Trends in Biotechnology, VoL 1, No.

4, 1983 113

The major impact of instability is on
the yield and efficiency of the fermenta-
tion process. However for pharma-
ceuticals which have to be manufactured
under GMP (Good Manufacturing
Practice) conditions the drug regu-
latory authorities consider instability to
be undesirable. Where the recombinant
protein is for industrial use, e.g.
enzymes for inclusion in detergents
etc., errors in the amino acid sequence
introduced by mistranslation are of
little importance unless they signifi-
cantly decrease the specific activity.
Again, however, they are of great sig-
nificance for drugs intended for human
use.
References
1 Hawley, D. K. and McClure, W. R.
(1983) Nucl. Acid. Res. 11, 2237-2255
2 de Boer, H. A., Comstock, L. J. and
Vasser, M. (1983)Proc. NatlAcad. Sci.
80, 21-25
3 Nugent, M. E., Primrose, S. B. and
Tacon, W. C. (1983) Develop. Ind.
MicrobioL 24 (in press)
4 Shepard, H. M., Yelverton, E. and
Goeddel, D. V. (1982) DNA 1,
125-131
5 Gelfand, D. H., Shepard, M.,
O'Farrell, P. N. and Polisky, B. (1978)
Proc. Natl Acad. Sci. 75, 5869-5873
6 Sninsky, J. J., Uhlin, B. E., Gustafsson,
P. and Cohen, S. N, (1981) Gene 16,
275-286
7 Uhlin, B. E. and Nordstrom, K. (1977)
Plasmid 1, 1-7
8 Gentz, R., Langer, A., Chang, A. C. Y.,
Cohen, S. N. and Bujard, H. (1981)
Proc. Natl Acad. Sci. 78, 4936-4940
9 Swamy, K. I-I. S. and Goldberg, A. L.
(1982)J. Bacterio1149, 1027-1033
10 Simon, L. D., Randolph, B., Irwin, N.
and Binkowski, G. (1983) Proc. Natl
Acad. Sci. 80, 2059-2062
11 Jones, I. M., Primrose, S. B., Robinson,
A. and Ellwood, D. C. (1980)214ol. Gen.
Genet. 180, 579-584
12 Kreft, J. and Hughes, C. (1982) in Gene
Cloning in Organisms other than E. coli
(P. H. Hofschneider and W. Goebel,
eds), pp. 1-17, Springer Verlag
13 Ehrlich, S. D., Niaudet, B. and Michel,
B. (1982) in Gene Cloning in Organisms
other than E. coil (P. H. Hofschneider
and W. Goebel, eds), pp. 19-29,
Springer Verlag
14 Jones, I. W., Primrose, S. B. and
Ehrlich, S. D. (1982)Mol. Gen. Genet
188, 486-489
15 Rood, J. I., Sneddon, M. K. and
Morrison, J. F. (1980)J. Bacterio1144,
552-559
16 Tomizawa, J. and Itoh, T. (1981)Mol.
Gen. Genet. 178, 525-533
17 Cesarini, G., Muesing, M. A. and
Polisky, B. (1982)Proc. NatlAcad. Sci.
79, 6313-6317
18 Tomizawa, J. and Itoh, T. (1982) Cell
31, 575-583
19 Stueber, D. and Bujard, H. (1982)
EMBOffournal 1, 1399-1404
20 Edelmann, P. and Gallant, J. (1977)
Cell 10, 131-137
21 Grosjean, H. and Fiers, W. (1982) Gene
18, 199-209
22 Chavancy, G., Daillie, J. and Garel,
J. P. (1971)Biochimie 53, 1187-1197
23 Nath, K. and Koch, A. L. (1971)
ft. Biol. Chem. 246, 6956-6967
24 Itakura, K., Hirose, T., Crea,
R., Riggs, A. D., Heyneker, H. L.,
Bolivar, F. and Boyer, H. W. (1977)
Science 198, 1056-1063

Mass transfer in fermentation where:

to= oxygen consumption rate (tool s -1)
Klaas van 't Riet r~ = CH20 consumption rate (mol s -1)
r x = biomass production rate (mol s-1)
An essential consideration in the design and operation of commercial
fermenters is to ensure adequate mass transfer. The complex By introducing the linear growth
composition of fermentation liquids makes it dittleult to predict equation:
accurately the mass transfer characteristics in large vessels. Here 1
various aspects of mass transfer are discussed and their relationships r~ = ~ rx + m~M~ (2)
examined. Strategies for predicting the most important type of mass
transfer - between gases and liquids - in large scale fermentations are where:
presented. Y~x = yield of biomass on substrate
(mol carbon in biomass per mol carbon
The changing environment of in substrate)
by comparing sets of calculated values
the organism with literature data for fermenter m s = maintenance coefficient (mol
At first sight, the relevant engineering tool-1 s-l)
engineering parameters and with the
parameters for the design of large-scale characteristics of the relevant organ- Mx = biomass in the reactor (mol)
bioreactors are heat transfer, mixing, ism. In this way 'engineering' and Eqn (1) can be modified to:
cell volume and mass transfer. Order of 'microbiology' are coupled. An aerobic
magnitude calculations show which is organism in a stirred or bubble-column OUR = { ( ~ x - 1.041/a + ms }Cx (3)
the restricting factor in most cases. type of fermenter is used as an example.
These calculations are based on the For specific applications the same pro- Fig. 1 shows the OUR values calcu-
application of elemental balances and cedure can be followed to check lated from Eqn 3 for Y~ -- 0.60 mol
the growth equations. These equations whether the result is very different. mo1-1 and ms = 10-s mol tool-1 s-1,
show the relationships between the Firstly the oxygen uptake rate, OUR taken from Ref. 1 as 'average' values. At
engineering parameters, the growth of (tool m -3 s -~) of a broth of given density steady state conditions the OUR value
the organism and the concentration of Cx (mol m-3), and a given growth rate/a is equal to the OTR value (oxygen
nutrients. The restricting engineering (s-l) is calculated. The application of transfer rate, m o l m -3 s-1). Later it will
parameter in each case can be estimated elemental balances for growth of be shown that from the viewpoint
Klaas van 't Riet is Professor of Food- and biomass (CH1.64N0.1600.52S0.0046P0.0054) limitations of mass transfer O T R =
Bio-Process Engineering, Department of on CH20, NH3, H3PO4 and H2SO4 2.8 x 10-5 mol m -3 s -1 (= 100 mol m -3
Process Engineering, Wageningen A g r i - leads to (Ref. 1): h -~) must be regarded as a rather high
c u l t u r a l University, De Dreijen 12, 6703 BC value for many types of fermenter. This
Wageningen, The Netherlands. ro = r~ - 1.04rx (1)
means that the growth of a culture can
1983,ElsevierSciencePublishersBN., Amsterdam 0166 9430/83l$01.00
114 Trends in Biotechnology, VoL 1, No. 4, 1983

be limited by inadequate mass transfer 10 -2 mol m -~ s -1. For small vessels, up OUR (mot m-3s-I)
even at intermediate growth rates and to about 1 m 3, the vessel wall provides Ex (rno[ m-3)
cell densities. Cell volume will not enough surface area for cooling. For
generally be limiting. The cell volume, larger vessels cooling coils or outside 100 _
derived from Fig. 1 for realistic OUR heat exchangers are needed. There is
values, does not exceed a value of 50% normally no engineering difficulty in
by volume, even at /~ - 0 where it providing the required surface of 1 m 2 10-1 _ 5000
reaches its maximum. It must be m -3 and thus considerations of heat
remembered that, in the case of fungal transfer will not restrict the design of
OUR= 2.8 x 10-2
growth, the viscosity is strongly related fermenters. i0 -2_
to cell density. This can lead to high Another restricting parameter in
viscosities2 and related problems in large scale fermenters might be mixing.
mixing and mass transfer, even for cell If the substrate is injected at only one 10-3_ so
densities corresponding to the region position into a continuous culture or
below OUR -- 2.8 x 10 -2 given in fed batch, then a volume element has to
Fig. 1. traverse the whole circulation loop I0 -~_
The rate of heat production H P R (W before it returns to the injection point.
m -3) is straightforwardly coupled to This circulation time can easily be 0 iC)-s 5x~O-S i~-~
OUR (Ref. 1): 10-100 s in large fermenters 5. The l.t (s -I)
concentration of substrate in the
H P R -- 4.7 x 105OUR (4) Fig. 1. Example of OUR values calculated
volume element will decrease during
The heat transfer rate H T R (W m -3) this circulation. A characteristic time by means of Eqn 3.
from the broth to the wall or cooling for substrate depletion, t~a, can be For a substrate consumption rate equal
coil can be calculated from: derived as follows. Assume a volume to that of the original one at t -- 0 with
HTR = a x 6T x A (5) element leaves the site where the Cs --- C,, the time needed to deplete the
substrate is added at t = 0. After a time substrate can be derived from Eqns 6
where: t, the decrease, ACa, of the substrate and 7, with AC, -- Cs introduced in Eqn
= heat transfer coefficient ( W m -2 concentration Cs, is given by 6:
K-') rat = ACs V (6) Y~C,+K~
A T = temperature difference (K) tcs - (8)
V = reactor volume (m 3) /amax Cx
A = cooling surface area (m2 m -3) When t < < tea then ACa < < Ca. At very
When maintenance is neglected, the
From Refs 3 and 4 the order of application of the Monod equation in low growth rates whereby Cs<< Ks, a
magnitude of a can be estimated as Eqn 2 leads to: minimum value of tcs is achieved, given
103Wm-2K-L Assuming AT = 10 K, by:
ra=l ~ CxV=~ Ca CxV(7 )
the surface area required for adequate ta _ YaxKa
cooling can be calculated to be of the Y,~ Ka+Ca Y,a /~m~x (9)
order of 1 m 2 m -3 for OUR = 2.8 x Ks = Monod constant (mol m -3) (, << ~ )
In many cases the time derived from
Symbols and their units Eqn 9 may be of the order of 1-100 s.
Symbol Definition Units Symbol Definition Units This means that, for large scale
A surface area m 2 m -3 OTR oxygen transfer rate mol m -3 fermenters with a single substrate
CoL,CoG concentration of oxygen s-~
P stirrer power
injection point, the organisms will
in the liquid (CoL) or gas
(CoG) phase mol m -3 consumption W experience severe variations in the con-
C~)L equilibrium COL value at ro oxygen consumption centration ofsubstrate in their environ-
the gas-liquid interface mol m 3 rate mol s- i ment. There is not much published
COL S concentration of oxygen rs CH20 consumption rate mol s information about the influence of fluc-
in the liquid in the stirrer rx biomass production rate mot s-j
region mol m 3 Sh Sherwood number no units
tuations in substrate concentration on
C biomass concentration mol m -3 tcO critical time for oxygen
the growth and metabolism of
/~L diffusion coefficient in the consumption s organisms. However, some articles6'7
liquid phase m 2 s -I tcs critical time for substrate show that there can be an effect.
db bubble diameter m consumption s Fluctuations in concentration,
g gravitational acceleration m s-2 vb bubble rise velocity m s-I
HPR heat production ratio W m -3
however, are inherently linked to large
vL liquid velocity m s-I
HTR heat transfer rate W m-3 Ysx yield of biomass on
scale operations and cannot be
kL liquid side mass transfer substrate mol eliminated completely by better
coefficient ms i mol -~ mixing. A simple method of decreasing
m partition coefficient of o~ heat transfer coefficient W m -2 the variations is to use multiple
oxygen between gas and K-I
liquid no units
injection points and well-defined
AT temperature difference K
ms maintenance coefficient mot gas hold up no units
circulation loops7.
mol -I I~ growth rate s- I Another restriction caused by poor
s-I blL liquid viscosity Pa s mixing is local oxygen depletion. This
OUR oxygen uptake rate mol m -3 60 density difference can happen when the transfer of oxygen
s-I between liquid and gas kg m -3
from the gaseous into the liquid phase
Trends in Biotechnology, Vol. 1, No. 4, 1983
~<
o tions the relevant mass transfer data
g~ COL
A
organism
bubble
diffusion
layer
Fig. 2. Schematic representation of oxygen
mass transfer from a bubble to an organism.
takes place in only one section of the
vessel (for instance, the stirrer region)
while long circulation loops transport
dissolved oxygen to other sections of
the fermenter. A critical oxygen deple-
tion time too can be derived as:
tcO- COLS (10)
OUR
where:
COLS = oxygen concentration in the
stirrer region (mol m -3)
Assuming CoLs to be the value that is
in equilibrium with air at atmospheric
conditions, then &o is of the order of
10 s. For large fermenters circulation
times can be of the order of 10-100 s, so
local zones with oxygen depletion may
be present. This phenomenon can be
reduced by improving the mixing
characteristics, increasing the static
pressure in the stirrer region, using
oxygen-enriched air and using multiple
impeller systems.
The order of magnitude calculations
presented so far show that heat transfer
and cell volume should not be regarded
as limiting factors in fermenters. The
distribution of substrate and oxygen
may be problematic, but some relative-
ly simple measures can solve these
problems. It should be noted that all
these calculations have been based on
an OTR of 2.8 x 10 -2 mol m -3 s -1,
assuming that to be a rather high value.
Fig. 1 shows, however, that at high
growth rates and moderate cell density
values much higher values of OTR are
possible. If indeed the OTR is deter-
mined by the capacity of the fermenter,
then this is the limiting parameter and
hence it largely determines the invest-
ment. Later it will alsobe shown that
115
effective mass transfer requires a con- bacteria) is beyond the scope of this
siderable power consumption and mass article*. In conclusion it can be said that
transfer determines part of the variable the kL and A values of the gas-liquid
manufacturing cost. In the next sec- interface in a fermentation broth play
the dominant role in oxygen transfer.
will be briefly reviewed and the costs The kL value is determined mainly by
worked out. The transfer of oxygen the character of the liquid interface
from air bubbles to the liquid phase will around the bubble. It can be 'rigid' or
be used as a model system. 'moving'. With a rigid interface there
exists a stagnant layer of liquid around
G a s - l i q u i d m a s s transfer the bubble. Oxygen is transferred by
The formula for the transfer of molecular diffusion along a concentra-
oxygen from the gaseous to the liquid tion gradient that can be assumed to be
phase is (see Fig. 2): linear, as shown schematically in Fig. 2.
OTR = kLA (C~L - COL ) (11) The thickness of the diffusion layer is of
the order of 10 -~ m. Outside this layer
where: there exists well-mixed bulk liquid.
COL, CoG = concentration of oxygen in The kL value will always be larger than
the liquid (L) or gas (G) phase (tool m -3) that for a bubble in completely
C~)L ---- CoG/m = equilibrium C O L value stationary surroundings, given by the
at the gas-liquid interface (molm -3) Sherwood number, Sh:
m = partition coefficient of oxygen
between gas and liquid (no units) Sh - kLdb _ 2 (12)
kL = liquid side mass transfer coeffi- DL
cient (ms -1) where
A = area of the gas-liquid interface (m2 db = bubble diameter (m)
m -3)
D L = diffusion coefficient in the liquid
This formula assumes that the limita- phase (m2 s -1)
tion to mass transfer is located in the
liquid layer around the gas bubble, kLA Experimentally determined kL values
is a reciprocal of the resistance to mass for bubbles with a rigid film and a well-
t r a n s f e r ; C~) L - C O L is the driving force
mixed bulk liquid are (Ref. 8):
for mass transfer. The resistance in the Sh - kLdb _ 2 + 0 31(db~33APg~'3 (13)
gas phase can be neglected due to the
large value o f m ( "-~35 for oxygen). The
where:
resistance in the liquid layer around
the organism can be neglected since the A# = density difference between liquid
organism is relatively small and thus and gas (kg m -3)
only an extremely small driving force is g = gravitational acceleration (m s -2)
necessary to transfer O2 from the bulk ~.1L = liquid viscosity (Pa s)
liquid into the organism. Mass transfer
to closely packed organisms (e.g. t Seep. 120forreviewon mouldsgrownin pellet
moulds in pellet form or immobilized form.
'0
E
J
2':/+-
~.:.:.:.:.:.:.: ............. eq. 1/-,
..............
..............
_ .,......
..........
..., ... i::i::iiiii::iii::iilregion wifh mectsu red
eq. 13 values
k~eq.12
, ~ , 1~)-2
d b m)
Fig. 3. k L values dependent of bubble diameter.
116
A bubble with a moving interface rises
through the liquid without any liquid
being entrained in the film layer. Mass
transfer is then described by penetra-
tion theory9. This describes non-
stationary mass transfer into an infinite
medium which initially has an oxygen
concentration equal to the bulk concen-
tration. The liquid is exposed to the gas
bubble for a very short time, equal to
that of the displacement of the gas
bubble over its diameter. It can theoret-
icaUy be derived that for such a system:
kL = /4_ JgL%~5 (14)
where:
% = bubble velocity
The relationships 12, 13 and 14 can
be verified by the vast amount of
measured data available from the
literature4; these are presented in Fig. 3.
Although the scatter is rather large, it
can be seen that the diameter has a
major influence on the ,/eL value. Very
small bubbles do have a rigid interface.
Eqn 13 describes the data; Eqn 12
predicts much lower values in
accordance with the difference between
absolutely static bubbles in a stagnant
liquid and the rigid diffusion layer of
limited thickness on which Eqn 12 is
based. Very large bubbles have a fully
moving interface and Eqn 14 predicts
the values rather well. For the region
between small and large bubbles the
data points are rather scattered. In this
region the bubble surface can be either
rigid or mobile, depending on the
presence of surface active materials in
the liquid. In general for large bubbles:
kL ~ 4 X 10-4 ms -1 and for small
bubbles k L --, 10-4 ms 1.
The surface area A is given by:
A-
6~ (15)
ab
where:
e = gas hold up (m3 m -3)
The hold up is dependent on the gas
superficial velocity vs and bubble
velocity % according to:
where:
VL = upward liquid velocity (due to
internal circulation phenomena in the
reactor).
The bubble rise velocity is mainly
dependent on the bubble diameter 4, as
'w 10 o-
IE
10-z , . , .
10-4 10-3 10-2
db(m)
Fig. 4. Bubble rise velocity.
shown in Fig. 4. It will be clear from the
kL, A, and vb formulas that the bubble
diameter plays a dominant role in
gas-liquid mass transfer. This is shown
also in Fig. 5 for an idealized system.
Unfortunately it is just this parameter
for which quantitative information is
lacking.
The bubble size is determined by two
separate processes: formation at the
sparger or stirrer and coalescence/
dispersion phenomena in the bulk
liquid. Air entering the reactor is
dispersed by the stirrer or the sparger.
The resulting bubble size can range
from very small (0.1-1 mm) up to large
(> 6 mm)4,u, depending on process
conditions. The collision frequency of
the bubbles will be high in the bulk of
the reactor, even at low gas flow rates
and hold up values. The result of the
collision is determined by the character
of the liquid. Two types of liquid can be
distinguished, coalescing and non-
coalescing. In a coalescing liquid each
collision leads to the formation of a
single bubble. In this way bubbles grow
A
I kL
.-J
I
I
I
i-~/--
QS- 20- 2~-
lo-
Fig. 5. The theoretical kLA value based on Eqns 15 and 16 (vL = 0) and Figs 3 and 4 for % =
0.1 ms-L
Trends in Biotechnology, VoL 1, No. 4, 1983
until they become large enough to be
dispersed by turbulent and shear
forces. An equilibrium bubble size is
established, resulting from continuous
coalescence and dispersion processes.
This equilibrium size is about 6 mm
for tap water. This means that within
0.1-1 m above sparger the bubble size
is independent of the bubble size near
sparger or stirrer.
In a non-coalescing liquid, the
bubbles remain at the size initially pro-
duced at the sparger or stirrer and thus
it is possible for much smaller bubbles
to exist in the bulk liquid than in a
coalescing liquid (Table 1). A typical
non-coalescing liquid is a strong salt
solution; a typical coalescing liquid is
distilled water. The addition of salt to a
vigorously stirred vessel with distilled
water decreases the bubble size
drastically"; in this case the very small
bubbles produced by the stirrer are
maintained in the bulk liquid.
Oxygen transfer in large scale
fermenters
Commercial fermentation liquids
have very complex compositions.
Besides a third phase, the ceils, there is
considerable amounts of medium, sub-
strates, products and cell debris. These
all influence coalescence characteristics
to some - mainly unknown - extent,
making it impossible to predict the
bubble size in the fermenter. This
uncertainty is increased by the presence
of chemicals often added to control the
foaminess of fermentation liquids. Even
traces of these chemicals can affect
coalescence and kLA considerably~z.
Although coalescence behaviour and,
--V b
d b ( m x 10 -3)
Trends in Biotechnology, VoL 1, No. 4, 1983
therefore, mass transfer cannot be
predicted in general terms for fer-
mentation liquids, it is possible to
arrive at an acceptable prediction. The
scale up and prediction method for kLA
will start from two extremes, two ideal
liquids: purely non-coalescing and
purely coalescing. These relations are
reviewed in Refs 4 and 13 as (see also
Fig. 6):
P/04
kLA = 2.6 x 10 -2 vs'5 (17)
(stirred vessel, water, coalescing)
kLA : 2.0 x 10 -3 v2 (18)
(stirred vessel, water with salts, non-
coalescing)
kLA = 0.32(vs) '7 (19)
(bubble column, water, coalescing)
where:
P -- power consumption of the stirrer
(w)
For a bubble column with a non-
coalescing liquid kLA depends on the
construction of the sparger and no
generalized relation can be presented.
It is clear that the differences in kLA
at one set ofP/Vand vs can be consider-
able, up to a factor 10 at high power
input in stirred vessels. For bubble
columns the differences are also con-
siderable for non-coalescing liquids
when combined with a sparger produc-
ing very fine bubbles. The character-
istics of a specific fermentation liquid
are unknown, but will, of course, lie
between these two extremes. A strategy
to overcome this problem is to take a
test run on an intermediate scale 0.1-
1 m 3 on the specific fermentation broth
and to determine kLA, P/V and vs,
preferably in a range relevant for the
large scale. Even these limited data,
when introduced into Fig. 6 and/or
eqns 17-19, will show the behaviour of
the specific fermentation liquid. These
data can be interpreted on the basis of
the two ideal liquids and kLA values at
scale up can be calculated based on P/V
and vs, with the assumption that the
composition of contaminants in the
liquid does not change. This strategy
using generalized correlations and
specific measured data gives the most
reliable scale up procedure until better
knowledge is acquired about the very
complex coalescence phenomena.
The influence of viscosity is not in-
cluded in eqns 17-19. In bubble
columns kLA decreases drastically
above ~ 0.05 Pa s. This type of reactor Fig. 6. kLA for a stirred vessel.
Table 1. Bubble size in the bulk fluid of the
reactor
Originalbubblesize (mm)
Liquidproperties at the spargerlstirrer
<<6 >6
coalescing ~ 6 ~6
non-coalescing <(<(6 ~6
is, therefore, useless for high-viscosity
fermentation liquids. For stirred
vessels kLA drops at viscosities about
~0.05 Pa s with/ar -5--'s (Ref. 3). To
date no generalized correlations are
available.
Several aspects complicate even
further the prediction of kLA for com-
mercial scale fermenters. One of these
is that the equations are based on kLA
data interpreted with the assumption
that kLA and P / V are both uniformly
distributed over the vessel. It is
common knowledge that, for viscous
media, the mass transfer mainly takes
place in the stirrer region and recently
the same was shown to be true even in
low viscosity liquids 14. This could com-
plicate the kLA prediction for com-
mercial fermenters (>10 m3). For
smaller vessels the formulas represent
empirical correlations that have been
proved valid.
Another aspect of large fermenters
(height (H) > 1 m) is the influence o f
static pressure differences on the value
of CoG and of oxygen depletion of the
bubbles. For small reactors the lack of
kLA (s -1)
10-
~f
~ 0.005
10 2
117
significant static pressure differences
and short mixing times result in CoG,
C~L and COLvalues that can be assumed
constant in the whole reactor. The CoG
value can be assumed to be equal to the
outlet gas concentration. Therefore the
calculation of COG, COL and C~L is not
difficult. However, most of the com-
mercial fermenters are considerably
larger. At H > 1 m static pressure dif-
ferences become relevant, the effect of
which is easily correlated with H. The
change in A due to pressure differences
can be considered in the same way. The
depletion of the bubbles cannot be
modelled simply by an ideally mixed
gas phase. For bubble columns correla-
tions for gas phase mixing are known 4,
but there is a lack of data for stirred
tanks. Here an estimation has to be
made, ranging from ideally mixed to
plug flow, on the basis of specific pro-
cess conditions and sound reasoning.
Finally a combined model for gas and
liquid phase must be made to calculate
O T R values ~4-~6.
The ratio between the concentration
of oxygen in the liquid and gas phases at
equilibrium is defined as m. The value
of m is independent of COG itself. The
solubility of oxygen in liquids decreases
with increasing temperature ~7 and this
is disadvantageous for thermophilic
fermentations due to the decreasing
driving force in eqn 10. This dele-
terious effect is partly- offset by the
increase of kLA with temperature 4.
vs (m s-I)
,,0.05
/ 0.005
f.'J .,, ~f
1
" / 0.05
d
non coal eq. 18
con[ eq. 17
163 10~
P/V (Wm-3)
118
Oxygen solubility also changes by up to
5070 with the addition of saks, sub-
strates and product to the liquid ~s. The
accurate measurement ofm is laborious
but can be omitted for many applica-
tions. In the scale-up procedure given
in this article a n u m b e r of measure-
ments of the specific fermentation
broth are incorporated, which can be
done by the static method 13. COL and
C~L can be measured by a polar-
ographic oxygen electrode as per-
centage of the saturation concentration.
In this way kLA/m can be calculated.
This value can be used for calculation
of O T R in large scale operations. This
method fails as soon as liquid or gas
flow modelling is required, because it is
then necessary to know the absolute
values of COL.
Economics of oxygen transfer
Eqns 17-19 show that kLA can be
increased by an increase of P/V in stir-
red vessels and ofvs in bubble columns.
T h e v~ dependency for bubbles can also
be regarded as a P / V dependency4.
Even the absolute values of P / V for
obtaining a kLA value are the same in
both types of fermenter 4. The question
is now: can we overcome the mass
transfer restriction by increasing P/V?.
In the introduction to this article an
O U R value of 2.8 x 10 -2 mol m -3 s -1
was stated as a relatively high value.
This corresponds to a P / V value of an
order of 2 kW m -3. It is shown now that
cost aspects exclude an order of magni-
tude increase of P/V. In eqns 17-19 kLA
is proportional to P / V with the
exponent 0.5, 0.7 and 0.6 respectively.
Let us assume:
kLA a ( p ) 0 6 (21)
According to Eqn. 11 this indicates that
O T R a (P/I0 .6.
For a given fermentation:
ro = O T R x V =
P/06
constant = CX x V (22)
Thus: p cz V-z3 (23)
E q n 23 shows that the total power
input increases with decreasing fer-
menter volume and fixed total oxygen
consumption. This can be regarded
as a general rule for each fermenter,
although the exponent may differ to
some extent. This means that the effi-
ciency of oxygen transfer decreases for
each type of fermenter with an increase
of the total amount transferred.
Trends in Biotechnology, VoL 1, No. 4, 1983
invesfrnenf + power cosf ( kf yr -I)
scenorio fclbte 2
1000-
500
0 I I I
0 100 200 300
votume (m 3)
Fig. 7. Total cost for the scenarios given in Table 2.
Numbers such as kg O2(kWh) -1 are only and cannot be used as absolute
meaningless without defining O T R figures. In addition to power and
values. investment costs, there are more
T o find out if power consumption is phenomena related to P / V and V, for
an important part of production costs example cell disruption at extreme P / V
we can look at an example based on a values and the relationship of cell con-
100 m 3fermenter with 2 kW m -3 power centration and broth volume to separa-
input and an investment cost that is tion costs. T h e trends, however, are
assumed to be proportional to V05 inevitable, showing that both very
(Table 2). These scenarios must be seen large, as well as very small volumes are
as indications only, but the range of the not feasible. More specifically, it can be
ratio between investment and power concluded that a low loaded reactor (.-. 1
costs of a factor 16 would point to kW m -3) without stirring device would
many commercial fermentations being be preferable for simple fermentations,
somewhere in this range. The summa- Table 2. Investment and power costs
tion of power and investment cost,
resulting from the three scenarios is Example used as basis
given in Fig. 7. In this figure the 100 m 3 Investment costs: Powercosts:
100 m3 fermenter 2 kW m-3
fermenter is used as standard. A 1000 kfinvestmenta 6 10-5 kfper kWh
decrease in volume to 10 m 3 lowers 300 kfper year 100 kfper year
investment costs but from Fig. 7 we can Scenarios
see that the increased power cost makes
Investment cost Powercost
the small fermenter feasible only for the (kfyr -2) (kf yr -1)
highest fermenter/power cost ratio. " V .o.s / V x-67 Like example
When the relative cost of the fermenter ,A) 300 ( ~ 0 ) 100~100}
is lower, large fermenters with inter-
mediate or even low P / V values
(B) 150 [_~V~o.s 200 I*//v'\ -0.67Power more
of 1-2 kW m -3 become feasible. t,100/ ,1100/ expensive.
Fermenters with P/Varound 1 kW m -3 Fermenter less
expensive.
are especially attractive because then
non-stirred vessels can be used and
I V ~0.5
these offer significant mechanical ad- (c) 600 ~1-~! t v $-o.s7Fermenter
50 ~100
vantages. The energy dissipated by the ] more .
expensive.
air bubbles suffices at superficial velo- Power less
cities of 0.1-0.2 m s-L expensive
The data in Fig. 7 indicate trends ~1 kf = 1000Dutch guilders ~ 350 US$.
Trends in Biotechnology, Vol. 1, No. 4, 1983 119

with a product separation that is rather scales, rather accurate data can be 6 Leegwater, M. P. M., Neijessel, O. M.
insensitive to broth density. This case is obtained by combining general and Tempest, D. W. ( 1981) SecondEur.
comparable to example B in Fig. 7. formulae for model liquids with a Congr. of Biotech. 5-10 April 1981,
Examples might be single cell protein limited number of measurements on Eastbourne England. Abstracts of
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7 Schiigerl, K. (1983) Chem. Ing. Teehn.
fermentations requiring expensive scale mass transfer may even be pre-
55, 123
fermenters with a recovery that is very dicted quite accurately in this way; 8 Calderbank, P. H. and Moo-Young, M.
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preferred. Even then P/Vvalues > > 10 the vessel can be expected. These can 365
kW/m 3 will rarely occur in practice. be partly overcome by modelling gas 10 Wallis, G. B. (1969) One-dimensional
The very high fermenter costs could be and liquid mixing. two-phase flow. McGraw Hill Book
caused by the necessity of expensive Oxygen transfer determines a signi- Company
construction materials, of (volume ficant part of manufacturing costs 11 Lee, J. C. and Meyrick, D. L. (1970)
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Eur. J. Appl. Microbiol. Biotechnol. 9,
fermenter is to be preferred. 249
Conclusion 13 Van 't Riet, K. (1979) Ind. Eng. Chem.
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