ADVANCED BIOREFINERY

ASSIGNMENT 1

FLORES AVILES, IRVING

MICHELET, ARTHUR
ROCA PUIGROS, MARTA
Wageningen University
April 2015

INDEX
Problem Statement....................

Description of flowsheet for the given problems/questions...

Flowsheet Batch process.

Flowsheet Continuous process

Description of all assumptions made (physical properties, operating conditions, cost/revenue

of reactant/product) for each question.......................................

Assumptions physical properties................

Assumptions/calculations for operating conditions...

Assumptions for cost/revenue of reactant/product

Description of results obtained for each question (mass and energy balances and money
balances/economy summary).

Acidic hydrolysis reactor optimization of temperature, pH and reaction times for batch
and continuous processes

Saccharification reactor Selection of optimal temperature, pH and reaction times for

batch and continuous reactors.....

Economic evaluation of continuous process and improvements...

Economic evaluation of batch process and improvements..

Mass and energy balance for the continuous process..

10

Mass and energy balance for the batch process

11

Conclusions.....................

11

References.

13

Annex.

13

Annex 1...

13

Annex 2

13

Annex 3

20

Annex 4

21

Annex5.

21

Problem Statement
This project aims to optimize the pre-treatment of lignocellulosic biomass for the production of
butanol. In order to obtain butanol, Clostridia micro-organisms will ferment glucose and xylose
monomers. These sugars are obtained from biomass, by acidic pre-treatment and enzymatic
hydrolysis (figure 1). More specifically in the acid pre-treatment the hemicellulose opens up
and is hydrolysed to xylose. The xylose in this first step can be degraded to 2-furfural, which
is a fermentation inhibitor and should be avoided as much as possible. During this first step,
the crystallinity of cellulose decreases and it opens up for enzymatic saccharification. At the
same time, some glucose is hydrolysed from cellulose during acidic pre-treatment and can be
degraded to 5-HydroxyMethylFurfural (5-HMF). Additionally in the acid treatment the lignin seal
is broken. All the reactions taking place in the acid pre-treatment increase the biomass surface
area. Finally during the enzymatic hydrolysis or saccharification process (2nd step), the
cellulose is hydrolysed to glucose.
On one hand, we are aiming to optimize both steps in order to get as much glucose and xylose
with the least fermentation inhibitors. Several parameters will be studied for optimization,
mainly process pH, temperature and reaction time.
On the other hand the optimized process is analysed from an economic point of view, to assess
its feasibility, strengths and weaknesses.

Lignocellulosic
material

Acid Pretreatment

Enzyme
hydrolysis

Product

Figure 1 Overall scheme of the process

Description of flowsheet for the given problems/questions

Flowsheet Batch process
Milled and clean biomass (in our case wheat straw) undergoes a dilute acid pre-treatment, at
a temperature of 200C, pressure of 16bar (this pressure is selected to have no water
evaporation), pH 2 (to have a high conversion yield) and using a residence time of 9min (in
later sections of this report these values are fully studied). In this first reactor, water and
sulphuric acid are also added to obtain a final pH of 2 and a slurry concentration of 100g/l of
solids. After this acidic treatment, the pressure in the reactor is dropped through a
depressurization valve to 1 bar and a cooling step takes place to reduce the temperature down
to 50C. Both processes (drop of pressure and cooling) take place inside the reactor itself. See
in the Annex 1 the composition of the stream S-102.

Afterwards a conditioning step is necessary to adjust the pH to 5 (the optimal pH for the
saccharification process) by adding sodium hydroxide.
The second step of the process consists in a saccharification using cellobiohydrolases and glucosidases (provided in the Celluclast mix, Novozymes). This step is performed at 50C, pH
5, 1atm and the reaction time is of 16h. The enzyme is used in a relatively high, 3% per mass,
concentration. See in the Annex 1 the composition of the stream S-103.

Figure 2 Flowsheet for a 2-stage batch process (acid pre-treatment and saccharification)

At last, a separation is required to have a product (glucose and xylose) as pure as possible.
To achieve that, a rotatory vacuum filtration is used, so the cake fraction (S-106) containing
the remaining solids: lignin, hemicellulose and cellulose, some water (not all the water can go
through the filtrate) and some solubles. The filtrate stream (product stream) contains mainly
glucose, xylose and water, however it contains a big portion of 2-furfural and salts from the
neutralization of NaOH and H2SO4. See composition of both streams S-106 and product
stream in Annex 1.
Flowsheet Continuous process

Figure 3 Flow sheet for a 2-stage continuous process (acid pre-treatment and saccharification)

The same process was designed for a continuous system. The conditions are similar to the
batch process, except that the residence times are longer in the continuous process for the
acid pre-treatment, being of 12min here. A counter-current heat exchanger is also added in
order to reduce heating and cooling costs and increase sustainability of the process, therefore
the stream will be preheated up to 147.8C before entering the acid pre/treatment step and

cool down to 50C before undergoing the saccharification. A depressurization valve is also
added to decrease the flows pressure down to 1bar before entering the saccharification
reactor.
See composition of all flows in Annex 2.
Description of all assumptions made (physical properties, operating conditions,
cost/revenue of reactant/product) for each question
Assumptions physical properties

Table 1 Physical properties assumptions

Compound
MW (g/mol)

The assumptions taken into account for the physical

2Furfural

96.09

properties of the compounds participating in the

5HMF

126.11

process are depicted in Table 1. The only physical

Glucose

180.16

Hemicellulose

132.12

Cellulose

162.14

Lignin

Xylose

150.13

properties needed for the process are the molecular

weights.
Assumptions/calculations for operating conditions
Table 2 Assumption for operating conditions for
both steps
1st Step
2nd Step
(Acid
Condition
(Saccharifi
treatme
cation)
nt)

The assumptions and calculated conditions for

each step of our process are depicted in Table2.
These calculated conditions are studied in detail
in the next section of this report.

Temperature

200C

50C

Pressure

16bar

1bar

The calculated values needed for sulphuric acid,

pH

sodium hydroxide and cellulase in order to obtain

Residence Time
Batch

9min

16h

a pH of 2 and 5, and a concentration of 3% of

Residence Time
Continuous

12min

16h

hemi (%) batch

84.2

--

hemi (%) continuous

71.32

--

xyl (%) batch

49.5

--

xyl (%) continuous

37.08

--

enzyme respectively are depicted in Table 3).

See the detailed calculations in Annex 1, table
A.1.
Table 3 Calculated values for sulphuric acid,
sodium hydroxyde and cellulase used in the
process

stands for conversion degree

Item

Batch

Continuous

H2SO4

490kg/batch

490kg/day

NaOH

398.60kg/batch

35.83kg/day

Cellulases

3ton/batch

3ton/day

Assumptions for cost/revenue of reactant/product

Milled and cleaned wheat straw was chosen as lignocellulosic material, it is a by-product from
wheat production and is considered a waste after being separated from the grain (1).
Moreover wheat straw has a high percentage of cellulose; 48.570.30% (50% was used in our
calculations), the following content of hemicellulose 27.70.12% (30% was used in our

calculations) and about 15% of

lignin

content

(1)

(2).

Table 4 Assumptions for costs and revenues of the items

Item

The

assumptions made in this project

regarding costs and revenues are
depicted in the table 4.

Price (euros)

Costs Milled Wheat

190euros/ton (3)

Costs Sulfuric Acid

211.6euros/ton (4)

Costs Water

0.10euros /ton

Costs Sodium Hydroxide

276euros/ton (5)

Costs enzymes (Celluclast)

Revenue sugars (Glucose and
Xylose)

91.08euros/ton (6)
460euros/ton

Description of the results obtained for each question (mass and energy balances and
money balances/economy summary)
The kinetics for the different reactions happening in both reactors have been established in
order to optimize the reactors conditions. The different reactions taking place are;
In the acidic hydrolysis reactor (AH);
k Xyl

kHemi

Hemicellulose water Xylose 2Furfural

k Cell

(Reaction 1)

k Glu

Cellulose water Glucose 5HMF

(Reaction 2)

In the saccharification reactor (SR);

+
+

Acidic hydrolysis reactor optimization of temperature, pH and reaction times for batch and
continuous processes.
The big issue for this process was to determine realistic reaction conditions in order to obtain
a good conversion degree of hemicellulose into xylose, with as little as possible xylose being
degraded into 2-furfural. The hydrolysis of cellulose and the degradation of glucose will be
neglected in further discussions for very little cellulose is being hydrolysed and therefore very
little glucose is being degraded into 5HMF comparatively to the amount of xylose and 2-furfural
formed. This is explained by the difference in reaction kinetics, indeed khemi is about 1000 times
higher than kcell (See Table A.3 and A.4 in Annex2).
In order to obtain the ideal conditions for reaction 1, the different reaction rate constants have
been calculated at different pH and temperatures. Furthermore, the conversion degrees of
hemicellulose and xylose are then compared for these different conditions.

Figures 4 and 5 (see values and calculations in tables A.12 and A.13 Annex 2) show the
different reaction constants
for

the

hydrolysis

cellulose

and

radation

of

tendencies

of

the

deg-

xylose.

Two

can

be

observed. First, khemi increases exponentially with

the temperature and with the
decrease in pH. Second, kxyl
increases exponentially with
the temperature and with the
pH.
Figure 4 Hemicellulose hydrolysis constant (1/min) in function of pH and
Temperature (C)

It can then be said that, in

order

to

favour

the

hydrolysis of hemicellulose
and limit the degradation of
xylose,

an

intermediate

temperature and a low pH

should

be

selected.

Indeed, Figure A.2, annex 2

shows

us

that,

at

temperature of 200C, the

pH has to be lower than 2.6
Figure 5 Xylose degradation constant (1/min) constant in function of pH and
Temperature (C)

for the xylose formed by

hydrolysis

to

accumulate

and to not be degraded straight away. Above pH 2.6, the degradation of xylose goes faster
than its formation, which means that all the xylose formed at a given time will be degraded and
no xylose will be present in the output stream of our AH reactor. The conditions we have
therefore selected for our AH reactor are 200 C and pH 2. However, once conditions are
selected, it is important to understand the influence of the reaction time on the formation of
products.

Therefore, the conversion degrees of hemicellulose and xylose in function of the

reaction/residence time have been calculated for both the batch and the continuous process.
These results are plotted in Figures 6 and 7 (See calculations and values in Table A.7,
Annex2). In order to retrieve the optimal reaction times for both our processes, the following
selection criteria have been posed;

We want our process to hydrolyse at least 70% of the hemicellulose,

We want the highest difference between the conversion degrees of hemicellulose and
xylose.

Figure A.1 in Annex 2, represents the difference between the conversion degrees of
hemicellulose and xylose in function of reaction time and different optimum reaction times can

Figure 6 Conversion degree of Hcell and Xylose in function

of reaction time in a batch process at conditions of 200C and
pH 2

Figure 7 Conversion degree of Hcell and Xylose in function of

reaction time in a continuous process at conditions of 200C and
pH 2

be retrieved for the batch and continuous process. Based on the two previous criteria, the
optimum reaction times for the batch and continuous processes are 9 and 12 minutes
respectively.
It must be precised that these points can be subject to discussion, as they represent an
optimum based on the amount of 2-furfural obtained and the amount of hemicellulose
hydrolysed. In a process where the hydrolysis of hemicellulose is a priority, longer reaction
times could be chosen, although the amount of xylose lost as 2-furfural quickly climbs to very
high percentages of 60-70% for hemicellulose conversions of 90%. With all these parameters
found, the AH reactor has been optimized for both batch and continuous processes, the final
results can be seen in Table 2 (in previous section).

Saccharification reactor Selection of optimal temperature, pH and reaction times for batch
and continuous reactors.
For this reactor, a cellulolytic enzyme had to be chosen and its optimal conditions selected for
the best saccharification yield. For further calculations, the reference cellulase selected was
CBHI from the fungi Trichoderma reesei, a cellobiohydrolase exo-active enzyme that cleaves
cellulose at the reducing end and produces cellobiose, a glucose dimer. The exo-active
cellobiohydrolase was selected as reference for the reactor kinetics because its activity, more
precisely its adsorption to the substrate cellulose, is the limiting step in the whole cellulolytic
activity. The other steps like the enzymatic catalysis of the formation of cellobiose and further
the cleavage of cellobiose into glucose are both faster than the adsorption of CBHI to the
cellulose.
In this case then, determining the kinetics of this limiting step determines the general kinetics
of the whole saccharification reaction.
Quantitative information about the adsorption equilibrium and kinetics have been found in Lynd
et al., (2002) (7). The reaction rate kcell calculated was of 8.14e-5 s-1 (or 4.89e-3 min-1) for a
concentration of 3% enzyme, a pH of 5 and at 50 C (see calculations in Table A.14 Annex 3).
To obtain the ideal residence time for our batch and continuous process, Figure 8 (see values
and calculations in table A.15, Annex 3) shows the conversion degree of cellulose in function
of reaction time. Once again, the batch process performs better than the continuous one, with
a conversion of 90% obtained after only 8 hours, whereas the continuous process needs 14
hours to get to a conversion of 80%.
To obtain the ideal reaction time,
Conversion degree of Cellulose in function of
time for batch and continuous process
batch

the

continuous

costs

and

revenues should be compared

100.00

for

80.00

(%)

operating

different

times

and

conversions. Furthermore, if the

60.00

conversion

40.00

is

not

complete,

some cellulose is lost as waste

20.00

or can be recycled for an extra

0.00
0

10

20

30

Time (h)
Figure 8 Conversion degree of Cellulose in function of time for batch
and continuous process

cost, which would be the case for

the continuous process where
reaction rates are much lower.
For the batch process it is

possible to avoid waste by applying a reaction time of 16 hours for a conversion of 99%. The

optimal solution here is attainable for the batch process (16h), and is not realistic for a
continuously stirred reactor who would need 2 weeks reaction time to get to 99% conversion.
In the idea of implementing a continuous process, a certain number of batches could be
implemented in parallel after a continuous acidic hydrolysis tank. That way, a continuous
process is achieved by taking advantage of the much better conversion achieved in batch
reactors.
Economic evaluation of continuous process and improvements
In the continuous process, as can be seen in Table 5, the operating costs represent the major
source of expenses and these costs are themselves linearly related to the reaction time in the
saccharification reactor. Raw materials represent only a fraction of the expenses, about 105
000$/yr for the enzymes, NaOH and H2SO4. The major optimization will therefore take place
on the saccharification time.
Labour represents 975 857$/yr, 5% of
total operating costs, which is not really

Table 5 Economic data of the continuous process, for a

saccharification time of 4h

an issue with room for optimization.

Utilities such as high pressure steam
and

cooling

water

represent

395

223$/yr, roughly 2% of operating costs,

once again not a bottleneck of the
process with room to optimize. The
enzyme loading does not represent such a bottleneck in the process neither, costing only
28,452$/yr, representing roughly 30% of raw materials expenses.
Compared to labour, utilities and raw materials, it is the operating costs that represent the real
bottleneck in this process, with an annual cost of 18 million $ for 4h saccharification time,
evolving linearly with that time.
Figure A.3, Annex4 shows clearly the optimum point for the saccharification time based on the
return on investment. This result brings a challenge, because at a saccharification time of 4
hours, the cellulose conversion degree achieved is around 60%. A recycling method should
then be developed to re-insert the non-hydrolysed cellulose back into the process. This would
add additional costs but would permit much shorter residence times, thus cut existing costs.
Economic evaluation of the batch process and improvements
For the batch process, as can be seen in Table 6, the operating costs represent the major
source of expenses and are also linearly related to the reaction time in the saccharification

reactor as the continuous process. Material costs represent 54% of the operating costs, and
enzymes about 30 000$/yr.
Labor represents 2,374,860$/yr, 16% of total operating costs, due to multiple units during the
operation. Utilities such as high pressure steam and cooling water represent a considerable
cost
Table 6 Economic data of the batch process, for a saccharification time of 16h

of
5,039,000$/yr,

roughly

34%

of

operating costs due

to the high amount
of

cooling

water

that is used during

the cooling step in
the

AH

reactor.

Here, there is room

for optimization.
Operating costs represent the real bottleneck in this process, with an annual cost of 15 million
$ for 16h saccharification time. Dependent facility costs were ignored to visualize a scenario
in which a gain is generated after 12.27 years of having made the investment.
Mass and energy balance for the continuous process
Table 7 Mass balance for continuous process, with an acidic hydrolysis time of 12
min and a saccharification time of 4h, all results are in tons/day

In Table 7 the mass

balance

for

our

continuous process
can

be

seen.

conversion of 72%
of the hemicellulose
is achieved, with a
degradation degree
of xylose of 37.5%.
Cellulose is hydrolysed to an extent of 57%, not much considering the achievable yields in a batch process.
However, the non-hydrolysed cellulose can be re-injected in the system.
Energy is saved with a counter-current heat exchanger, see its specifications in Table A.16,
Annex 5. The cold out stream is then heated to 200C consuming 2.7 ton/h of high pressure
steam.

10

Mass and energy balance for the batch process

In

Table

the

mass

balance

for

the

batch

process

can

be

seen.

A conversion of 86% of the hemicellulose is achieved, with a degradation degree of xylose of

25%. Regarding cellulose, it is shown in Table 8 a 100% of hydrolysis (solids out). The batch
cooling process in the acid pretreatment

consumes

56467.80

Table 8 Mass balance for batch process, with an acidic hydrolysis time
of 9 min and a saccharification time of 16h, all results are in tons/day

Mt/h of cooling water to cool from

200C to 50C. The temperature is
maintained at 50C the rest of the
process

up

after

the

acidic

hydrolysis reactor (Table A.17,

Annex 5).
Conclusion
When designing the processes to
extract sugars from lignocellulosic material, the first challenge we encountered was the amount
of 2-furfural formed after the acid pre-treatment. In order to reduce its concentration the optimal
conditions were calculated keeping a relatively high xylose production, however after
performing such an approach, a xylose degradation of 37.5% was obtained for our continuous
process and of 25% for our batch process. Taking into account the parameter of xylose
degradation, a batch process would be chosen over a continuous one. Moreover the
conversion of hemicellulose is higher for a batch process than in a continuous process.
Regarding cellulose, a full hydrolysis is achieved with a batch process, whereas only 57%
hydrolysis is achieved with our continuous process. It seems that, as such, a batch process
woud be more interesting than a continuous one, regarding yields and inhibitor formation.
However, a continuous process has many advantages and if the process had to be
industrialized, special attention should be paid to make the process continuous, using batch
reactors
Economically, the batch process has a much higher capital investment than the continuous
process and lower operation costs; since the batch process provides more product its gross
margin is less negative than the continuous process. Hence economically the batch process
is a better option than the continuous.
Another interesting parameter to optimize in the future is the valorisation of the solids waste
stream, containing mainly lignin. The merchandising of this stream could not only reduce process costs but could also provide a valuable and interesting source of bio-based chemicals for
the industry.
11

References
(1) Saha, B. C.; Iten, L. B.; Cotta, M. A.; Wu, X. V. Diluted acid pretreatment enzymatic
saccharification and fermentation of wheat straw to ethanol. 2005. Process
Biochemistry 40:3693-3700.
(2) Qureshi N; Saha, B. C.; Cotta M.A. 2007. Butanol production from wheat Straw
hydrolysate using Clostridium beijerinckii. Bioprocess Biosyst Eng 30:419-427
(3) http://www.hgca.com/markets.aspx
(4) http://www.alibaba.com/trade/search?fsb=y&IndexArea=product_en&CatId=&Search
Text=sulfuric+acid
(5) http://www.alibaba.com/trade/search?fsb=y&IndexArea=product_en&CatId=&Search
Text=sodium+hydroxide
(6) Klein-Marcuschamer, D; Oleskowciz-Popiel, P; Simmons, B. A.; Blanch, H. W. 2011.
The challenge of enzyme cost in the production of Lignocellulosic Biofuel.
Biotechnology and Bioengineering.
(7) Lynd, L.R., P.J. Weimer, W.H. van Zyl, and I.S. Pretorius. 2002. Microbial cellulose
utilization: fundamentals and biotechnology. Microbiol Mol Biol Rev 66:506577

12

Annex
Annex 1
Table A.1 Calculations of amount of acid sulphuric and sodium hydroxide needed

H2SO4
Mw

98

g/mol

Mw

NaOH
39.9

Volume

1.00E+06

pH

pH

[NaOH]

0.00999

mol/l

[H+]

0.01

[NaOH]

0.398601

g/l

[H2SO4]

0.005

mol/l

mNaOH

398.60

kg/"batch"

[H2SO4]

0.49

g/l

mH2SO4

490

kg/"batch"

g/mol

Annex 2: Calculations for the acid pre-treatment step

Table A.2 Data needed for Kcell and Kglu calculations

Cellulose data
kcell0
pH
Ecell
mcell
Slurry Concentrations
T
R
Cs,0 (mol/m3)
Glucose data
Eglu
T
R
pH
Slurry Concentrations

4.44E+18
2
165000
1.28
100
473
8.31
308.6666667
121000
473
8.3
2
100

Table A.3 Determination of reaction kinetic constants for cellulose and glucose at different temperatures

T(C)

T(K)

kcell (1/min)

kglu (1/min)

Acell

Aglu

120

393

1.39785E-06

5.5746E-05

2.03813E+14

11450000000

130

403

4.89694E-06

0.000139794

1.06963E+13

16250000000

140

413

1.61444E-05

0.000335294

150

423

5.03066E-05

0.000771615

160

433

0.00014874

0.001708662

170

443

0.00041877

0.003650265

180

453

0.001126355

0.007541152

190

463

0.002902776

0.015098687

200

473

0.007187332

0.029355683

210

483

0.017140255

0.055524999

220

493

0.039459766

0.102342462

230

503

0.087880525

0.184104258
13

Table A.4 Calculation of conversion degree of cellulose in function of reaction time at a pH of 2 and temperature of
200C

k(min-1)

res
time
(min)

Cell
concentration
batch

Cell
concentratio
n CSTR

batch

CSTR

Glucose
Formed
(Batch)

Glucose
Formed
(CSTR)

0.00718
7

308.667

308.667

0.000

0.000

0.000

0.000

306.456

306.464

0.716

0.714

2.211

2.203

304.261

304.293

1.427

1.417

4.405

4.374

302.082

302.152

2.133

2.111

6.584

6.515

299.919

300.041

2.834

2.795

8.748

8.626

297.771

297.959

3.530

3.469

10.896

10.708

295.639

295.906

4.221

4.134

13.028

12.761

293.521

293.881

4.907

4.790

15.145

14.786

291.419

291.884

5.588

5.437

17.247

16.783

289.332

289.913

6.264

6.076

19.334

18.753

10

287.260

287.969

6.935

6.705

21.406

20.697

11

285.203

286.051

7.602

7.327

23.464

22.615

12

283.161

284.159

8.263

7.940

25.506

24.508

13

281.133

282.291

8.920

8.545

27.534

26.376

14

279.119

280.447

9.573

9.142

29.547

28.219

15

277.120

278.628

10.220

9.732

31.546

30.039

16

275.136

276.832

10.863

10.314

33.531

31.835

17

273.165

275.059

11.502

10.888

35.501

33.608

18

271.209

273.308

12.135

11.455

37.458

35.358

19

269.267

271.580

12.765

12.015

39.400

37.087

20

267.338

269.873

13.389

12.568

41.328

38.793

30

248.798

253.917

19.396

17.737

59.868

54.750

For the batch calculation the equation used is the following one:
Table A.5 Data needed for KHemi and Kxyl calculations

dCs/dt = -k*Cs
Cs,t=Cs,0 * exp(-kt)

HemiCelullose data
kcell0

1.47E+12

pH

Ecell

104000

mcell

0.68

Slurry Concentration

100

120

R
hcel conc

8.31
(mol/m3)

227.268

Xylose Data
Exyl

141000

120

8.3

pH
Slurry Concentration

2
100

14

Table A.66 Determination of reaction kinetic constants for hemicellulose and xylose at different temperatures

T(C)

T(K)

khcell (1/min)

kxyl (1/min)

Ahcell

Axyl

120

393

0.00094898

5.11708E-05

1069463789

4.8E+12

130

403

0.002091388

0.00014938

1069463789

4.8E+12

140

413

0.00443602

0.000414029

150

423

0.009080553

0.001093542

160

433

0.017982939

0.002761582

170

443

0.034531214

0.006688316

180

453

0.064424792

0.015578115

190

463

0.117002409

0.034982542

200

473

0.207194971

0.075915795

210

483

0.358333022

0.159543818

220

493

0.606098249

0.32534393

230

503

1.003976235

0.644912442

Table A.7 Calculation of conversion degree of hemicellulose and xylose degradation in function of reaction time at
a pH 2 and temperature of 200C
res
HCell
HCell
Xylose Xylose
Xylose
Xylose

k(min-1)
time
concentration concentration
formed formed degraded degraded
(batch) (CSTR)
(min)
batch
CSTR
(batch) (CSTR) (batch) % (CSTR) %
0.207194
0
227.27
227.27
0.00
0.00
0.00
0.00
0.00
0.00
(khcell)
0.075915
1
184.74
188.26
18.71
17.16
42.53
39.01
7.31
6.47
(Kxyl)
2
150.17
160.68
33.93
29.30
77.10
66.59
14.09
11.38
3

122.06

140.15

46.29

38.33

105.20

87.12

20.37

15.36

99.22

124.27

56.34

45.32

128.05

102.99

26.19

18.74

80.65

111.63

64.51

50.88

146.62

115.64

31.59

21.70

65.56

101.32

71.15

55.42

161.71

125.95

36.59

24.38

53.29

92.75

76.55

59.19

173.98

134.52

41.22

26.83

43.32

85.52

80.94

62.37

183.95

141.75

45.52

29.12

35.21

79.33

84.51

65.09

192.06

147.94

49.50

31.27

10

28.62

73.98

87.41

67.45

198.65

153.29

53.19

33.30

11

23.27

69.31

89.76

69.50

204.00

157.96

56.62

35.24

12

18.91

65.19

91.68

71.32

208.36

162.08

59.79

37.08

13

15.37

61.53

93.24

72.93

211.90

165.74

62.73

38.85

14

12.50

58.26

94.50

74.36

214.77

169.01

65.45

40.54

15

10.16

55.32

95.53

75.66

217.11

171.94

67.98

42.17

16

8.26

52.67

96.37

76.83

219.01

174.60

70.32

43.72

17

6.71

50.25

97.05

77.89

220.56

177.01

72.49

45.22

18

5.46

48.05

97.60

78.86

221.81

179.21

74.50

46.65

19

4.43

46.04

98.05

79.74

222.83

181.23

76.36

48.03

20

3.60

44.18

98.41

80.56

223.66

183.09

78.09

49.35

30

0.45

31.50

99.80

86.14

226.81

195.77

89.75

59.98

40

0.06

24.47

99.97

89.23

227.21

202.80

95.20

67.14

50

0.01

20.01

100.00

91.20

227.26

207.26

97.75

72.18

60

0.00

16.92

100.00

92.55

227.27

210.35

98.95

75.89

70

0.00

14.66

100.00

93.55

227.27

212.61

99.51

78.73

15

80

0.00

100.00

94.31

227.27

214.34

99.77

80.98

90
0.00
11.57
100.00
For the batch calculation the equation used is the following one:

94.91

227.27

215.70

99.89

82.79

Batch

40.00

hemi - xyl (%)

12.93

dCs/dt = -k*Cs
Cs,t=Cs,0 * exp(-kt)

Continuous

30.00
20.00
10.00
0.00
0

10
15
Incubation time (min)

20

25

Figure A.1 Difference in conversion degree between the formation

and degradation of xylose at 200C and pH 2
Table A.8 Calculations of reaction kinetic constant for cellulose and glucose in function of pH

pH

kcell (1/min) kglu (1/min)

Acell

Aglu

2 0.007187332 0.029355683 2.03813E+14 11450000000

2.1 0.005352636 0.028557428 1.51786E+14 11138645493
2.2 0.003986279 0.028164944

1.1304E+14 10985559695

2.3 0.002968709 0.028157331 8.41844E+13 10982590231

2.4 0.002210893 0.028534183 6.26948E+13 11129578964
2.5 0.001646523 0.029315568 4.66908E+13 11434353582
2.6 0.001226218

0.0305431 3.47722E+13 11913144444

2.7 0.000913204 0.032282147 2.58959E+13 12591448909

2.8 0.000680092 0.034625321 1.92855E+13 13505389163
2.9 0.000506486 0.037697404 1.43626E+13 14703635858
3 0.000377197 0.041661996 1.06963E+13 16250000000
3.1

0.00028091 0.046730225 7.96584E+12 18226831118

3.2 0.000209203 0.053171994 5.93242E+12 20739402677

3.3

0.0001558

0.06133035 4.41806E+12 23921518276

3.4 0.000116029 0.071639755 3.29027E+12 27942637173

3.5 8.64107E-05 0.084649223 2.45037E+12 33016898613
3.6 6.43528E-05 0.101051556 1.82487E+12 39414525521
3.7 4.79256E-05 0.121720237 1.35904E+12 47476214859
3.8 3.56917E-05 0.147755949 1.01212E+12 57631280981
3.9 2.65808E-05 0.180545188 7.53758E+11 70420518189
4 1.97956E-05 0.221834102 5.61347E+11 86525000000

16

khcell (1/min)

0.6

kxyl (1/min)

0.5

0.4
0.3
0.2
0.1
0
2 2.1 2.2 2.3 2.4 2.5 2.6 2.7 2.8 2.9 3 3.1 3.2 3.3 3.4 3.5 3.6 3.7 3.8 3.9 4
pH
Figure A.2 Reaction constants for hemicellulose and xylose in function of pH at 200C

Table A.9 Determination of cellulose converted and glucose formed in batch and CSTR in the acid pre-treatment
at pH of 3 (similar as table A.4 in different ranges of residence time and pH)

K
(min-1)
0.000377

res
time
(min)
0

Cell
converted
batch
185.2

Cell
converted
CSTR
185.2

Glucose
Formed(Batch
)
0

Glucose
Formed(CSTR
)
0

10

184.5027479

184.5040571

0.37648

0.697252123

0.695942883

20

183.8081208

183.8133251

0.75155

1.391879189

1.386674931

30

183.1161089

183.1277455

1.12521

2.083891082

2.072254451

40

182.4267024

182.4472611

1.49746

2.773297646

2.75273888

50

181.7398913

181.7718152

1.86830

3.460108691

3.428184807

60

181.055666

181.101352

2.23776

4.144333989

4.098647986

70

180.3740167

180.4358167

2.60582

4.825983273

4.76418335

80

179.6949338

179.775155

2.97249

5.505066244

5.424845028

90

179.0184074

179.1193136

3.33779

6.181592562

6.080686361

100

178.3444281

178.4682401

3.70171

6.855571852

6.731759911

110

177.6729863

177.8218825

4.06426

7.527013705

7.378117481

120

177.0040723

177.1801899

4.42544

8.195927674

8.019810126

For the batch calculation the equation used is the following one:

dCs/dt = -k*Cs
Cs,t=Cs,0 * exp(-kt)

Table A.10 Calculations of reaction kinetic constant for hemicellulose and xylose in function of pH

kxyl (1/min)

Ahcell

Axyl

khcell
(1/min)
0.207194971

0.075915795

1069463789

4.8E+12

2.1

0.177165523

0.077144328

914462887

4.87768E+12

2.2

0.151488341

0.07869096

781926774

4.97547E+12

2.3

0.129532638

0.080638054

668599557

5.09858E+12

2.4

0.110759047

0.083089299

571697225

5.25357E+12

2.5

0.094706374

0.086175235

488839267

5.44868E+12

2.6

0.080980268

0.090060197

417990185

5.69432E+12

2.7

0.069243532

0.094951075

357409494

6.00356E+12

2.8

0.059207839

0.101108326

305608961

6.39287E+12

pH

17

2.9

0.050626652

0.108859846

261316050

6.88298E+12

0.043289165

0.11861843

223442656

7.5E+12

3.1

0.037015124

0.130903761

191058377

8.27678E+12

3.2

0.031650401

0.146370075

163367658

9.25468E+12

3.3

0.027063204

0.165841012

139690247

1.04858E+13

3.4

0.023140845

0.190353468

119444480

1.20357E+13

3.5

0.019786966

0.221212823

102132999

1.39868E+13

3.6

0.016919176

0.260062449

87330527.7

1.64432E+13

3.7

0.014467024

0.30897123

74673427.2

1.95356E+13

3.8

0.012370271

0.370543737

63850762

2.34287E+13

3.9

0.010577407

0.448058932

54596661.2

2.83298E+13

0.009044389

0.545644779

46683787.6

3.45E+13

Table A.11 Determination of hemicellulose converted and xylose formed in batch and CSTR in the acid pretreatment at pH of 3 (similar as table A7 in different ranges of residence time and pH)
res
time
(min)
0

378.78

HCell
converted
CSTR
378.78

10

245.6883615

264.3465746

35.1369

133.0916385

114.4334254

20

159.3610301

203.0139294

57.9278

219.4189699

175.7660706

30

103.366467

164.7818884

72.7106

275.413533

213.9981116

40

67.04667064

138.6676784

82.2993

311.7333294

240.1123216

50

43.48853332

119.6982134

88.5187

335.2914667

259.0817866

60

28.20800067

105.2941775

92.5523

350.5719993

273.4858225

70

18.29657708

93.98444088

95.1696

360.4834229

284.7955591

80

11.8677228

84.86862965

96.8668

366.9122772

293.9113703

90

7.697770131

77.36480312

97.9677

371.0822299

301.4151969

100

4.993010535

71.08011517

98.6818

373.7869895

307.6998848

110

3.238620247

65.73977972

99.1449

375.5413798

313.0402203

120

2.100668731

61.14581994

99.4454

376.6793313

317.6341801

130

1.362558368

57.1519837

99.6402

377.4174416

321.6280163

140

0.883797278

53.64788686

99.7666

377.8962027

325.1321131

150

0.573258105

50.54865279

99.8486

378.2067419

328.2313472

160

0.371832844

47.78794698

99.9018

378.4081672

330.992053

170

0.241182222

45.31317582

99.9363

378.5388178

333.4668242

180

0.156438209

43.08210387

99.9587

378.6235618

335.6978961

190

0.101470636

41.06042348

99.9732

378.6785294

337.7195765

200

0.065816976

39.21997807

99.9826

378.714183

339.5600219

210

0.042690916

37.53744271

99.9887

378.7373091

341.2425573

220

0.027690641

35.99333039

99.9926

378.7523094

342.7866696

230

0.017961002

34.57123384

99.9952

378.762039

344.2087662

240

0.011650058

33.25724021

99.9969

378.7683499

345.5227598

250

0.007556586

32.03947445

99.9980

378.7724434

346.7405256

260

0.004901434

30.90773932

99.9987

378.7750986

347.8722607

K
(min-1)
0.0432

HCell converted
batch

Xylose formed
(batch)

Xylose formed
(CSTR)

18

270
0.003179221
29.85322926
99.9991 378.7768208
348.9267707
Table A.12 Determination of hemicellulose hydrolysis constant in function of pH and temperature
T
pH
2
2.1
2.2
2.3
2.4
2.5
2.6
2.7
2.8
2.9
3
3.1
3.2
3.3
3.4
3.5
3.6
3.7
3.8
3.9
4

120

130

140

150

160

170

180

190

200

210

220

230

0.0009
49
0.0008
11

0.0020
91
0.0017
88

0.0044
36
0.0037
93

0.0090
81
0.0077
64

0.0179
83
0.0153
77

0.0345
31
0.0295
26

0.0644
25
0.0550
87

0.1170
02
0.1000
45

0.2071
95
0.1771
66

0.3583
33
0.3063
99

0.6060
98
0.5182
54

1.0039
76
0.8584
67

0.0032
43

0.0066
39

0.0131
48

0.0252
47

0.0471
03

0.0855
45

0.1514
88

0.2619
91

0.4431
42

0.7340
46

0.0027
73
0.0023
71
0.0020
28
0.0017
34
0.0014
82
0.0012
68
0.0010
84
0.0009
27
0.0007
92
0.0006
78
0.0005
79
0.0004
95
0.0004
24
0.0003
62
0.0003
1
0.0002
65
0.0002
26
0.0001
94

0.0056
77
0.0048
54
0.0041
51
0.0035
49
0.0030
35
0.0025
95
0.0022
19
0.0018
97
0.0016
22
0.0013
87
0.0011
86
0.0010
14
0.0008
67
0.0007
42
0.0006
34
0.0005
42
0.0004
64
0.0003
96

0.0112
42
0.0096
13
0.0082
2
0.0070
28
0.0060
1
0.0051
39
0.0043
94
0.0037
57
0.0032
13
0.0027
47
0.0023
49
0.0020
08
0.0017
17
0.0014
68
0.0012
56
0.0010
74
0.0009
18
0.0007
85

0.0215
88
0.0184
59
0.0157
84
0.0134
96
0.0115
4
0.0098
68
0.0084
37
0.0072
15
0.0061
69
0.0052
75
0.0045
1
0.0038
57
0.0032
98
0.0028
2
0.0024
11
0.0020
62
0.0017
63
0.0015
07

0.0402
77
0.0344
39
0.0294
48
0.0251
8
0.0215
3
0.0184
1
0.0157
42
0.0134
6
0.0115
09
0.0098
41
0.0084
15
0.0071
95
0.0061
53
0.0052
61
0.0044
98
0.0038
46
0.0032
89
0.0028
12

0.0731
47
0.0625
45
0.0534
8
0.0457
29
0.0391
02
0.0334
34
0.0285
89
0.0244
45
0.0209
02
0.0178
73
0.0152
83
0.0130
68
0.0111
74
0.0095
54
0.0081
69
0.0069
85
0.0059
73
0.0051
07

0.1295
33
0.1107
59
0.0947
06
0.0809
8
0.0692
44
0.0592
08
0.0506
27
0.0432
89
0.0370
15
0.0316
5
0.0270
63
0.0231
41
0.0197
87
0.0169
19
0.0144
67
0.0123
7
0.0105
77
0.0090
44

0.2240
2
0.1915
52
0.1637
9
0.1400
51
0.1197
53
0.1023
97
0.0875
56
0.0748
66
0.0640
16
0.0547
38
0.0468
04
0.0400
21
0.0342
21
0.0292
61
0.0250
2
0.0213
94
0.0182
93
0.0156
42

0.3789
16
0.3239
99
0.2770
4
0.2368
88
0.2025
55
0.1731
98
0.1480
96
0.1266
32
0.1082
79
0.0925
86
0.0791
67
0.0676
93
0.0578
82
0.0494
93
0.0423
2
0.0361
86
0.0309
42
0.0264
57

0.6276
59
0.5366
9
0.4589
06
0.3923
95
0.3355
24
0.2868
95
0.2453
15
0.2097
6
0.1793
59
0.1533
64
0.1311
36
0.1121
3
0.0958
79
0.0819
83
0.0701
01
0.0599
41
0.0512
53
0.0438
25

0.0006
94
0.0005
93
0.0005
07
0.0004
34
0.0003
71
0.0003
17
0.0002
71
0.0002
32
0.0001
98
0.0001
7
0.0001
45
0.0001
24
0.0001
06
9.06E05
7.75E05
6.63E05
5.67E05
4.84E05
4.14E05

0.0015
29
0.0013
07
0.0011
18
0.0009
56
0.0008
17
0.0006
99
0.0005
98
0.0005
11
0.0004
37
0.0003
74
0.0003
19
0.0002
73
0.0002
34
0.0002
0.0001
71
0.0001
46
0.0001
25
0.0001
07
9.13E05

Table A.13 Determination of xylose degradation constant in function of pH and temperature

T
pH
2
2.1
2.2
2.3
2.4
2.5
2.6

120

130

140

150

160

170

180

190

200

210

220

230

5.12E05
5.2E05
5.3E05
5.44E05
5.6E05
5.81E05
6.07E05

0.0001
49
0.0001
52
0.0001
55
0.0001
59
0.0001
63
0.0001
7
0.0001
77

0.0004
14
0.0004
21
0.0004
29
0.0004
4
0.0004
53
0.0004
7
0.0004
91

0.0010
94
0.0011
11
0.0011
34
0.0011
62
0.0011
97
0.0012
41
0.0012
97

0.0027
62
0.0028
06
0.0028
63
0.0029
33
0.0030
23
0.0031
35
0.0032
76

0.0066
88
0.0067
97
0.0069
33
0.0071
04
0.0073
2
0.0075
92
0.0079
34

0.0155
78
0.0158
3
0.0161
48
0.0165
47
0.0170
5
0.0176
83
0.0184
81

0.0349
83
0.0355
49
0.0362
61
0.0371
59
0.0382
88
0.0397
1
0.0415

0.0759
16
0.0771
44
0.0786
91
0.0806
38
0.0830
89
0.0861
75
0.0900
6

0.1595
44
0.1621
26
0.1653
76
0.1694
68
0.1746
2
0.1811
05
0.1892
7

0.3253
44
0.3306
09
0.3372
37
0.3455
82
0.3560
87
0.3693
12
0.3859
61

0.6449
12
0.6553
49
0.6684
88
0.6850
29
0.7058
52
0.7320
67
0.7650
71

19

2.7
2.8
2.9
3
3.1
3.2
3.3
3.4
3.5
3.6
3.7
3.8
3.9
4

6.4E05
6.82E05
7.34E05
8E-05
8.82E05
9.87E05
0.0001
12
0.0001
28
0.0001
49
0.0001
75
0.0002
08
0.0002
5
0.0003
02
0.0003
68

0.0001
87
0.0001
99
0.0002
14
0.0002
33
0.0002
58
0.0002
88
0.0003
26
0.0003
75
0.0004
35
0.0005
12
0.0006
08
0.0007
29
0.0008
82
0.0010
74

0.0005
18
0.0005
51
0.0005
94
0.0006
47
0.0007
14
0.0007
98
0.0009
04
0.0010
38
0.0012
06
0.0014
18
0.0016
85
0.0020
21
0.0024
44
0.0029
76

0.0013
68
0.0014
56
0.0015
68
0.0017
09
0.0018
86
0.0021
08
0.0023
89
0.0027
42
0.0031
86
0.0037
46
0.0044
51
0.0053
38
0.0064
54
0.0078
6

0.0034
54
0.0036
78
0.0039
6
0.0043
15
0.0047
62
0.0053
24
0.0060
33
0.0069
24
0.0080
47
0.0094
6
0.0112
39
0.0134
79
0.0162
99
0.0198
49

0.0083
65
0.0089
08
0.0095
91
0.0104
5
0.0115
33
0.0128
95
0.0146
11
0.0167
7
0.0194
89
0.0229
12
0.0272
21
0.0326
46
0.0394
75
0.0480
72

0.0194
84
0.0207
48
0.0223
38
0.0243
41
0.0268
62
0.0300
36
0.0340
31
0.0390
61
0.0453
93
0.0533
65
0.0634
02
0.0760
37
0.0919
43
0.1119
68

0.0437
54
0.0465
91
0.0501
63
0.0546
6
0.0603
21
0.0674
48
0.0764
21
0.0877
16
0.1019
36
0.1198
39
0.1423
76
0.1707
49
0.2064
69
0.2514
37

0.0949
51
0.1011
08
0.1088
6
0.1186
18
0.1309
04
0.1463
7
0.1658
41
0.1903
53
0.2212
13
0.2600
62
0.3089
71
0.3705
44
0.4480
59
0.5456
45

0.1995
48
0.2124
88
0.2287
79
0.2492
87
0.2751
06
0.3076
1
0.3485
3
0.4000
45
0.4648
98
0.5465
44
0.6493
31
0.7787
31
0.9416
36
1.1467
21

0.4069
21
0.4333
09
0.4665
29
0.5083
5
0.561
0.6272
82
0.7107
26
0.8157
77
0.9480
27
1.1145
21
1.3241
24
1.5879
98
1.9201
97
2.3384
09

0.8066
19
0.8589
26
0.9247
76
1.0076
76
1.1120
41
1.2434
29
1.4088
36
1.6170
72
1.8792
26
2.2092
57
2.6247
42
3.1478
07
3.8063
06
4.6353
08

Annex 3: Calculations for the saccharification step

Table A.14 Calculations of enzyme kinetics constant for saccharification, using the formula below the table

Parameters

Value

Units

Sigma

0.01733

kmols/kmole-s

Kd

3.57E-03

kmols/m3

Sigma/kd
Enzyme
concentration
k=reaction rate

4.85E+00

m3/kmol-s

1.68E-05

kmol/m3

8.14E-05

s-1

Remarks/reference
Lynd et al, Trichoderma reesei CBHI/BMCC

for 3% concentration or 0.96ton/day

Table A.15 Determination of conversion degree of Cellulose in function of time for batch and continuous process

Time (h)

Cell concentration

(batch)

(CSTR)

185.20

0.00

0.00

138.14

25.41

22.67

103.04

44.37

36.96

76.85

58.50

46.80

57.32

69.05

53.97

42.76

76.91

59.45

31.89

82.78

63.76

23.79

87.16

67.24

17.74

90.42

70.11

13.23

92.85

72.52

10

9.87

94.67

74.57
20

11

7.36

96.02

76.33

12

5.49

97.03

77.87

13

4.10

97.79

79.22

14

3.06

98.35

80.41

15

2.28

98.77

81.47

16

1.70

99.08

82.43

17

1.27

99.32

83.29

18

0.95

99.49

84.07

19

0.71

99.62

84.78

20

0.53

99.72

85.43

21

0.39

99.79

86.03

22

0.29

99.84

86.58

23

0.22

99.88

87.09

24

0.16

99.91

87.56

Annex 4: Economic evaluation

Operating costs (*e5 $/yr)

gross margin (%)

250
200
150
100
50
0
-50 0

-100
-150
-200
Residence time (h)
Figure A.3 Operating costs and Return on Investment in
function of saccharification time for a continuous process

Annex 5: Energy balances

Table A.16 Energy balances for continuous process
Hot in Hot out Cold in

Cold out

Temperature (C)

200

51

25

147.88

Enthalpy (TJ/day)

2818

207

101

2712

Table A.17 Energy balances for batch process

In

Out

Temperature (C)

25

50

Enthalpy (kJ/day)

65,370,195.7

1,307,322,599.05

21