Overall yield
acel/ml; b recombinant yeast; *hemicellulose hydrolysate (FOGEL et al., 2005)
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