Presentation of Marcos S. Buckeridge for the Workshop on Hydrolysis Route for Cellulosic Ethanol from Sugarcane.
Apresentação de Marcos S. Buckeridge realizada no "Workshop on Hydrolysis Route for Cellulosic Ethanol from Sugarcane"
Date / Data : February 10 - 11th 2009/
10 e 11 de fevereiro de 2009
Place / Local: Unicamp, Campinas, Brazil
Event Website / Website do evento: http://www.bioetanol.org.br/workshop1
protein chemistry, Biochemistry
the different level of organisation of the protein .
detail on individual structure and the bonds stabilising the structure of the protein.
protein chemistry, Biochemistry
the different level of organisation of the protein .
detail on individual structure and the bonds stabilising the structure of the protein.
Event / Evento: II Workshop on Sugarcane Physiology for Agronomic Applications
Speaker / Palestrante: Gaspar H. Korndörfer (Federal University of Uberlândia)
Date / Data: Oct, 29-30th 2013 / 29 e 30 de outubro de 2013
Place / Local: CTBE/CNPEM Campus, Campinas, Brazil
Event Website / Website do evento: www.bioetanol.org.br/sugarcanephysiology
Event / Evento: II Workshop on Sugarcane Physiology for Agronomic Applications
Speaker / Palestrante: Jorge Donzeli (Sugarcane Research Center - CTC)
Date / Data: Oct, 29-30th 2013 / 29 e 30 de outubro de 2013
Place / Local: CTBE/CNPEM Campus, Campinas, Brazil
Event Website / Website do evento: www.bioetanol.org.br/sugarcanephysiology
Event / Evento: II Workshop on Sugarcane Physiology for Agronomic Applications
Speaker / Palestrante: Renato Vicentini (University of Campinas - Unicamp)
Date / Data: Oct, 29-30th 2013 / 29 e 30 de outubro de 2013
Place / Local: CTBE/CNPEM Campus, Campinas, Brazil
Event Website / Website do evento: www.bioetanol.org.br/sugarcanephysiology
Event / Evento: II Workshop on Sugarcane Physiology for Agronomic Applications
Speaker / Palestrante: Frederick C. Botha (Sugar Research Australia)
Date / Data: Oct, 29-30th 2013 / 29 e 30 de outubro de 2013
Place / Local: CTBE/CNPEM Campus, Campinas, Brazil
Event Website / Website do evento: www.bioetanol.org.br/sugarcanephysiology
Apresentação de Rosana Di Giorgio realizada no "Workshop sobre Procedimentos que Regem o Relacionamento do CTBE com a Indústria"
Data: 1 de junho de 2010
Local: CTBE, Campinas, Brasil
Website do evento: http://www.bioetanol.org.br/workshop6
Apresentação de Naldo Dantas realizada no "Workshop sobre Procedimentos que Regem o Relacionamento do CTBE com a Indústria"
Data: 1 de junho de 2010
Local: CTBE, Campinas, Brasil
Website do evento: http://www.bioetanol.org.br/workshop6
Apresentação de Marco Aurélio P. Lima realizada no "Workshop sobre Procedimentos que Regem o Relacionamento do CTBE com a Indústria"
Data: 1 de junho de 2010
Local: CTBE, Campinas, Brasil
Website do evento: http://www.bioetanol.org.br/workshop6
Apresentação de Gilson Spanemberg realizada no "Workshop sobre Procedimentos que Regem o Relacionamento do CTBE com a Indústria"
Data: 1 de junho de 2010
Local: CTBE, Campinas, Brasil
Website do evento: http://www.bioetanol.org.br/workshop6
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Data: 1 de junho de 2010
Local: CTBE, Campinas, Brasil
Website do evento: http://www.bioetanol.org.br/workshop6
Presentation of Arnaldo Walter for the "2nd Workshop on the Impact of New Technologies on the Sustainability of the Sugarcane/Bioethanol Production Cycle"
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Date / Data : Novr 11th - 12th 2009/
11 e 12 de novembro de 2009
Place / Local: CTBE, Campinas, Brazil
Event Website / Website do evento: http://www.bioetanol.org.br/workshop5
Presentation of Celso Manzato for the "2nd Workshop on the Impact of New Technologies on the Sustainability of the Sugarcane/Bioethanol Production Cycle"
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Date / Data : Novr 11th - 12th 2009/
11 e 12 de novembro de 2009
Place / Local: CTBE, Campinas, Brazil
Event Website / Website do evento: http://www.bioetanol.org.br/workshop5
Presentation of Thelma Krug for the "2nd Workshop on the Impact of New Technologies on the Sustainability of the Sugarcane/Bioethanol Production Cycle"
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Date / Data : Novr 11th - 12th 2009/
11 e 12 de novembro de 2009
Place / Local: CTBE, Campinas, Brazil
Event Website / Website do evento: http://www.bioetanol.org.br/workshop5
Presentation of Robert Boddey for the "2nd Workshop on the Impact of New Technologies on the Sustainability of the Sugarcane/Bioethanol Production Cycle"
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Date / Data : Novr 11th - 12th 2009/
11 e 12 de novembro de 2009
Place / Local: CTBE, Campinas, Brazil
Event Website / Website do evento: http://www.bioetanol.org.br/workshop5
Presentation of Manoel Regis Leal for the "2nd Workshop on the Impact of New Technologies on the Sustainability of the Sugarcane/Bioethanol Production Cycle"
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Date / Data : Novr 11th - 12th 2009/
11 e 12 de novembro de 2009
Place / Local: CTBE, Campinas, Brazil
Event Website / Website do evento: http://www.bioetanol.org.br/workshop5
Presentation of Manoel Regis Leal for the "2nd Workshop on the Impact of New Technologies on the Sustainability of the Sugarcane/Bioethanol Production Cycle"
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Date / Data : Novr 11th - 12th 2009/
11 e 12 de novembro de 2009
Place / Local: CTBE, Campinas, Brazil
Event Website / Website do evento: http://www.bioetanol.org.br/workshop5
Presentation of Martin Junginger for the "2nd Workshop on the Impact of New Technologies on the Sustainability of the Sugarcane/Bioethanol Production Cycle"
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Date / Data : Novr 11th - 12th 2009/
11 e 12 de novembro de 2009
Place / Local: CTBE, Campinas, Brazil
Event Website / Website do evento: http://www.bioetanol.org.br/workshop5
Presentation of Marcia Azanha for the "2nd Workshop on the Impact of New Technologies on the Sustainability of the Sugarcane/Bioethanol Production Cycle"
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Date / Data : Novr 11th - 12th 2009/
11 e 12 de novembro de 2009
Place / Local: CTBE, Campinas, Brazil
Event Website / Website do evento: http://www.bioetanol.org.br/workshop5
Presentation of Marcel Gomes
for the "2nd Workshop on the Impact of New Technologies on the Sustainability of the Sugarcane/Bioethanol Production Cycle"
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Date / Data : Novr 11th - 12th 2009/
11 e 12 de novembro de 2009
Place / Local: CTBE, Campinas, Brazil
Event Website / Website do evento: http://www.bioetanol.org.br/workshop5
Presentation of Dr Mairi J Black
for the "2nd Workshop on the Impact of New Technologies on the Sustainability of the Sugarcane/Bioethanol Production Cycle"
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Date / Data : Novr 11th - 12th 2009/
11 e 12 de novembro de 2009
Place / Local: CTBE, Campinas, Brazil
Event Website / Website do evento: http://www.bioetanol.org.br/workshop5
Builder.ai Founder Sachin Dev Duggal's Strategic Approach to Create an Innova...Ramesh Iyer
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Date / Data: Oct, 29-30th 2013 / 29 e 30 de outubro de 2013
Place / Local: CTBE/CNPEM Campus, Campinas, Brazil
Event Website / Website do evento: www.bioetanol.org.br/sugarcanephysiology
Event / Evento: II Workshop on Sugarcane Physiology for Agronomic Applications
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Date / Data: Oct, 29-30th 2013 / 29 e 30 de outubro de 2013
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Event Website / Website do evento: www.bioetanol.org.br/sugarcanephysiology
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Date / Data: Oct, 29-30th 2013 / 29 e 30 de outubro de 2013
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Date / Data: Oct, 29-30th 2013 / 29 e 30 de outubro de 2013
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Local: CTBE, Campinas, Brasil
Website do evento: http://www.bioetanol.org.br/workshop6
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Data: 1 de junho de 2010
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Routes to Cellulosic Ethanol: biotechnological solutions for the sustaineble improvement of cell wall degradation
1. Routes to cellulosic ethanol: biotechnological
solutions for the sustainable improvement of
cell wall degradation
Marcos Buckeridge
Departamento de Botânica
Instituto de Biociências – USP
msbuck@usp.br
2. The wall in the context of plant composition
Obtained from CO2 and water
Carbon...............................................45% Cellulose,
96% Oxigen...............................................45% hemicelluloses & pectins
Hidrogen..............................................6% 96-10%=86%
Macronutrients
Proteins and
Nitrogen.............................................1.5% X 6.25 = 9.4% (10%) Nucleic acids
Potassium..........................................1.0% X
Calcium..............................................0.5%
Pectins = 0.7%
3.6% Magnesium.........................................0.2%
Phosphorous......................................0.2% X
Sulfur..................................................0.1% X
Silicium...............................................0.1% Pectins? = 0.7%
Micronutrients
Boron..................................
Pectins = traces
Manganese.........................
Chloride.............................. X
Iron..................................... X Lipids are approximately 15% of plant tissues
0.4% Sodium............................... X
Thus, the wall corresponds
Zinc.................................... X
Copper............................... X to ca. 70 % of the plant
Nickel................................. X In sugarne = leaves contain 68% and
stem 50% plus 18% of sucrose
Molibdenium.......................... X
3. V
A Cell walls from
PC leaves (left) and
ML
root (below) of
legumes
A
2µm
C
B
PC
S1 S3
S2 ML
PP
Buckeridge et al. 2008. Parede Celular, Cap 9 in Kerbauy G.B. Fisiologia Vegetal. Guanabara Koogan
4. A B
Glycosidic linkage beta-(1,4)
H bridges paper
Cellulose: the most abundant polymer on Earth.
Photograph by Cesar Gustavo Lisboa e Marcos Buckeridge, 2005
5.
6. alpha-(1,4) HOMOGALACTURONNAN
GLC
AGA AGA AGA AGA AGA AGA AGA
MAN AGA
methyl A
Egg boxes divalent
ion, maily calcium
and magnesium
induce the formatio
of gels in regions
- - that are not
- - methylated of
homogalacturonan
- - - - -
B
Buckeridge et al. 2008. Parede Celular, Cap 9 in Kerbauy G.B. Fisiologia Vegetal. Guanabara Koogan
7. GAL
beta (1,3) GAL
ARA
GAL
GAL GAL alpha (1,6)
beta (1,6) arabinogalactan I
GalA RHA
GAL GAL GAL
alpha (1,5)
GalA RHA ARA
beta (1,4)
GalA RHA
alpha (1,2)
alpha (1,4) GalA RHA
GalA RHA
GalA
Buckeridge et al. 2008. Parede Celular, Cap 9 in Kerbauy G.B. Fisiologia Vegetal. Guanabara Koogan
11. microfibril
Hemicellulose
strongly likd to
cellulose
Hemicelluluse
loosely bound to
cellulose
Pectins
Proteins
Ferulic acid
Type I Type II
Buckeridge et al. 2008. Parede Celular, Cap 9 in Kerbauy G.B. Fisiologia Vegetal. Guanabara Koogan
20. DIA
NOITE
Buckeridge et al. 2008. Parede Celular, Cap 9 in Kerbauy G.B. Fisiologia Vegetal. Guanabara Koogan
21. A
Cell expansion in
papaya during
development
50µm
PC
B
50µm
Buckeridge et al. 2008. Parede Celular, Cap 9 in Kerbauy G.B. Fisiologia Vegetal. Guanabara Koogan
22. A B
expansin
New Microfibril
Microfibril 2
Microfibril 1
Microfibril 1
Microfibril 2
ATACKS OF XTH AND EXPANSIN RELINK OF XYG BY XTH AND INTUSSUCEPTION
Buckeridge et al. 2008. Parede Celular, Cap 9 in Kerbauy G.B. Fisiologia Vegetal. Guanabara Koogan
23. Cotyledons of
Hymenaea courbaril (jatobá)
Storage walls can be
very complex
M1 antibody binds specifically to fucosylated XGs, which are present only in primary cell walls
Tiné, Braga, Hahn, Freshhour & Buckeridge, unpublished results
25. Cotyledons of Lupin: one enzyme
does the job
Buckeridge et al. 2005. Annals of Botany, Vol.96:435.
26.
27.
28.
29.
30. Sugar composition of sugarcane leaves
70
69,88
60
50
40
30
20
12,58
10 14,52
0,24
0,21
0 2,32
fuc
rham
ara
gal
glc
xyl
Collaboration with EMBRAPA Bioenergy and INCT do Bioetanol
Maria Thereza Bazzo Martins, Amanda P. De Souza, Hugo Molinari & Marcos Buckeridge
31. Parede
celular e
1
senescência
3 2
foliar
4
5
6
7 8 EMBRAPA agroenergia
Plantas de RB867515
coletadas
Maria Thereza Bazzo
33. Do composition change along the leaf blade?
Fuc % Rham % Ara %
0,40 0,30 13,5
0,25 13,0
0,30
0,20 12,5
0,20 0,15
12,0
0,10
0,10 11,5
0,05
0,00 0,00 11,0
Base Middle Tip Base Middle Tip Base Middle Tip
Gal % Glc % Xyl%
2,7 16,0 74
2,5 72
15,0
2,3
70
2,1 14,0
68
1,9
13,0 66
1,7
1,5 12,0 64
Base Middle Tip Base Middle Tip Base Middle Tip
Collaboration with EMBRAPA Bioenergy and INCT do Bioetanol
Maria Tereza Bazzo Martins, Amanda P. De Souza, Hugo Molinari & Marcos Buckeridge
34. From 1999 to 2001, the SUCEST genome program
produced 238,000 ESTs from various tissues of the
sugar cane plant.
Since then we found:
1) 469 cell wall related genes in different cane tissues
(Lima et al. 2001, GMB)
1) Determined the chemical composition and structure of the cell wall
polymers of different sugarcane tissues
35. How to modify the wall to obtain energy?
Microorganisms Change synthesis Control of hydrolysis
Change polymer
structure Activation of
Hydrolytic enzymes Increase wall and endogenous
decrease sucrose hydrolysis
Change wall
Action on the architecture
bagasse wall
MODIFIED WALL
“Papaya Cane”
Increase accessibility Increase accessibility
or” Energy cane”
Free fermentable sugars
Fermentation
Ethanol
36. THANK YOU
Marcos Buckeridge
Departamento de Botânica
Instituto de Biociências – USP
msbuck@usp.br
Hugo Molinari
EMBRAPA agroenergia
http://bioethanolbrazil.wordpress.com
37.
38. Sesbania virgata, a galactomannan storing seed
ex
me ex
en
e
Barra=34µm
Barra=84µm me
1 2A 2B
end
sc
Barra=40µm
Barra=22µm
e e
3 4
42. ROTAS PARA O ETANOL LIGNOCELULÓSICO
Quais genes? CO2
Expressão gênica Sinalização
(sequenciamento Celular Temperatura
completo)
Água
Seqüências e Mitigação e
Proteômica
propriedade das adaptação
proteínas
B Estudar a
variabilidade ETANOL
D
genética
Fermentação
Controlar a arquitetura Hidrolases fúngicas
da parede B (modo de ação, Xilose ?
B
cristalografia e
D
D genética)
CANA ENERGIA
43. A parede celular é Considerando uma sequência..
composta por A-B-C-D-E-
polímero que são
uma combinação Ácidos Nucléicos:
de Quatro pares de bases ......45 = 1024
monossacarídeos
Pode-se comparar Proteinas:
o número de
Vinte aminoácidos....205 = 3,200,000
combinações
possíveis em
relação a outras Carboidratos:
macromoléculas
Dez monossacarídeos.... 105 = 100,000
Quatro hidroxilas (hexoses) ou três (pentoses).... x 3.55 = 525
Ligações alfa ou beta... X 25 = 32
NÚMERO TOTAL DE POSSIBILIDADES = 1,680,700,000
44. Estrutura e
conformação da
celulose
cellulose
amilose glucanos
dextranos
46. Microfibrila de celulose
Ramnogalacturonano I
Ponto em que foi proposta ligação
covalente entre extensina e pectina
Extensina com tetrassacarídeos
de arabinose (losangos)
Arabinogalactano
Ponto em que foi proposta ligação
covalente entre xiloglucano e
pectina
Xiloglucano
Figura 10 . Modelo de parede celular proposto em 1973 por Peter Albersheim e colaboradores. Neste, as ligações covalentes
(exemplos circundados) seriam a principal forma de manter os diferentes polissacarídeos em interação. Note porém, que a
interação não covalente entre xiloglucano e celulose já havia sido proposta neste modelo.
47. Legumes, Ferns Legumes, Ferns
G G G
MMMMM
(Csl n)
G G Evolution of the
Cellulose-Xg binding control ?
JG cellulose-hemicellulose
FG XG
domain in plants
SG
Galactosyl tranferase
Water retention
LG G
Alpha galactosidase
MMMMM
Hardness
PP,PT,TP
G
(Csl 6)
XXXG
XXGG XXXXG MMMMM
G
Bromeliads
Epiphytism
Orchids,
XG
A A
MMMMM MMMMM
(Csl 5)
Beta glucan
Coffee,
GAX A A
Palms
Csl 4 Acetyl transferase
(cellulose)
Ara tran
Bromeliads,
Asteraceae
GGGG MGMGM
G G G
Orchids,
CesA
? Csl 3
Ferns,
Xylan MGMGM
Galactosyl transferase
48.
49. beta-(1,4) CELLULOSE
A
GLC GLC GLC GLC GLC GLC GLC GLC
beta (1,2) alpha (1,2) B
XYL XYL GAL XYL XYL
FUC
GLC GLC GLC GLC GLC GLC GLC GLC
alpha (1,6)
XYL XYL
Action of XTH e cellulases
52. beta-(1,4) PURE MANNAN
MAN MAN MAN MAN MAN MAN MAN MAN
beta-(1,4) GLUCOMANNAN
MAN GLC MAN GLC MAN GLC MAN GLC
acetyl
beta-(1,4) GALACTOMANNANO
GAL
MAN MAN MAN MAN MAN MAN MAN MAN
GAL GAL
alpha-(1,6)
53. Pectinas Celulose-Hemicelulose Proteínas
DOMÍNIOS
PAREDE
Figura 12. Modelo descontínuo da parede
celular. Neste desenho a parede é mostrada
em “corte transversal” ao eixo das microfibrilas.
Ao invés de se sustentar por ligações
covalentes, a parede celular primária e
composta por três domínios independentes:
Pectinas, Celulose-hemicelulose e Proteínas.
Os três coexistem independentemente, ou seja
sem ligações químicas covalentes, mas por
interações fracas (pontes de hidrogênio).
54. Cell wall deposition is not uniform
Buckeridge et al. 2004, Cereal Chemistry, Vol. 81 pg. 115