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Journal of Food Engineering 87 (2008) 136–147

www.elsevier.com/locate/jfoodeng

Modelling gas–liquid and liquid–gas transfers in vinegar

production by genetic algorithms
José-Marı́a González-Sáiz a,*, Consuelo Pizarro b, Diego Garrido-Vidal a
a
Chemical Engineering, Department of Chemistry, University of La Rioja, C/Madre de Dios 51, 26006 Logroño, La Rioja, Spain
b
Analytical Chemistry, Department of Chemistry, University of La Rioja, C/Madre de Dios 51, 26006 Logroño, La Rioja, Spain

Received 30 August 2007; received in revised form 10 November 2007; accepted 20 November 2007
Available online 28 November 2007

Abstract

The transfer of oxygen from aeration flow to the fermentation medium and the transfers of ethanol, ethyl acetate, acetoin, acetic acid
and water from the fermentation medium to aeration flow were modelled for the industrial production of vinegar. The study was devel-
oped in a pilot fermentor, working with continuous processes and industrial fermentation mediums. The cores of the proposed models
were the semi-empiric correlations for the volumetric transfer coefficients kLa for oxygen transfer and kA,Ga for the liquid–gas transfers.
The main novelty was the introduction of the effect of pressure on the fermentation medium, together with the expression proposed for
kA,Ga, with the same structure as the expression for kLa. A genetic algorithm was developed to calculate the parameters of the models,
with a four composed desirability function as the evaluation function. The optimised sets of parameters provided models with good pre-
dictive ability. The models can be applied for any fermentor with the same agitation and aeration system design and the same geometry.
Ó 2007 Elsevier Ltd. All rights reserved.

Keywords: Fermentation; Genetic algorithms; Mass transfer; Modelling; Oxygen transfer; Vinegar production

1. Introduction – Models based on theoretical predictions: These equations

The efficiency of the industrial production of vinegar
depends mainly on the efficiency of the contact between
the gas phase and the liquid fermentation medium, since vin-
egar is obtained by the aerobic fermentation of an alcoholic
medium in an aerated fermentor. If bulk liquid composition
is uniform and the bubbles are uniformly dispersed through-
out the fermentation vessel, the mass transfer rates through
the gas–liquid interphase are explained by Whitman’s two-
film theory, as shown in Fig. 1.
Oxygen is essential for the survival of Acetobacter aceti
and the study of the oxygen transfer is critical for model-
ling this industrial process. The volumetric oxygen transfer
coefficient in the liquid-side interphase, kLa, has been
modelled using three main approaches:
are based on theoretical or ab initio models for kL and a.
Recently examples of this approach include studies by
Dhanasekharan et al. (2005) and Gómez et al. (2006).
– Semi-empiric correlations based on dimensional analyses.
Dimensionless parameters like the Sherwood number,
Sh, or the modified Sherwood number, Sh0 , have been
used to model kL and kLa, respectively. The specific
interfacial area, a, can be obtained from Calderbank’s
expression (Aiba et al., 1973). Table 1 summarises some
published works based on this approach.
– Semi-empiric correlations based on process variables.
Cooper et al. (1944) were pioneers in the measurement
of the volumetric oxygen constant coefficient in aerated
vessel using the sulphite-oxidation method. They pro-
posed correlations with the following form, which has
been widely applied (Aiba et al., 1973):

  z2
Pg
*
Corresponding author. Tel.: +34 941299634; fax: +34 941299621. k L a ¼ z1   lzS3 ð1Þ
E-mail address: josemaria.gonzalez@unirioja.es (J.-M. González-Sáiz). VL

0260-8774/$ - see front matter Ó 2007 Elsevier Ltd. All rights reserved.
doi:10.1016/j.jfoodeng.2007.11.020
 J.-M. González-Sáiz et al. / Journal of Food Engineering 87 (2008) 136–147 137

Nomenclature

a gas–liquid interfacial area per liquid + gas vol- OTR oxygen transfer rate, g L1 h1
ume, dm1 P pressure in fermentation medium, atm
ak constant for aerated mechanical power input in Pa atmospheric pressure, atm
correlations for kLa and kA,Ga PA,G partial pressure of component A in gas phase, atm
B number of blades on the impeller P A;G partial pressure of component A in equilibrium
bk constant for superficial air velocity in correla- with C A;L , atm
tions for kL a and kA,Ga PA,i partial pressure of component A in gas-side
CA,i concentration of component A in liquid-side interphase, atm
interphase, g L1 Pg aerated mechanical power input, w
CA,L concentration of component A in liquid phase P H2 O;T water vapour pressure at temperature T, atm
in equilibrium with PA,G, g L1 Po non-aerated mechanical power input, w

C A;L concentration of component A in liquid phase P O2 ;G partial pressure of oxygen in oxygen saturated
before mass transfer, g L1 gas phase, atm
Co global constant in correlations for kLa and kA,Ga P O2 ;G partial pressure of oxygen in gas phase after
C O2 ;i concentration of oxygen in the liquid-side inter- mass transfer, atm
phase, g L1 P O2 ;i partial pressure of oxygen in gas-side inter-
C O2 ;L concentration of oxygen in liquid phase, g L1 phase, atm
C O2 ;L concentration of oxygen in the oxygen-saturated QO volumetric gas flow rate at the exhaust line of
liquid phase in equilibrium with P O2 ;G , g L1 fermentor, m3 h1, dm3 h1
D impeller diameter (m) QS volumetric gas flow rate through liquid med-
DV fermentation vessel diameter, m ium, m3 s1, dm3 h1
DO2 molecular diffusivity of oxygen in liquid med- R number of baffles
ium, m2 s1 R2A adjusted correlation rate2
g gravitational constant, 9.8 m s2 Re Reynolds number, qLgND
L
J width of baffles, m RG gas constant, 0.082 atm dm3 mol1 K1, or
kA,G local liquid phase transfer coefficient for com- 1.9872 cal mol1 K1
ponent A, dm h1 RMS root mean square of the predicted values versus
kA,Ga volumetric component A transfer coefficient in experimental values
gas-side interphase, h1 SC Schmidt number, q gDLO
L 2
kA,L local liquid-phase transfer coefficient for com-
Sh Sherwood number, DkLOD
ponent A, dm h1, or m s1 2 2
Sh0 modified Sherwood number, kDL aD
KA,L overall transfer coefficient for component A, O2

dm h1, or m s1 TL liquid phase temperature, °C or K

kG local gas-phase transfer coefficient for oxygen, Tout average temperature of the air after passing
dm h1 through the fermentation medium, K
kL local liquid-phase transfer coefficient for oxy- VL liquid phase volume, dm3, m3
gen, dm h1, or m s1 vvm air flow rate divided by the reactor working
KL overall transfer coefficient for oxygen, dm h1, volume, h1
or m s1 W width of the impeller
2 3
blades, m
kLa volumetric oxygen transfer coefficient in liquid- We Weber number, qL Nr D
side interphase, h1 zx empiric constant number x
L length of the blades, m
LGTRA liquid–gas transfer rate for component A, Greek letters
g L1 h1 gG gas medium viscosity, Pa s
M O2 oxygen molecular mass, 32 g mol1 gL liquid medium viscosity, Pa s
N impeller rotation speed, rpm or s1 lS superficial air velocity through the liquid med-
Na aeration number, NlD ium, m s1
S
qL liquid medium density, kg m3
Ngf gas flow number, gqLl
L S hP pressure constant in correlations for kLa and
Np power number
kA,Ga
O2(o) oxygen effluent stream, g L1 h1
hT temperature constant in correlations for kLa
O2(i) oxygen feed stream, g L1 h1
and kA,Ga
OUR oxygen uptake rate, g L1 h1
r surface tension, g s2
138 J.-M. González-Sáiz et al. / Journal of Food Engineering 87 (2008) 136–147

GAS PHASE LIQUID PHASE is an invaluable tool for scale up and fermentor design
GAS SIDE LIQUID SIDE (Micheletti et al., 2006; Wong et al., 2003). Oxygen transfer
PA*,G INTERPHASE INTERPHASE C A* ,L models have been also applied to improve oxygen transfer
PA,i C A,i systems (Fregapane et al., 1994; Ory et al., 2004) in vinegar
PA,G C A, L industry.
Liquid–gas transfers in acetic fermentation have been
PO*2 ,G less studied, although some studies have been published
CO* 2 ,L
on ethanol loss in ethanol fermentation (Caro et al., 1992)
PO2 ,i CO2 ,i and acetic fermentation (Romero and Cantero, 1998a,b).
These models are based on the estimation of the compound
PO2 ,G CO2 ,L activity coefficients which are applied to calculate the par-
tial pressure of each component in the gas phase. Romero
and Cantero (1998b) applied the UNIFAC (UNIquac
Fig. 1. Mass transfers through the gas–liquid interphase in acetic
fermentation. Functional Group Activity Coefficients) method to calcu-
late the activity coefficients. The control of losses due to
liquid–gas transfers is very important for the economical
Table 1
Semi-empirical correlations for kLa based on dimensional analyses
yield of fermentation. In fact, one line of research into acetic
fermentation consists in the design of fermentation systems
Correlation Author
which recover volatile compounds (Ory et al., 2004).
z1 z2
Sh / Re Sc Rushton (1951) The aim of this study was the development of a new
Sh ¼ 0:13Sc1=3 Re3=4 Calderbank (1967)
  z4  z5  model for gas–liquid and liquid–gas transfers in industrial
gG
Sh0 ¼ z1 Rez2 Scz3 N1gf Sideman et al. (1966)
g L acetic fermentation. The variables of the models were the
0 z2 z3 z4
Sh ¼ z1 ðRe Na We Þ Garcı́a-Ochoa and Gómez (1998) process variables of acetic fermentation. The effects of
these process variables on the acetification process were
Table 2 shows examples of semi-empirical expressions studied in a previous paper published by the authors, Garr-
based on operational conditions, which could be applied ido-Vidal et al. (2003). A genetic algorithm was developed
to acetic fermentation. These types of correlations are the to optimise the parameters of the models. Genetic algo-
most useful since process variables such as agitation or rithms have been applied in parameter estimation and opti-
temperature are commonly monitored in fermentation pro- misation of bioprocesses in food industry, as reported by
cesses and can be scaled to different fermentation sizes. kLa González-Sáiz et al. (2003) or Goñi et al. (2008).

Table 2 2. Materials and methods

Semi-empirical correlations for kLa based on process variables
Correlation Author 2.1. Bioreactor design
 0:95
P
k L a ¼ 0:0635 V gL l0:67 vaned disk impeller Cooper
S
et al. The studies were performed in an Aerated Stirred Reac-
 0:53 (1944) tor (ASR) New Brunswick-BioFlo IV fermentor with a
P
k L a ¼ 0:038 V Lg l0:67
S paddle impeller maximum working volume of 10 L. The fermentor was
 0:4 equipped with a sparger, which diffused the air homoge-
P
k L a / V Lg lS0:5 N 0:5 Richards
(1961) nously into the vessel, and a mechanical agitation system
 0:4
k L að20  CÞ ¼ 0:026 V gL
P
l0:5
S coalescing systems Van’t Riet with two Rushton impellers. The sparger was a ring type,
 0:7 (1983) with section 6.6 mm and forming a 6-cm diameter ring with
P
k L að20  CÞ ¼ 0:002 V gL l0:2
S non coalescing systems 7, 2-mm homogeneously distributed holes. The sparger was
k L a ¼ hT L
ðT 20Þ
 kL a Oosterhuis located 2 cm from the bottom of the vessel and 1.45 cm
and from the first impeller. If the sparger were closer to the
Kossen impeller, it could cause bubble coalescence and flooding.
(1983)
If it were far away from the impeller, it could impair the
kL a
k L að20  CÞ ¼ 1:0 þ 0:02103ðT L  20Þ þ 8:261  105 ðT L  20Þ2 Gulliver
et al. break-up of the bubbles. Agitation was controlled by
(1990) PID, with a sensitivity of ±1 rpm, and temperature was
k L a ¼ 8:27  104 N 2:01 l0:259
S Moucha maintained by a jacket of water and controlled by PID,
et al. with a sensitivity of ±0.1 °C. Air flow measures were pro-
 0:35 (1995)
Pg vided by a mass flow sensor and pressure in the head space
k L a ¼ 3:2  103 VL ðlS Þ0:41 Montes
et al. of the fermentor was controlled by a valve installed in the
(1999) air exhaust line and measured with a barometer. Online
k L a ¼ ½0:0461 lnðQS Þ þ 0:1165N Aguiar data of air flow, rpm, temperature, pH and dissolved oxy-
et al. gen were obtained by the probes and stored in the database
(2002)
of the control software provided by New Brunswick. An
 J.-M. González-Sáiz et al. / Journal of Food Engineering 87 (2008) 136–147 139

Ingold sterilizable dissolved oxygen electrode was installed D

in the vessel. The electrode was fitted with an NTC therm- DLi
istor, which compensated the effect of temperature within
the range 0–70 °C. Pressure did not affect the signal under
3 atm and the hydrodynamics of the medium affected the
signal within the range 3–5%, which was assumed within
experimental error.
Fig. 2 shows the bioreactor design and Table 3 presents
the fermentor dimensions. The working volume was estab-
lished within the range proposed by New Brunswick, i.e.
[2–7] L. This design prevented flooding effects, configura-
tions of impellers that were too close or too separated
and uncontrollable hold-up effects. Medium height vs ves-
sel diameter was 1.3733. When this rate is within range W
L
[1–1.8], two turbines are necessary (Perry et al., 2001).
Therefore, the design included two flat blade Rushton tur-
bines, with six blades fixed onto the disk. Fig. 3 and Table 4 Fig. 3. Scheme of the Rushton impeller.
presents the dimensions of both Rushton impellers.
Table 4
Dimensions of the Rushton impeller
Parameter Dimension
D = agitator diameter 0.08135 m
DLi = internal impeller diameter 0.06135 m
W = width of the impeller blades 0.017 m
DV L = length of the blades (m) 0.021 m
B = number of blades on impeller 6

J
Air inlet
HT 2.2. Experimental methodology

Hm The experimental kLa values were obtained by continu-

ous fermentation processes developed in the pilot fermen-
H I2
LB tor. In this study, 35 steady states were reached,
according to a Doehlert design (Doehlert, 1970; Massart
et al., 1997) applied to the five process variables of fermen-
H I1
HB tation, i.e. hydrostatic pressure, ethanol concentration,
vvm, agitation and temperature (Garrido-Vidal et al.,
HS
2003). Hydrostatic pressure is the sum of the pressure
due to the column of medium and the head space pressure
of the fermentor. As regards the dimensions of the pilot fer-
Fig. 2. Scheme of the bioreactor.
mentor, the contribution of the height of the medium was
despised, and the pressure in the fermentation medium
was controlled by the pressure of the head space. Table 5
Table 3 shows the levels and the range studied for each variable.
Dimensions of the bioreactor Then, 6 additional continuous fermentations were devel-
Parameter Dimension oped to obtain the external validation set.
The oxygen transfer rate (OTR) was defined by the well-
DV = fermentor diameter 0.19 m
HB = baffle height 0.0419 m known balance:
HI1 = height of impeller 1 0.0345 m OTR ¼ k L a  ðC O2 ;L  C O2 ;L Þ ð2Þ
HI2 = height of impeller 2 0.194 m
Hm = liquid medium height 0.2609 m In each steady state, kLa was calculated by the equilibrium
Hs = sparger height 0.02 m reached between the oxygen transfer rate and the oxygen
HT = fermentor vessel height 0.3825 m
J = baffle width 0.02175 m
consumed, or oxygen uptake rate (OUR):
LB = baffle length 0.275 m k L a  ðC O2 ;L  C O2 ;L Þ ¼ OUR ð3Þ
R = number of baffles 3
VL = working volume 6.5 L This method is known as the gas balance method and it is
VT = total vessel volume 10 L very useful because fermentation is not altered. The results
140 J.-M. González-Sáiz et al. / Journal of Food Engineering 87 (2008) 136–147

Table 5
Range and levels of the studied variables
Variable Range Levels
Oxygen transfer Liquid–gas transfer
Pressure 1–2 atm 1–2 atm 5
Ethanol concentration 1–50 g L1 3–50 g L1 7
Acetic acid concentration 58–115 g L1 59–124 g L1
Ethyl acetate concentration 2–4 g L1 0.3–3 g L1
Acetoin concentration 0.75–1.2 g L1 0.03–0.50 g L1
Water 875–891 g L1
vvm 3–37 h1 3–35 h1 7
Agitation 200–1000 rpm 200–1000 rpm 7
Temperature 26–33 °C 26–33 °C 3

obtained with this method are not affected by the lag time
in response of the oxygen electrode, as in the electrode
momentum method.
The initial inoculums to start fermentation were
obtained from one of the industrial fermentors of a vinegar
firm from La Rioja. The wine used as feed was the same
white wine used by this firm. Inoculums and wine were
introduced into the fermentor without any pre-treatment.
Each experiment was subject to the same process condi-
tions for at least 168 h; the feed stream was controlled by
a PID control to keep ethanol concentration constant
within the set point of each experiment. When constant
values of ethanol, acetic acid, biomass concentrations,
dissolved oxygen and flow rate were obtained, it was con-
sidered that a stationary state had been reached, and the
conditions were maintained for 72 h more. Each stationary
state was well assessed and robust measures of the wine
flow rate and concentrations in the medium were obtained,
compensating the effect of the response time of the dis-
solved oxygen electrode. Therefore, robust values of kLa
were obtained.
When the steady state was reached, the balance between
the oxygen effluent stream – O2(o) – and the oxygen feed
stream – O2(i) – was due only to the oxygen consumed:
OUR ¼ O2ðoÞ  O2ðiÞ
O2(i) was calculated as
P a P H2 O;25 C
RG T out
 QO  0:2095  M O2
O2ðiÞ ¼
VL
where Pa = 1 atm, Tout = 298.15 K is the average tempera-
where oxygen and nitrogen were retained, and through a
packed column, with support carbopack B 80/20, to sepa-
rate ethanol, acetic acid, ethyl acetate and acetoin. Oxygen
was quantified by a TCD detector and polar compounds
were quantified by a FID detector. As standard for oxygen
was used air from laboratory, i.e. the same air which was
introduced into the fermentor by a compressor.
C O2 ;L values were calculated from the signal provided by
the dissolved oxygen probe, which is proportional to the
oxygen partial pressure in the gas phase in equilibrium with
C O2 ;L . In this study, C O2 ;L values for different temperatures
at 1 atm were obtained by saturating with oxygen a fermen-
tation medium in the pilot fermentor at different tempera-
tures and 1 atm, instead of taking the values for water from
literature. When the medium was saturated and the dis-
solved oxygen provided a constant reading, oxygen concen-
tration was assessed by an azide modification of the
Winkler Method called the Pomeroy–Kirshman–Altsberg
Method, AOAC method 973.45. This method is recom-
mended for water saturated with oxygen containing
organic compounds. Fig. 4 represents ln C O2 ;LðT L ;1 atmÞ vs
1/TL, together with the regression parameters and the
adjusted correlation rate, R2A. Data fitted well the linear
model, and, as regards Raoult’s Law, the vaporization
ð4Þ enthalpy of the medium, medium, 3140 cal mol1 – was
calculated from the slope of the linear regression. This
value was close to other oxygen–water values reported by
ð5Þ
2.25

ture of the air after passing through the fermentation med- 2.2
y = (1580 ± 20)x - (3.18 ± 0.08)
ium, R = 0.082 atm dm3 mol1 K1, P H2 O;25  C is the water
2
2.15 R A= 0.9991

pressure for 298.15 K and 0.2095 is the molar fraction of 2.1

Ln (C*O2.L)

oxygen in air, both taken from Perry’s Handbook (Perry 2.05

et al., 2001). O2(o) was calculated by a Hewlett Packard 2
5890 series II gas chromatograph analysing on line the 1.95
gases of the exhaust line. The chromatograph was equipped 1.9
with two chromatographic columns, two detectors and one 1.85
electro valve. Firstly, the samples were homogenised in a 1.8
0.00315 0.0032 0.00325 0.0033 0.00335 0.0034 0.00345
2 ml loop, prior to the column, with a 150 °C pre-heating
1/TL
program to avoid condensations. After injection, the sam-
ples passed through a molecular sieve (13  45/60 mesh), Fig. 4. Correlation plots of ln C O2 ;LðT L ;1 atmÞ vs 1/TL.
 J.-M. González-Sáiz et al. / Journal of Food Engineering 87 (2008) 136–147 141

other authors such as Dean (1992) or Wilhelm et al. (1977),
Finally, the C O2 ;LðT L ;P Þ values for different P pressures were
obtained by applying Henry’s Law to the C O2 ;LðT L ;1 atmÞ val-
ues provided by the linear regression.
The liquid–gas transfer rates for ethanol, acetic acid,
ethyl acetate, acetoin and water were evaluated by batch
experiments. A Doehlert design for 5 variables and 35
experiments was developed for each component and 6
additional experiments were developed to obtain the exter-
nal validation set. In each experiment, process variables,
which were equal in the five experimental matrices, were
established, and component concentrations were assessed
by adding synthetic ethanol, acetic acid, ethyl acetate, acet-
oin and distilled water to the medium at the beginning of
the experiment to adjust the concentrations and maintain
the working volume, 6.5 L. The medium was a real medium
used in industrial fermentation. Fermentation was not
started to avoid the changes in the concentrations due to
the biological process. Table 5 shows the levels and the
range studied for each variable. Experimental conditions
were maintained during at least 8 h for each experiment.
The transfer rates from well-dissolved components in liquid
phase to the gas phase can be considered so low that losses
of the studied components during the experiment were neg-
ligible compared to the experimental error and concentra-
tions were considered as constant during each experiment.
The fermentor’s gas outlet was connected to a condenser
to collect and quantify the mass of the compounds trans-
ferred to the gas phase. Concentrations of ethanol, acetic
acid, ethyl acetate and acetoin in fermentation medium
and in the liquid condensed from the gas stream were ana-
lysed by a Hewlett Packard 5890 series II gas chromato-
graph. The features of the chromatographic method have
been described previously (Garrido-Vidal et al., 2004).
The condenser’s gas outlet was connected to the Hewlett
Packard 5890 series II gas chromatograph and analyses
were performed on line to quantify the traces of com-
pounds which were not condensed, using the method
described for the oxygen transfer study. The sum of etha-
nol, acetic acid, ethyl acetate and acetoin in the condensed
liquid and the condenser’s gas outlet provided the
liquid–gas transfer rates (LGTRA). The liquid–gas transfer
rate of water was calculated by a mass balance with the rest
of components.
3. Results and discussion
3.1. Genetic algorithm developed for parameter optimisation
The parameters of the volumetric transfer coefficients
were calculated using a genetic algorithm. The algorithm
was an evolution of the basic genetic algorithm (Lucasius
and Kateman, 1993, 1994), programmed in Matlab
6.1.0.450 (The MathWorks Inc.). Table 6 describes the
main features of the algorithm. The genetic algorithm did
not require an initial estimate of the parameters but rather
a range of possible values. The uniform crossover increased
the genetic diversity of the offspring and the exploration of
new areas of the domain, thus avoiding convergence to a
relative minimum. The methodology was stochastic, thus
enabling an optimised set of parameters to be obtained
after running the programme several times. The effects of
each variable on the volumetric transfer coefficients were
not imposed ab initio. The restrictions on the range of val-
ues were only imposed to avoid negative values for the vol-
umetric transfer coefficients, without physical sense.
Finally, a critical point of the algorithm was the function
working as the evaluation function. The goal was to
optimise the predicted vs measured correlation of the
calibration set. A model which fits the experimental data
provides a predicted vs measured correlation with:
– a slope close to 1, including 1 within a 95% confidence
interval;
– a y-intercept value close to 0, including 0 within a 95%
confidence interval;
– an adjusted correlation rate close to 1;
– a low value of the root mean square.
Therefore, given the multi-criteria nature of optimisa-
tion, a desirability function was developed as the evalua-
tion function to maximize:

Table 6
Features of the genetic algorithm
Parameter Description
Coding Binary. Each chromosome is a different set of the parameters to optimise
Population 100 chromosomes. The population is constant. Initial population is randomly generated
responseðiÞ
Select-copy The probability of a chromosome being selected for cross-over is probðiÞ ¼ sumðresponseÞ
Cross-over Uniform cross-over. Cross-over probability 50%
Mutation 2%
Elitism 20 best chromosomes of each generation
Filters Out-of-range parameters and twins are not allowed
Stoppage Ten generations without changes higher than 5% in the mean response of the elitist chromosomes
condition
Heats and final The algorithm is completed 5 times. A final run is performed where the initial population is composed of the 20 best chromosomes
found in each of the 5 runs
142 J.-M. González-Sáiz et al. / Journal of Food Engineering 87 (2008) 136–147
   
1 1 800
 (a) kLa - Calibration
1 þ RMS ð1 þ absð1  slopeÞÞ 700

k La predicted (1/h)
   14 600
y = (1.04 ± 0.07 )x - (16 ± 24 )
2
1 1 R A= 0.9616
  ð6Þ 500
1 þ absðy interceptÞ 1 þ absð1  r2A Þ
400
The desirability function was scaled between 0 and 1, 300
allowing for an objective comparison of the predictive abil- 200
ity of each set of parameters. The four single functions had 100
to provide maximum values to obtain a maximum global
0
value, conferring more selective power on the evaluation 0 100 200 300 400 500 600 700 800
function, particularly between sets of parameters with sim- kLa measured (1/h)
ilar RMS. In this case, the set of parameters providing a 800
graphic closer to a linear correlation with slope 1 and (b) kLa - Validation
700
y-intercept 0 were rewarded because the values provided

kLa predicted (1/h)

600
for the parameters were not only close to the experimental y = (1.1 ± 0.2 )x - (83 ± 90 )
2
500
ones but also explained better the trend displayed by the R A= 0.9755

experimental data. 400

300

3.2. Model for oxygen transfer 200

100
A new semi-empiric correlation for KLa based on pro- 0
0 100 200 300 400 500 600 700 800
cess variables is proposed in this study:
 ak kLa measured (1/h)
P TL Pg
k L a ¼ C o  ðhP Þ  ðhT Þ   ðlS Þbk ð7Þ Fig. 5. Correlation plots of predicted vs measured concentrations for kLa:
VL (a) calibration, (b) validation.
where kLa is expressed in h1, p in atm, TL in °C, Pg in W;
VL in m3 and lS in m s1. The parameters were optimised
power input, Po, and obtain the aerated mechanical power
using the genetic algorithm and the results are shown in
input, Pg:
Table 7. Fig. 5 shows the predicted vs measured correlation
!1=5  1=4
plots for the calibration and validation set. The experimen- N 2  D4 QS
tal values of the external validation were not used to obtain P g ¼ P o  0:1  2=3
ð8Þ
gW VL N VL
the parameters of the model. Both plots show good linear
correlation and the 95% confidence interval of slope in- Po was calculated by the correlation proposed by Richards
cludes 1 and the 95% confidence interval of y-intercept in- (1963) for the power number of a Rushton turbine in the
cludes 0. Therefore, the plots show the good correlation of turbulent regime:
the model with the experimental data and the predictive
Po  g
ability of the model. NP ¼
The value of parameter aK is positive, so kLa increases qL  N 3  D5
  1:5  0:3  0:56  0:4
when aerated mechanical power input vs liquid volume W L J B R
increases, as it has been reported by other authors (Table ¼ 336:5 ð9Þ
D D D 6 4
2). Mechanical power transferred to the medium reduces
bubble size and thereby increases interfacial area. This is Po of the two Rushton impellers was calculated as two
mainly due to the high dynamic pressure near the impeller times the power number calculated by Eq. (9), since the
tip, which produces small bubbles. This effect is enhanced impellers were separated 1.96 times the diameter of the
by the baffles, which help break the bubbles and produce impeller, i.e. within the range [1.5–2] times the diameter
small bubbles. The correction proposed by Hughmark of the impeller (Rushton et al., 1950). The effect of vessel
(1980) has been used to correct the non-aerated mechanical geometry and impeller design on Pg and kLa was consid-
ered by these expressions. This influence has been reported
in literature, particularly the influence of DDV rate (Prasher
Table 7 and Wills, 1973; Garcia-Ochoa and Gómez, 2004).
Optimised set of parameters for kLa In terms of superficial air velocity, the value of parame-
Parameter Value ter bK is positive, in line with other models reported in
ak 0.363
Table 2, so kLa increases when superficial air velocity
bk 1.439 increases. It is important to mention that the air superficial
Co 1.0386  104 velocity measured at the exhaust line of the fermentor has
hP 1.319 to be corrected for the effect of pressure and temperature of
hT 1.107 the medium on the air flowing through the fermentation
 J.-M. González-Sáiz et al. / Journal of Food Engineering 87 (2008) 136–147
medium, before being used in Eq. (7). Parameter bK is
higher than parameter aK, thus showing that the effect of
aeration is more efficient in increasing oxygen transfer than
agitation. This effect should be taken into account in scale
up an optimisation of industrial processes, especially in
large-scale fermentors, since the economical cost of agita-
tion is higher than the economical cost of aeration. It
should be noted that aeration has a counteracting effect
on kLa because it decreases the mechanical power trans-
ferred to the medium. Furthermore, if the impeller were
unable to disperse the incoming air, it could flood, i.e.
the air-flow would dominate the flow pattern, thus making
the impeller work only in the gas phase.
The effect of temperature is explained by the coefficient
hT, directly related to temperature and the other parame-
ters, without considering a reference value of kLa at
20 °C, as considered by other authors (Table 2). In a
well-mixed and temperature-homogenised fermentor, bub-
ble air temperature can be considered to be equal to fer-
mentation temperature and remains constant due to heat
transfer from the medium. Temperature affects the diffu-
sion coefficient, viscosity and bubble diameter. The value
of hT is higher than unit, so temperature increases kLa, as
reported by Gulliver et al. (1990), Oosterhuis and Kossen
(1983).
The effect of hydrostatic pressure on kLa in biological
reactions has not been studied in as much depth in Aerated
Stirred Reactors, where most of models have been obtained
at atmospheric pressure. Instead, it has been studied more
in Bubble Column Reactors (Letzel et al., 1999; Lai et al.,
2004), where the positive effect of hydrostatic pressure over
kLa has been reported. In large-scale fermentors, medium
column height increases hydrostatic pressure, which must
be considered for scale up, and some authors have
described the use of the group kLap, which includes hydro-
static pressure (Bailey and Ollis, 1986). For this reason, the
coefficient hP, was introduced in the expression, with a
mathematical structure similar to coefficient hT, taking into
account that pressure affects the diffusion coefficient, vis-
cosity, bubble diameter and interfacial area, like tempera-
ture. The introduction of hydrostatic pressure enhances
the applicability of the model since it explains fermentation
processes in which it increases due to the height of the med-
ium and/or the overpressure of the head space.
The value hP is also higher than unit, demonstrating that
kLa increases with hydrostatic pressure. The bubbles rise
from the sparger at a pressure equal to the hydrostatic
pressure, and pressure into the bubble remains in equilib-
rium with the pressure of the medium while the bubble rise
from the sparger to the surface. When pressure rises,
bubble volume decreases to equilibrium, increasing the
interfacial area, a, and decreasing surface tension. Increas-
ing hydrostatic pressure enhances gas solubility, which
decreases surface tension. Density increases with pressure
and both effects reduce the bubble diameter (Hinze,
1955). The sum of these effects is the enhancement of kLa
with hydrostatic pressure. On the other hand, increasing
143
hydrostatic pressure also increases the viscosity of
ungassed liquid, which has counteracting effects on kLa.
Decreasing viscosity increases the Reynolds number and
therefore mechanical power input, but it increases ascend-
ing bubble velocity, and therefore residence time and gas
hold up decreases, leading to a smaller interfacial area.
However, it also enhances kL (Levich, 1962). Due to the
molecular structure of water, it is self compressed by
hydrogen bonding and hydrostatic pressure has almost
no effect on viscosity in water-similar mediums, such as
acetic fermentation. Therefore, the effect of pressure on vis-
cosity in these mediums is negligible. All in all, it is deduced
that hydrostatic pressure enhances kLa, thus explaining the
value obtained for parameter hP.
3.3. Models for liquid–gas transfers
In this study, liquid–gas transfer is explained by Whit-
man’s two-film theory, as proposed in Fig. 1. In terms of
oxygen transfer, kLa corresponds to the local liquid-phase
transfer coefficient kL multiplied by a, the gas–liquid inter-
facial area per total volume. It is generally assumed that kL
approximates the value of the global coefficient,KL, since
the solubility of oxygen in a water similar medium, such
us vinegar, is very low. Therefore, the dynamic of the gas
molecules in the gas side interphase is favoured and the
numerical value of the local gas-phase transfer coefficient,
kG, is very large compared to kL. However, related to
liquid–gas transfers, the solubility of component A in a
water-similar medium is very high, while the partial pres-
sure of component A in the gas phase is very low and the
dynamic of the liquid molecules of component A in the
liquid-side interphase is favoured. It is concluded that resis-
tance to the transfer of component A through the liquid-
gas interphase is mainly due to the gas-side interphase
inside the bubble. Therefore, the numerical value of kA,L
is very large compared to kA,G, which approximates the
value of the global coefficient KA,L. A balance similar to
the oxygen transfer rate is proposed for the liquid–gas
transfer rate:
LGTRA ¼ k A;G a  ðC A;L  C A;L Þ ð10Þ
where kA,Ga is defined as the volumetric component A
transfer coefficient in the gas-side interphase.
The moles of component A passing from the liquid
phase to the gas phase are negligible compared with molar
concentration in the liquid phase. Therefore, CA,L, is negli-
gible in comparison with C A;L , and the balance is
simplified:
LGTRA ¼ k A;G a  ðC A;L Þ ð11Þ
Both sides of the gas–liquid interphase are similar
enough to assume the same mathematical structure for
the models which explain the local liquid phase transfer
coefficients and the local gas phase transfer coefficients.
The same mathematical structure proposed for kLa is
proposed for kA,Ga:
144 J.-M. González-Sáiz et al. / Journal of Food Engineering 87 (2008) 136–147
 ak
p TL Pg bk 0.20
k A;G a ¼ C o  ðhP Þ  ðhT Þ   ðlS Þ ð12Þ (a) Ethanol - Calibration
VL

kA.Ga predicted (1/h)

y = (1.0 ± 0.1)x - (0.002 ± 0.006)
0.15
where kA,Ga is expressed in h1, in atm, in °C, Pg in W; VL 2
R A= 0.9242

in m3 and in m s1. This approach is one of the main con-

tributions of this study. Pg and lS are calculated as ex- 0.10

plained for kLa.The parameters were optimised using the

genetic algorithm and the results are shown in Table 8. 0.05
Figs. 6–10 show the predicted vs measured correlation
plots for the calibration and validation set. The experimen- 0.00
tal values of the external validation were not used to obtain 0.00 0.02 0.04 0.06 0.08 0.10 0.12 0.14 0.16 0.18
kA.Ga measured (1/h)
the parameters of the models. In each figure, both the cal-
ibration and the validation plot show good linear correla- 0.20
tion and the 95% confidence interval of slope included 1 (b) Ethanol - Validation

kA.Ga predicted (1/h)

and the 95% confidence interval of y-intercept included 0. 0.15
y = (1.0 ± 0.3) x - (0.00 ± 0.03 )
2
R A= 0.9627
Therefore, the plots show the good correlation of each
model with experimental data and the predictive ability
0.10
of the models.
It is noteworthy that parameter CO is lower in kA,Ga the
models than in the kLa model, and the volumetric 0.05

0.00
0.00 0.05 0.10 0.15 0.20
Table 8 kA.Ga measured (1/h)
Optimised set of parameters for kA,Ga
Fig. 7. Correlation plots of predicted vs measured concentrations for
Parameter Ethyl acetate Ethanol Acetoin Acetic acid Water ethanol kA,Ga: (a) calibration, (b) validation.
ak 0.019 0.030 0.154 0.167 0.102
bk 1.106 0.910 1.251 1.404 1.335
Co 565.2 0.590 0.061 0.173 0.892
hP 0.3730 0.4346 0.6006 0.5126 0.3028
hT 1.008 1.060 1.087 1.087 1.070
2.5E-04

(a) Acetoin - Calibration

0.35 2.0E-04 y = (1.0 ± 0.1)x - (0 ± 1E-05)
kA.Ga predicted (1/h)

2
(a) Ethyl Acetate - Calibration R A= 0.8395
0.3
kA.Ga predicted (1/h)

y = (0.98 ± 0.08)x + (0.005 ± 0.007)

2 1.5E-04
0.25 R A= 0.9450

0.2 1.0E-04

0.15
5.0E-05
0.1

0.05 0.0E+00
0.0E+00 5.0E-05 1.0E-04 1.5E-04 2.0E-04 2.5E-04
0
0 0.05 0.1 0.15 0.20 0.25 0.3 0.35 kA.Ga measured (1/h)
kA.Ga measured (1/h)
1.2E-04
0.35 (b) Acetoin - Validation
(b) Ethyl Acetate - Validation 1.0E-04
0.30 y = (1.0 ± 0.4 ) x - ( 4E-06 ± 2E-05 )
kA.Ga predicted (1/h)

y = (0.9 ± 0.2) x + (0.01 ± 0.03) 2

kA.Ga predicted (1/h)

2
R A= 0.9183
0.25 R A= 0.9821 8.0E-05

0.20 6.0E-05

0.15
4.0E-05
0.10
2.0E-05
0.05

0.00 0.0E+00
0.00 0.05 0.10 0.15 0.20 0.25 0.30 0.35 0.0E+00 2.0E-05 4.0E-05 6.0E-05 8.0E-05 1.0E-04 1.2E-04
kA.Ga measured (1/h) kA.Ga measured (1/h)

Fig. 6. Correlation plots of predicted vs measured concentrations for ethyl Fig. 8. Correlation plots of predicted vs measured concentrations for
acetate kA,Ga: (a) calibration, (b) validation. acetoin kA,Ga: (a) calibration, (b) validation.
 J.-M. González-Sáiz et al. / Journal of Food Engineering 87 (2008) 136–147 145

0.06 the rates are negligible compared to the acetification rate.

(a) Acetic Acid - Calibration However, this approach has provided process models
0.05
kA.Ga predicted (1/h)

y = (1.0 ± 0.1)x - (0.001 ± 0.003)

2
R A= 0.8422
which cannot consider the sensorial impact of losing com-
0.04 pounds such as ethyl acetate and acetoin, nor yield losses
due to the transfer of ethanol, acetic acid and water to
0.03
the gas phase.
0.02 The parameters ak and bk are within the same magnitude
order of the parameters ak and bk of the kLa model. The
0.01
enhancement of the gas–liquid interfacial area due to the
0.00 mechanical power transferred to the liquid medium also
0.00 0.01 0.02 0.03 0.04 0.05 0.06
kA.Ga measured (1/h)
favours the transfer of components from the liquid phase
to the gas phase. Aeration is also critical in this transfer
0.07 and parameter bk is higher than parameter ak for each
(b) Acetic Acid - Validation model, as in the case of the kLa model. This finding should
0.06
kA.Ga predicted (1/h)

y = (0.8 ± 0.2) x + (0.002 ± 0.006)

2
be taken into account when optimising the acetification
0.05 R A= 0.9646
process, since high aeration rates improve oxygen transfer
0.04
but also increase compound loss due to liquid–gas
0.03 transfers.
0.02
In terms of temperature, the parameter hT is higher than
unit in each model, and the values are similar to the value
0.01
for the kLa model. The effects of temperature on the viscos-
0.00
0.00 0.01 0.02 0.03 0.04 0.05 0.06 0.07
ity of the liquid medium, bubble diameter and interfacial
kA.Ga measured (1/h) area also increase the transfer of liquid compounds to the
gas phase. Furthermore, the molecular diffusivity of com-
Fig. 9. Correlation plots of predicted vs measured concentrations for pound A in the gas phase is also expected to improve with
acetic Acid kA,Ga: (a) calibration, (b) validation.
temperature since the internal energy of the molecules
increases with temperature and molecule dynamics through
0.7 the gas-side interphase are favoured. The viscosity of the
(a) Water - Calibration gas-side interphase increases with temperature due to the
0.6
y = (1.0 ± 0.2)x - (0.01 ± 0.04) different behaviour of viscosity of gases and liquids and it
kA.Gap redicted (1/h)

2
0.5 R A= 0.8028
should block the transfer of molecules from the liquid
0.4 phase to the gas phase. However, air viscosity increases
0.3 very slightly with temperature, hence its effect is compen-
sated by the other effects.
0.2
Lastly, the values obtained for parameter hP are less
0.1
than unit in all the liquid–gas transfer models. Therefore,
0
0 0.1 0.2 0.3 0.4 0.5 0.6 0.7
the effect of hydrostatic pressure is contrary to the effect
k A.Ga measured (1/h) reported for kLa. Although the effect of the pressure of
the medium over the bubble diameter and gas-liquid inter-
1.0
facial area should increase kA,Ga, and the effect on the vis-
(b) Water - Validation
0.8
cosity of the liquid medium is negligible, the effect of
kA.Ga predicted (1/h)

y = (1.3 ± 0.3)x - (0.1 ± 0.1 )

2
R A= 0.9633 pressure on the viscosity and density of the gas-side inter-
0.6 phase must be considered. In the case of viscosity, pres-
sures above 2 atm should be applied to detect increases in
0.4 viscosity. However, gas density increases with pressure
and the gas-side interphase offers more resistance to the
0.2
transfer of molecules from the liquid phase. Therefore, it
0.0
is logical to expect a reduction in the value of the diffusion
0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 coefficient kA,G, thus compensating for the other effects and
kA.Ga measured (1/h) decreasing kA,Ga.
Fig. 10. Correlation plots of predicted vs measured concentrations for
water kA,Ga: (a) calibration, (b) validation. 3.4. Application of the models

liquid–gas transfer coefficients are lower than kLa. The models explain the transfers through the whole
Liquid–gas transfer is less favoured than oxygen transfer, fermentation medium. No compartmental models for dif-
and it is normal despite losses of volatile compounds in ferent sections of the fermentor have been proposed and
modelling the acetification process since the magnitude of predictive ability will improve when mixing conditions
146 J.-M. González-Sáiz et al. / Journal of Food Engineering 87 (2008) 136–147
are closer to the ideal perfect mixed reactor. The correla-
tions do not include a specific parameter for viscosity since
the acetic fermentation medium is a Newtonian medium
and viscosity is not affected by agitation rate or aeration.
The models can be applied to aqueous fermentation medi-
ums similar to the industrial fermentation medium studied.
The model proposed for kLa can be applied for large-
scale vinegar fermentations with the same geometry dimen-
sion rates as the pilot fermentor and the same agitation and
aeration system. Therefore, the same kLa value may be
achieved in different-sized vessels by adjusting the process
variables. Modelling kLa enables the oxygen transfer rate
to be predicted. As it will be reported in future research,
it will be used together with an oxygen uptake rate model
to evaluate aeration-agitation configurations of fermenta-
tion systems for industrial production of vinegar.
Liquid–gas transfer models are very useful for optimis-
ing industrial-scale fermentation processes. One of the
main causes of reduced yields during acetic fermentation
processes is the loss of ethanol and acetic acid. Given the
oxygen requirements of fermentation, a large amount of
aeration and agitation is required, increasing the amount
of volatiles that leave the fermentor within the gas outlet.
Know-how in the vinegar industry shows that a reduction
in stoichiometric yield of about 5% can occur due to losses
of ethanol and acetic acid through the gas outlet. Ethyl ace-
tate and acetoin are very important for the sensorial profile
of vinegar. The typical flavour and taste of ethyl acetate is a
key factor for aged quality vinegar with a high content of
residual ethanol, which stimulates the formation of the
ester. Therefore, losses of ethyl acetate during acetic
fermentation deplete the sensorial features of the final
product. Acetoin has a biological origin by secondary met-
abolic routes of bacteria. The presence of acetoin demon-
strates the chemical and microbiological integrity of
vinegar and also makes a positive contribution to its senso-
rial profile, especially in terms of flavour. Lastly, water
losses have an important effect on concentration profiles
and the total working volume, which affects hydrodynamic
variables during fermentation.
4. Conclusions
Gas–liquid and liquid–gas transfers were modelled in
the industrial production of vinegar by applying Whit-
man’s two-film theory. The core of the models were the
expressions for the volumetric transfer coefficients kLa
and kA,Ga. The expression developed
h for ikLa included a
Pg
widely-used correlation between ; lS and kLa, and
VL
introduced the effects of temperature and pressure in the fer-
mentation medium, thus extending the range of applicability
of the model. The expression developed for kA,Ga was based
on similar fundamentals of kLa. The effects of the process vari-
ables on liquid–gas transfer were similar to the effects on gas-
liquid transfer, except for the effect of hydrostatic pressure,
which enhanced kLa but reduced kA,Ga.
The Genetic Algorithm optimised the parameters of the
mathematical models thanks to the desirability function
developed as the evaluation function. The expressions pro-
posed fitted the experimental data very well and were vali-
dated with external validation sets.
The models can be applied for acetic fermentation in
any fermentor with the same agitation and aeration system
design and the same geometry. The fundamentals and
mathematical form of the models proposed and the exper-
imental methodology can be applied to other systems. The
evaluation of specific coefficients for each fermentor design
and fermentation medium provides models with better
prediction ability and better applicability for scaling up.
In future research, the fundamentals and results of this
article will be a part of a global model for the industrial
process of vinegar production.
Acknowledgements
The authors thank the Ministry of Science and Technol-
ogy – INIA (Project No. CAL01-053) and the Autonomous
Government of La Rioja – Consejerı́a de Educación, Juven-
tud y Deportes (Research grant FPI-1999) for their finan-
cial support.
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