4 Continuous Reactors
257
Run 3: 2021 steps in 0.15 seconds
120
0.008 0.007
100
1 I
80 I I  60
1
1
\ . ll ', 1
1
. !i
40
\\
v i
120
1 i
20
\ i I i '\ * \ \! \l ! \\ \
1
20
40
60
80
100
140
160
180
200
TIME Figure 3. Influence of initial KLa value from 100 to 160 h"^ on the S and O profiles.
8.4
8.4.1
System
Continuous Reactors
SteadyState Chemostat (CHEMOSTA)
The steady state operation of a continuous fermentation having constant volume, constant flow rate and sterile feed is considered here (Fig. 1).
Biological Reaction Engineering, Second Edition. I. J. Dunn, E. Heinzle, J. Ingham, J. E. Pfenosil Copyright 2003 WILEYVCH Verlag GmbH & Co. KGaA, Weinheim ISBN: 3527307591
258
D,S
Model
The dynamic balance equations may be modified to apply only to the steady state by setting the time derivatives equal to zero. The corresponding equations are then: For biomass,
0 =  D X + rx
For substrate,
0 = D (S0  S) + rs
Growth kinetics,
rx = ^ X
259
The productivity of the reactor for biomass is X D. The above equations represent the steady state model for a chemostat with Monod kinetics. Using them it is possible to calculate the values of S and X, which result from a particular value of D, and to investigate the influence of the kinetic parameters.
Program
In Madonna programs, time can be used as a variable which will increase from the starting time. Here it is renamed D. Thus equations will be solved for increasing values of the dilution rate. Fortunately X and S can be explicitly solved for in this problem. If not, the ROOT FINDER facility of Madonna can be used. The program is found on the CDROM.
Nomenclature
The nomenclature is the same as the example CHEMO, Sec. 8.1.2.
Exercises
260
Results
The steady state curves of X, S, and XD versus D are given Fig. 2. The results in Fig. 3 were obtained by varying K$ in each run. An interesting effect can be observed on the position of the washout point.
Run 1:113 steps in 0 seconds
10 9876v5
f*r
4
3.5
i
S" //
0
0.1 0.2 0.3 0.4
m ^r
*S**~\ ! I \ Y.I 1i
3
2.5
~s;i
mm
\!
"
1 /{
0.5 0.6 0.7 0.8 0.9
2
1.5
43
1
0.5
210
0 1
261
Run 5:113 steps in 0 seconds
8.4.2
System
Inhibitory substrates at high concentrations reduce the specific growth rate below that predicted by the Monod equation. The inhibition function may be expressed empirically as
where KI is the inhibition constant (kg/m3). If substrate concentrations are low, the term S2/Kj is lower in magnitude than KS and S, and the inhibition function reduces to the Monod equation. In batch cultures the term S2/Kj may be significant during the early stages of growth, even for higher values of K[. The inhibition function passes through a maximum at Smax = (Kg Ki)5. A continuous inhibition culture will often lead
262
to two possible steady states, as defined by the steady state condition JLI = D and as in shown Fig. 1.
D=
One of these steady states (A) can be shown to be stable and the other (B) to be unstable. Thus, only state A and the washout state (S = SQ) are possible.
Model
A model of a chemostat with its variables is represented schematically in Fig. 2.
F,S 0
+
F,S,X
263
Program
When the system equations are solved dynamically, one of two distinct steady state solutions are obtained, the stable condition A and the washout condition. The initial substrate and organism concentrations in the reactor will determine the result. This is best represented as a phaseplane plot X versus S. All results indicate washout of the culture when the initial cell concentration is too low; higher initial substrate concentrations increases the likelihood of washout.
Nomenclature Symbols
D KI KS Dilution rate Inhibition constant Saturation constant 1/h kg/m3 kg/m3
264
S
Smax X Y
Substrate concentration Maximum in S for inhibition function Biomass concentration Yield coefficient Specific growth rate
Indices
0 I m
Exercises
265
Results
Run 1:2000 steps in 0 seconds 112
10
15
20 TIME
25
30
35
40
5 i
4.5 
4
3.5
3
e/> 2.5
2
1.5
1
0.5
0
0.5 2.5
Figure 4. Phaseplane plot of X versus with varying ST from 0 to 5 kg/m3 using Batch Runs with overlay.
266
54.5
X,
4 3.5
^^^>
*" 3
</) 2 . 5 
^ *t %%
\
"
t**
* * C^1 v
"% % V
\ ^ x i ! l
2
1.5
1
0.5
/ /; ,^, f/ // / ^' . *x
/
J
3:2 (0.5) 3:3 (0.5857) _   3:4(0.6714) 3:5(0.7571) 3:6(0.8429) 3:7(0.9286) 3:8(1.014) 3:9(1.1)
j* S ^
f f / t / ,f !
rf'^T
"^ s^^
"^"^>*i'^' ** ^
^^^*^^$^S? ** ^^*^^ **
0.5
Figure 5. Phase plane plot of influence of the initial biomass Xi from 0.5 to 1.1 for Steady states upper left and lower right.
Run 20:2000 steps in 0.0167 seconds
= 0.0.
Reference
Edwards, V.H, Ko, R.C. and Balogh, S.A. (1972). Dynamics and Control of Continuous Microbial Propagators Subject to Substrate Inhibition Biotechnol. and Bioeng. 14, 939974.
267
8.4.3
System
Nitrification is the process of ammonia oxidation by specialized organisms, called nitrifiers. Their growth rate is much slower than that of the heterotrophic organisms which oxidize organic carbon, and they can be washed out of the reactors by the sludge wastage stream (Fs). In an activated sludge system (Fig. 1) when the organic load (F So/V) is high, then the high biomass growth rates require high waste rates. Nitrification will not be possible under these conditions because the concentration of nitrifiers (Ni) will become very low.
O,F O
2, F4
Reieto*
2, F3
Figure 1. Configuration and streams for the activated sludge system.
Model
The dynamic balance equations can be written for all components around the reactor and around the settler. The settler is simplified as a wellmixed system with the effluent streams reflecting the cell separation. Organic substrate balance for the reactor: = F 0 So + F 2 S 2 
268
R2Vt
= F 2 N 2  F i N i + R2Vi
V 2 dS 2
= F i S i  F3S2  F4S2
3t
V 2 dA 2
= F A
I I 
 F4A2
= Fl l  F3 02
V2 dN2 34 = F i N i  F 3 N 2
The equations for the flow rates are given below. Recycle flowrate:
F2 = F 0 R
where R is the recycle factor. Reactor outlet flow: Flow of settled sludge:
FI = F2 + F0 = F O R + FO
269
where C is the concentration factor for the settler. Flow of exit substrate:
F4 = FI  F3>
Flow of exit sludge wastage:
F5 = F3  ?2.
Note that C and R must be chosen so that F5 is positive. Monodtype equations are used for the growth rates of the two organisms.
l^2max
2 = ^Ni =
Program
The program is given on the CDROM.
Nomenclature Symbols
A C F Fo5 KI K2 N O R Ammonia substrate concentration kg/m3 Concentrating factor for settler Flow rate m3/h Flow rates, referring to the figure m3/h Saturation constant of heterotrophs kg/m3 Saturation constant of nitrifying organisms kg/m3 Concentration of nitrifiers kg/m3 Concentration of heterotrophs kg/m3 Recycle factor 
270
Rl
R2
V Y Hi
Growth rate of heterotrophics kg/m3h Growth rate of nitrifying organisms kg/m3h Organic substrate concentration kg/m3 Volumes m3 Yield coefficients kg/kg Specific growth rate of heterotrophs 1/h Specific growth rate of nitrifying organisms 1/h
Indices
Flow and concentration indices referring to Fig. 1 are as follows: 0 Refers to feed and initial values 1 Refers to reactor and organic oxidation 2 Refers to settler and ammonia oxidation 3 Refers to recycle 4 Refers to settler effluent 5 Refers to sludge wastage m Refers to maximum
Exercises
271
Results
The results in Fig. 2 demonstrate the influence of flow rate on the effluent organics 82 The ammonia in the effluent A2 is seen, in Fig. 3, to respond similarly to FQ, but for a very high value of FQ = 1000 m3/h the nitrification ceases, and A2 becomes the same as the inlet value AQ. This corresponds to washout of the nitrifiers, which would be seen by plotting NI versus time.
Run 4: 405 steps in 0.0167 seconds
0.9 , 0.8 I ,/"
M if
0.3 rr
02.
If J II / It
J II " 01 JM
" . 6
10
12
14
16
18
20
TIME Figure 2. Transient of S2 at various flow rates F0 (20 to 500m3/h, bottom to top).
0.090.08
3 0.06 _005 J* 1
0.040.03
S.
0.02J
0 2 4 6 8 10 TIME 12 14 16 18 20
Figure 3. Ammonia in the effluent (A2) at various flow rates F0 (5 to lOOOm^/h, bottom to top).
272
8.4.4 System
A tubular, packedbed, immobilizedenzyme reactor is to be investigated by simulation. The flow is assumed to be ideal plug flow. The distribution of the enzyme is not uniform and varies linearly from the inlet to higher values at the outlet, as shown in Fig. 1.
Distance along reactor, Z Figure 1. Distribution of enzyme along the tubular reactor.
Model
The equations for steady state operation are given below. Substrate balance,
dS dZ
=
1 ~v
Kinetics,
The linear flow velocity is increased by the presence of the solid enzyme carrier particles according to
273
V7 L
F Ae
~
Program
The model is solved by renaming the independent variable, TIME, to be the reactor length coordinate Z. The program is given on the CDROM.
Nomenclature Symbols
A F K KM m r S vm vz Z e E Reactor tube cross section Flow rate Rate constant MichaelisMenten constant Enzyme distribution constant Reaction rate Substrate concentration Maximum reaction velocity Linear flow velocity Reactor length Void volume fraction of packing Enzyme concentration m2 m3/h 1/h kg/m3 kg/m3 m kg/m3 h kg/m3 kg/m3h m/h m kg/m3
274
Indices
0 S
Exercises
Results
Flow rate is the primary operating variable, along with enzyme loading and inlet concentration. In Fig. 2 the influence of F is seen in the steadystate, axial, substrate profile.
275
Run 6:1000 steps in 0.05 seconds
10
12
14
16
18
20
8.4.5
System
In definednutrient growth media, one substrate can usually be made to be limiting by adjusting its concentration relative to those of the other medium components. In general, however, more than one substrate may limit the cell growth rate. In this case the yield coefficients for the various components, Yxsi> may vary depending upon the growth regime. This situation was discussed by Egli et al. (1989), who examined results at steady state with dual nutrient limitation. The present mathematical model simulates the transient behaviour of such a dual (Si carbon, 82 nitrogen) nutrientlimited system when carried out in a chemostat. The model assumes that the yield coefficients are each a function of the ratio 81/82, i.e. the ratio of the carbonnitrogen substrate concentrations in the vessel. The original paper took the carbonnitrogen ratio in the feed stream as the controlling parameter. Here the concentrations in the reactor are assumed to be controlling.
Model
Assuming a perfectly mixed, constant volume continuousflow stirredtank reactor, the mass balance equations for the cells and for the two limiting substrates are as follows:
276
= D
(SlFeed  Si) 
D
where D = F/V.
/O
O \
I ^__i^^_ I i i "V
The yield coefficients are assumed to vary with the carbonnitrogen ratio in the reactor. Si RATIO = ^ The yield coefficients are varied according to RATIO using the following logic:
Y X Sl=Yi m i n where, _ Y2min Y 2max Bi = \r 1  and Bo = 1v~,T"
Imax  1mm
and and
if if
The boundaries of the three growth regimes in Fig. 1 are defined by the quantities BI and B2.
277
C limitation
Double limitation
N limitation
10
XSi
r
0.8
XS1
B2
S2
Figure 1. Limitation regions for carbon and nitrogen showing influence on yield.
The yield coefficients for biomass on nitrogen and carbon take maximum or minimum values when only one substrate is limiting and vary linearly with opposing tendencies in the doublelimitation region.
Program
Note that the programing of this example is rather more complicated than usual owing to the need to allow for the logical conditions of carbon limitation, nitrogen limitation or both substrates together causing limitation. A partial listing is seen below and the full program is on the CDROM.
(CALCULATION OF YIELD VALUES)
YXSl=if (RATIO < Bl) then YlMAX else ( if (RATIO > B2) then Y1MIN else (Y1MAX+(RATIOB1)/(B2B1)*(Y1MINY1MAX)) ) YXS2 = if (RATIO < Bl) then Y2MIN else ( if (RATIO > B2) then Y2MAX else (Y2MIN+(RATIOB1)/(B2Bl)*(Y2MAXY2MIN)) )
278
Nomenclature Symbols
Bi
B2 Cc Cn D F
Ratio of Y 2min /Yi max Ratio of Y 2max /Yi min Carbon source concentration Nitrogen source concentration Dilution rate Volumetric feed rate Affinity constant Reaction rates Carbon source concentration Nitrogen source concentration Biomass concentration Yield coefficient Specific growth rate
_

kg/m3 kg/m3
1/h
m3/h kg/m3 kg/m3 h kg/m3 kg/m3 kg/m3 kg/kg
KS
R
Si S2 X Y H
1/h
Indices
1
2
Exercises
279
Results
The startup of a continuous culture is shown in Fig. 2. Note that the nitrogen level 82 in the reactor drops to a low level after 15 h and causes a change in the yield coefficients. The influence of dilution rate on the system was investigated by varying D from 0 to 1.5 as shown in Fig. 3.
3c
X 1.5
280
Reference
Egli, Th., Schmidt, Ch. R. (1989). "On DualNutrientLimited Growth of Microbes, with Special Reference to Carbon and Nitrogen Substrates", in Proceed. Microb. Phys. Working Party of Eur. Fed Biotech. Eds. Th. Egli, G. Hamer and M. Snozzi, HartungGoree, Konstanz, 4553. This example was developed by S. Mason, ETHZurich.
8.4.6
System
The process involves the removal of dichloromethane (DCM) from a gas stream and the subsequent degradation by microbial action. The reactor consists of biofilm sand bed column with circulation to an aeration tank, into which the substrate and oxygen enters in the gas phase, or the substrate can be fed in a liquid stream, as shown in Fig. 1. The column is approximated by a series of six stirred tanks. The reaction is treated with homogeneous, double saturation kinetics with dichloromethane (DCM) inhibition. The oxidation of one mole of DCM produces 2 moles of HC1, making a hydrogen ion balance for pH important. The yield with respect to oxygen is 4.3 mg DCM/mg 62. In practice, care must be taken to prevent stripping of DCM to the air stream.
281
SR6>
SRin C jn , pH jn
Model
The model does not include a gas balance on the aeration tank, since it is assumed that the gas phase dynamics are comparatively fast and hence an equilibrium with the inlet concentration of oxygen and DCM may be assumed. The biomass is assumed to grow slowly, and growth rates are therefore also not modelled. The model for pH changes does not include buffering effects. For the inlet section 1 at the bottom of the column the balances are as follows: O2 balance,
dCQ1 ^ dt
282
DCM balance,
dC
Srl _ dt
Srin~ C Srl
i CHI
2r S i 84900
Here T is the residence time of the liquid in one section of the column. The constant 84,900 converts grams to moles and includes the stoichiometry. pHi = 0.434 log Cm Evaluation of rates for the inlet section 1:
maxCSrl
01
KI )
~ = (CSr6  CSrin )
dC
at
R V
DCM (Cs2eq 
Program
The program constants describe DCM entering the reactor in the gas stream. The DCM concentration in the liquid feed is set to zero. The program is on the CDROM.
283
Nomenclature
H+ ion concentration in section n kg mol/m3 Inlet dissolved oxygen concentration g/m3 Oxygen saturation constant g/m3 DCM saturation constant g/m3 DCM inlet concentration g/m3 DCM concentration in section n g/m3 Oxygen concentration in section n liquid g/m3 DCM concentration in feed g/m3 DCM gas concentration g/m3 Feed rate m3/h Inhibition constant g/m3 Transfer coefficients for DCM and 2 1/h Saturation constants g/m3 pH in n section n pH units Recirculation rate m3/h Oxygen uptake rate in section n g/m3 h Substrate uptake rate in section n g/m3 h Reactor volume m3 Volume of aeration tank m3 Maximum degradation rate g/m3 h Yield coefficient for DCM/oxygen Liquid residence time in one section h
oin
srin
CSFO
CSG F
KI KLa KS
pHn R
VR
VT
Vmax YSO
Exercises
1 1
284
Results
The concentrations in the stream leaving the top of the column (CSr6) during startup of the fluidized bed are shown in Fig. 2 for four values of F (0.5 to 10) The change of the pH for one flow rate (F = 0.5) is shown in Fig. 3.
285
Run 4: 55 steps in 0.0167 seconds
0.05
0.1
0.15
0.2
0.25
0.3
0.35
0.4
0.45
0.5
TIME
Figure 2. Fluidized bed startup for four values of F (0.5 to 10, bottom to top).
3
2.5
2
1.5
1
0.5
0 0
0.05
0.1
0.15
0.2
0.25
0.3
0.35
0.4
0.45
0.5
TIME
Figure 3. Change of carbon substrate and pH in the top section 6 during startup.
Reference
D. Niemann Ph.D. Dissertation 10025, ETH, 1993.
286
8.4.7
System
Two chemostats are arranged in series (Fig. 1) with the intention that the first operates at a relatively high rate of cell growth, while the second operates at low growth rate, but high cell density, for secondary metabolite production. Additional substrate may be fed to the second stage.
, 810
X1.S!
Hi US
Model
The balance equations are written for each component in each reactor. Stage 1 with sterile feed,
= F[S O S!] 
287
Stage 2 with additional substrate feed and an input of cells and substrate from Stage 1,
V2
 [F + Fi]X2
v Y
Program
The program is on the CDROM.
Nomenclature Symbols
F
KS
Prod S V X Y
Volumetric feed rate Saturation constant Productivity for biomass Substrate concentration Reactor volume Biomass concentration Yield coefficient Specific growth rate
288
Indices
0 1 2 10 m
to tank 1 inlet to tank 1 and inlet of tank 2 to tank 2 and outlet of system to separate feed for tank 2 to maximum
Exercises
Results
The results in Fig. 2 give biomass concentrations and productivities for both tanks during a startup with a constant feed stream to the first tank (F = 0.5). In Fig. 3 the influence on X2 of feed to the second tank Fl (0 to 1.0) with constant F is shown.
289
Run 1: 805 steps in 0.0333 seconds
T5
35
40
Figure 2. Biomass (Xj X2) and productivities for both tanks (F = 0.5).
5
4.5
4
3.5
32.5
2
1.5
1
0.5
0 10 15 20 25 30 35 40
TIME
Figure 3. Influence on X2 of feed to the second tank (Ft = 0 to 1.0, curves right to left).