optimization and modeling of phosphoric-acid-pretreated
TRANSCRIPT
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Optimization and Modeling of Phosphoric-Acid-Pretreated
Sugarcane Bagasse
Ismael U. Nieves, Claudia C. Geddes, Ralph W. Hoffman, Mike Mullinnix, and Lonnie O. Ingram
http://fcrc.ifas.ufl.edu/
April 30th, 2012
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Introduction
• Lignocellulosic biomass as energy source – Abundant resource
– High sugar content
• Resistant to degradation
• Pretreatment is essential for downstream processes
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Introduction
Composition Analysis of Raw Sugarcane Bagasse
Sugar Percentage of
Dry Weight
Glucan 43.2 ± 0.8
Xylan 22.8 ± 0.8
Galactan 2.2 ± 1.4
Arabinnan 2.1 ± 0.6
Total Sugars 70.3 ± 1.9
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Introduction
Curr. Opin. Biotechnol. 22, 312-319
SHF – Separate Hydrolysis and Fermentation
SSF – Simultaneous Saccharification and Fermentation
SHCF – Simultaneous Hydrolysis and Co-Fermentation
L+SScF – Liquefaction +
Simultaneous Saccharification and Co-Fermentation
SScF –Simultaneous Saccharification and Co-Fermentation
CBP – Consolidated Bioprocess
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Process Simplification
Research Advances Required for Process Simplification:
1. Replace Sulfuric with less aggressive acid
2. Solve mixing and pumping issues with high fiber solids
3. Develop biocatalysts with improved resistance to hemi toxins
4. Develop process using only fertilizer chemicals a. (N, P, K, Mg, S, trace metals)
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Process Simplification
Replace sulfuric acid with phosphoric acid
– Eliminate Zirconium
– Lower the level of inhibitors
– Gypsum piles replaced with crop fertilizer
150C Verenium Sulfuric Acid Hydrolysate
0 20 40 60 80 1000
2
4
6
8
10
Furfural
HMF
0.0
0.5
1.0
1.5
Acetate
Concentration of Hydrolysate (%)
Aceta
te (
g/L
) Fu
ran
s (g
/L)
160C Phosphoric Acid Hydrolysate
0 25 50 75 1000
2
4
6
8
10
HMF
0.0
0.5
1.0
1.5
Furfural
Acetate
Concentration of Hydrolysate (%)
Aceta
te (
g/L
) Fu
ran
s (g
/L)
Bioresour. Technol. 101, 1851–1857
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Process Simplification
Research Advances Required for Process Simplification:
1. Replace Sulfuric with less aggressive acid
2. Solve mixing and pumping issues with high fiber solids
3. Develop biocatalysts with improved resistance to hemi toxins
4. Develop process using only fertilizer chemicals a. (N, P, K, Mg, S, trace metals)
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Process Simplification
0 1 2 3 4 5 60
2000
4000
6000
8000
10000
12000
0.5%
19500
20000
20500
60C 0.5-10% BioW pHz 10% Solids
1%
10%
23 24
Time (h)
Vis
co
cit
y (
cen
tip
ois
e,c
P)
0.25 FPU/g fiber 0.50 FPU/g fiber 5.0 FPU/g fiber
5 FPU 99% reduction in viscosity
Bioresour. Technol. 101, 9128–9136
Liquefaction with cellulases
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Process Simplification
Less steps
Bioresour. Technol. 102, 2702-2711.
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Process Simplification
Research Advances Required for Process Simplification:
1. Replace Sulfuric with less aggressive acid
2. Solve mixing and pumping issues with high fiber solids
3. Develop biocatalysts with improved resistance to hemi toxins
4. Develop process using only fertilizer chemicals a. (N, P, K, Mg, S, trace metals)
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Process Simplification
Strong biocatalyst developed through metabolic evolution
– Derived from KO11 strain – Increased tolerance to inhibitors – High ethanol yields (> 95%) – Co-fermentation of pentose and hexose sugars
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Process Simplification
Research Advances Required for Process Simplification:
1. Replace Sulfuric with less aggressive acid
2. Solve mixing and pumping issues with high fiber solids
3. Develop biocatalysts with improved resistance to hemi toxins
4. Develop process using only fertilizer chemicals a. (N, P, K, Mg, S, trace metals)
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Process Simplification
Fermentation scale-up to 80 L
Bioresour. Technol. 102, 6959-6965
Glu
Xyl
EtOH
Furfural
Yields;
0.27 g EtOH/g bagasse
342 L/tonne
90 gal/tonne
82 gal/US ton
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Process Simplification
• Still need further process simplification
– Minimize need for nutrients
• Modify AM1 media
– Eliminate need added sugar
• Use sugars present in hydrolysate
– Need to grow seed in hydrolysate
• Use the hydrolysate generated during pretreatment
• Start with pretreatment optimization
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Pretreatment
• Temperature
– 180 °C, 190 °C, 200 °C
• Acid concentration
– 1%, 0.5%, 0.25%, 0.1%, 0.05%, 0.01%
• Residence time
– 5, 10, and 30 min
• Measure sugars and inhibitors
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Pretreatment
Thanks to Dr. Guido Zacchi, Lund University
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Pretreatment Results
Dimers Glucose Xylose Galactose Arabinose HMF Furfural Acetic Formic Total Inhib Total Sug
5 180 0.5 50.13 5.40 84.22 4.55 12.27 0.12 1.04 7.01 0.87 9.04 156.57
5 190 0.5 41.31 13.71 181.53 8.27 16.42 0.07 4.15 18.59 2.16 24.97 261.24
5 200 0.5 22.50 19.65 188.37 8.39 14.43 0.52 7.32 26.09 2.96 36.90 253.34
10 180 0.01 10.30 1.37 4.91 1.43 10.50 0.37 5.88 1.65 7.90 28.50
10 180 0.05 19.78 1.23 9.40 1.31 9.35 0.42 4.30 0.86 5.58 41.06
10 180 0.1 38.69 1.71 25.35 1.58 9.23 0.77 4.88 2.43 8.07 76.57
10 180 0.25 56.59 5.26 104.56 4.45 8.05 2.13 10.91 0.94 13.98 178.92
10 180 0.5 44.70 8.21 125.06 6.07 13.54 3.26 14.59 1.78 19.62 211.06
10 180 1 20.30 17.19 187.82 6.87 15.27 0.39 6.91 29.66 2.39 39.35 247.45
10 190 0.01 39.91 2.92 17.01 2.68 12.75 0.66 10.84 1.97 13.46 75.27
10 190 0.05 30.00 1.60 21.18 1.75 7.00 1.56 5.62 0.93 8.11 61.53
10 190 0.1 40.05 3.07 51.22 2.77 8.21 2.34 7.07 1.09 10.49 105.32
10 190 0.25 30.28 6.47 110.49 3.53 10.79 6.81 16.44 1.31 24.56 161.56
10 190 0.5 19.69 17.64 188.30 8.37 15.38 8.79 30.37 2.97 42.13 249.38
10 190 1 6.87 27.40 185.53 5.58 12.18 1.09 14.86 39.19 2.74 57.87 237.56
10 200 0.01 57.49 3.38 47.45 2.97 7.77 6.00 17.09 3.63 26.73 119.06
10 200 0.05 53.97 5.24 92.41 4.03 10.22 7.50 18.83 2.55 28.88 165.86
10 200 0.1 37.27 10.11 131.84 6.37 11.71 9.47 24.03 2.43 35.93 197.30
10 200 0.25 22.50 23.58 185.33 9.10 9.03 14.22 36.85 2.80 53.87 249.54
10 200 0.5 12.00 26.81 178.64 7.90 15.40 0.42 17.69 40.85 4.07 63.02 240.75
10 200 1 6.55 45.01 166.42 4.85 12.65 1.41 19.71 47.94 3.47 72.54 235.48
30 180 0.1 46.46 6.58 112.20 5.70 13.51 0.46 11.03 34.96 0.71 47.16 184.46
30 190 0.1 17.26 12.81 119.83 5.81 10.47 1.45 25.42 21.61 1.90 50.38 166.18
30 200 0.1 14.38 12.94 63.45 3.49 7.78 2.31 25.27 30.51 2.74 60.82 102.04
30 200 0 21.52 9.65 66.57 3.25 6.08 0.03 22.71 38.88 7.47 69.10 107.07
Concentration (g/kg DW)Duration
(min)
Temp
(°C)
Acid
Conc (%)
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Pretreatment Model
• Combine process parameters of T, t, and C
• Relationship between severity and hemicellulose solubilization
• Model based on derivation by Chum, et al. (1990). Appl. Biochem. Biotechnol. 24-25, 1-14
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Pretreatment Model
𝑅𝑜 = 𝑡 ∗ 𝑒𝑇𝑟−10014.75 ln − ln 1 − 𝛼 = log 𝑅𝑜 − 𝑝𝐻
Ro = Severity coefficient; t = Time (min); Tr = Pretreatment temperature; α = Fraction of hemicellulose remaining
R² = 0.8751
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Pretreatment
180
185
190
195
200
40
80
120
160
200
240
280
0.0
0.2
0.4
0.6
0.81.0
10 min Pret.
(color grid)
30 min Pret.
5 min Pret.
Tota
l S
ugar
Concentr
ation (
g/k
gD
W)
Acid C
oncentra
tion (%
)
Temperature ( oC)
180
185
190
195
200
0
15
30
45
60
75
0.0
0.2
0.4
0.6
0.81.0
10 min Pret.
(color grid)
30 min Pret.
5 min Pret.
Tota
l In
hib
itors
Concentr
ation (
g/k
gD
W)
Acid C
oncentra
tion (%
)
Temperature ( oC)
180185
190
195
200
0
5
10
15
20
25
0.0
0.2
0.4
0.6
0.8
1.0
10 min Pret.
(color grid)
30 min Pret.
5 min Pret.F
urf
ura
l C
oncentr
ation (
g/k
gD
W)
Acid
Con
cent
ratio
n (%
)
Temperature ( oC)
A) Total Sugars, B) Total Inhibitors, and C) Furfural
A
B
C
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Pretreatment
180185
190
195
200
2
4
6
8
10
12
14
16
18
0.0
0.2
0.4
0.6
0.8
1.0
190oC,
0.5% acid,
5 min
180oC, 0.5% acid, 10 min 10 min Pret
(color grid)
30 min Pret
5 min Pret
Tota
l S
ugars
/Tota
l In
hib
itors
Acid
Con
cent
ratio
n (%
)
Temperature ( oC)
180oC, 0.5% acid, 5 mind e m o d e m o d e m o d e m o d e m o
d e m o d e m o d e m o d e m o d e m o
d e m o d e m o d e m o d e m o d e m o
d e m o d e m o d e m o d e m o d e m o
d e m o d e m o d e m o d e m o d e m o
d e m o d e m o d e m o d e m o d e m o
d e m o d e m o d e m o d e m o d e m o
Temperature (°C)
Acid Concentration
(%)
Residence Time (min)
Total Sugars (g/kg)
Ratio Sugars to Inhibitors
180 0.5 5 157 17.3
190 0.5 5 261 10.5 180 0.5 10 211 10.8
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Fermentation
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Fermentation
10% Solids 180C, 0.5% H3PO4, 10 min (0.5 mM SMB)
0 24 48 72 96 120 1440
5
10
15
20
25
0.0
0.1
0.2
0.3
0.4
0.5
0.6
Xylose
Cellobiose
Glucose
Arabinose
Furfural
Ethanol
Galactose
Time (h)
Concentr
ation (
g/L
)
Furfu
ral (g
/L)
10% Solids 190°C, 0.5% H3PO4, 10 min (0.5 mM SMB)
0 24 48 72 96 120 1440
5
10
15
20
25
0.0
0.1
0.2
0.3
0.4
0.5
0.6
Cellobiose
Glucose
Legend
Arabinose
Furfural
Ethanol
Galactose
Time (h)C
oncentr
ation (
g/L
)
Furfu
ral (g
/L)
190 °C, 0.5% acid, 5 min 180 °C, 0.5% acid, 10 min
Slightly better ethanol yields for the 190 °C, 0.5% acid, 5 min bagasse.
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Process Simplification
• Still need further process simplification
– Minimize need for nutrients
• Modify AM1 media
– Eliminate need added sugar
• Use sugars present in hydrolysate
– Need to grow seed in hydrolysate
• Use the hydrolysate generated during pretreatment
• Continue with seed train development
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Seed Train Development
Hydrolysate conditioning using NH4OH
Only add trace metals, MgSO4, and sodium metabisulfate
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Seed Train Development
Max Yields;
0.277 g EtOH/g bagasse
351 L/tonne
93 gal/tonne
84 gal/US ton
Sugarcane Bagasse
Max Yields;
0.309 g EtOH/g bagasse
392 L/tonne
103 gal/tonne
94 gal/US ton
Sorghum Fiber
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Stan Mayfield Biorefinery
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Location of Stan Mayfield Biorefinery Pilot Plant
Buckeye Technologies, Perry, Florida
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Stan Mayfield Biorefinery
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18,500 ft2 (1720 m2) Total Space
3,500 ft2 (325 m2) Lab Area
6,000 ft2 (560 m2) Client Space
Stan Mayfield Biorefinery
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Stan Mayfield Biorefinery
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Stan Mayfield Biorefinery
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Stan Mayfield Biorefinery
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Acknowledgements
• Lorraine Yomano
• Sean York
• David Walker
• William Sagues
• Dr. Jim Peterson
• Dr. Zhihong Fu
• Dr. Zhaohui Tong
• Dr. K. T. Shanmugam
Thank you!