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6.10. You are going to cultivate yeast, Saccharomyces Cerevisiae, by using a 10m3 fermenter
your company owns. You want to find out the amount of ethanol the fermenter can produce.
Therefore, a chemostat study was carried out and the Monod kinetic parameters. For the
microorganism grown in the glucose medium at 30oC, pH= 4.8, were found to be Ks= 0.025 g/L
and umax= 0.25h-1. The ethanol yield (Yp/s) is 0.44 (g/g) and cell yield (Y X/S) is 0.019 (g/g). The
inlet substrate concentration is 50 g/l.
a. What flowrate will give the maximum total ethanol production in the continuous fermenter and
what is the maximum ethanol production rate?
b. If you want to convert 95% of the incoming substrate, what must be the ethanol production
rate for the continuous fermenter?
c. If you have two 5m3 fermenters instead of one 10m3 fermenter, what is your recommendation
for the use of these fermenters to convert 95% of the incoming substrate? Would you recommend
connecting two fermenters in series to improve the productivity? Why or why not?
Given: V=10m3; Ks= 0.025 g/L; umax= 0.25h-1; Yp/s= 0.44; YX/S= 0.019 g/g; Cso= 50 g/l
Required:
(a) F
(b) F @ 95% Cso conversion
Solution:
(a) =
Ks+Cso
=
Ks
0.025 g 50 g
+
L
l
=44.73
0.025 g / L
Cso
Cs= CS opt = +1 = 50gL-1/ (44.73 +1) = 1.0934g/L
D= umax.CS/ (Ks+Cs)= 0.25h-1(1.0934g/L)/ (0.025 g/L+1.0934g/L)= 0.2444/h
F= DV= 0.2444/h (10m3)= 2.444 m3/h
(b) For 95% substrate conversion Cs= 0.05Cso = 4.75g/l.
0.25
g
4.75
h
l
D= umax.CS/ (Ks+Cs) =
= 0.0249 /h
g
g
0.025 + 4.75
l
l
( )( )
( )( )
44.73
Cx1= YX/SCso( +1 = (0.019)(50)( 4.573 )=
0.9292g/L
Cso
Cs1= +1 =
0.8598
50 g/l
(44.73+1) = 1.0934g/L
Cs 1
+1 =
1.0934
45.73 = 0.0239 g/L
0.02080.9292
YX/S= 0.02391.0934
0.8494
Therefore; a two- 5 cubic meters of fermenters in series will increase the Growth
yield for 95% Ethanol conversion
max
K S+ S
Where:
= specific growth rate
max =maximum specific growth rate
Ks = Monod constant
S = substrate concentration
For the above process, show how the time required to reach the maximum population of
cells can be estimated.
If the concentration of cells at the start of the exponential growth is 0.08g/L and the
essential substrate concentration is 36g/L. Find the time where the maximum population
of cells is reached given that max = 0:55h1, Ks = 1:2g/L and k = 1:4gsubstrate/(gcellsh)
Soln:
Substrate consumption during the growth can be described by;
dS
=kX
dt
1 dX
dt
Substitute X into
dS
dt and integrating gives;
dS
1 dX
=k (
)
dt
dt
dS=
But
=
k
dX
max
K S+ S
hence;
dS=
k
dX
max
K S+ S
rearranging gives;
0
XS
S
k
K + S dS= dX
S
max X
S
0
max
KS
(S0 + K S ln
)
k
K S+ S0
The average can is determined by dividing the specic growth rate with the total
substrate in the reactor, which gives;
0
dS
avg=
S0
dS
S0
avg=
K S+S dS
S0
dS
S0
avg=
max [ S 0+ Ksln
KS
]
K S +S0
S0
Therefore, the new cell growth expression should be in the form of;
dX
= avg X
dt
and integrating,
XS
1
1
dt= X dX
avg X
0
0
gives;
t=
X
1
ln S
avg X 0
max
KS
X
(S 0 + K S ln
) ]ln S
k
K S + S0
X0
S
t= 0
k
1.2
36+1.2
36+1.2 ln
0.55
X S=0.08+
1.4
XS=12.604
and using the expression for avg;
0.55[36+1.2 ln
avg=
1.2
]
36+ 1.2
36
avg=0.487
thus, the time taken for the cell population to reach a maximum value;
X
1
t=
ln S
avg X 0
t=
1
12.604
ln
0.487
0.08
t = 10.34 hrs.
Given: E. coli
td=0.5 h
Cn=10 Cno=0.1
Reqd: a)
b) t
Soln:
a)
ln 2
td=
ln 2
= 0.5
=1.3868 h-1
b)
Cn
ln (
)
Cno =(t-to)
t=
ln (
Cn
)
Cno
10
)
.1
t=
1.3863
t= 3.3219 h
ln (
1. Assuming the exponential growth during this period, evaluate the specific growth rate.
2. Evaluate the doubling time during the exponential growth phase.
3. How much time would be required to grow from 0.10 kg-dry cell/m3 to 1.0 kg dry cell/m3?
You may assume the exponential growth during this period.
Given: E. coli
Cn= 0.50 kg dry cell/m3 Cno=0.10 kg dry cell/m3
t=1 h
Reqd: a)
b) td
c) t (from 0.1 to 1)
Soln:
a)
ln
Cn
Cno
t
.5
)
.1
1
ln (
=
=1.6094/h
b)
td=
ln 2
td= ln 2 / 1.6094
td= 0.4307 h = 25.8412 min
c)
ln
Cn
Cno
1
.1
=
1.6094
ln
t =
CHAPTER
6: CELL KINETICS AND FERMENTER DESIGN ; ADDITIONAL PROBLEM
Bioprocess
1.4307 h Engineering Principles by Pauline M. Duran, pp. 364- 367
Yxs=0.06
g/g;
Ypx=7.7
g/g,
umax=0.3/h;
ms=2.2g/g.h;
Cx =
D ( CsoCS )
D
=
+ms
Yxs
= 0.42 g/L
(c) Assuming ethanol is not present in the feed, Cpo=0. Steady-state product concentration is
3.4
g
0.42
qpx
h
L
Cp= Cpo+ D = 0 +
= 5.5 g/L
0.26
(
)
h
( )(
( )( )
( )( )
Qx= D (Cso-
KsD
-1
3
umaxD Yxs = 0.32h (20kg.m -
3.17 kg.m-3.h-1
Maximum biomass @ Dopt
Dopt= umax (1-
KS
KS+Cso
)= umax(1- -1)
( 0.8m3kg )( 0.32h )
(
0.45 0.32
)
h
h
)(0.55kg/kg)=
Dopt= 0.45h-1(1-
0.8 kg
m3
0.8 kg
kg
+20
m3
m3
) = 0.36h-1
KsD
umaxD Yxs
)
h
h
3.17
3.33 x100 = 95%
h . The lactose concentration in the feed stream is 9.5 kg.m -3; a substance conversion 98% is
required. The column is operated with plug Flow for a total of 310 days a year.
a. At what flowrate should the reactor be operated?
b. How many tonnes of gluctose are produced a year?
Given: Cs= 0.02Cso= 0.19 kg.m-3; Cso= 9.5 kg.m-3; umax= 45 kg.m-3h-1; Km= 1.32 kg.m-3;
VPFR=0.5 m3
Required: a. F, b. mass glucose (Tons)
Solution:
(a) For 98% substrate onversion:
Km
Cso CsoCs
= umax ln Cs + umaz = 0.32 h
F=
V
3
= 1.56 m /h
(b) The rate of lactose conversion is equal to the difference between inlet and outlet mass
flow rates of lactose= F(Cso-Cs)= 1.56 m3(0.5-0.19) kg/m3= 14.5 kg/h
kg 24 h 310 d 1 kmol
kmol
Lactose converted= 14.5 h . 1d . 1 yr . 342 kg =315 yr
The enzyme reaction is:
Lactose+ H20 --) glucose + galactose
Therefore, from reaction stoichiometry, 315 kmol glucose are produced a year. The
molecular weight of glucose is 180; thus,
Mass glucose= 315
kmol 180 kg
1T
Tonnes
.
.
=56.7
yr 1 kmol 1000 kg
yr
(mg/l)
(mg/l)
0.79
36.9
487
1.03
49.1
490
1.31
64.4
489
1.78
93.4
482
2.39
138.8
466
2.68
164.2
465
Solution
The accumulation = input output + rate of formation,
which for the biomass gives:
V(dX/dt) = FX0 FX + V(mSX)/(Ks + S) kdXV
(equation 5.126)
(equation 5.127)
(i)
(ii)
From equation (i), it is seen that a plot of X/D(S0 S) and 1/S will produce a straight line of
slope KsY/m and intercept Y/m.
From equation (ii), it is seen that a plot of (S0 S)/X against 1/D will produce a straight line of
slope kd/Y and intercept 1/Y.
The data are calculated as follows:
Feed
Exit
1/S
flowrate
substrate
(l/mg)
(F l/h)
(S mg/l)
0.79
36.9
0.0271
5.19
1.03
49.1
0.0204
4.05
1.31
64.4
0.0155
3.22
1.78
93.4
0.0107
2.40
2.39
138.8
0.0072
1.80
2.68
164.2
0.0061
1.64
The data are then plotted in Figure 5b from which:
KsY/m = 170,
1/D
(h)
12.41
9.51
7.48
5.51
4.10
3.66
(h
)
S
0
S
(
D
/
6
5
Y/ m= 0.59
4
3
X
2
1
0
(
X
/)
S
2.4
slope,kd / Y = 0.0133/h
2.3
(
X
2.2
1/ Y = 2.222
2.388
2.349
2.322
2.296
2.277
2.228
0 2 4 6 8 10 12
1/D (h)
m = (0.45/0.59) = 0.76 h1
and:
with m = 0.25 h1,Ks = 0.12 g/l, and the yield coefficient is 0.42, calculate the substrate and
biomass concentrations in the effluent from the second vessel. What would happen if the flow
were from the 50 l fermenter to the 100 l fermenter?
Solution
A material balance for biomass over the first fermenter, leads to the equation:
D1 = 1
(equation 5.131)
where D1 is the dilution rate in the first vessel and 1 is the specific growth rate for that vessel.
Assuming Monod kinetics to apply:
D1 = mS1/(Ks + S1)
(equation 5.132)
(equation 5.133)
(equation 5.134)
(equation 5.167)
where D2 is the dilution rate in the second vessel, 2 is the specific growth rate in that vessel and
X2 the steady-state concentration of biomass.
The yield coefficient to the second vessel is then:
Y = (X2 X1)/(S1 S2)
where S2 is the steady-state concentration of substrate in that vessel.
Thus:
X2 = X1 + Y(S1 S2)
= 1.97 + 0.42(0.309 S2)
= 2.1 + 0.42S2
Substituting this equation for X2 into equation 5.167, together with values for D2,2 and X1 leads
to a quadratic equation in S2:
0.
from which:
S2 = 0.0113 and:
g/l
When the tanks are reversed, that is with fresh feed entering the 50 litre vessel, then the dilution
rate for this vessel will be as before, 0.36 h1, although the critical dilution rate will now be:
Dcrit = mS0/(Ks + S0)
(equation 5.148)
concentration of substrate