University of San Carlos

School of Engineering
Talamban, Cebu City, Philippines

CHE 423N
Engineering of Homogeneous Chemical Reactions

A Case Study on the

Kinetics of Cellulose Saccharification by Trichoderma reesei Cellulases

submitted to

Alchris W. Go, PhD

CHE 423N Instructor

by

Carl John Louie G. Navalta

Von Adrian O. Raboy

March 2, 2017
I. Introduction

In the enzyme industry specially for the biomass production, Trichoderma reesei (T.
reesei) is now considered a major cell factory and a benchmark organism for the production of
cellulases. It is a telemorph of the Hypocrea jecorina and a mesophilic filamentous fungus
(Paloheimo et al., 2016). This fungus has been used as a model organism in the degradation of
cellulose studies for a very long time, and its potential to produce cellulases was discovered not
until the demand for alternative fuel was very high. Cellulases are used to hydrolyze cellulose-
rich biomass and convert it into a fermentable sugar, and this fungus can naturally produce a
complete set and high levels of cellulases that can cleave -1,4-glycosidic bonds present in
cellulose and its derivatives (Ortega, Busto, & Mateos, 2000). The enzyme classes commercially
produced by T. reesei include amylase that is widely used in the processing of starch,
glucoamylase which is used in the production of first generation biofuel, protease useful in
detergent production, cellulase for textile manufacturing, beta-glucanase for brewing and
production paper and pulp and many others.
In the presence of a cellulase and cellulose, which is a soluble catalyst and an insoluble
reagent respectively, the enzymatic hydrolysis is a heterogeneous kind of a catalytic reaction
(Martin et al., 1988). The advantages for this type of hydrolysis include high specificity, the
reaction can be carried out at low temperatures from 30 oC to 50oC and at atmospheric pressure,
and it does not require the use of dangerous chemicals so that the deterioration of equipment can
be avoided. Aside from the availability of raw materials, the hydrolysis is very dependent on the
optimization of the steps required to transform cellulose to glucose and other reducing and
fermentable sugars. The steps in this process are as follows: pretreatment, production of enzyme,
and then enzymatic conversion of cellulose to reducing sugars. The fundamental step among
these is the conversion, wherein the pretreated raw materials are hydrolyzed using cellulases
from enzymes. Based on the paper being studied, the enzymes used for hydrolysis are those that
came from T. reesei and Aspergilus niger (A. niger).

II. About the Article

The article, Kinetics of Cellulose Saccharification by Trichoderma reesei Cellulases

(Ortega, et al), aims to optimize the enzymatic hydrolysis of the three celluloses under study
(carboxymethyl cellulose, powdered cellulose and microgranular cellulose) by investigating the
effect of varying enzyme and substrate concentration, temperature, and pH. Hydrolysis versus
percent saccharification curves are presented in order to show the trend of the hydrolysis in
separate variations of temperature, pH, and substrate concentration. Hydrolysis versus reducing
sugar release curves are also presented to show the same trend of hydrolysis in separate
variations of enzyme concentration.
 Based on the results of the study, the degree of saccharification of carboxymethyl
cellulose (CMC) under the influence of temperature and time of incubation is higher than the
other two celluloses. The optimal hydrolysis increases with increasing temperature at constant
pH, substrate concentration, and enzymes concentration. The same behavior is noted in which
the optimal hydrolysis of CMC increases with increasing acidity (decreasing pH) at constant
temperature, substrate concentration, and enzymes concentration. Overall, CMC exhibited higher
conversion compared to the other two celluloses.

III. Description of the Reactive System to be Modeled

Out of the three celluloses, CMC is the substrate of interest in this case study since it
shows the highest degree of saccharification with time, and is the best choice for industrial
processes, though the time required to achieve a certain conversion still is extremely high. The
conversion of the other two celluloses with time is so low that they may require even greater
sizing of the reactors in order to achieve higher conversion.

The hydrolysis time versus percent saccharification curves are provided as shown in
Figure 1. The hydrolysis of CMC was carried out at a concentration of 2.5% in a total volume of
100mL. The initial concentrations of the two participating enzymes coming from T. reesei and
Aspergilus niger are both 0.25mg/mL. Enzymes ratio of 1:1, pH of 5.0, and pressure of 1atm are
kept constant at all temperatures. The set inlet volumetric flowrate for all reactor schemes are
80L/h. The initial concentration of CMC is equal for all reactor schemes.

Figure 1. Percent saccharification of CMC with respect to time at varying temperature.

 It is desired to obtain a conversion of 6.8386% for CMC in large-scale continuous reactor
tanks. Such value is chosen because this is approximately the maximum conversion that can be
achieved within 24 hours. For practicalitys sake, 24 hours is considered to be the operation time.

IV. About the Products

In the enzymatic hydrolysis of CMC, the products are glucose and cellobiose. The
desirable product is the glucose due to its many practical uses and industrial importance. Glucose
is a carbohydrate, and is the most important simple sugar in the human metabolism. It is used as
nutritional supplement to improve nutritional value in infant formula, sweetener in confectionery
and other food production, supplement in oral tablets, buffering agent in injection for veterinary
medicine, main ingredient in dextrose in the production of various vitamins, fermentation
substrate in manufacturing of various organic substance to provide energy, and antioxidant in
leather production.
Cellobiose is simultaneously produced with glucose. Cellobiose is a reducing sugar and a
disaccharide. It is used against some inflammatory responses. It can be further converted to
glucose by enzymatic or acidic hydrolysis. It is commonly used as a reagent in bacteriology
studies, especially as an indicator carbohydrate for intestinal permeability in Crohns disease and
malabsorption syndrome.

V. Objectives

The objectives of this case study are to

propose a mechanism of the reaction, derive its respective rate expression, and fit the
experimental results in order to calculate for the rate constants;
select the best system temperature that would optimize the conversion of the
reactants;
propose and develop three multiple reactor schemes where the reaction can take
place;
select the best multiple reactor scheme using appropriate selection criteria; and
calculate the energy required in order to bring the system to a desired temperature.

VI. Reaction Mechanism and Rate Expression

If we assume that the two enzymes coming from T. reesei and Aspergilus niger act on the
hydrolysis of CMC synergistically, meaning their activity is the same, then their concentrations
are simply additive. Changes of the concentration of water as the reaction proceeds is negligible
since the water is in excess. The following mechanism is proposed.
 2G + E3

E3C
Figure 2. Reaction scheme in the hydrolysis of CMC assuming synergistic activity of enzymes.
where A = CMC polymer chain
E = Enzymes (endoglucanase + exoglucanase)
R = Products (glucose +cellobiose)
A Michaelis-Menten rate expression can be derived based from the kinetics. However, an
extremely poor fit is obtained upon processing the given data. Hence, the said assumption is
invalid. Another mechanism is proposed.
+
The generally accepted mechanism in enzymatic hydrolysis is as follows: the
endoglucanase forms an intermediate compound with the long-chain cellulose and hydrolyzes
E3
them at random. With this, the cellulose is converted into a less polymerized chain and reducing
sugars. In this soluble fraction, cellubiose and glucose can be found. Then, the exoglucanase
C + E2 +A
binds
G+ with
E1the
+ non-reducing
A ends of the chain and with the oligosaccharides derived from them.
This separates the non-reducing end and oligosaccharides away from the cellubiose. Lastly, in a
homogeneous phase the -1,4-glucosidase hydrolyzes the cellubiose into glucose.
E2A
E1A
The following shows the reaction scheme in the hydrolysis of CMC.

A
E1
+
E2 E1 S
+
E2
A
A

Figure 1. Reaction scheme in the hydrolysis of CMC by cellulases from T. reesei and A. niger

where:
A CMC polymer chain
A less polymerized CMC chain (shorter chains)
E1 endoglucanase
E2 exoglucanase
 E3 -1,4-glucosidase
C cellubiose
G glucose
The assumptions in the determination of the rate expression for enzymatic hydrolysis is
that the mechanism includes no inhibition and the enzymes acting on the cellulose act
separately the synergistic effect that could exist between the endo- and exoglucanase is not
taken into account. Also, it is assumed that water is in excess so that its changes in concentration
as the reaction proceeds are negligible.
The first enzyme endoglucanase hydrolyzes the CMC (A) at random thereby producing a
cellubiose (C) and glucose (G) and as well as the less polymerized chain (shorter CMC chains
A and A):
A + E1 E 1 A C + E1 + A' (1)

and
''
A + E1 E 1 A G+ E 1+ A (2)

Simultaneously, the second enzyme exoglucanase binds to the non-reducing end of the CMC
chain producing cellubiose and shorter chains:
A + E2 E 2 A C + E2 + A' (3)

Then lastly, the third enzyme -1,4-glucosidase hydrolyzes the produced cellubiose into glucose:
C+ E 3 E3 C 2 G+ E3
(4)

Assuming that the concentration of the intermediate products does not change through
time (stationary state), the change of concentration of cellobiose and glucose are presented
below.
dC k 1 C E 1 C A k 3 C E 2 C A k 5 C E 3 CC
= +
dt 1+k 2 C A 1+ k 4 C A 1+k 6 C C

(5)
and
'
dG k 1 C E 1 C A k C C
= +2 5 E 3 C
dt 1+ k 2 C A 1+k 6 C C

(6)
 The overall product (P) of hydrolysis consists of the cellobiose and the glucose formed in
which the cellobiose can be expressed as glucose equivalent. The change in the concentration of
the total product formed with time is equal to twice the amount of cellobiose plus the amount of
glucose (2C+G):
dP dC dG
=2 +
dt dt dt (7)

Substituting equations (5) and (6) to this equation,

dP (2 k 1 +k '1 )C E 1 C A 2 k 3 C E 2 C A
=r P = +
dt 1+ k 2 C A 1+k 4 C A

(8)
r P =r A
and since , then the final form of the rate expression for the enzymatic hydrolysis of
CMC is
(2k 1+ k '1) C E 1 C A 2 k 3 C E 2 C A
r A= +
1+k 2 C A 1+ k 4 C A

(9)
VII. Determination of Kinetic Parameters
The values of percent saccharification with respect to time are extracted from the graph
using Plot Digitizer software (see Annex A.1). The degree of saccharification is defined to be
amount reducing sugars formed ( g ) 0.89
degree of saccharification=
initial amount of CMC(g)

(10)
Since the amount of reducing sugars formed is equal to the amount of CMC converted, therefore
the degree of saccharification is equal to the conversion of substrate CMC, XA, in units of g/mL.
The concentration of the CMC, CA, in every XA is calculated using the formula
C A =C A 0 (1 X A ) (11)

assuming constant density since the hydrolysis takes place in the liquid phase. The initial
CA 0
concentration is calculated from the data 2.5mL CMC in a total volume of 100mL. The
mass of substrate is calculated by multiplying it to its density. Since the mixture is in excess of
water, the density can be assumed to be that of water. The density of water at each temperature is
taken from literature (Geankoplis, 2003). Calculation for the initial concentration at each
temperature is shown in Annex A.2.
 CA versus time curve is generated and a tangent line is drawn in each point. The slope of
dC A dC A
the tangent line is equal to . For a constant-volume batch reactor, r A = . The
dt dt

r A
obtained values from each slope are based from the experimental results (see Annex
A.2). Since it is desired to fit Equation 9 to the experimental results, in order to obtain the kinetic
parameters, the method of least square regression is used due to the complexity of the rate
expression that integral analysis becomes rigorous. Sample regression and the graphical
representation of the fit of Equation 9 to the experimental results are shown in Annex B.1.
Table 1 shows values of the rate constants at each temperature. The activation energy for
each rate constant is calculated using Arrhenius Equation. Calculation for the activation energies
and frequency factors are shown in Annex C.1.

Table 1. Rate Constants as a Function of Temperature, Activation Energy, and Frequency Factor per
Kinetic Parameter.
Rate Constants k1 (h-2) k1' (h-2) k2 (h-1) k3 (h-2) k4 (h-1)
30 2.4674 1.2919 1.0415 2.4692 1.0237
40 2.6449 1.3514 1.0410 2.6460 1.0234
T (oC)
50 2.7393 1.3724 1.0407 2.7404 1.0233
60 2.7420 1.3994 1.0400 2.7439 1.0228
Frequency Factor, ko 3.1006 1.5119 1.0388 3.1019 1.0222
Activation Energy, E
55.5225 38.8330 -0.6510 55.4610 -0.3783
(J/mol-K)

VIII. System Temperature

The rate of conversion is affected by temperature. Therefore, setting the system
temperature is vital in order to achieve optimum conversion. Based from Figure 1, higher
conversion was achieved at 60. In the design part, the reaction is employed at 60. The rate
constants at this temperature are used. At this temperature, the initial concentration of CMC is
0.024581g/mL. If one decides to set the temperature lower or higher, the Arrhenius equation is
E

used: k =k o e RT .

IX. Performance of Each Reactor Type

 The performance equations for PFR and CSTR are derived. After which, the performance
curves are shown. This way, the progress of conversion and products formation is visually
observed if the reaction is to take place in a CSTR or a PFR.

Continuous Stirred-Tank Reactor (CSTR)

For CSTR, the following assumptions are:
- Steady-state operation. There is no accumulation. Volumetric flow rate is
constant.
- The system is well-mixed
- Constant density since the reaction is in the liquid phase ( A=0). The mixture
is in excess of water as well.
- Isothermal reaction. Temperature of 60 is maintained throughout the
reaction.
- No pressure difference along the reactor.
- No partial conversion in the feed. At t=0, XA=0.

V, XA
CA, (-rA)

Reactant A (CMC) is selected for consideration. The mass balance is

Figure 4. Schematic diagram of a mixed flow reactor.
Input = output + disappearance by reaction + accumulation (12)

The operation is done in steady-state; therefore, the accumulation term is zero. The remaining
terms are
input of A (g/h) = FA0
output of A (g/h) = FA = FA0 (1-XA)
disappearance of A by reaction (g/h) = (-rA)V
Introducing these known terms into equation 11 and simplifying, the following equation is
obtained:
 F A 0 X A=( r A ) V (13)

V X
= A
F A 0 (r A )

(14)
F A 0=C A 0 v 0 v 0 =v f =v
where . At steady-state, .

Since negligible density changes are assumed (A=0),

C A 0C A
X A=
CA 0

(15)
FA 0 XA
Substituting and from equation 12 and simplifying, the following equation is
obtained:
V C C A
= = A0 (16)
v r A

where is the performance measure for both CSTR and PFR. It is defined to be the time required
to process one reactor volume of feed measured at specified conditions. Finally, the derived rate
expression in equation 9 is substituted to equation 15 to get the performance equation for CSTR.
V C A 0C A
= = '
v (2 k 1 +k 1 )C E 1 C A 2 k 3 C E 2 C A
+
1+ k 2 C A 1+k 4 C A

(17)

Plug Flow Reactor (PFR)

For PFR, the following assumptions are:
- Steady-state operation. There is no accumulation. Volumetric flow rate is
constant.
- Fluid flow is only along the length of the PFR.
- There are no radial mixing and back mixing.
- Fluid composition varies along the flow path.
- Constant density since the reaction is in the liquid phase ( A=0). The mixture
is in excess of water as well.
 - Isothermal reaction. Temperature of 60 is maintained throughout the
reaction.
- No pressure difference along the reactor.
- No partial conversion in the feed. At t=0, XA=0.

dV

CA0 CA0
FA0 FA FA + dFA FA0
XA0 = 0 XA XA + dXA XAf

Figure 2. Schematic diagram for a plug flow reactor.

Similar to CSTR, mass balance of reactant A is similar to equation 11. At steady-state, the
accumulation term becomes zero. For PFR, the remaining terms are
input of A (g/h) = FA
output of A (g/h) = FA = dFA
disappearance of A by reaction (g/h) = (-rA)dV
Introducing these three terms into equation 11, the following equation is obtained:
F A =( F A +d F A ) + (r A ) dV (18)

Taking the derivative of FA = FA0 (1-XA) to get

dF A=d [ F A 0 ( 1 X A ) ]=F A 0 d X A (19)

Substituting equation 18 to 17 and simplifying, the following equation is obtained:

dV d X A
=
F A 0 r A

(20)
Integrating equation 19,
XA
V d XA
=
F A 0 0 r A

(21)
 d C A
F A 0=C A 0 v 0 d X A=
where . At constant density (A=0), C A 0 . At steady-state,

v 0 =v f =v
. Substituting to equation 20 and simplifying:
CA
V d CA
= = (22)
v0 C
r A A0

Substituting the rate expression from equation 9 to 21 to get the performance equation for PFR.
CA
V dCA
= =
v0 C
'
(2 k 1 +k 1 )C E 1 C A 2 k 3 C E 2 C A
A0
+
1+ k 2 C A 1+k 4 C A

(23)

Performance Curves

Figure 6. Substrate and total products concentration as a function of time for mixed flow and plug flow reactors

Figure 6 shows the performance of PFR and CSTR. It can be observed that a great
amount of residence time is required in order to achieve complete conversion. Nevertheless, PFR
requires relatively lower residence time compared to CSTR. By initial judgement, a multiple
reactor scheme involving PFR can be proposed due to PFRs better performance. Nevertheless, a
CSTR-CSTR scheme is also proposed in order to establish a difference and to verify the initial
interpretation of the Figure 6.
X. Development of Multiple-Reactor Schemes
Two Equal-Sized CSTRs in Series
Two-equal sized reactors are being proposed to hydrolyze CMC, as shown in Figure 7.
The feed stream is labeled as 0, and the stream exiting the 1 st CSTR is labeled as 1.
Furthermore, the exiting stream is labeled as 2. The feed enters the first reactor at a volumetric
flow rate 80L/h with an initial concentration of 0.024581g/mL. In order to determine the volume
required in order to achieve a conversion of 0.068386 or a final concentration of 0.02290g/mL,
analytical method is used. The same assumptions are considered, which are mentioned earlier in
this paper.

CA0, CA1, XA1

XA0 = 0

V1 1 V2 2

CA2, XA2

Figure 3. Scheme 1: two equal-sized mixed flow reactor or continuously stirred tank
reactor (CSTR).

By analytical method:
1= 2 (24)

Substitute equations 15 to equation 23

r
r
( A )2
C C
( A)1= A 1 A 2
C A 0C A 1

(25)
 Substitute equation 9 to equation
C A 0C A 1 C A 1 C A 2
'
= '
(2k 1+ k ) C E 1 C A 1 2 k 3 C E 2 C A 1
1 (2 k 1+ k )C E 1 C A 2 2 k 3 C E 2 C A 2
1
+ +
1+k 2 C A 1 1+ k 4 C A 1 1+k 2 C A 2 1+ k 4 C A 2

(26)
C A 0=0.024581 g /mL C E 1=C E 2=0.00025 C A 2=0.02290 g/mL
Where , , , and using rate
CA 1
constants at 60 tabulated in Table 1, is calculated.

C A 1=0.0237 g/mL

T VT
Solving for the total residence time and the total volume ,

T = 1+ 2

C A 0 C A 1 C A 1C A 2
T = '
+ '
(2 k 1+ k )C E 1 C A 1 2 k 3 C E 2 C A 1
1 ( 2 k 1+ k ) C E 1 C A 2 2k 3 C E 2 C A 2
1
+ +
1+k 2 C A 1 1+ k 4 C A 1 1+k 2 C A 2 1+ k 4 C A 2

T =24.3 h

V T =1968.0 L

The volume of the two CSTRs is 984.0L each. The progress of the reaction in the two CSTRs is
shown in Figure 8 below.

CSTR 1 CSTR 2
CSTR 1 CSTR 2

Figure 8. Reactant and total products concentration as a function of time for two equal-sized CSTRs in series at
CSTR and PFR in Series
The second proposed scheme is using a CSTR followed by a PFR in series. The same
nomenclature for the streams is followed as in scheme 1. The feed enters the first reactor at the
same volumetric flow rate of 80L/h with an initial concentration of 0.024581g/mL. The same
feed conditions are used. The same conversion (XA2=0.068386) is desired.

CA0,
XA0 = 0
V1

CA1
XA1 CA2,
V2 XA2

Figure 9. Scheme 2: CSTR and plug flow reactor in series.

In order to compare the effect of changing the 2 nd reactor from a CSTR into a PFR, the
volume of the CSTR in this scheme is purposely set as the same volume of the first CSTR in the
previous scheme. Therefore, the concentration of the reactant exiting the first reactor the same in
the first scheme.

V 1=984.0 L

C A 1=0.0237 g/mL

X A 1=0.03584

1=12.15 h
 In order to calculate the volume of the PFR, graphical method is used since the integral of
the rate expression in equation 23 is complicated. As shown in Figure 10, the area of the

rectangle is equal to the space time 1 of the CSTR while the area under the curve from XA1 to
2
XA2 is equal to for the PFR. In order to relate the volume required for the given conversion,
rate of reaction, and the volumetric flow rate, the performance equation for the PFR is that of
equation 23.
CA 2
V d CA
2= 2 =
v '
C (2 k 1 + k 1) C E 1 C A 2k 3 C E 2 C A
A1
+
1+ k 2 C A 1+ k 4 C A

(27)

Figure 4. Graphical design procedure for CSTR and PFR in series using the 1/(-
rA) versus conversion (XA) plot.

From the plot, the area under the curve for the PFR is 11.53h. The volume of the PFR and
is calculated using equation 23. Finally, the total volume is obtained.

V T =1881.39 L
 T =23.68 h

Figure 10. Reactant and total products concentration as a function of time for CSTR and PFR connected in serie
Figure 11 shows the progress of the hydrolysis of CMC and the formation of products for

Two Equal-Sized PFR in Parallel

The third proposed scheme is using a two-equal sized PFR in parallel to hydrolyze CMC.
For parallel schemes, each branch must have the same for efficiency. Since the volume of each
reactor is equal, the feed is split into two equal streams in order to maintain equal for both
branches. Thus, the feed enters the both reactors at a volumetric flow rate of 40L/h each for the
same total volumetric flowrate of 80L/h, with the same initial concentration of 0.024581g/mL.
The same final concentration of 0.02290g/mL (XA=0.068386) is desired.

V1

CA0 CA
XA
XA0 = V2
0

Figure 5. Schematic diagram for two plug flow reactors in parallel

configuration.

Figure 6. Graphical design procedure for two PFR in parallel using the 1/(-r A)
versus conversion (XA) plot.
 For a PFR, the total shaded area under the curve is the ratio of the space time with the
initial concentration of the reactant. For a total volumetric flow rate of 80L/h, it has been
determined that a total volume of 1876.08L would be required to achieve a final concentration of
X A =0.068386
0.0229 ( ), then the total volume is halved. At the same time, the flow rates
going into the reactors are also halved thus maintaining the same .

Reactant Products
 Figure 7. Reactant and total products concentration as a function of time for two equal-sized PFRSs
connected in parallel at 60.

Figures 14 and 15 show the comparison of the hydrolysis of CMC and formation of the
total products, respectively, in the three multiple reactor schemes. The two are presented
separately in order to clearly establish the differences of the three schemes. If the two figures are
merged into one, no differences will be observed due to their very small variations. Likewise, the
progress of the reaction and products formation is shown from 15h to 25h in order to stretch out
the curves and to visually observe the differences of the three schemes.

Figure 14. Performance curves for the hydrolysis of the reactant (CMC) at different multiple reactor schemes.

Figure 15. Performance curves for the formation of total products at different multiple reactor schemes.
XI. Selection
The basis for selecting the appropriate reactor scheme is the set inflow rate (80 000
X A =0.068386
L/min) and conversion ( ) in a 24-h operation. The objective is to solve the
residence time needed for each scheme and compare the values. The residence time computed
corresponds to a volume, and the lesser the residence time the smaller is the volume needed to
obtain the conversion required. In this manner, the volume is minimized yet the same conversion
is achieved.

Table 2 summarizes the calculated residence time and total volume required. The scheme
that has the smallest residence time is the 3 rd scheme where two PFRs are installed in parallel.
The residence time for this reactor scheme is 23.46 hours and the total volume of reactors needed
to affect the conversion is 1876.08L.

Table 2. Calculated Residence Time and Corresponding Reactor Volume for Each Reactor Scheme
T (h) VT (L)
Scheme Orientation
CSTR-CSTR Series 24.30 1968.00
Series (CSTR first
CSTR-PFR 23.68 1881.40
followed by PFR)
PFR-PFR Parallel 23.46 1876.08

Aside from the performance of each reactor scheme, the operating conditions must also
be taken into account. These include temperature, pH and the pressure. As the paper in study
suggests, the optimum temperature where conversion of CMC is highest is at 60 oC (333.15K).
Since the basis of calculations in this case study is the conversion vs. time data at 60 oC and a pH
of 5.0, this pH will also be used for the modeled scheme. The pressure will be set constant at 1.0
atm.
 Therefore, the best reactor scheme for the hydrolysis of CMC is two PFRs installed in
parallel with operating conditions set at T=60oC, pH=5.0 and P=1atm.

XII. Energy Requirement

1atm 1atm 1atm

303.15 K 333.15 K 333.15 K

Reactor
PFR in Parallel 3
1 1a

H-01

Figure 8. Schematic diagram for the reactor design and energy requirement.

Assumptions/Basis

1. The basis for the calculation of the energy requirement is that the reactors is operating at
24 hours.
2. The reference temperature used to calculate for the enthalpy of a specific stream is 0oC or
273.15 K.
3. The streams going in and out of the reactor are in steady state.
4. The concentration of water is assumed to be constant because the water is in excess.
5. No reaction along the pipes reactions only occur at the reactors.
6. The reactors and pipes are well-jacketed so that there are no heat losses.

Table 3. Process Stream Summary Around the Reactors

Stream Number <1> <1a> <3>
Feed CMC-H2O Heated Feed CMC-H2O
Stream Name Products
Solution Solution
Component g/h g/h g/h
CMC 76 692.72 76 692.72 1 832.00
H2O 1 966.48 1 966.48 1 966.48
Cellobiose - - 66.88
Glucose - - 70.4
Enzymes (from T. reesei
- - -
and A. niger)
Volumetric flowrate, mL/h 80 000 80 000 80 000
Temperature, K 303.15 333.15 333.15
Pressure, atm 1 1 1
Phase Liquid Liquid Liquid
Calculations

Overall balance around heat exchanger (H-01)

dH
= H 1 H 1 a + Q
dt (28)

dH
=0
at steady state, dt

0= H 1 H 1 a + Q

H 01= H 1 a H 1
Q

(29)

where QH 01 is the heat required to raise the temperature of the entering stream

H1
Determining at 303.15K

303.15
H 1 = n i C p ,i dT (30-a)
i 273.15

303.15 303.15
H 1 =n H o 2
C p , H o dT + nCMC
2
C p ,CMC dT (30-b)
273.15 273.15

Summarized in the table below are the data for the heat capacity and the calculated
enthalpy of each of the components in stream <1> and <1a>.

Calculation of heat capacity, Cp

The heat capacity of water is estimated using the following equation:

Cp L ,i
[ kJ
kmol ]
K =R ( A + BT +C T 2 )
and the heat capacities of the cellulose and glucose are estimated using the group contribution
method by Rzicka and Domalski (1993). The method is expressed by:

[ ( )]
2
T T
Cp L ,i [ kJ /kmol K ]=R A+ B +D
100 100

(31)

where R is the gas constant and T is temperature in K. Parameters A, B, and D are obtained from

A= ni ai B= n i b i D= ni di (31a)

Sample calculations and the heat capacity parameters are tabulated in Annex D.

Table 4. Heat Capacity for Each Components (Constant Pressure) and Calculated Enthalpies
Molar flowrate Enthalpy
Stream Component Mass flowrate (g/h) Cp (kJ/kmol-K)
(kmol/h) (kJ/h)
CMC 1966.48 0.0075 12 166.76 2 737.52
<1>
H2O 76692.72 4.2607 2 259.05 288 754.03
CMC 1 966.48 0.0075 25 415.04 11 436.77
<1a>
H2O 76 692.72 4.2607 4 526.64 1 157 199.30

H1
Calculating using eq (26-b),

kJ kJ
H 1 =288754.03 +2 737.52
h h

kJ 1h
H 1 =291 491.55 (
h 3600 s )
H 1 =80.97 kW

H1a
Determining at 333.15K

333.15
H 1 = n i C p ,i dT (32-a)
i 273.15
 333.15 303.15
H 1 =n H o
2
C p , H o dT + nCMC
2
C p ,CMC dT (32-b)
273.15 273.15

H1a
Calculating using eq (27-b),

kJ kJ
H 1 a =1157 199.30 +11 436.77
h h

kJ 1 h
H 1 a =1168 636.07 (
h 3600 s )
H 1 a =324.62kW

Then, using eq (25)

H 01=324.62 kW 80.97 kW
Q
H 01=243.65 kW
Q

Overall balance around the reactor:

Since it is assumed that the operation in the reactor is adiabatic, hence there are no heat
losses from the system to the surroundings. Inside the reactor hydrolysis of CMC occurs and the
reactions are shown in stoichiometric equations (33) (35) with enzymes (endoglucanase,
exoglucanase and -1,4-glucosidase) as catalysts. The heat of reaction rxnH is determined at the
operating temperature of the reactor which is 60oC or 333.15 K. To calculate the
rxnH(T=333.15K), a hypothetical process path, represented by the dashed line, is followed. The
solid line represents the actual path of the reaction. This is shown in figure 15.

'
CMC+ H 2 O E 1 G+CMC + E1
(33)

'
CMC+ H 2 O E 2 C+ CMC + E2
(34)
 C E3 2G+ E 3

(35)

REACTANTS PRODUCTS
T=333.15 K T=333.15 K

REACTANTS PRODUCTS
T=298.15 K T=298.15 K

Figure 9. Process path on determining the heat of reaction at 333.15K.

From the figure, the enthalpy change is independent to the actual path of the reaction, and
therefore the rxnH(T=333.15K) can be calculated from the following equation:

rxn H ( T =333.15 K )= H s 1 + rxn H ( T =298.15 K )+ H s 2 (36)

Hs1 Hs2
where and are enthalpy change of the input and the output stream from
333.15K to 298.15K and from 298.15K to 333.15K respectively, and these enthalpy changes are
due to sensible heat. To calculate for the enthalpy changes due to sensible heat, the following
equations are used:

298.15
H s 1= n i Cpi dT (37)
i 333.15

and

333.15
H s 2= n i Cpi dT (38)
i 298.15
 Hs1 Hs2
Determining the and

298.15 298.15
H s 1= n H 2 O C p H O dT + n CMC
2
C p CMC dT
333.15 333.15

and

333.15 333.15
H s 2= n CMC C p CMC dT + n glucose C p glucose dT
298.15 298.15

Summarized in the table below are the data for the heats of formation, and the calculated
sensible heats and enthalpy of each of the components in the stream entering (reactants) and
exiting (products) the reactor.

Table 4. Data for the Heat Capacity and Calculated Enthalpy of each Component
Sensible Heat
Stream Component Molar flowrate (kmol/h) Enthalpy (kJ/h)
(kJ/kmol)
Entering CMC 0.0075 -889 526.38 -6 671.45
(Reactants) H2O 4.2607 -92 564.31 -394 388.76
Exiting CMC 6.9873 889 526.38 6 215 387.68
(Products) Glucose 0.7465 368 395.47 275 007.22

Hs1 Hs2
Calculating and using eq (32) and (33) respectively,

kJ kJ
H s 1=6 671.45 394 388.76
h h

kJ 1 h
H s 1=401060.21 (
h 3 600 s )
H s 1=111.41 kW
 kJ kJ
H s 2=6 215387.68 +275 007.22
h h

kJ 1 h
H s 2=6 490 394.90 (
h 3600 s )
H s 2=1802.89 kW

Determining rxn H ( T =298.15 K )

H rxn ,298.15 K = H products H reactants (39)

H rxn ,298.15 K = v H of 298.15 ( products ) v H of 298.15 ( reactants ) (40)

The standard heats of formation of each component and the calculated enthalpies are
shown in table 7.

Table 5. Standard Heats of Formation and Calculated Enthalpies of Each Component

o
Molar Flowrate - f , 298.15
Component Mass flowrate (g/h) Enthalpy (kJ/h)
(mol/hr)
(kJ/mol)
CMC 1 966.48 7.50 963.00 -7222.69
H2O 76 692.72 4260.71 285.84 -1217881.35
Cellobiose 127.75 0.37 2.43 -0.90
Glucose 70.40 0.75 1275.00 -956.25

Since there are three simultaneous reactions in the system (indicated by equations 33-35),
each of the heat of reaction is calculated. This is done by using eq (39):

H rxn ,298.15 K = v H of 298.15 ( products ) v H of 298.15 ( reactants )

For reaction 1

kJ kJ kJ kJ
(
H rxn ,298.15 K = 956.25
h
7222.69
h )(
1217881.35 7222.69
h h )
 kJ 1 h
H rxn ,298.15 K =1 216 925.10 (
h 3600 s )
H rxn ,298.15 K =338.03 kW

For reaction 2

kJ kJ kJ kJ
(
H rxn ,298.15 K = 7222.69
h
0.90 )(
h
1217881.35 7222.69
h h )
kJ 1 h
H rxn ,298.15 K =1 217 880.45 (
h 3600 s )
H rxn ,298.15 K =338.30 kW

For reaction 3

kJ kJ kJ
(
H rxn ,298.15 K =2 956.25
h)(
0.90 1217881.35
h h )
kJ 1 h
H rxn ,298.15 K =1 215 967.95 (
h 3600 s )
H rxn ,298.15 K =337.77 kW

The total heat of reaction at 298.15K is:

H rxn ,298.15 K =1014.10 kW

Calculating the rxn H ( T =333.15 K ) using eq. (31)

rxn H ( T =333.15 K )= H s 1 + rxn H ( T =298.15 K )+ H s 2

rxn H ( T =333.15 K )=111.41 kW +1 014.10 kW + 1802.89 kW

rxn H ( T =333.15 K )=2705.58 kW

Total Energy Requirement

The total energy requirement takes into account both the energy to be supplied by the
heat exchanger (H-01) to heat up the entering solution to 60oC and the energy required to
compensate the heat that is absorbed due to endothermic reaction. That is,
tot =Q
Q H 01+rxn H ( T =333.15 K )

tot =243.65 kW + 2705.58 kW

Q

tot =2 949.23 kW
Q

XIII. Conclusion
The concentration of the products formed has been monitored and a kinetic model was
established from the batch reaction data for the hydrolysis of CMC at a temperature of 60 oC (333.15K).
The kinetic parameters corresponding to the activity of enzyme were calculated through nonlinear least
square regression. It can be concluded that the production of glucose and as well as cellobiose can be
modeled as three simultaneous reactions by three enzymes, specifically the endoglucanase, exoglucanase
and the -1,4-glucosidase, with each one having to contribute in the conversion of CMC into reducing
sugars.

After the reaction kinetics was determined, reactor schemes were proposed and the best reactor
scheme that would yield a set conversion at a lesser residence time and requires minimum volume is two
plug flow reactors installed in a parallel configuration. The operating conditions of the reactor is set the
same with the batch operating conditions in which high conversion of CMC is achieved. And in order to
attain the set temperature, an energy balance was set-up and the energy required to reach and maintain the
temperature of the reacting mixture was calculated.

XIV. Recommendations
Enzyme recovery

Enzymes are costly due to the difficulty in the processing to have high specificity, low
impurities and a stable structure and function. Enzyme manufacturers will have to encounter
significant costs during the production, from isolation to purification and evaluation steps which
is part of the product evaluation.
 There are many industrial applications that includes enzyme in the process, and usually
high enzyme purity is required. That is why it is recommended that the enzymes must be
recovered at each operation. An enzyme recovery system must be developed in order to save
money and time to produce a new set of enzymes as catalysts. Recovery (separation from the
broth and purification) can involve several steps, and new techniques and methods are already
developed, an example of which is liquid-liquid extraction which is commonly and widely
employed in chemical industries due to the simplicity of the process and will incur low costing
and as well as its ease for scale-up. For very sensitive enzymes, extraction with aqueous two-
phase systems can be employed which offers mild environment so that biomolecules
denaturation can be prevented (Silvrio et al., 2012).

Effect of enzyme concentration

An enzyme has the ability to convert a substrate into a desired product. This is referred to
as an enzyme activity since if the enzyme activity is increased, more substrate is converted into
product per unit time. The enzyme activity can be increased so that there are more enzyme
molecules that are available to react with the substrate and convert them into desirable products
per unit time. In addition, increasing the enzyme activity, hence increasing the enzyme
concentration, will prevent enzyme saturation specially when the substrate concentration is high.

Effect of pH

As the paper in study suggests, % saccharification is increased when the pH is set to 4.0
(when the substrate in use is CMC). The pH on the reactor scheme selected can be set to 4.0
instead of 5.0 and an increase in the percentage of substrate converted to product can be
expected.

XV. References

Geankoplis, C. (2003). Principles of Transport Processes and Separation Processes. New Jersey:
Pearson Education, Inc.
Kabo, G. J., Voitkevich, O. V., Blokhin, A. V., Kohut, S. V., Stepui, E. N., & Paulechka, Y. U.
(2013). Thermodynamic properties of starch and glucose. J. Chem. Thermodynamics, 87-
93.

Levenspiel, O. (1999). Chemical Reaction Engineering, 3rd Edition. New York: John Wiley &
Sons.

Martin, C., Negro, M. J., Alfonsel, M., & Sez, R. (1988, July). Enzymatic Hydrolysis of
Lignocellulosic Biomass From Onopordum Nervosum. Biotechnology and
Bioengineering, 32.

Ortega, N., Busto, M. D., & Mateos, M. P. (2000). Kinetics of Celluose Saccharification by
Trichoderma reesei Cellulases. Elsevier.

Paloheimo, M., Haarmann, T., Mkinen, S., & Vehmaanper. (2016). Production of Industrial
Enzymes in Trichoderma reesei. Gene Expression Systems in Fungi: Advancements and
Applications. Switzerland: Springer International Publishing .

Poling, B. E., Prausnitz, J. M., & O'Connell, J. P. (2004). The Properties of Gases and Liquids.
The Mcgraw-Hill Companies.

Rodriguez, V., Duenas, M., Requena, A., & El-Hadj, M. (2001). The Influence of pH upon the
Hydrolysis of Carboxymethylcellulose with Cellulases from Trichoderma reesei. The
Canadian Journal of Chemical Engineering, Volume 79, 289-295.

Ruzicka, V., & Domalski, E. (1993). Estimation of the Heat Capacities of Organic Liquids as a
Function of Temperature using Group Additivity II Compounds of Carbon, Hydrogen,
Halogens, Nitrogen, Oxygen, and Sulfur. Journal of Physical and Chemical Reference
Data, 619-657.

Ruzicka, V., & Domalski, E. S. (1993). Estimation of the Heat Capacities of Organic Liquids as a
Function of Temperature using Group Additivity I Hydrocarbon Compounds. Journal of
Physical and Chemical Reference Data, 597-618.

Semmar, N., Tanguier, J., & Rigo, M. (2002). Specific heat of carboxymethyl cellulose and
Carbopol aqueous solutions. Thermochimica Acta, 225-235.

Silverio, S., Rodriguez, O., Tavares, A., Teixeira, J., & Macedo, E. (2012). Lacasse recovery with
aqueous two-phase systems: Enzyme partitioning and stability. Journal of Molecular
Catalysis B: Enzymatic, 37-43.

Tewari, Y. B., & Goldberg, R. N. (1989). Thermodynamics of Hydrolysis of Disaccharides. The

Journal of Biological Chemistry, 3966-3971.
XVI. Annex

ANNEX A

A.1 Data Gathering

 From the paper studied, the data used were from the temperature vs. hydrolysis time plot
for CMC. Data points were obtained through PlotDigitizer v2.6.8.

Sample digitization of data points:

CalibrationData points selection Digitized points

Table A.1-6. Digitized Data Points Using the % Saccharification versus Hydrolysis Time Plot (T=60 oC)
Hydrolysis Time (h) % Saccharification
0.0000000 0.000000
2.2622929 4.667918
5.0384927 5.202085
12.081143 5.687840
24.146793 6.249169
36.206665 6.600473
48.000000 6.760847
A.2 Processing of Data

C Ao
Determination of (calculated from volume of substrate and density of water
which is assumed to be in excess)

V A H O (T =6 0 o C)
C Ao = 2

VT

o g
H O ( T =6 0 C )=0.98324 V A =2.5
from literature data, 2
mL , and dry basis in a 100 mL

volume

g
C Ao =
(
( 2.5 mL ) 0.98324
mL )
100 m L

C Ao =0.02458

Sample calculation of CA

C A =C Ao (1X A )

C Ao =0.02458 g /mL X A =0.0467

where and at t=2.2623h,

g
C A =0.02458 (10.0467)
mL
 Determination of -rA (CA vs. time at T=60oC)

The calculated
o
substrate concentrations at 60 C were plotted versus hydrolysis time. Tangent lines were drawn
at each point, and the slope of the tangent line represents the -r A. Tabulated below are the
substrate concentrations and the manually determined -rA values.

Table A.2-7. Calculated Value of the Concentration of CMC After 48-h Hydrolysis and Experimental
Determination of -rA
Experimental
t(h) % Saccharification XA CA (g/mL)
slope= - rA
0.0000 0.0000 0.0000 0.024581000 5.90E-04
2.2623 4.6679 0.0467 0.023433579 1.80E-04
5.0385 5.2021 0.0520 0.023302275 2.22E-05
12.0811 5.6878 0.0569 0.023182872 1.38E-05
24.1468 6.2492 0.0625 0.023044892 9.23E-06
36.2067 6.6005 0.0660 0.022958538 4.83E-06
48.0000 6.7608 0.0676 0.022919116 1.67E-06
ANNEX B

B.1 Nonlinear Least Square Regression

Proposed reaction kinetics

(2k 1+ k '1) C E 1 C A 2 k 3 C E 2 C A
r A= +
1+k 2 C A 1+ k 4 C A

Initial values for each parameter were selected, and minimization of the difference of the -rA values were done to get the
correct values of the kinetic parameters. Tabulated below are the regressed data.

Table 8. Nonlinear Least Square Regression on -rA values to Determine Kinetic Parameters (T=30oC)
Experimental Theoretical
t(h) XA CA (g/mL) (-rA,exp - -rAtheo)2 (-rA,exp - -rAexp,ave)2
slope= - rA -rA
0.0000 0.0000 0.024892000 5.47E-04 0.00052387 0.000000000544 1.85027E-07
2.3064 0.0171 0.024466727 2.21E-04 0.00021239 0.000000000066 1.07014E-08
4.9571 0.0224 0.024333758 2.21E-05 0.00002869 0.000000000043 9.01505E-09
12.0026 0.0283 0.024186704 1.40E-05 0.00001489 0.000000000001 1.06137E-08
24.1917 0.0332 0.024065994 9.57E-06 0.00000900 0.000000000000 1.15532E-08
36.1276 0.0373 0.023964284 4.90E-06 0.00000755 0.000000000007 1.25782E-08
48.0000 0.0408 0.023876837 1.07E-06 0.00000602 0.000000000025 1.34527E-08
0.000117053 0.000000000686 2.52941E-07
2
R =9973
Table B.1-9. Nonlinear Least Square Regression on -rA values to Determine Kinetic Parameters (T=40oC)
Experimental Theoretical
t(h) XA CA (g/mL) (-rA,exp - -rAtheo)2 (-rA,exp - -rAexp,ave)2
slope= - rA -rA
0.0000 0.0000 0.0248062500 2.61E-04 0.00027177 0.000000000125440 3.6976E-08
2.1882 0.0198 0.0243161296 1.22E-04 0.00013035 0.000000000073102 2.86454E-09
4.9673 0.0261 0.0241575733 4.68E-05 0.00004583 0.000000000000884 4.6249E-10
12.0312 0.0387 0.0238452564 3.08E-05 0.00002992 0.000000000000835 1.40195E-09
23.9816 0.0481 0.0236136496 1.30E-05 0.00001345 0.000000000000233 3.05893E-09
35.9168 0.0519 0.0235193947 4.20E-06 0.00000423 0.000000000000001 4.10606E-09
48.0000 0.0526 0.0235009301 8.00E-07 0.00000250 0.000000000002890 4.55336E-09
6.83E-05 0.000000000203386 5.34233E-08
2
R =9962

Table B.1-10. Nonlinear Least Square Regression on -rA values to Determine Kinetic Parameters (T=50oC)
Experimental Theoretical
t(h) XA CA (g/mL) (-rA,exp - -rAtheo)2 (-rA,exp - -rAexp,ave)2
slope= - rA -rA
0.0000 0.0000 0.024701750 2.14E-04 0.00021170 0.000000000006250 2.07895E-08
2.3080 0.0177 0.024265579 1.55E-04 0.00015850 0.000000000014900 7.16149E-09
4.8497 0.0290 0.023985056 7.05E-05 0.00006994 0.000000000000291 2.11206E-13
12.0462 0.0442 0.023610390 2.91E-05 0.00003104 0.000000000003861 1.67587E-09
23.9945 0.0528 0.023398624 1.25E-05 0.00001228 0.000000000000054 3.30676E-09
36.0570 0.0572 0.023288263 6.00E-06 0.00000672 0.000000000000514 4.09785E-09
48.0000 0.0591 0.023241574 3.20E-06 0.00000320 0.000000000000000 4.46417E-09
7.00E-05 0.000000000025869 4.14958E-08
2
R =9994
Table B.1-11. Nonlinear Least Square Regression on -rA values to Determine Kinetic Parameters (T=60oC)
Experimental Theoretical
t(h) XA CA (g/mL) (-rA,exp - -rAtheo)2 (-rA,exp - -rAexp,ave)2
slope= - rA -rA
0.0000 0.0000 0.024581000 5.90E-04 0.00059629 0.0000000000369664 2.23491E-07
2.2623 0.0467 0.023433579 1.80E-04 0.00025935 0.0000000062568100 3.94236E-09
5.0385 0.0520 0.023302275 2.22E-05 0.00002185 0.0000000000001340 9.07214E-09
12.0811 0.0569 0.023182872 1.38E-05 0.00001332 0.0000000000002550 1.07397E-08
24.1468 0.0625 0.023044892 9.23E-06 0.00000899 0.0000000000000588 1.17143E-08
36.2067 0.0660 0.022958538 4.83E-06 0.00000487 0.0000000000000011 1.26852E-08
48.0000 0.0676 0.022919116 1.67E-06 0.00000257 0.0000000000008100 1.34085E-08
0.0001175 0.0000000062950350 2.85053E-07
 2
R =9779 Calculating the R2 value (sample, T=60oC)

2
2 ( (r exp
A ) ( r A ) )
theo

R =1 2
( (r exp
A ) (r A ,ave ) )
theo

2
R =1
( 0.0000000062950350 )2
( 2.85053E-07 )2
2
R =0.9779

Fitting of experimental and modeled kinetics

30oC

Model Experimental

40
 40oC

Model Experimental

50oC

Model Experimental

41
 60oC

Model Eperimental

42
ANNEX C

C.1 Activation Energy

Using the Arrhenius equation:

E
k =k o e RT

E 1
ln k =ln k o ( )
R T

This is in the form of y=mx + b where y=ln k , m=

E
R , ( T1 )
x=
,
b=ln k o
. Plotting

y against x:

k1
f(x) = - 6.68x + 1.13
R = 0.96

43
 k'

f(x) = - 4.67x + 0.41

R = 0.99

44
 k2

f(x) = 0.08x + 0.04

R = 0.89

k3
f(x) = - 6.67x + 1.13
R = 0.96

45
 k4

f(x) = 0.05x + 0.02

R = 0.89

Sample calculation of the activation energy

From the plot for k1, the equation of the line is y = -6.6782x + 1.1316 with an R value of
0.95564, the slope corresponds to E/R:

46
 E
m= =6.6782
R

J
(
E=6.6782 K 8.314
mol K )
J
E=55.5225
mol

Sample calculation of the frequency factor

b=ln k o =1.1316

k o =e 1.1316

k o =3.1006

47
ANNEX D

D.1 Reactor Performance (CSTR vs. PFR)

V C A 0C A
CSTR = = '
v (2 k 1+ k 1 )C E 1 C A 2 k 3 C E 2 C A
+
1+k 2 C A 1+k 4 C A

CA
V d CA
PFR= =
v0 '
C (2k 1+ k 1)C E 1 C A 2k C C
A0
+ 3 E2 A
1+k 2 C A 1+ k 4 C A

Table D.1-12. Performance of Continuously Stirred Tank Reactor and Plug Flow Reactor in the
Hydrolysis of CMC
CA, g/mL , h , h
0.024581 0.0000 0.0000
0.022581 29.3054 28.1070
0.020581 64.1766 58.7610
0.018581 106.4110 92.4820
0.016581 158.6729 129.9710
0.014581 225.0887 172.1990
0.012581 312.4084 220.5690
0.010581 432.4858 277.2139
0.008581 608.2257 345.6220
0.006581 890.3762 432.0906
0.004581 1418.3096 549.8913
0.002581 2763.3929 736.0700
0.000581 13364.3496 1218.8800

D.2 Determination of Reactor Volumes

v0
The volumetric flowrate, , was set to 80 000 mL/min and the total volume of the
reactors in each of the proposed multiple reactor scheme was determined.

Sample calculation, scheme 1: two equal sized CSTR in series

48
 1= 2

C A 0C A
where = r A

r
r
( A )2
C A 1 C A 2
( A)1=
C A 0C A 1

(2k 1+ k '1) C E 1 C A 2 k 3 C E 2 C A
since r A= 1+k 2 C A
+
1+ k 4 C A , then

C A 0C A 1 C A 1 C A 2
=
(2k 1+ k '1) C E 1 C A 1 2 k 3 C E 2 C A 1 (2 k 1+ k '1 )C E 1 C A 2 2 k 3 C E 2 C A 2
+ +
1+k 2 C A 1 1+ k 4 C A 1 1+k 2 C A 2 1+ k 4 C A 2

where

C A 0=0.024581 g /mL

C E 1=C E 2=0.00025 g /mL

C A 2=0.02290 g/mL

k 1=2.7420 , k '1=1.3994 , k 2=1.0917 , k 3 =2.7439 and k 4=1.0520

substituting known values, and solving for

CA 1

49
 0.024581C A 1 C A 10.02290
=
( 2 ( 2.4720 ) +1.3994)(0.00025)C A 1 2(2.7439)( 0.00025) C A 1 (2 ( 2.4720 )+ 1.3994)(0.00025)(0.02290) 2(2.
+ +
1+1.0917 C A 1 1+1.0520 C A 1 1+1.0917 (0.02290)

C A 1=0.0237 g/mL

ANNEX E

E.1 Energy Requirement

Table E.1-13. Heat Capacity Parameters for Each Component (at Constant P)
Component A 103B 106C
Water 8.712 1.25 -0.18
CMC Group ai bi di
-CH2 2.7972 -0.054967 0.10679
-COOH 13.1180 16.120 -5.1273
-OH 12.9520 -10.1450 2.6261
-COC -3.3182 2.6317 -0.44354
Glucose Group
-OH 12.9520 -10.1450 2.6261
-COC -3.3182 2.6317 -0.44354

Determination of heat capacity of each component

o
CpH O , l kJ
kmol
2
K [ ]
Cp H 2
O, l =R ( A + BT +C T 2 )

50
where A=8.712, B=1.25E-3, C=-0.18E-6 and R=8.314 kJ/kmol-K

kJ
Cp H 2 O, l (
= 8.314
kmol K )
( 8.712+1.25 103 T 0.18 106 T 2 )

The heat capacities of CMC and glucose are determined via group contribution (the
structure of the glucose will be the ring form so that the functional group present will be 5 OH
(hydroxyl) group and COC (ether) group.

o Cp CMC
[ kJ
kmol
K
]
[ ( )]
2
T T
Cp CMC [kJ /kmol K ]=R A+ B +D
100 100

where A= ni ai , B= n i b i , D= ni di and R=8.314 kJ/kmol-K

A= ni ai=2.7972+13.1180+ 5 ( 12.952 )3.3182=77.457

B= n i b i=0.054967+16.120+5 (10.1450 ) +2.6317=32.0283

D= ni di =0.106795.1273+5 ( 2.6261 )0.44354=7.6665

)[ ( )]
2
kJ T T
Cp CMC [kJ /kmol K ]= 8.314 ( kmol
77.45732.0283
100
+7.6665
100

o Cpglucose
[ kJ
kmol
K ]
A= ni ai=5 ( 12.952 )3.3182=61.4418

B= n i b i=5 (10.1450 ) +2.6317=48.0933

51
 D= ni di =5 (2.6261 ) 0.44354=12.6870

)[ ( )]
2
kJ T T
Cp glucose [kJ /kmol K ]= 8.314 ( kmol
61.441848.0933
100
+12.6870
100

H1
Sample calculations of enthalpies (for )

303.15
H H o=n H
2 2 o C p , H o dT
2
273.15

303.15
kmol kJ
(
H H o= 4.2607
2
h
8.314
kmol K )( ) (8.712+1.25 10
273.15
3
T 0.18 106 ) dT

kJ
H H o=288 754.03
2
h

303.15
H CMC =nCMC C p ,CMC dT
273.15

303.15

[ ( ) ] dT
2
kmol kJ T T
(
H CMC = 0.0075
h
8.314
kmol )( )
273.15
77.45732.0283
100
+7.6665
100

kJ
H CMC =2 7 3 7.52
h

H 1 = H H o+ H CMC
then, 2

kJ kJ
H 1 =288754.03 +2 737.52
h h

52
 kJ 1h
H 1 =291 491.55 (
h 3600 s )
H 1 =80.97 kW

Calculation of the sensible heat

o For H s 1

298.15 298.15
H s 1= n H 2 O C p H O dT + n CMC
2
C p CMC dT
333.15 333.15

298.15
kmol kJ kmol
(
H s 1= 4.2607
h
8.314
kmol K )( ) (8.712+1.25 10
333.15
3
(
T 0.18 106 ) dT + 0.0075
h )(
8.31

kJ 1 h
H s 1=401060.21
h 3600 s ( )
H s 1=111.41 kW

o Hs2
333.15 333.15
H s 1= n CMC C p CMC dT + nglucose C p glucose dT
298.15 298.15

303.15

[ ( ) ] dT +(0.7465 kmh
2
kmol kJ T T
(
H s 2= 6.9873
h
8.314
kmol K )( )
273.15
61.441848.0933
100
+12.6870
100

kJ 1 h
H s 2=6 490 394.90
h 3600 s ( )
H s 2=1802.89 kW

53
ANNEX F

F.1 Density of Water

T () (g/mL)
30 0.99568
40 0.99225
50 0.98807
60 0.98324

54