Optimal Conditions for Batch Tube Pretreatment Hot water only, 210 o C, 6 min -Total xylose yield is 52.1%. -49.3% xylose and 106% glucose overall mass

Optimal Conditions for Batch Tube Pretreatment

• Hot water only, 210oC, 6 min - Total xylose yield is 52.1%.

- 49.3% xylose and 106% glucose overall mass balance closure.- Cellulose digestibility at enzyme loading of 60FPU/g glucan is 57.6 % at 48 hours

and 15FPU/g glucan is 19.5% at 48hours.

• 1wt% sulfuric acid, 140oC, 40min - Total xylose yield is 95% (87% from pretreatment and 8% from enzymatic

hydrolysis); Total glucose yield in pretreatment is 8%.

- 101.7 % xylose and 97.2 % glucose overall mass balance closure.

- Cellulose digestibility at enzyme loading of 60FPU/g glucan is 94% at 48 hours and 15FPU/g glucan is 82.4% at 72 hours;

- Cellulose digestibility by SSF at the 60 FPU/g glucan enzyme loading is 94.8 % after 3 days and 15FPU/g glucan enzyme loading is 86% after 7 days.

Hydrolysis

Enzyme@ 60 FPU/g of glucan, 48h

Treated Solids

HydrolyzateLiquidSteam

Explosion

H2O

Corn Stover

100 lb(dry basis)

21.4 lb xylan

23.1 lb glucose (G)

2.1 lb xylose (X)

70.7 lb

57.6% Glucan to glucose conversion

83.4% mass balance closure (solids + G + GO + X + XO)

36.1 lb glucan

Mass Balance for Water-Only Pretreatment (60 FPU/ g of glucan)

Fermentation

2.68galethanol

Ethanol

92% theoretical ethanol yield from glucose + xylose

0.6 lb glucose (G)9.3 lb xylo-oligomer (XO)

1.1 lb gluco-oligomer (GO)

47.9 lbResidual Solids

Washing 1.66 gal ethanol

1.03 gal ethanol

9.7 lb xylan33.8 lb glucan

Hydrolysis


Treated Solids

HydrolyzateLiquidSteam

Explosion

H2O

Corn Stover

100 lb(dry basis)

21.4 lb xylan

7.8 lb glucose (G)

2.1 lb xylose (X)

70.7 lb

19.5% Glucan to glucose conversion

83.4% mass balance closure (solids + G + GO + X + XO)

36.1 lb glucan

Mass Balance for Water-Only Pretreatment (15 FPU/ g of glucan)

Fermentation

1.59 galethanol

Ethanol


0.6 lb glucose (G)9.3 lb xylo-oligomer (XO)


47.9 lbResidual Solids

Washing 0.56 gal ethanol

1.03 gal ethanol

9.7 lb xylan33.8 lb glucan

Hydrolysis


Residual Solids

HydrolyzateLiquidBatch Tube

Pretreatment

1 wt% H2SO4

Corn Stover

100 lb(dry basis)

21.4 lb xylan


21.8lb xylose (X)

64.0 lb

95.9% Glucan to glucose + 73.9% xylan to xylose conversion89.8% mass balance closure (all solids + G + GO + X + XO)

36.1 lb glucan

Mass Balance for Dilute Acid Pretreatment (60 FPU/ g of glucan)

Fermentation

4.45 galethanol

Ethanol


2.1 lb xylose (X)35.2 lb glucose (G)

0.1 lb xylo-oligomer (XO)3.0 lb glucose (G)

27.4 lbTreated Solids2.5 lb xylan33 lb glucan

2.66 gal ethanol

1.79 gal ethanol

Hydrolysis


Residual Solids

HydrolyzateLiquidBatch Tube

Pretreatment

1 wt% H2SO4

Corn Stover

100 lb(dry basis)

21.4 lb xylan


21.8lb xylose (X)

64.0 lb

84.2% Glucan to glucose + 45.8% xylan to xylose conversion89.7% mass balance closure ( all solids + G + GO + X + XO)

36.1 lb glucan

Mass Balance for Dilute Acid Pretreatment (15 FPU/ g of glucan)

Fermentation

4.08 galethanol

Ethanol


1.3 lb xylose (X)30.9 lb glucose (G)


32.4 lbTreated Solids2.5 lb xylan33 lb glucan

2.29 gal ethanol

1.79 gal ethanol

SSF

Enzyme+ D5A@ 60 FPU/g of glucan, 3 days

Batch Tube Pretreatment

Corn Stover

100 lb(dry basis)

21.4 lb xylan


21.8lb xylose (X)

64.0 lb

94.8% glucan to glucose conversion

36.1 lb glucan

Mass Balance for Dilute Acid Pretreatment w/SSF (60 FPU/ g of glucan)

4.48 galethanol

Ethanol

92.1% ethanol yield from glucose


Treated Solids

2.5 lb xylan33 lb glucan

2.48 gal ethanol from glucan by D5A

1 wt% H2SO4

(1.79 gal ethanol)

(0.20 gal ethanol from xylan)


SSF

Enzyme+ D5A@ 15 FPU/g of glucan, 3 days

Batch Tube Pretreatment

Corn Stover

100 lb(dry basis)

21.4 lb xylan


21.8lb xylose (X)

64.0 lb

86 % glucan to glucose conversion

36.1 lb glucan

Mass Balance for Dilute Acid Pretreatment w/SSF (15 FPU/ g of glucan)

4.02 galethanol

Ethanol

83.2 % ethanol yield from glucose


Treated Solids

2.5 lb xylan33 lb glucan

2.03 gal ethanol from glucan by D5A

1 wt% H2SO4

(1.79 gal ethanol)

(0.20 gal ethanol from xylan)


Optimal Conditions for Flowthrough Pretreatment

• How water only, 200oC, 10mL/min, 16-24min (depending on the biomass loading)

- Total xylose yield is 96.2%;

- 98% xylose and 100% glucose overall mass balance closure.

- Cellulose digestibility at enzyme loading of 60FPU/g glucan is 99.2% at 48 hours and 15FPU/g glucan is 96.5% after 72h hours;

• 0.05wt% sulfuric acid, 190oC, 10mL/min, 16-20min (depending on the biomass loading)

- Total xylose yield is 94.7%;

- 96% xylose and 100% glucose overall mass balance closure.

- Cellulose digestibility at enzyme loading of 60FPU/g glucan is 99.5% at 48 hours and 15FPU/g glucan is 97.8% after 72 hours;

Mass Balance For Water-Only Flowthrough Pretreatment (60 FPU/ g

of glucan)

FL reactorTreatedSolids

Hydrolysis

Enzyme @60FPU/ g of glucose, 48h

Water

Corn Stover

ResidualSolids

HydrolyzateLiquid

Fermentation

43.4 lb34.3 lb glucan

0.4 lb xylan

100 lb(dry basis)

36.1 lb glucan21.4 lb xylan

37.8 lb glucose0.4 lb xylose

4.53 gal ethanol

7.2 lb

99.2% glucan to glucose, 88% xylan to xylose at 60 FPU/ g of glucan97% overall glucan and xylan conversion92% theoretical ethanol yield from glucose + xylose97.5% overall mass balance closure (total dissolved mass+residual solids + G + X)

Dissolved mass

52.1 lb23.4 lb xylose2.1 lb glucose

2.72 gal ethanol

1.81 gal ethanol

Mass Balance For Water-Only Flowthrough Pretreatment (15 FPU/ g

of glucan)


Hydrolysis

Enzyme @15FPU/ g of glucose, 72h

Water

Corn Stover

ResidualSolids

HydrolyzateLiquid

Fermentation

43.4 lb34.3 lb glucan

0.4 lb xylan

100 lb(dry basis)



4.45 gal ethanol

7.2 lbDissolved mass

96.5% glucan to glucose, 68.2% xylan to xylose at 15 FPU/ g of glucan97% overall glucan and xylan conversion92% theoretical ethanol yield from glucose + xylose96.3% overall mass balance closure (total dissolved mass+residual solids + G + X)


1.81 gal ethanol

2.64 gal ethanol

Mass Balance For Very Dilute Acid Flowthrough Pretreatment (60FPU/g

of glucan)

FL reactorTreatedStover

Hydrolysis

Enzyme@60FPU/ g of glucan, 48h

0.05wt %wt H2SO4

Corn Stover

ResidualSolids

HydrolyzateLiquid

Fermentation

46.6 lb33.2 lb glucan0.43 lb xylan

100 lb(dry basis)



13.4 lb

99.5% glucan to glucose, 83% xylan to xylose at 60FPU/g of glucan97.5% overall glucan and xylan conversion92% theoretical ethanol yield from glucose + xylose 98.7% overall mass balance closure (total dissolved mass+residual solids + G + X)

Dissolved mass


4.55 gal ethanol

2.64 gal ethanol

1.91 gal ethanol

Mass Balance For Very Dilute Acid Flowthrough Pretreatment (15FPU/g

of glucan)


Hydrolysis

Enzyme@15FPU/ g of glucan, 72h

0.05wt %wt H2SO4

Corn Stover

ResidualSolids

HydrolyzateLiquid

Fermentation

46.6 lb33.2 lb glucan0.43 lb xylan

100 lb(dry basis)



13.4 lbDissolved mass

4.5 gal ethanol

97.8% glucan to glucose, 80% xylan to xylose at 15FPU/g of glucan97.5% overall glucan and xylan conversion92% theoretical ethanol yield from glucose + xylose 98.1% overall mass balance closure (total dissolved mass+residual solids + G + X)


1.91 gal ethanol

2.59 gal ethanol

Fermentability of Hydrolyzate from Flowthrough Pretreatment of Corn Stover

Pretreatment conditions

Hydrolyzate

(% of the medium)

Time

(hours)Ethanol Conc.

(g/L)Fermentability

(%)

Water only, 200oC,

10mL/min, 20min 80

24

72

9.8

10.0~100

Water only, 220oC,

10mL/min, 10min 80

24

7210.8

11.0

~100

0.05wt% sulfuric acid, 190oC,

10mL/min, 20min 80

24

72

11.1

11.2

~100

Control

0

( Just DI water)

24

729.6

9.8

~100

Note: Fermentability was measured and calculated according to the attached procedure

Documents

Optimal Conditions for Batch Tube Pretreatment Hot water only, 210 o C, 6 min -Total xylose yield is 52.1%. -49.3% xylose and 106% glucose overall mass