BIOTECHNOLOGY AND BIOENGINEERING,

VOL. XVIII, PAGES 1001-1016 (1976)

Fed Batch Culture of Saccharomyces cerevisiae:

A Perspective of Computer Control to Enhance
the Productivity in Bakers Yeast Cultivation
SHUICHI AIBA, SHIRO NAGAI,* and YOSHINORI NISHIZAWA, Institute of Applied Microbiology, University of Tokyo,
Tokyo, J a p a n

Summary
A means t o avoid the glucose effect in the production of bakers yeast from
glucose and/or molasses in a fed batch culture by controlling the feed rate of
fresh medium with an ad hoc measurement of the respiratory quotient, RQ, is
presented. The feed rate is changed stepwise here such that the value of RQ
ranges from 1.0 to 1.2 throughout the cultivation. Thus far, the specific growth
rate based on the total cell mass and the growth yield obtained throughout
are 0.24 hr-l and 0.55 g cell/g glucose.
Prior to the experimental run mentioned a,bove, equations to predetermine
the feed rate and concentration of glucose in the feed are derived from the mass
balance of limiting substrates (glucose). Since values of either RQ or l o ,
(Qo2 2, oxygen consumption rate with respect to the total cell mass in the fermenter) can be measured quite easily and reliably, computer control of the fermentation in light of this information is discussed.

INTRODUCTION
The well-known phenomenon termed glucose effect cannot be
prevented in the aerobic cultivation of Xaccharom yces cerevisiae in a
glucose medium. This phenomenon is the so-called aerobic fermentation. When glucose concentration in an aerobic culture medium
reaches 70 mg/liter,2 glucose tends to be partly metabolized during
the fermentation to ethanol and COz; the fermentation is claimed to
cease if glucose concentration in the medium is less than the specific
level of 70 mg/liter.
Whereas a larger than expected yield of cells in the absence of aerobic fermentation deteriorates the specific growth rate, the fact that the
* Present address: Department of Fermentation Technology, Hiroshima
University, Hiroshima, Japan.
1001
@ 1976 by John Wiley & Sons, Inc.

1002

AIBA, NAGAI, A N D NISHIZAWA

increase of the specific growth rate is accompanied by necessity by a

lower yield of cells is a characteristic of the aerobic cultivation of
baker's yeast. From the viewpoint of securing the most favorable
productivity of an aerobic cultivation of the yeast, there should be a
mediation in uncompromising phenomena between fermentation and
respiration.
Maxon and Johnson3 reported earlier a diauxic growth of the X.
cerevisiae on glucose in an aerobic and batch culture; the maximum
value of the specific growth rate and growth yield reported therein
was 0.41 hr-1 and 0.14 g cell/g glucose when fermentation prevailed.3
Von Meyenburg' published his work on a chemostat culture with S.
cerevisiae using glucose as the limiting substrate, in which the value
of the growth yield deteriorated from 0.50 to 0.145 'g cell/g glucose,
depending on the dilution rates ranging from 0.24 to 0.45 hr-1.
However, the data on the residual concentrations of glucose in the
culture medium are not described in the original paper;' it could
easily be envisaged that the residual concentration of glucose increased when the dilution rate was enhanced.
It is considered significant a t this point to minimize the glucose
effectin order to produce a higher yield of cells and, in fact, empiricism
has contributed to the development of the fed batch culture in the
production of baker's yeast. I n addition, Pirt presented some discussion on the theoretical aspect of the fed batch culture,4 although
a formula t o define the feed rate required for negating the glucose
effect has not been established.
I n this connection, either the ethanol concentration in exit gas from
the aerobic cultivation or the respiratory quotient (RQ)value with
respect to the exhaust gas may be employed as parameters in the fed
batch culture. If the concentration of ethanol in the exit line becomes detectable, the feed rate of fresh medium into the culture might
be squeezed down and vice versa; indeed, this idea has already been
materialized.5 However, the installation of gas chromatograph on
the exit line is not always allowable from the viewpoint of economy.
I n this work, the RQ value which can be determined by measuring the partial pressures of oxygen and carbon dioxide in the exhaust
gas will be used as a parameter to control and minimize the aerobic
fermentation. The purpose of this work is, first, to demonstrate
the controllability of this fermentation via RQ values after formulating the feed rate and concentration of glucose in the feed and second,
to discuss a perspective of computer control of baker's yeast production which guarantees the maximal yield and productivity.

F E D BATCH CULTURE OF S. CEREVISIAE

1003

THEORETICAL CONSIDERATION
In line with the experimental setup appearing later on, suppose
that a fresh feed is charged into a culture vessel at a constant rate of
Fi during Ati (= ti+* - ti). The subdivision of the cultivation period
of time from 1 = to to t = t p into At; (i = 0 to i = n), culminating in
the increase of broth volume from V = Vo to V = V p as shown in
Figure 1, is only for the convenience of experimentation as well as
the derivation of some equations which follow.
Assuming a complete mixing of the medium in a well-agitated and
aerated fermenter, the rate of change in concentrations of the cell
mass and growth-limiting subst,rate (glucose) are :

AX;
=
At;

p i x i

Fi
)Xi
Vo; Fi Ati
1 d(X;V;) _
d X_
;
- 1_
X i dt
X i V ; dt

1dx;
--

PI,
a -

X;

=pi

to

dt

+--V1i ddtV ;

~~

+ Voi +1F ; At; Fi

tl

kAto+-Atq

tl

72
4

------

tl+l

bat14

---

tn

tnti

C A t n 4

t
Fig. 1. Schematic diagram of parameters in each time interval in the fed batch
culture.

AIBA, NAGAI, AND NISHIZAWA

1004

where X is the concentration of cell mass in culture medium (g/liter),

J: is the total cell mass in fermenter (g), V is the broth volume (liter),
P is the feed rate of fresh medium (liter/hr), p is the specific growth
rate based on cell mass concentration (hr-l), p' is the specific growth
rate based on total cell mass (hr-l), t is the time (hr).
The subscript i is the ith interval of time (see Fig. 1) and the subscript 0 is the initial of the ith interval.
The mass balance for the limiting substrate during Ato (cf. Fig. 1)
yields,

Si { Vo

+ Fo(ti -

to)) -

Sovo

S~Po(ti- t o )

(eccumuhtion)

(input)

- [ X i { vo

Yo

Fo(t1

- to)

1 - XoVoI

(3)

(consumption due to cell growth)

Rearranging eq. (3),

(Si - So) { Vo

+ Fo

(ti

to)) =

Po(ti - t o ) ( S ~- So)

provided
Po=

1 dX
1 ( X , - XO)
--=Xo dt
Xo (tl - t o )

Where S is the concentration of glucose in the culture medium

(g/liter), SR is the concentration of glucose in fresh feed (g/liter),
Y is the growth yield (g cell/g glucose).
Modifying eq. (4) so that SI - SO= AS0 and tl - t o = Ato,
AS0
--

Ate

Fo

Vo

+ Po Ato

(SR

- SO)
1

- - POX0

Yo

Yo Vo

+FoPo Ato xo

(5)

A general expression for eq. ( 5 ) is:

Now the concentration of glucose, S R , in a fresh feed and the feed

rate, Fi of the fresh medium are predetermined. In quasi-steady

FED BATCH CULTURE OF S. CEREVISIAE

state, where ASi/At; v 0 and assuming that SR

equation is derived from eq. (6).

F;SR =

Fix;

Yi

1005

>> S;, the following

1
AX.
+(Vo; + F i At;)
Yi
At;

(7)

If Y ; is constant throughout the fed batch culture,

FOSR(ti - to)

ti)

= 1, F ~ S R
(tz -

1
(xi -

= -

XO)

(9)

( x Z- x l )

( 10a)

1
7
(Xn+1

(&+I

tn)

2F i AtiSR

- (Xn+l

i = n, FnSR
i =O

Xn)

(lob)

- 20)

therefore
S R =

Xn+1

ZO -

2 F i At;

XF

( V F

XO

VO)

(11)

i =O

where X F is the final value of the total cell mass, xn+lin the fermenter
(g), V F is the final value of the broth volume (liter).
Equation (11) suggests that the value of S R in the fresh medium
can be estimated once the target of production is established, i.e.:
X F starting from zo in the total cell mass and the broth volume increases, VF - V oreach expected values in the fed batch culture, and
the value of Y is given, respectively. The feed rate F ; for the ith
time interval can also be assessed as shown below from eqs. (2) and
(6) when the quasi-steady state and the specific case in which
SR >> are dealt with.

MATERIALS AND METHODS

Organism and Medium Composition
The strain used in this work was bakers yeast (8.cereerisiae) from
Oriental Yeast Co., Ltd., Tokyo. The two kinds of media used,

1006

AIBA, NAGAI, A N D NISHIZAWA

semisynthetic and molasses, were composed of the following. Semisynthetic medium: glucose, 20 g; (NH2)&O, 2.15 g; NaHP04.2Hz0,
1.0 g; MgS04-7Hz0, 0.38 g; KC1, 0.22 g; sodium citrate, 2.5 g; yeast
extract, 0.5 g; 1 ml of vitamin solution (biotin, 0.04 g; vitamin B1,
0.08 g; vitamin B6, 2.0 g; calcium pantothenate, 1.0 g and inositol,
20 g/liter), 1 ml of mineral solution (CuS04.5H@, 0.05 g, ZnS04.7Hz0,
0.8 g; and Fe(S04)z(NH4)2-6Hz0,
0.3 g/liter), tap water, 1,000 ml;
and p H = 5.0 adjusted with an aqueous solution of HzS04 (2N).
Molasses medium: molasses was treated with steam for 1 hr a t about
80C and centrifuged for 1 min at 4,000 x g to be free from solid
ingredients and then diluted to about a 30% sugar content (as
glucose). Urea was supplemented to the medium with a ratio of
0.5 g (urea) t o 30 g (glucose).
Fed Batch Culture
A cell suspension obtained from the Oriental Yeast Co. was inoculated into a bench-scale fermenter (nominal volume = 10 liters,
initial working volume = 3.5 liters, L. E. Marubishi Co., Ltd.,
Tokyo, Model MD-500) in order to have 4 g/liter in the cell concentration just prior to the start of the fed batch culture. After 0.5 hr
in the batch culture (initial glucose concentration = 1 g/liter), the
feeding of the fresh medium into the fermenter was begun at a n
adequate rate using two peristaltic pumps (A and B, Taiyo Kagaku
Co., Tokyo), where the feed rate of Pump B was about 10 to 20% of
that of Pump A. Pump A, as a staple control, was operated continuously t o feed the fresh medium a t a feed rate which was changed
stepwise periodically to achieve a cell growth such that the RQ value
did not fluctuate too much from 1.0. Pump B, as a fine control,
was operated intermittently ; the time of operation depended on the
value of RQ, i.e., this latter pump was switched on when the RQ
value was below the datum of 1.0 and/or turned off when the RQ
value exceeded another datum of 1.10. Usually, about 60 sec was
needed to change the feed rate after confirming the varying RQ values.
Obviously, using two pumps made it convenient to substitute for
a single one which could vary the feed rate continuously in light of
information originating from the experimentation (see Figs. 2 and 3).
As was mentioned earlier, the improvised use of this pump (or pumps)
necessitated the change in the feed rate stepwise rather than continuously.
The rotation speed of an impeller and the temperature of the cultivation were kept a t 600 rpm and 30C, respectively. The air flow
rate was controlled with a specific miniflow valve set a t 2.45 and/or

FED BATCH CULTURE OF S. CEREVISIAE

1007

3.27 liters/min with respect to the cell growth; the value of p H was
also controlled a t 4.5 with an aqueous solution of NaOH (2N).

Analytical Methods
The optical densities measured a t 610 nm in wavelength with a
spectrophotometer (Hitachi Works, Model 101) were converted to
dry cell mass concentration after establishing a calibration chart.
The calibration was made by filtering the cells through a Millipore
filter (pore size = 1.2 pm) and drying the cake at 105C for 1 hr in a
semisynthetic culture medium of glucose. In the molasses medium,
the cell mass concentration was determined directly by filtering a
sample broth through the Millipore filter (pore size = 1.2 pm),
followed by drying a t 105C for 1 hr.
The concentration of glucose in the glucose medium was determined with the Glucostat reagent (Fujisawa Medical Supply Co.,
Ltd., Osaka), whereas the molasses was first hydrolyzed with a
concentration of HC1 in a boiling water bath for 40 min and then
analyzed by the Somogyi method to determine the concentration of
glucose.
The concentration of ethanol in the culture medium was determined by the microdiff usion method. The dissolved oxygen concentration was measured occasionally with a membrane electrode (L. E.
Marubishi Co., Ltd.) ; actually, the oxygen concentration was well
above the level which might have limited the cell growth.
The respiration rate, Qo2, the total oxygen consumption rate, I o 2 ,
the specific rate of carbon dioxide evolution, &con,and the respiratory
quotient, RQ, were estimated by calculating the difference of the
partial pressures of oxygen and carbon dioxide in air between input
and output through the fermenter, with a Beckman oxygen analyzer
(Type 777) and an infrared gas analyzer (Shimadzu Works, Tokyo,
Model URA-2), respectively. The response times of these analyzers
were quite short, approximately 30 sec in preliminary experiments.

RESULTS AND DISCUSSION

Fed Batch Culture of Glucose Medium
An example of fed batch culture of glucose medium is shown in
Figure 2. The total cell mass, 5 , the total oxygen consumption rate,
I o 2 , and the respiratory quotient, RQ, are on the left-hand side of
the ordinate, while concentrations of glucose, S, and ethanol, P in the
culture medium, the specific rate of respiration, Qo2,the specific rate

1008

AIBA, NAGAI, AND NISHIZAWA

0
0

Ihr)

Fig. 2. Growth patterns of bakers yeast in the fed batch culture of the
glucose medium. The differences in partial pressures of oxygen, Ape, and
carbon dioxide, A p c o , between input and output air were recorded continuously.
RQ values as the ratio of A ~ c o , / A ~ o ,and l o , (Ape, times air flow rate) are
then recorded continuously. Since the total cell mass, x, was determined intermittently, Qo, (Zo,/z) and QCO,(Qo, RQ) were both observed in discrete rather
than continuous terms. The flow rate of fresh feed, F , was observed directly
by using a measuring cylinder, but the data in the figure are described scheand Y (r/180 Y) values were assessed
matically. M ( l / z . A z / A t ) , Y (FiS~/zi),
from the curve drawn through the data points of x, respectively. ( 0 )z, (A)S ,
(0)
P , (0)
Qo,, ((3)Qco,.

of carbon dioxide evolution, Qco,, the specific rate of glucose consumption, v, the specific rate of increase in the total cell mass, p, the feed
rate of the fresh medium, F , and the growth yield, Y , are on the righthand side of the ordinate. The data observed are shown in the lower
diagram of the figure, whereas in the upper diagram the characteristic
of the fed batch culture is demonstrated by respective calculations
from the data given in the lower diagram.

FED BATCH CULTURE OF S. CEREVISIAE

1om

Prior t o the experimental run, the concentration of glucose, S R in

the feed was calculated from eq. (11) since S R = 24 g/liter, assuming
x0 = 14 g, Vo = 3.5 liters, X F = 26 g, V F = 4.5 liters, and Y = 0.5 g
cell/g glucose. The actual data of S R prepared thus far was S R =
23.2 g/liter.
Yeast cells inoculated into the fermenter were cultivated for 0.5 hr
in batch as stated earlier, followed by the fed batch culture at t = 0
(see Fig. 2). The initial feed rate, F o = 0.22 liters/hr at t = 0 was
derived from eq. (la), assuming SR = 23.2 g/liter, 5 0 = 12.8 g,
pa' = 0.20 hr-l, and Y = 0.5 g cell/g glucose. The feed rate which
should have been periodically changed was arbitrarily manipulated
stepwise such that the RQ values in situ would be within the preset
1.10) (see Materials and Methods). It is clear
boundaries (1.0
from the figure that RQ values, though oscillated, could be controlled
within the preset range by changing the feed rate stepwise; the
response of RQ to the feed-rate change was fairly rapid.
Thus far, as concerns this experimental set-up (analyzer per se plus
dead-space above the culture medium in the fermenter, etc.), the
time required for the analyzer to respond to the change in C02
emergence from the yeast was estimated to be of the order of 60 sec.
Since the time required for manipulating the pump(s) was also of the
same order of magnitude as referred to earlier, the change in RQ
values observed could be commensurable with that of the feed rate.
I n fact, when RQ > 1, the change in F entails fairly rapid emergence of QCO,(see Figs. 2 and 3). When RQ < 1, on the other hand,
the change in F was accompanied by a short delay (see Figs. 2 and 3),
implying that the yeast utilized, most probably ethanol, formed due
to the lack of glucose in the culture medium. The delay was hardly
relevant considering the instrument that was used.
The fact that concentrations of glucose, S , and ethanol, P , were
rather high during the early period of the fed batch culture might
have been attributed to the glucose initially added (about 1,000
mg/liter) in the batch culture and the ethanol that then emerged.
However, the value of S was kept low a t about 10 mg/liter throughout
the rest of the fed batch culture. Therefore, from this experimental
fact, the quasi-steady state assumed earlier in deriving relevant
equations appears to have been warranted.
Values for the specific rate of respiration, Qo, ( I ~ , / xin
) Figure 2
remained nearly unchanged a t about 7.8 mmol 0 2 /g cell hr throughout. Accordingly, the pattern of the t.otal oxygen consumption rate,
lo,most likely represented the rate of increase in the total cell mass
and the oscillation of the RQ values was caused principally be Qco,.

1010

AIBA. NAGAI, A N D NISHIZAWA

For the oscillation of Q C O ~it had also been confirmed from preliminary
experiments that the construction of the infrared analyzer used here
was not responislbe a t all; the change in feed rate followed by the
physiological activities of the yeast was considered to reflect the
oscillation.
Values for the specific growth rate, p', the specific glucose consumption rate, v', and the growth yield, Y , shown in the upper
diagram of the figure were assessed here as follows: pr (1/x . A z / A t )
is the curve through the data points of the total cell mass, Y' is
F;SR/x; (therefore, S R >> S) (cf. eq. (la)), and Y is p'/v' . 180 (on a
gram basis) derived from p' and v' assessed earlier.
Values of p' increased from 0.20 to 0.24 hr-' in Figure 2, while V'
values were between 2.0 and 2.4 mmol glucose/g cell hr, depending
on the feed rate. Accordingly, values of Y increased most likely
from 0.48 t o 0.55 g cell/g glucose, although they were a bit modulated.
These values of pr and Y observed in this fed batch culture are nearly
the same as the maximum values reported for aerobic growth
(RQ = 1.0) of the S . cerevisiae in glucose-limited chemostat cultures.'
Fed Batch Culture of Molasses Medium
The experimental conditions employed in this run were as follows:
xo = 15 g, V O= 3.5 liters, XF = 35 g, and V F = 5.0 liters. If Y is
taken as 0.5 g cell/g glucose, S R is assumed t o be 26.7 g/liter from
eq. (11). However, the actual data of SR prepared and xo were 28.4
g/liter and 14.1 g, respectively.
By and large, the results of this run in Figure 3 resembled that of
glucose in Figure 2 except for S , which will be elaborated below.
The higher concentration of residual sugar might have originated from
the nonfermentative sugar which accumulated as the fed batch
culture progressed. Values of p', v', and Y were assessed as shown in
the upper portion of Figure 3 exactly by the same procedure mentioned earlier in Figure 2. Incidentally, p' ranged from 0.20 to 0.22
hr-l, while the growth yield, Y fluctuated around 0.48 to 0.52 g cell/g
glucose.
Y,I, and RQ

It was confirmed from Figures 2 and 3 that the feed rate of the
fresh medium could be controlled solely by the observed value of RQ.
However, it is deemed more desirable when considering computer
control t o formulate the feed rate in close connection with the metabolic activity of baker's yeast represented by the value of RQ.

FED BATCH CULTURE OF S. CEREVISIAE

0
0

1011

(hr)

Fig. 3. Growth patterns of bakers yeast in the fed batch culture of the molasses
( 0 )Qo,, ((3)Qco,.
medium. ( 0 )2, (A)3, (O)P,

With respect to the aerobic fermentation, another yield, Yp,ais

defined as follows:
Qp

YPIS

= V

1012

AIBA, NAGAI, AN11 NISHIZAWA

provided that Q, is the specific rate of ethanol production (mol

ethanol/g cell hr), v is the specific rate of glucose consumption (mol
glucose/g cell hr), kl is the stoichiometric constant which correlates
C 0 2evolution with ethanol production in the fermentation of glucose
(1 mol ethanol/mol CO,), k2 is the stoichiometric constant which
correlates 0 2 consumption with C 0 2 evolution in complete oxidation
of glucose (1 mol C02/mol 0 2 ) , l o , is the total oxygen consumption
rate (Qo,a)(mol 02/hr).
Equation (13) can be translated to the ith interval of time as
follows :
k~ ( I 0 , ) i (RQi - 1)
( y p / s ) = ( d i X i / Y ;) ( 1/ 180)
Assuming again the quasi-steady state and S R >> Si, the following
equation which assesses the value of F i is derived from eqs. (12)and
(14).
(15)

Equation (15) clearly denotes that the information in Io2 and RQ

stemming from the analysis of the exhaust gas from the fermenter at
each time-interval defines the feed rate, if and only if the values of
Yplaand S R given earlier from eq. (11) are made available.
The relationship between Y p l aand RQ calculated from the experimental data in Icigurcs 2 and 3 is shown in Figure 4. The linear
correlation could be assumed to be in the range of RQ (RQ = 1.0
1.2) which was employed here in each fed batch culture. Then

= K ( R Q i - 1)

(16)
where K is the proportionality constant (empirical) (mol ethanol
mol 02/mol glucose mol C02).
The values of K given from the slope of the lines in Figure 4 were
3.SO for glucosc and 2.65 for molasses media. Substituting eq. (16)
into cq. (15).
( Y p / a )i

Equation (17) means that the feed rate in order to maintain the
quasi-steady state can be defined exclusively with I,,, values at th a t
time.
Procedures to Control the Feed Rate
Two ways t o control the feed rate in the f(.d batch culture were
already proposed here in this work. On(. is to change the rate

FED BATCH CULTURE O F S. CEREVISIAE

"O

1013

0.8

0.6

K = 3.80
(semi - synthetic 1

Fig. 4. The relationship between Y,,, and RQ in the fed batch culture of
baker's yeast in semisynthetic (glucose) and molasses media.. Y,,* was assessed
) eq. (13));
from the data of QCO,(Qo, RQ), Qo, (ZoJz), and Y' ( F i S ~ / z i (cf.
the data points do not correspond exactly to those of z in the original figures
(Fig. 2 or 3), because the interpolated data (see curves drawn through the data
of z) pertaining to RQ > 1 were ta.ken. ( 0 )Semisynthetic medium; (A)molasses medium.

referring to the values of RQ, the other is by affecting l o 2 using eq.

(17).
The feed rate, Fc,l, estimated from l o , (eq. (17)) and another
rate, F,,,, employed in the experimental runs which refer to instantaneous values of RQ are sporadically compared in Figure 5a (semisystematic medium) and b (molasses medium). The value of Fcal
should define the feed rate once the value of RQ, approximately between 1.0 and 1.15 or 1.20 (see Fig. 4) in these examples which
warrant the linearity between Ypl8and (RQ - 1)) is given. In other
words, control based solely on l o z is essentially provided with freedom
t o choose an appropriate value of RQ within the specific range.
Conversely, the control of the feed rate based on l o 2 should be
coupled with a sophisticated mechanism for controlling the feed
pump, keeping the value of RQ as closely as possible around a par-

AIBA, NAGAI, A N D NISHIZAWA

1014

0.6

(0)

0.4

semi - synthetic

4
LL

rn

Fig. 5. The feed rate, Fexpreproduced from Figs. 2 and 3 is compared with
the rate, Fa!,as assessed from eq. (17): (a) semisynthetic medium, (b) molasses
medium. In the assessment of Fa,, K values shown in Fig. 4 were used, respectively. For ease of discussion, ZZQ values are also reproduced from previous
figures.

ticular value employed; otherwise, the RQ values oscillate greatly

around RQ = 1.0 as illustrated in the figure. If the above situation
where a precise and instantaneous control of the pump had been
understood, the variations of RQ in the figure would have been
minimized.
In carrying out the experiments in this work, thc feed rate, which
was far above the value of F,,,,was acceptable so far as the requirement to minimize the glucose effect in terms of RQ from 1.0
1.15
was considered.
Briefly, i t goes without saying th at the value of Fcz,lserves as datum
to define the flow rate of the medium, leaving the exact value of RQ
undefined. I t is shown from Figure .5 that the upper and lower

F E D BATCH CULTURE OF S. CEREVISIAE

1015

diagrams dealt with F,,, values rather than with the Fcalvalues,
respectively, as a whole, even in the absence of any automatic control of the feed pump where the deviation between Fcsl and F,, in
both diagrams in Figure 5 seems to support the appropriate range
of RQ from the viewpoint of minimizing the glucose effect.
The possibility and advisability of controlling the fed batch
culture using either lorand/or RQ coupled with a computer is now
self-evident.

Nomenclature
feed rate of fresh medium (liters/hr)
total oxygen consumption rate (QOZz) (mol Ot/hr)
proportionality constant (empirical) (mol ethanol mol 02/mol glucose mol
COr)
stoichiometric constant to correlate COZ evolution with ethanol production in fermentation of glucose (1 mol ethanol/mol COZ)
stoichiometric constant t o correlate 0 2 consumption with COZevolution
in complete oxidation of glucose (1 mol COr/mol 0 2 )
specific rate of COZevolution (mol CO2/g cell hr)
specific rate of respiration (mol 02/g cell hr)
specific rate of ethanol production (mol ethanol/g cell hr)
respiratory quotient (mol COn/mol 0 2 )
concentration of glucose in culture medium (g/liter)
concentration of glucose in fresh feed (&liter)
time (hr)
broth volume (liter)
final value of broth volume (liter)
concentra.tion of cell mass in culture medium (g/liter)
total cell mass in fermenter (9)
final value of total cell mass in fermenter (g)
growth yield (g cell/g glucose)
yield coefficient of ethanol produced to glucose consumed (mol ethanol/
mol glucose)
Greek Letters
p
p
Y

specific growth rate based on cell mass concentration (hr-I)

specific growth rate based on total cell mass (hr-1)
specific rate of glucose consumption (mol glucose/g cell hr)

Subscript

i
0

i t h interval of time
initial of ith interval

The authors are indebted to Dr. M. Ohashi, Oriental Yeast Co., Ltd. for the
bakers yeast and molasses used and to Mr. T. Yamagata, L. E. Marubishi Co.,
Ltd. for some instruments used throughout this work. They are also grateful
to Mr. T. Karasawa, Oriental Yeast Co., Ltd. and Mr. H . Sakuma, L. E. Marubishi Co., Ltd. for their technical assistance.

1016

AIBA, NAGAI, AND NISHIZAWA

References
1. H. K. von Meyenburg, Arch. Mikrobiol., 66, 289 (1969).
2. F. J. Moss, P. A. D. Rickard, F. E. Bush, and P. Caiger, Biotechnol. Bioeng.,
13, 63 (1971).
3. W. I>. Maxon and M. J. Johnson, Znd. Eng. Chem., 45,2554 (1953).
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Accepted for Publication February 18, 1976