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FAPESP BIOENERGY PROGRAM BIOEN
Brazilian External Dependence on Oil
2
Brazilian Cane, Sugar and Ethanol Production
First period: “necessity” Now: a great “opportunity”!!
-
100.000
200.000
300.000
400.000
500.000
600.000
74/75 78/79 82/83 86/87 90/91 94/95 98/99 02/03 06/07
Su
ga
rca
ne
(1
0³
t)
-
5.000
10.000
15.000
20.000
25.000
30.000
35.000
Eth
an
ol (1
06 lit
res
) &
Su
ga
r (1
03 t
)
Sugar
Sugarcane
Ethanol
Brazil increased ethanol production and the same time that increased its sugar production
3Source: UNICA
0%
10%
20%
30%
40%
50%
60%
Non-Renewable Renewable
En
erg
y s
ou
rces i
n B
razil,
2006
47% of Brazil’s energy comes from renewable sources (2009)
4
cane
18%
Renewables in Brazil: 47%; World: 13%; OECD: 7,2%
08082010; Programa FAPESP BIOEN
Energy from renewable sourcesSome industrialized countries
5
Source: IEA, Renewables Factsheet, 2007
10052010;bioenergy-in-brazil-20100510.pptx;CHBritoCruz & BIOEN
Energy CaneSugarcane Mill
Brazilian Bioethanol production costs are the cheapest in the world
Industry estimates the cost of producing ethanol from sugarcane at approximately US$ 0.29/L (1 gallon = US$ 1.00).
(Goldemberg, Nigro e Coelho, 2008).
Total bioethanol production for 2010/11 is projected at 28.5 billion L
Total sugarcane production is estimated to be 664,33 ton/ha for 2010/2011
54,6% (362,8 million tons) for ethanol (20,14 billlion L hydrated and 8,4 billion L anyhidride)
45,4% (301,6 million tons) for sugar (38,7 million tons)
405 plants of which 157 are exclusive to ethanol
Sugarcane for ethanol uses 0.4% of total area
9
Area used for sugarcane for ethanol (3.4Mha, 0.4%)
Area used for agriculture (76.7 Mha, 9%)
Rural properties area (355 Mha, 42%)
Total country area (851 Mha, 100%)
Source: Horta Nogueira e Seabra (2008)
Small bioenergy footprint
Pasture 200 MhaSoya 22 Mha
Average yield 80 ton/hectare
High yield variety: 260 ton/ha in 13 months (commercial at Agrovale, Bahia) and 299 ton/ha (experimental at Fazenda Busato, Bahia)
Sugarcane, the most productive of cultivated plants - a large biomass
Commercial sugarcane is vegetatively propagated through stem cuttings
In 12 months the plant will reach 4-5 meters with the extractable culm measuring 2-3 meters
After harvest, underground buds will sprout giving rise to a new crop (6 harvests)
C4 carbohydrate metabolism - large amount of carbon partitioned into sucrose (up to 42%
of the stalk dry weight, around 0.7 M in mature internodes)
Fonte: International Energy Agencia (2005). *ISPA
Biofuel yield per hectare
12
FAO, “The state of agriculture 2008”
08082010; Programa FAPESP BIOEN
Ethanol
Biodiesel
Produ�‹ o de cana-de-a�œcar no Brasil (milh› es
de toneladas)
0,00
100,00
200,00
300,00
400,00
500,00
1975
1978
1981
1984
1987
1990
1993
1996
1999
2002
2005
Ano
(mil
h›e
s t
on
)
Cultivar Biomass and Ethanol Production in Brazil
70’s
Hoje
82 %
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
até
1990
1992 1993 1995 1996 1998 1999 2000 2001 2002 2003 2005 2006
Biomass/ha Sugar
Sugarcane improvement is lagging behind
Cellulosic Ethanol
70’s
Today
82 %
In 10 years?
40-60 % 12.000/h
a
BIOEN DIVISIONS
BIOMASSContribute with knowledge and technologies for Sugarcane ImprovementEnable a Systems Biology approach for Biofuel Crops
PROCESSING AND ETHANOL TECHNOLOGIESIncreasing productivity (amount of ethanol by sugarcane ton), energy saving, water saving and minimizing environmental impacts
ENGINESFlex-fuel engines with increased performance, durability and decreased consumption, pollutant emissions
BIOREFINERIES AND ALCOHOL CHEMISTRYComplete substitution of fossil fuel derived compoundsSugarchemistry for intermediate chemical production and alcoholchemistry as a petrochemistry substitute
IMPACTSStudies to consolidate sugarcane ethanol as the leading technology path to ethanol and derivatives productionHorizontal themes: Social and Economic Impacts, Environmental studies and Land Use
6 Calls for Projects
55 projects granted118 fellowshipsR$ 57 million (FAPESP)
2 9
5
13
3
10
30
46
118 Fellowships
Fellowship (Visiting schollar)
Fellowship (PhD)
Fellowship (Direct PhD)
Fellowship (Scientific Initiation)
Fellowship (Young Researcher)
Fellowship (Masters)
Fellowship (Post-doc)
6
8
25
16
55 Grants
Grant (Young Researcher)
Grant (PITE)
Grant ("Temático")
Grant (Regular)
30% of the projects are related to Chemistry
0 5 10 15 20 25 30 35
Agronomy
Biophysics
Biochemistry
Botany
Information Technologies
Food Technologies
Ecology
Economy
Materials Engineering
Mecanical Engineering
Nuclear Engineering
Chemical Engineering
Environmental Engineering
Pharmaceutical sciences
Physics
Genetics
Geosciences
Interdisciplinary
Mathematics
Microbiology
Chemistry
Série1
6 Calls for Projects
5 DivisionsR$ 57 million (FAPESP)
13.066.846
30.538.839
3.327.769
3.325.014
Budget grants
Biofuel Technologies
Biomass
Biorefinery
Impacts
19
20
6
8
1
DIvisions
Biofuel Technologies
Biomass
Biorefinery
Impacts
Engines
314 researchers under the BIOEN Program
84%
8%
2%
BRAZIL
USA
FRANCE
GERMANY
THE NETHERLANDS
DENMARK
SPAIN
AUSTRALIA
AUSTRIA
CHINA
FINLAND
PORTUGAL
BRAZIL 262
USA 26
FRANCE 7
GERMANY 4
THE NETHERLANDS 4
DENMARK 3
SPAIN 3
AUSTRALIA 1
AUSTRIA 1
CHINA 1
FINLAND 1
PORTUGAL 1
52 foreign researchers from 36 institutions
BIOEN Foreign Partners (11 countries)
AUSTRALIA2%
AUSTRIA2%
CHINA 2%
DENMARK6%
FINLAND2%
FRANCE13%
GERMANY7%
PORTUGAL2%
SPAIN6%
NETHERLANDS9%
USA50%
AUSTRALIA
AUSTRIA
CHINA
DENMARK
FINLAND
FRANCE
GERMANY
PORTUGAL
SPAIN
THE NETHERLANDS
USA
USA 26
FRANCE 7
GERMANY 4
THE NETHERLANDS 4
DENMARK 3
SPAIN 3
AUSTRALIA 1
AUSTRIA 1
CHINA 1
FINLAND 1
PORTUGAL 1
TOTAL 52
State of São Paulo Bioenergy Research Center
• Proposed by FAPESP, USP, Unicamp and UNESP to the State of São Paulo government
• BIOEN Research Areas
• Investment (R$ 165 million)
– R$ 55 million from the State Government (R$ 45 M already committed)
– R$ 55 million from the Universities
– R$ 55 million from FAPESP
23092009; FAPESP BIOEN22
BIOMASS DIVISION
Improvement of Biomass (Agronomy, Breeding, Biotechnology)Identify new paths to genetically manipulate the energy metabolism of cultivated
plants, creating new biofuel alternatives
Contribute with knowledge and technologies for Sugarcane ImprovementEnable a Systems Biology approach for Biofuel Crops
• Uncover metabolic networks related to the production of carbohydrates and sucrose through the use of “omics” technologies• Integrate the results in a single platform and develop bioinformatic tools to assess the information• Discovery of genes associated with agronomic characteristics of interest• Development of new sugar cane cultivars• Signaling, regulation of gene expression and regulatory networks• Genetic transformation of sugarcane and other grasses • Molecular markers, statistical-genetics, mapping and breeding• Sequencing, physical, genetic and molecular mapping of genomes • Understand cell wall structure, architecture and biological function• Discover new cellulolytic fungi species capable of degrading biomass• Refine field practices for enhancing crop production including soil management, fertilization and precision agriculture• Improve control of weed, pests, and diseases though chemical or biological control, resistant varieties and field practices
Brazil has great tradition in sugarcane cultivation
(since 1532)
• 8.0 million ha
• World leader
• Several public and private institutions
dedicated to P&D
• Very competitive costs and production
technology
• Breeding Programs (since 1910)IAC (1934)
Campos (1946-1972)
CTC (ex Copersucar, 1968)
Ridesa (1971)
Canavialis (2003)
Fertilizer experiments. IAC, 1938
Sugarcane production
Domestication and early evolution of sugarcane
Saccharum
officinarum
Chewing
Saccharum sinence
Saccharum barberi
(crosses to wild relatives
natural hybrids)
Sugar extraction
SE Asia
Pacific Islands
Intertropics Persia
Manufacturing
Mediterranean
Spain
Canary
Madeira
West Africa
Cottage
industries
World
trade
plantation
factories
Saccharum
officinarum
clones
Dominican
Republic
Brasil
S. officinarum X
S. spontaneum
(interspecific
hybrids)
Java
India
Modern
Breeding
Mo
dern
su
garc
an
e c
ult
ivars
Interspecific breeding: a major breakthrough in modern sugarcane breeding
Solved some of the disease problems but also provided increased yields,
improved ratooning ability and adaptability for growth under various stress
conditions
Genome organization of a
modern cultivar
Each bar represents a
chromosome
Chromosomes in the same
column are homologuesS. officinarum
S. spontaneum
Contributing genera: Saccharum, Erianthus, Miscanthus, Sclerostachya and Narenga
Saccharum genus (six polyploid taxonomic groups):
Wild species Early cultivars Marginal species
S. spontaneum (2n=40 to 128) S. officinarum (2n= 80) S. edule (2n = 60 to 122))
S. robustum (2n= 60, 80 and up to 200) S. barberi (2n=81-124)
S. sinense (2n=116-120)
Giant Genome (n 750-930 Mpb) Polyploid (2n = 70-120 cromossomos) ~10 Gb
50 labs
200 researchers
238000 ESTs
43000 Transcripts
The SUCEST EST Sequencing Project
26 libraries - 13 cultivars - Over 90% of sugarcane genes tagged
Defining Initiatives for Sugarcane Genomics and Biotechnology in Brazil
R570 BAC Library
R570 BAC library (HindIII)= 103,296 clones(Tomkins et al, 1999, TAG)1.3x total genome equivalent14x basic genome equivalent
http://bacman.sourceforge.net
Screening for gene-rich regions by membrane hybridization, overgo or 3D-pools
Statistics of Align - Sequences = 15
Identical Sites = 133,845 (97.4%) - Pairwise % Identity = 67.6% - Freq % of non-gaps = A: 43,107 28.5%; C: 33,665 22.3%; G: 34,215 22.6%; T: 40,174 26.6%; N: 3 0.0% GC: 67,880 44.3%
Finding the 5´ and 3´ UTRs and promoter regions using sorghum as reference
BAC by BAC Structural Annotation
[ BAC_NAME] Shcrba_022_o20
[Annotation_Ref_Gene_1] Replication protein A 70kDa
[Synonyms] REPA1, Replication factor A protein 1, Replication protein A 70 kDa DNA-binding
[Zea_mays_gene] GRMZM2G115013 [Sorghum_bicolor_Genomic_Blastn] Chr 4 [Sorghum_Bicolor_Gene] Sb04g034780 [SEQ_GENE]
gtggtagtt... [SEQ_5Kb_5’UTR] tgg... [SEQ_5Kb_3’UTR] tgg... [ORIENTATION] (+/-) [LINK_GBROWSE] http://...
dec/10 assembly
A
B C D
EF
G
H
Shotgun sequencing of sugarcane genome (SP80-3280)
Leaf DNA from the sugarcane variety SP80 3280 was broken into 500 to 1000 bp fragments (average ~650 bp) and sequenced using 454
10 runs were made using 2 gaskets
~11 million reads were obtained
~3.5 Gbp were sequenced
8 contigs align to SbGI; 96% identity 71% coverage
A draft of the sugarcane monoploid genome (1 Gb)
Genome organization of a
modern cultivar
Each bar represents a
chromosome
Chromosomes in the same
column are homologuesS. officinarum
S. spontaneum
Giant Genome (n 750-930 Mpb) Polyploid (2n = 70-120 cromossomos) ~10 Gb
Area Crossings
Bi-parental Crossings
Multiple Crossings
Breeding
Seedlings
New Varieties
13 clones in
final phase
168 clones in
Experimental Trials
619 clones in T3
6.792 clones in T2 (1.349 Clones Brix > RB855156)
398.477 Seedlings in T1
Total: 406,069 genotypes
2006
12 years
Selection
Sugarcane Map incorporating double and triple dose markers (SSR, EST-SSR, RFLP)
Mollinari, M; Silva, RR; Margarido, GRA; Marconi, TG; Souza, AP; Garcia, AAF
SP80-180 x SP80-4966, 200 individuals
934 markers
Final map with 347 linked markers329 single dose (239 1:1 and 90 3:1)16 double dose 2 triple doseAssembled in 102 linkage groupsThe total map length 2,880.3 cM (4,361.3) with a density of 7.6 cM (11.6)
Setting up the priorities
1 – Gene DiscoveryGenes associated to agronomic traits of interestGene function evaluationC4 model plant system2 – PhysiologySucrose metabolism, carbon partitioning, photosynthesis, stress responses3 - Genome SequenceFull length transcriptsSurveys of several genomes Gene enrichment methods for regions to be sequenced BAC library with 10x coverageBAC screening mthods4 - International Bioinformatics International Consortium5 – TransgenicsExpression vectorsComplete ORFeomePromotersScreening methods for phenotyping (metabolomics, qPCR, biochemical assays, cell wall)6 - Marker identificationIntegrated databases and translation of transcriptome data to marker assays
Biotechnological Roadmap for Sugarcane Improvement
How far can we go?
Waclawovsky, A. J., Sato, P. M., Lembke, G. M., Moore, P. H. and Souza, G. M. Sugarcane for Bioenergy Production: an assessment of yield and regulation of sucrose content (Plant Biotech. J. 2010)
High yield variety: 260 ton/ha in 13 months (commercial at Agrovale, Bahia) and 299 ton/ha (experimental at Fazenda Busato, Bahia)
Waclawovsky, A. J., Sato, P. M., Lembke, G. M., Moore, P. H. and Souza, G. M. Sugarcane for Bioenergy Production: an assessment of yield and regulation of sucrose content (Plant Biotech. J. 2010)
Potential yield of sugarcane
Teoretical potential of sugarcane crop production
Approach
Over 50 cultivars and ancestor genotypes are being evaluated at the sequencing, expression and physiological level
SP80-3280 as a reference cultivar
Contrast:High vs LowPrecocious vs LateTolerant vs Susceptible
RB867515
S. officinarumS. robustum
S. sinensisS. spontaneum
Genetical Genomics of Traits of interest
Progeny 1
Genotypes Brix
Sucrose
% m/m
Glucose
% m/m
Fructose
% m/m
Progeny 2
Genotypes Brix
CTC98-241 18.00 7.311 1.322 0.988 C158 18.3
CTC98-242 18.60 9.183 1.430 1.014 C121 18.8
CTC98-243 19.20 10.956 0.649 0.602 C171 16.8
CTC98-244 14.60 11.161 0.633 0.646 C496 17
CTC98-246 18.80 10.974 0.709 0.545 C11 19.2
CTC98-252 18.00 6.370 0.840 0.579 C6 18.2
CTC98-253 19.60 11.120 0.660 0.643 C113 21
CTC98-258 18.00 6.739 1.116 0.865
CTC98-261 7.00 1.14 0.878 0.755 C436 13.9
CTC98-262 7.40 1.37 0.968 0.823 C292 15
CTC98-265 6.40 0.49 1.200 1.090 C231 13.9
CTC98-268 4.80 0.70 0.342 0.326 C38 12.9
CTC98-271 6.00 0.92 1.098 0.992 C250 11.5
CTC98-272 6.80 1.07 0.725 0.632 C405 15.2
CTC98-277 7.40 1.58 0.774 0.716 C144 13.2
CTC98-279 7.80 1.74 1.318 1.066
SnRK1
Phosphorylates sucrose phosphate synthase (SPS) and nitrate reductase (NR), which together with binding of 14-3-3 proteins inhibits their activity
category sub category 1 sub category 2 HB vs LB MIn vs IIn Drought ABA Sucrose Glucose
adapter 14-3-3 protein GF14 1 3adapter 14-3-3 protein GF14 4adapter 14-3-3 protein GF14 2adapter 14-3-3 protein GF14 1protein kinase SNF-like kinase caneSnRK1-2 1 2 3
A B
C D
R
R
fStorage parenchyma (R), Phloem (f), Fiber and bundle parenchymal cells (arrows), Bundle distal
cells (ce), Bundle proximal cells (ci)
SNF-like Protein kinases
Boudsocq & Lauriere (2005)Plant Physiol. 138,1185-1194
Rocha et al. (2007). BMC Genomics 8, 71
Physiology of sucrose and biomass accumulation
Waclawovsky, A. J., Sato, P. M., Lembke, C. G., Moore, P. H and Souza, G. M.Sugarcane for Bioenergy Production: an assessment of yield and regulation of sucrose content. Plant Biotechnology Journal (2010)
Regulation of sucrose accumulation and biomass
Hemicellulose and pectin synthesis
Cellulose synthesis
Sucrose synthesis
Sucrose synthesis
Drought responses
Drought responses
Waclawovsky, A. J., Sato, P. M., Lembke, C. G., Moore, P. H and Souza, G. M. Sugarcane for Bioenergy Production: an assessment of yield and regulation of sucrose content. Plant Biotechnology Journal (accepted)
BIOEN - HC 03-2010 GSB Project 46
Sugarcane crop production
• Southeastern BR: > 1100 mm rain– Rainfed– “Salvage” irrigation to ensure
germination in dry periods– Irrigation is option for higher
yields
• Northeastern BR: irregular ou insufficient rain– Part of the area is irrigated
GSB Project
BIOEN - HC 03-2010 GSB Project 47
• In most regions irrigation is not necessary
(except salvage irrigation in some areas)
• Breeding programs: search for drought tolerant varieties: dry winter and soils with low water holding capacity
• Industry: Legislation restricts water use in new projects to maximum 1 m3/t cane.
– Great water use efficient improvements in the last 20 years (5.6 to 1.8 m3/t cane)
– Model mill: 0.46 m3/t
• More strict laws for surface water protection
Water use in sugarcane in Brazil
Sensors (30, 60, 90 cm)
Irrigation
Drought Field Experiments with 10 genotypes: Goianésia, Goiás (CW)
10 genotypes (pre-selected based on performance over 2009 from a group of 100 clones)
IAC
Drought Field Experiments with 6 cultivars: São Paulo, Pernambuco, Alagoas (SE, NE)
irrigated Non-irrigated USP, UNICAMP, UFV, UFAL, UFPE, UFRPE
Plant Information
Variety, Location, Sample, Replicate
Sugarcane Physiology and Technological Data
Measures
IRGA, LAI, SPAD,Fluorometer, Brix, etc
Treatment
Condition, Field/Vase, Date/Time
Statistical MeasuresMicroarray Expression
Phenotype Explanation/Interested Genes
Statistical Analysis
Pathways/Annotation
Sugarcane Transgenics
Explants: Immature Leaves
CEBTEC and IQ - USP
Callus Induction
Regeneration Selective Medium
Greenhouse
PCR and qPCRRooting Shoot Growth
Production of transgenic sugarcane plants
0
0,02
0,04
0,06
0,08
0,1
0,12
4A
-07
4A
-18
4B
-09
5A
-07
5A
-02
6A
C-3
A
6A
C-3
B
6A
C-0
6
10
-03
11
-10
11
-29
11
-34
11
-40
11
-41
12
-01
12
-20
12
-21
18
A-0
4
19
B-0
7
19
B-0
8
20
B-0
3
Nív
el d
e e
xp
res
sã
opKannVazio
pKannPP2c
A (Photosynthetic rate)
0.00
5.00
10.00
15.00
20.00
25.00
30.00
irrigated drought rew atering
µm
ol m
-2 s
-1
6AC-3
5A-07
4B-09
18A04
18A-01
11-34
11-29
silenced
control
Sugarcane transgenic plants with increased sucrose content:3 CIPKs gene silencing using RNAi
Papini-Terzi, F. S., Rocha, F. R., Vêncio, R. Z. N., Felix, J. M., Branco, D., Waclawovsky, A. J., Del-Bem, L. E. V., Lembke, C. G., Costa, M. D-B. L., Nishiyama-Jr, M. Y., Vicentini, R., Vincentz, M., Ulian, E. C., Menossi, M., Souza, G. M. (2009). Genes associated with sucrose content. BMC Genomics 10, 120. doi:10.1186/1471-2164-10-120
Genes associated to sucrose content, sugarcane with increased sucrose levels USPTO Patent US 11/716,262. PCT/BR2007/000282. Gláucia Mendes Souza, Flávia Stal Papini-Terzi, Flávia Riso Rocha, Alessandro Jaquiel Waclawovsky, Ricardo Zorzetto Nicollielo Vêncio, Josélia Oliveira Marques, Juliana de Maria Felix, Marcelo Menossi Teixeira, Marcos Buckeridge, Amanda Pereira de Souza, Eugênio César Ulian.Universidade de São Paulo, Unicamp, Centralcool, CTC and FAPESP
Gene Expression
Physiology
Sequencing
Gene Categorization Functional Annotation
TFs
Kinases
Cell Wall
Phosphatases
cDNA Chip
qRT-PCR miRNA
oligo Chip
Gene Ontology
Pathways
GRASSIUS
Molecular Markers
KEGG
Genome
PromotersSUCEST
Functional Genomics
The SUCEST-FUN DB is based on five main topics: Gene Annotation, Gene Expression, Public Resources, Sequencing Projects and Functional Genomics.
Transgenic Plant
SUCEST-FUN DB (http://sucest-fun.org)
http://sucest-fun.org
Lignin Biosynthesis is associated to sucrose content
Genes associated to sucrose content, sugarcane with increased sucrose levels USPTO Patent US 11/716,262. PCT/BR2007/000282. Gláucia Mendes Souza, Flávia Stal Papini-Terzi, Flávia Riso Rocha, Alessandro Jaquiel Waclawovsky, Ricardo Zorzetto Nicollielo Vêncio, Josélia Oliveira Marques, Juliana de Maria Felix, Marcelo Menossi Teixeira, Marcos Buckeridge, Amanda Pereira de Souza, Eugênio César Ulian.Universidade de São Paulo, Unicamp, Centralcool, CTC and FAPESP
PROCESSING DIVISION
Engineering, processing and equipment design aspects of bioethanol productionCellulosic Ethanol
Increasing productivity (amount of ethanol by sugarcane ton), energy saving, water saving and minimizing environmental impacts
• Identify and improve bottlenecks in ethanol production chain at the mill (reception, juice extraction, hydrolysis process, among others)• Improve fermentation process yield• Optimization of water recycling and energy efficiency• Improve separation processes especially to dehydrated ethanol• Develop cellulosic ethanol technology focusing the following objectives:
Characterization and development of physical-chemical pretreatment of bagasse for ligninocellulose hydrolysisDevelopment of acid catalyzed and biocatalyzed saccharification Development of high performance cellulases and hydrolases Reduction of the impact of fermentation inhibitorsDevelopment of microorganism strains capable to efficient fermentation of pentoses and hexosesByproducts recovery
Energetic Potential of Sugarcane
Co-regeneration
Co-generation in 2009/10 = 1.800 MW (3% of electricity matrix)In 2020 = 14.000 MW (14% equals 1 Itaipu) Investments of R$ 45 billion until 2015 on Co-generation (boilers from 21 bar to 92 bar)Eolic = 602 MW (2009)UN approved 140 new Clean Energy Generation Projects for Carbon CreditsWater usage decreased from 5 to 1 million m3/processed ton
RB92579
RB98710RB855113
RB863129RB931003
SP79-1011
RB93509
RB961552
RB9629
VAT90-212RB72454
RB951541
RB931566RB867515
15,0 15,5 16,0 16,5 17,0 17,5 18,0 18,5 19,0 19,5 20,0 20,5
BRIX
200
220
240
260
280
300
320TC
H
65 60 55 50 45 40 35 30
Fazenda Busato – Bom Jesus da Lapa - BA
BIOREFINERIES DIVISION
Industrial units near sugar and alcohol industrial facilities that take advantage of renewable raw materials available (ethanol, sugar, CO2, bagasse, trash, yeasts) to produce high added value products
Sugarchemistry for intermediate chemical productionAlcoholchemistry as a petrochemistry substitute
SYNTHETIC BIOLOGY
Complete substitution of fossil fuel derived compounds
• Consorted bioethanol-biodiesel-biokerosene production • Develop products from ethanol via acetaldehyde and ethylene route • Perform Chemical synthesis of intermediate oxygenated chemicals (alcohols, acid ketones), polymers (PHA, lactic acid), nutraceuticals directly from sucrose• Develop biocatalysis for transformations of carbohydrates in valuable chemicals• Bio-based chemicals using a synthetic biology approach
Industrial microbiology Bioprospection and directed evolution of microorganisms
Automation, robotics for screening and selection of organisms Metabolic engineering Design of proteins and enzymesDNA synthesis, expression systems Systems biologySynthetic chemistry Consolidated bioprocessing
Third Generation Bioethanol is produced from algal/glycerin/bagasse biomass either from catalytical or biological fermentation of syngas.
A branch of Third Generation Bioethanol has a major appeal of consuming carbon dioxide produced in the First and Second Generation processes :
great impact of almost zero CO2 emission within the
whole integrated process.
Project GoalsZero Carbon Emission Biorefinery Concept
Rubens Maciel (Unicamp)
Project GoalsZero Carbon Emission Biorefinery Concept
Rubens Maciel (Unicamp)
Project GoalsZero Carbon Emission Biorefinery Concept
Rubens Maciel (Unicamp)
From microorganism model to large scale plant operation, with proposition of integrated alternative routes and alternative processes of production, fermentation and bioethanol separation high efficient plants.
Sugar
Glycose
Sacarose
FE
RM
EN
TA
TIO
N
Ethanol
Acetic acid
Lactic Acid
butanol Acetone ethanol
CH
EM
ICA
L S
YN
TH
ES
IS
Acetaldhyde
Anhydride acetic
Ethyl Acetate
Vinyl Acetate
Crotonaldhyde
Paraldhyde
Butanol
Buthyl Acetate
Piridine
Nicotinamide
Glycol
Butadiene
Glyoxalate
Succinic acid
Biorefinery Concept: Some Chemical Products via fermentation
Learning curve already competitive
Etilene
Ethanol
Acetaldehyde
Acetic acid
Propene
Propylene
___Acrylic Acid
Glycerol
Lactic acid
Butadiene
Butanodiol
Succinic acid
BIOMASSH
YD
RO
LY
SIS
Sugar
Glycose
Sacarose
Xylose
Arabynose
FE
RM
EN
TA
TIO
N
Other Products to be obtained from biomass
Learning curve costs have to be reduced
Sugarcane sucrose
Microorganism Selection
(bioprospection and
collection analysis)
Culture Medium Optimization
Mathematic modeling and
metabolic route selection
(structured model)
Fermentation
Lactic Acid L- and D-Separation/Purification
ProductionKi
netics
ProcessesDehydration reductionAcrylic Acid Propionic
Acid
Process
OptimizationKinetics Modeling Process Control
New Products: Green Acrylic Acid
AÇÚCAR
metionina Acetil-CoA
Piruvato
DMSP
glicerol
Lactato
metilcitrato
propanoil-CoA
Ácido Acrílico
Acrilil-CoA
lactoil-CoA
3-HP-CoA 3-HP 3-hidroxipropanol
-alanina
propanoato
malonil-CoA
aspartato
oxaloacetato
-alanina
metilmal-CoA
-alanina-CoA
mal.semiald
ENGINES DIVISION
Research to consolidate ethanol as the renewable substitute for gasoline on a short to medium term (10 to 20 years), with the evolution of internal combustion engines, and on a long term with fuel cells.
Flex-fuel engines with increased performance, durability, less fuel consumption and less pollutant emissions
Adjust the engine compression ratioSolve the cold-starting problem when using only ethanolUnderstand the role of ethanol in new engine configurations (direct fuel injection together with port injection and turbo-chargers to decrease fuel consumption and CO2 emission) Study compatibility with after-treatment devices used for emissions controlDevelop ethanol or sugarcane products with physical-chemical properties adequate for compression ignition engines
IMPACTS DIVISION
Ethanol as a global strategic fuel
Studies to consolidate sugarcane ethanol as the leading technology path to ethanol and derivatives production
Monitoring biofuels expansion, new technologies and its impacts on sustainability (economic, social and environmental)
Providing/generating data and tools for establishing measurable indicators of sustainability of bioenergy Developing methodologies/models to integrate areas related to sustainability from a global perspective
Land use changesGHG emissions
Biomass and soil carbon stocksWater use
BiodiversityRegional income generationJob creation and migration
Integrating tools
Where can we plant sugarcane?
Amazon Rainforest
Pantanal
Atlantic Forest
Other important preservation areas
Above 12% slope area
High
Average
Low (World average)
Inapropriate
69
Source: Leite et al. 2009
POTENTIAL FOR SUGAR CANE PRODUCTION: SOIL AND CLIMATE - WITHOUT IRRIGATION
615 GL of Ethanol will substitute for 30% of the world’s gasoline
2004 2025
Gasoline consumption 1,200 GL 1,700 GL
Ethanol consumption 30 GL
Ethanol substituting 30% gasoline 615 GL
70
Área disponível no Brasil: 60 Mha (Zoneamento para Cana, Set 2009, aptidão
Alta e Média)
@ 10 kL/Ha (usando somente sacarose da cana) 600 GL por ano
@ 20kL/Ha (usando sacarose e celulose) 1.200 GL/ano
10052010;bioenergy-in-brazil-20100510.pptx;CHBritoCruz & BIOEN
Expansion of sugarcane to new land areas
Southwest: dry winter
Marginal land, pastureland, and poor soils
Research:
• Drought resistance
• Crop breeding to new environments
• Soil/chemical management for deep
rooting (addition of calcium)
• Chemical/fertilizer supply to compensate
for deficiencies in new land areas
• Revise nutritional needs: (inorganic
nutrients are 5% of plant dry matter)
• Optimize the use of fertilizers and
chemicals (Sugarcane: 13% of fertilizer
used in Brazil)
• Recycle nutrients of crop and industry
residues
Agro-environmental Protocol for burning phasing-out
BIOEN - HC 03-2010 GSB Project 72
UNICA, 2010
Manual harvestingNumber of workers to cut cane manually:
1.000.000.000 t (projection)6 t cane / man.day180 days / seasonapproximately 1000 t cane / man.seasonMeans 1.000.000 men cutting cane
Cane needs to be burned - EnvironmentalLower yields, higher costsor mechanization with less, but better jobs?
Social challenges: labour
Manual harvesting482 total men
US$ 6,56 / t
Mechanical harvest66 total men
US$ 4,08 / t
Burned X Green Cane
Burning phasing out in 2014/2017 in São Paulo
Green cane:Thick mulch of plant residues (8-14 t/ha DM)better soil protection and nutrient cycling (C, N)
Challenges: Problems with some insectsDifficult to incorporate fertilizers (lower efficiency, nutrient losses)Some varieties have reduced sprouting
Research needs: Assess environmental gains due to cycling, soil protection, C accumulation in soilCreate varieties adapted to green caneAdequate management practices to green cane
Pests and Diseasesmay compromise cane production
Disease resistant varietiesBiological Control (viable for several pests in sugarcane)Chemical control and crop management: combination of best management practices to minimize use of pesticides
BIOEN - HC 03-2010 GSB Project 75
• Unburned sugarcane (green cane)– +50% of sugarcane area (and
increasing)
– Deposition of large amounts of plant material on soil surface (8 to 16 t/ha DM)
– Large root system
– Soil and water conservation
– Potential to increase soil C
Sugarcane: Soil C
Depends on the referenceForest into sugarcane: loss of soil CDegraded pasture, row crops into sugarcane: soil accumulates C
gCO2eq/Mj
RFS a: Preliminary rule; b: final rule, no trash for energy and marginal electricity credit; c: final rule, trash for energy and marginal electricity credit (substitute thermal plant); LCFS a: no electricity credit; b: average electricity credit and mechanically harvested. Source: EPA; CARB; European Commission; Macedo (2009) all trash, marginal electricity credit and improved co-generation. RED: still deciding on ILUC, less credit for co-generation, sustability criteria under discussion
Sugarcane Ethanol GHG Emissions
Trash: preserve soil or use for co-generation and 2nd. generation ethanol?
ARA 003, 2007(Penatti, 2004)
Probably not
Research data in Brazil (CTC) shows that leaving 40% of trash is enough
Amounts may vary with soil type and region/climate
More research needed
Constant addition of OM needed to maintain soil chemical and biological quality
Is all the trash necessary to maintain soil fertility?
Land
Sugarcane
Pasture
Crop rotation with fallow lands
(Soybean, corn, peanuts, etc.)
Distillery(Sugar, Alcohol, Energy,
Livestock Feed)
Feedlot
Cattle
Biodigester
Lot sales
Manure
Biogas
Fertilizer
Feed
Source: NIPE-CGEE (2009)LuísCortez-June14,2010
Bioethanol and Livestock Integration
Recycling of residues
Improve recycling of residues (vinasse, filter cake, ashes, plant residues etc)
BIOEN - HC 03-2010 GSB Project 81
Filter cake and vinasse are regularly applied to the field
FAPESP BIOENERGY PROGRAM BIOEN
http://bioenfapesp.org
Program CoordinatorGlaucia Mendes SouzaDepartamento de BioquímicaInstituto de QuímicaUniversidade de São Paulo
CommitteeMarie-Anne VanSluysInstituto de Biociências - Universidade de São Paulo
Rubens MacielFaculdade de Engenharia QuímicaUniversidade Estadual de Campinas
Heitor CantarellaInstituto AgronômicoSecretaria de Agricultura e Abastecimento do Estado de São Paulo
Andre Nassar ICONE
Biomass Feedstock DevelopmentEthanol and Biofuel Technologies
Ethanolchemistry and BiorefineriesConversion technologies: Engines, Turbines, Fuel Cells
Sustainability and ImpactsBioenergy Market: Clean Tech Opportunities
Renewable Energy Policy
http://bbest.org.br
