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J. Ind. Eng. Chem., Vol. 13, No.

7, (2007) 1054-1061
Determination of the Process Parameters Relative Influence on k
L
a
Value using Taguchi Design Methodology
Marko TRAMEK and Andreja GOREK

University of Maribor, Faculty of Chemistry and Chemical Engineering, SI-2000 Maribor, Slovenia
Received January 3, 2007; Accepted November 22, 2007
Abstract: This article describes experimental determination of the relative impact of significant process parame-
ters that influence volumetric oxygen mass transfer coefficient (k
L
a) using Taguchi design methodology. For this
purpose an automated RC1 reaction calorimeter (Mettler-Toledo), which was originally developed for chemical
processes, was modified for the bioprocesses. Simple fermentation using Bakers yeast was studied to illustrate
the design procedure. Orthogonal array L
25
was selected for the proposed design and ANOVA method was used
for recognizing the relative influence of the process parameters. Within the observed range of temperature (),
fermentation media volume (V
FM
), and yeast mass concentration (
Y
), these process parameters were found to be
unimportant compared to the volumetric air flow rate (q
V,a
) and rotational frequency of the stirrer (f
m
). The q
V,a
had a substantial effect on the k
L
a value (89.2 %) and the f
m
had just a small one (3.6 %), meanwhile the remain
fraction to 100 % represents error. The results refer strictly to the selected case study. Anyhow, the proposed pro-
cedure shows that application of the Taguchi approach for analyzing the oxygen mass transfer based on the ex-
perimental data obtained from a highly-automated laboratory reactor appears to have potential usage in general
biopharmaceutical process design.
Keywords: volumetric oxygen mass transfer coefficient, Taguchi method, process parameters, bioprocess, analy-
sis of variance

Introduction
1)
The pharmaceutical industry is the professional and
business activity containing the most important scientific
potential of the humans. The fast development of in-
dustrial biotechnology as a sustainable alternative to tra-
ditional chemical production has caused significant
changes in the pharmaceutical industry [1,2]. For in-
stance, highly molecular biopharmaceuticals (recombin-
ant therapeutic proteins, monoclonal antibody-based
products, nucleic acid-based medicinal products) repre-
sent as much as one-fourth of all newly developed phar-
maceuticals [3]. Due to increasingly demanding regu-
latory requirements in the global market, their produc-
tion needs to fully follow the principles of current Good
Manufacturing Practice (cGMP) [4]. GMP envisages the
use of optimally planned processes ensuring the highest

To whom all correspondence should be addressed.


(e-mail: andreja.gorsek@uni-mb.si)
possible product quality, safety and effectiveness.
The basic production process of biopharmaceuticals is
fermentation [4]. The oxygen mass transfer (OMT) at
the gas-liquid interphase is of decisive importance for
the rate of the aerobic fermentation course, consequent-
ly, the volumetric oxygen mass transfer coefficient, k
L
a,
is considered to be one of the most important factors
when planning bioreactors [5,6]. Despite numerous well
known theoretical equations for the assessment of its
value [7,8] in fermentation processes it is usually de-
termined experimentally [9]. It was established that
OMT was influenced by different process parameters
[10,11]. In order to provide the optimum performance of
the fermentation process, complying with cGMP [12],
should also be necessary to determine the relative influ-
ence of those different process parameters affecting the
k
L
a value.
Traditionally, the influence of parameters on process
performance has been determined experimentally th-
rough planning and implementing experiments on large
Determination of the Process Parameters Relative Influence on kLa Value using Taguchi Design Methodology 1055
Figure 1. Block scheme of adjusted RC1 (Bio-RC1).
industrial devices. With the technological development,
however, the process parameters analyzing has been
transferred to laboratory scale, which resulted in in-
creased effectiveness and reduced planning costs.
The technique for the determination and investigation
of the influential experiment parameters at different lev-
els is called the design of the experiment [13]. The se-
lection of relevant planning techniques depends on the
number of parameters influencing the product quality,
and the type of problem. However, conventional techni-
ques involve altering of one parameter at a time keeping
all other parameters constant. When we want to study
any given system with a set of independent variables
(process parameter) over a specific region of interest
(levels region) and intend to improve the process plan-
ning strategy and quality optimization of the process pa-
rameters at the same time, we use the so-called Taguchi
methodology [13]. The use of its algorithm is observed
in various optimization problems, starting with opti-
mization of diesel engine parameters [14], the leaching
of non-sulphide zinc ore in the ammonium-sulphate sol-
ution [15], to the production of laccase by Pleurotus os-
treatus 1804 [16], etc. However, considering available
literature, it can be stated that this method was not used
for experimental determination of relative impact of the
significant process parameters that influence the k
L
a
value. In contrast to the traditional design of experiment
(DOE), the standardized Taguchis methodology for two
independent problem solution plans usually brings the
same results, which enables determination of individual
process parameters relative influence on the final result.
This methodology envisages implementation of a mini-
mum number of experiments, which are defined by spe-
cific standard orthogonal arrays (OA). Selection of rele-
vant OA is conditioned by the number of parameters and
levels.
We exclude unnecessary additional experiments by im-
plementing experiments using the technically perfect
and computer supported laboratory equipment, thus en-
abling full repeatability. Among such type of equipment,
there is also a highly automated RC1 reaction calo-
rimeter (Mettler Toledo) intended for investigation of
temperature changes in basic chemical and physical
processes. By using the specific modifications in hard-
ware and software, first introduced by Marison and oth-
ers [17,18], it could also be used for investigations in bi-
oprocesses [19-21].
This research is aimed at studying the possibility of ex-
perimental determining the relative influence of various
process parameters on the k
L
a value in biotechnological
processes, by using the Taguchis process planning
methodology. Simple fermentation of glucose with the
Bakers yeast (Saccharomyces cerevisiae) in the ad-
justed RC1 reactor (Bio-RC1) was used as a case study.
We investigated the effects of stirrer rotational fre-
quency, f
m
, volumetric air flow rate, q
V,a
, fermentation
media volume, V
FM
, temperature, , and the Bakers
yeast mass concentration,
Y
.
Materials and Methods
Equipment
The RC1 reaction calorimeter is a computer-controlled
batch laboratory reactor (V = 2 L) for the performance of
isothermal and adiabatic reactions, basic physical oper-
ations, and the determination of thermal data and
constants. Its detailed description can be found in pre-
vious work [22].
For the purpose of bioprocess investigations, a hard-
ware modification to the original RC1 was introduced,
which was different from the one introduced by Marison
and others [17,18]. Thus, RC1 was modified to Bio-
RC1. Modifications (air compressor, oxygen electrode)
are represented by the two shaded blocks in Figure 1.
Bottom valve of the glass reactor was replaced by a sep-
arately constructed aeration module, which is connected
to the air compressor (Senco; model: PC1010) through
the volumetric air flowmeter. This modification enables
aeration of the fermentation medium.
A dissolved oxygen sensor (Mettler Toledo; lnPro6800/
12/320) was installed on the glass cover. Through the
transmitter (Mettler Toledo; model: O2 4100e) it was
connected to an RD10 controller. After this connection,
dynamic measurements of the dissolved oxygen concen-
tration were possible using the PC.
The optimal mixing regime at the aeration of the fer-
mentation mixture was achieved by replacement of the
existing stirrer (Mettler Toledo; model: anchor) with the
intermig one (Ekato; model: intermig).
Marko TRAMEK and Andreja GOREK 1056
Table 1. ANOVA Equations and Their Relationships
Eq. No. Expression Eq. No. Expression
(4)

(10)

(5)

(11)


(6)

(12)

(7)

(13)

(8)

(14)

(9)

Determination of Volumetric Oxygen Mass Transfer


Coeficient
The k
L
a in the fermentation system was determined us-
ing the dynamic method proposed by Mignone and
Ertola [23], which is based on a step change in the f
m

during cultivation. The method considers the liquid film
effect on the diffusion along the membrane of the oxy-
gen electrode, and the effect of oxygen electrode re-
sponse time resulting from electrode diffusion resi-
stance. Both effects can be neglected by the application
of a top-level dissolved oxygen sensor (Mettler Toledo)
with quick response, and by assuring effective fermenta-
tion media mixing.
During aerobic fermentation with constant air flow rate
and at a rotational frequency of the stirrer, f
m,1
, the first
steady state is reached:
OTR = /

a
1

*
-
O

(1)
where: OTR - oxygen transfer rate (g/(L s))

O

- dissolved oxygen mass concentration in the
first steady state (g/L)
k
L
a
1
- volumetric oxygen mass transfer coeffici-
ent at f
m, 1
(s
-1
)

*
- saturated oxygen mass concentration (g/L)
The step change of the stirrers rotational frequency to
f
m,2
,

causes approaching to the second steady state. When
defining the dimensionless concentration variable, D

(
O

-
O

.1
),(
O

.2
-
O

.1
) (2)
then transition from the first to the second steady state
can be described as follows [23]:

1-c
-(/

)t
(3)
where:
O

- dissolved oxygen mass concentration (g/L)

2
- dissolved oxygen mass concentration in
second steady state (g/L)
k
L
a
2
- volumetric oxygen mass transfer co-
efficient at f
m,2
(s
-1
)
t - time (s)
The oxygen-dissolving kinetics in liquid phase at a ro-
tational frequency of the stirrer, f
m,2
, is described by Eq.
(3). Therefore, the experimental measurements (
O

f(t)) during the approach to the steady state after the step
change of stirrers rotational frequency from f
m,1
to f
m,2
,
enable determination of k
L
a value at f
m,2
.
Taguchis Experiment Planning Methodology
Taguchi has defined the optimization criterion quality
(in our case, namely, k
L
a) as a consistency in achieving
the desired value through minimization of the deviation
[13]. This goal is connected with the performance of a
series of experiments with different process parameters
at different levels. The process parameter is a factor af-
fecting the optimization criterion quality, and its value is
called the level. The number of experiments and their
sequence are determined by standard OA. When plan-
ning the experiments using six process parameters at
five levels, we use the OA L
25
. Such a plan envisages the
performance of 25 experiments, which is significantly
less when compared to the traditional factorial method-
ology with 5
6
= 15625 experiments.
Due to performing only a part of the envisaged experi-
ments by using the traditional factorial methodology we
have to include an analysis of the results confidence.
The standard statistical technique is used for this pur-
pose, the so-called analysis of variance (ANOVA), which
Determination of the Process Parameters Relative Influence on kLa Value using Taguchi Design Methodology 1057
Table 2. Process Parameters and Their Levels
Process
Parameter
Level
1 2 3 4 5
A: fm/min
1
80 120 160 200 240
B: qV,a/(L/min) 1 2 3 4 5
C: Y/(g/L) 7.50 5.00 3.75 3.00 2.50
D: VFM/L 1.1 1.2 1.4 1.6 1.8
E: /C 24 26 28 30 32
recognizes the relative influence of the process parame-
ters for the optimization criterion value.
The mathematical algorithm of the ANOVA statistical
technique is based on calculation of the variance, which
is an indicator of the optimization criterion quality. The
ratio between the variance of the process parameter, and
the error variance shows, whether the parameter has af-
fected the products quality. The equations required for
calculating the relative influence of the process parame-
ters affecting the optimization criterion are presented in
Table 1. The meanings of symbols indicated in Table 1,
are described in the chapter Nomenclature.
We compare F
j
to the statistical value F
m,n
for certain
confidence rate, which is obtained from the standard F
Tables [13], whereby m stands for the f
j
and n means the
f
e
, and thus determine the process parameter influence
accordingly. In the case where the F
j
falls below F
m,n
,
the process parameter has no effect on the optimization
criterion, therefore, it is pooled and ignored in the
calculations. Consequently, the V
e
changes, as the S
j
and
f
j
of the pooled process parameter are added to the S
e
and
f
e
respectively. By using the adjusted V
e
, we determine
new F
j
and compare them again by the F
m,n
. The process
of pooling is sequential, which means that the parameter
having the smallest effect on the optimization criterion
should be pooled first, then we recalculate the F
j
and
continue pooling until each process parameter meets the
condition F
j
> F
m,n
. When the pooling procedure is com-
pleted, the X
j
and X
e
can be calculated using Eqs. (13)
and (14).
Chemicals
Simple fermented medium water-glucose was used for
the research purposes with a mass concentration of
3D-(+) Glucose anhydrous (Fluka),
G
= 5 g/L, purchas-
ed from Sigma-Aldrich Co (St. Lous, MO). Ferment-
ation was performed with the Bakers yeast, in the fresh
compressed form, obtained from the local grocery store.
It was composed by natural yeast (Saccharomyces cer-
evisiae) and water. The dry matter was more then 30 %.
As the shelf life of the fresh compressed yeast is short (t
= 45 d), the proper storage conditions, clean and dry
place between = (0-4)
o
C, were taken into con-
sideration.
Experimental Work
Determining the relative influence of various process
parameters on the k
L
a in bioprocesses based on the
Taguchis approach, requires the performance of a series
of experiments. A Bio-RC1 reactor was involved in the
research for this purpose. As a bioprocess test example,
we used a simple fermentation of glucose into ethanol
by the Bakers yeast. It is well known that the opti-
mization criterion (k
L
a) would be the most significantly
affected by f
m
, q
V,a
,
Y
, V
FM
, and . We examined the
relative influence of the selected process parameters at
five different levels, as shown in Table 2. Usually the se-
lection of levels and their ranges depends on how opti-
mization criterion is affected due to different levels
settings. However, we defined levels ranges according to
optimal process conditions (
Y
and ) and equipment
range-capacity (f
m
, q
V,a
, V
FM
). The boundary levels (1 and
5) were selected first followed by selection of others (2,
3, and 4).
During the first stage of the experimental work, it is
necessary to prepare the experiment performance plan.
The plan envisages determining the number of experi-
ments, their performance conditions, and their sequence.
Based on the assumption that the k
L
a would be affected
by five process parameters being considered at five lev-
els, we chose the L
25
array as the most adequate OA re-
quiring the performance of 25 experiments [13]. The OA
L
25
is usually intended for the investigation of six proc-
ess parameters at five levels; however, it may also be
used in our case (five parameters at five levels) by ignor-
ing the process parameter F. The experiment perform-
ance plan is presented in Table 3.
The first column of Table 3 presents the experimental
serial number. Each experiment was defined by the proc-
ess parameters (A, B, C, D, E, and F) marked at specific
levels by numbers from 1 to 5.
During the second stage of the experimental work, we
implemented the proposed plan by performing the ex-
periments in the Bio-RC1 reactor. The performance pro-
cedure was the same for all experiments. Individual ex-
periments were implemented by means of first charging
the reactor by the fermentation medium quantity de-
termined by the plan and heating it up to the working
temperature under the experimental conditions (q
V,a
and
f
m
). After establishing the steady state (temperature and
saturation by oxygen), we charged the reactor by the
Bakers yeast mass defined by the plan and reduced the
rotational frequency of the stirrer to f
m
= 50 min
-1
. The
addition of the yeast caused a launch of the aerobic fer-
mentation and consumption of oxygen. The concen-
tration of the dissolved oxygen decreased until the first
steady state was reached; then we increased f
m
back to
the original value. Consequently, there was a second
Marko TRAMEK and Andreja GOREK 1058
Table 3. Experiment Performance Plan OA L25
Experiment
Process parameter
b
A B C D E F
a
1 4 1 4 2 5 3
2 3 4 1 3 5 2
3 3 1 3 5 2 4
4 1 4 4 4 4 4
5 4 2 5 3 1 4
6 1 3 3 3 3 3
7 3 5 2 4 1 3
8 5 2 1 5 4 3
9 2 5 1 2 3 4
10 5 3 2 1 5 4
11 1 2 2 2 2 2
12 5 1 5 4 3 2
13 1 1 1 1 1 1
14 4 3 1 1 2 5
15 2 4 5 5 2 3
16 2 3 4 4 1 2
17 5 4 3 3 1 5
18 3 3 5 5 4 1
19 2 2 3 3 5 1
20 3 2 4 4 3 5
21 4 4 2 2 3 1
22 2 1 2 2 4 5
23 5 5 4 4 2 1
24 4 5 3 3 4 2
25 1 5 5 5 5 5
a
In our case process parameter F is not considered.
b
Process parameters are indicated in Table 2.
Table 4. Experimental Values of kLa OA L25
Experiment 1 2 3 4 5 6 7 8 9
kLa s
-1
0.0134 0.0260 0.0134 0.0237 0.0133 0.0221 0.0277 0.0139 0.0250
Experiment 10 11 12 13 14 15 16 17 18
kLa s
-1
0.0192 0.0149 0.0137 0.0131 0.0172 0.0237 0.0196 0.0221 0.0249
Experiment 19 20 21 22 23 24 25
kLa s
-1
0.0152 0.0144 0.0197 0.0121 0.0261 0.0254 0.0253
steady state reached. The o
2
was monitored. Based on
the experimental data during the transition from the first
to the second steady state, we used the Eq. (3) to calcu-
late the k
L
a value at the conditions determined by the ex-
periment performance plan.
Results and Disscussion
The k
L
a values experimentally determined under differ-
ent conditions proposed by the experimental perform-
ance plan (Table 3), are presented in Table 4. All k
L
a
values are of a similar size rank, as was reported by
Badino and coworkers [24]. Thus, we have basically
confirmed the relevance of the adjusted RC1 (Bio-RC1)
reactor for further investigations the OMT from the gas
to the liquid phase in bioprocesses. Table 4 shows the
lowest value of k
L
a = 0.0121 s
-1
, and the highest one, k
L
a
= 0.0277 s
-1
. Both values were achieved at the lowest
and highest q
V,a
, respectively, which indicates that the
q
V,a
might be the most important parameter.
Moreover, the average effects of the process parame-
ters along with interactions at the assigned levels on the
k
L
a value are shown on Figure 2. The difference between
levels of each process parameters indicates their relative
influence [16]. The larger the difference, the stronger is
the influence. It can be observed from Figure 2 that
among process parameters studied, q
V,a
showed the
stronger influence. However, we defined the actual rela-
tive influence of the process parameters effects by the
ANOVA statistical method.
The results of the variance analysis by the ANOVA
statistical method are shown in Table 5. The f
j
and f
e

equaled (f
j
= f
e
= 4) in all cases. At 95 % confidence
(level of importance 0.05), the value F
4,4
= 6.3883 was
determined through standardized Tables of F-statistics.
Determination of the Process Parameters Relative Influence on kLa Value using Taguchi Design Methodology 1059
Figure 2. Individual process parameters performance at different levels on kLa value.
Table 5. Analysis of Variance OA L25
Process
Parameter
Sj10
5
fj Vj10
5
Fj
A: fm 3.249 4 0.812 2.495
B: qV, a 61.026 4 15.257 46.868
C: Y 0.601 4 0.150 0.461
D: VFM 0.906 4 0.226 0.695
E: 0.453 4 0.113 0.348
Error 1.302 4 0.326 1.000
Total 67.537 24
Table 5 shows that the F
j
of the f
m
,
Y
, V
FM
, and the
fell below F
4,4
. In accordance with the Taguchis method
algorithm, we pooled from further statistical consid-
eration as the least important process parameter, i.e.,
with the lowest F
j
value compared to F
4,4
.
Pooling of the as an insignificant process parameter
requires a repeated variance analysis, whereby the S
j
and
the f
j
of the pooled process parameter is added to the S
e

and the f
e
, respectively. The results in Table 6 show that,
consequently, the F
j
of the remaining process parameters
increase. In spite of this, a repeated comparison of F
j
in-
dicated in Table 6 with the F-statistics value, F
4,8
=
3.8378, shows that only q
V,a
meets the F
j
> F
4,8
condition.
In a similar way, we further sequentially pooled the
Y

and the V
FM
from the statistical consideration. The final
result of the variance analysis is shown in Table 7. By
sequential pooling of insignificant process parameters,
the f
e
increased to the value 16. The F
j
of both remaining
Table 6. Analysis of Variance with Pooled Temperature OA L25
Process
Parameter
Sj10
5
fj Vj10
5
Fj
A: fm 3.249 4 0.812 3.702
B: qV,a 61.026 4 15.257 69.532
C: Y 0.601 4 0.150 0.684
D: VFM 0.906 4 0.226 1.032
E: Pooled
Error 1.755 8 0.219 1.000
Total 67.537 24
process parameters (f
m
and q
V,a
) met the conditionF
j
>
F
4,16
= 3.2389, therefore, we can use Eq. (13) to calcu-
late the X
j
.
Table 7 shows that the k
L
a value in a simple fermenta-
tion of glucose with the Bakers yeast was the most sig-
nificantly affected by the q
V,a
(X = 89.2 %). A sub-
stantially smaller share of the effect may be attributed to
the f
m
(X = 3.6 %) while the share of the error effect was
accounted for X
e
= 7.2 %.
Results obtained by this study are not directly com-
parable with some previously known results from the lit-
erature [25,26], where f
m
was established as the process
parameter with the highest effect on k
L
a. Many other
equipment setup factors exist (i.e., impeller and aeration
configuration, distribution and size of air bubble, mixing
delay time, bioreactor characteristics), which are highly
specific for each aeration and agitation system, and also
have an influence on k
L
a value. However, these factors
Marko TRAMEK and Andreja GOREK 1060
Table 7. Final Results of Variance Analysis OA L25
Process Parameter Sj10
5
fj Vj10
5
Fj Xj
A: fm 3.249 4 0.812 3.980 3.6
B: qV, a 61.026 4 15.257 74.789 89.2
C: Y Pooled
D: VFM Pooled
E: Pooled
Error 3.262 16 0.204 1.000 7.2
Total 67.537 24 100.0
were not considered in our case.
Nevertheless, a relatively small impact of the f
m
regard-
ing to q
V,a
may be explained by the replacement of the
existing anchor stirrer by the intermig one providing
equally-intensive distribution of small air bubbles within
the entire reactor volume at a low rotational frequency of
the stirrer (f
m
= 80 min
-1
). The further increase in the f
m

within levels range therefore, has not caused consid-
erable increase in the active gasliquid surface; con-
sequently, its share of effect on the k
L
a value was small.
Furthermore, it is wellknown that the impact of com-
pared to the q
V,a
on the k
L
a is small [27]. In our study,
within the range of (24-32)
o
C it has also been found that
the effect of the compared to the f
m
and the q
V,a
was
irrelevant. A similar statement can be applied for other
insignificant process parameters.
Conclusion
The k
L
a value, as one of the most important factor im-
plied on the design, control and optimization of bio-
process, is affected by many process parameters. Their
influence on k
L
a value is usually determined by conven-
tional one-parameter-at-a-time techniques, which do
not provide any insight into the behavior of the system.
Moreover, these procedures are time consuming cumber-
some and requires more experimental data. The Taguchi
method of experimental design is an immediate replace-
ment of the conventional techniques. After all, with a
standard OA as a main part of this method the number of
experiments is significantly reduced when compared to
the traditional factorial experimental design methodo-
logy.
Using experimental measurements in the adjusted RC1
(Bio-RC1), we examined the possibility of determining
the relative influence of various process parameters on
the k
L
a in bioprocesses by the Taguchis design me-
thodology. A simple case of glucose fermentation with
the Bakers yeast was used as a case study. The effect of
the f
m
, q
V,a
, V
FM
, , and the
Y
at five different levels
were considered.
The experimentally determined k
L
a values basically
confirm the relevance of the RC1 reactor modification
into the Bio-RC1 reactor, and the relevance of its further
use in the examinations of OMT from the gas to the liq-
uid phase in bioprocesses. By analysis of the ex-
perimental data variance (ANOVA), we established that
the relative impacts of the, , V
FM
, and the
Y
on the
k
L
a, compared to the q
V,a
and the f
m
, were negligibly
small or insignificant. The q
V,a
share of influence on the
k
L
a was 89.2 %, the f
m
contributed by 3.6 %.
However, the results are specific for Bio-RC1 and for
described fermentation system. Therefore, they cannot
be presented as a general for all bioprocesses and for all
equipment configurations. Nevertheless, our study con-
firms that the Taguchis planning methodology is appro-
priate for the quantitative identification of various proc-
ess parameters impacts on the k
L
a in bioprocesses using
different equipment setup. We have successfully linked
the state-of-the-art laboratory equipment, the Taguchis
planning methodology and the bioprocess, thus con-
tributing to an economically and dynamically more ef-
fective planning of biopharmaceutical applications.
Nomenclature
cGMP current Good Manufacturing Practice
DOE Design of Experiments
D

dimensionless concentration variable (1)


f
e
degree of freedom of error variance (1)
F
j
variance ratio of process parameter j (1)
f
j
degree of freedom of process parameter j (1)
F
m,n
standardized value from the F Tables at defined
level of significance (1)
f
T
total degree of freedom of the result (1)
f
m
rotational frequency of the stirrer (min
-1
)
k
L
a volumetric oxygen mass transfer coefficient (s
-1
)
L number of levels (1)
M number of process parameters (1)
N total number of experiments (1)
N
k
number of experiments on k level (1)
OA orthogonal array
Determination of the Process Parameters Relative Influence on kLa Value using Taguchi Design Methodology 1061
OMT Oxygen Mass Transfer
OTR Oxygen Transfer Rate (g/(L s))
q
V,a
volumetric air flow rate (L/min)
S
e
error sum of squares ( / )
S
j
sum of squares of process parameter j ( / )
S
T
total sum of squares ( / )
t time (s)
V
e
variance error (1)
V
j
mean square (variance) of process parameter j ( / )
V
FM
fermentation media volume (L)
X
e
relative influence of error on optimization crite-
rion (%)
X
j
relative influence of process parameter j on opti-
mization criterion (%)
Y
i
i value of optimization criterion ( / )

G
glucose mass concentration (g/L)

Y
Bakers yeast mass concentration (g/L)

dissolved oxygen mass concentration (g/L)

*
saturated oxygen mass concentration (g/L)
temperature (
o
C)
References
1. M. Gavrilescu and Y. Chisti, Biotechnol. Adv., 23,
47 (2005).
2. A. Rajapakse, N. J. Titchener-Hooker, and S. S.
Farid, Comput. Chem. Eng., 29, 1357 (2005).
3. G. Walsh, 2005, Trends Biotechnol., 23, 553 (2005).
4. M. Narhi and K. Nordstrom, Euro. J. Pharm.
Biopharm., 59, 397 (2005).
5. S. Hiraoka, Y. Kato, Y. Tada, S. Kai, N. Inoue, and
Y. Ukai, J. Chem. Eng. Jpn., 34, 600 (2001).
6. J. A. Williams, Chem. Eng. Prog., 98, 31 (2002).
7. F. Garcia-Ochoa and E. Gomez, Chem. Eng. Sci.,
59, 2489 (2004).
8. K. Vant Riet and J. Tramper, Basic bioreactor de-
sign, p. 236, Marcel Dekker, New York (1991).
9. M. Tobajas and E. Garcia-Calvo, Heat Mass
Transfer, 36, 201 (2000).
10. M. Gavrilescu, R. V. Roman, and V. Efimov, Acta
Biotechnol., 13, 59 (1993).
11. R. Lemoine and B. I. Morsi, Int. J. Chem. R. Eng.,
3, 35 (2005).
12. M. Thiry and D. Cingolani, Trends Biotechnol, 20,
103 (2002).
13. K. R. Ranjit, A primer on the Taguchi method, Van
Nostrand Reinhold, New York (1990).
14. M. Nataraj, V. P. Arunachalam, and N. Dhandapani,
Indian. J. Eng. Mater S., 12, 505 (2005).
15. J. Moghaddam, R. Sarraf-Mamoory, Y. Yamini, and
M. Abdollahy, Ind. Eng. Chem. Res., 44, 8952
(2005).
16. K. K. Prasad, S. V. Mohan, R. S. Rao, B. R. Pati,
and P. N. Sarma, Biochem. Eng. J., 24, 17 (2005).
17. I. Marison, M. Linder, and B. Schenker, Ther-
mochim. Acta, 310, 43 (1998).
18. I. Marison, J. S. Liu, S. Ampuero, U. Von Stockar,
and B. Schenker, Thermochim. Acta, 309, 157
(1998).
19. M. Janssen, R. Patino, and U. Von Stockar, Ther-
mochim. Acta, 435, 18 (2005).
20. F. Aulenta, C. Bassani, J. Ligthart, M. Majone, and
A. Tilche, Water Res., 36, 1297 (2002).
21. P. Vellanki, J. Guhan, I. W. Marison, J. S. Liu, and
K. Jayaraman, Thermochim. Acta, 309, 105 (1998).
22. M. Hvalec, A. Gorsek, and P. Glavic, Acta. Chim.
Slov., 51, 245 (2004).
23. C. F. Mignone and R. J. Ertola, J. Chem. Technol.
Biotechnol. B., 34, 121 (1984).
24. A. C. Badino, P. I. F. Almeida, and A. J. G. Cruz,
Chem. Eng. Educ., 38, 100 (2004).
25. M. S. Puthli, V. K. Rathod, and A. B. Pandit,
Biochem. Eng. J., 23, 25 (2005).
26. C. Bandaiphet and P. Prasertsan, Carbohydrate
Polymers, 66, 216 (2006).
27. T. Kaskiala, Miner. Eng., 15, 853 (2002).