Optimization, Modeling, and Online

Monitoring of the Enzymatic Extraction of

Banana Juice
Vrani Ibarra-Junquera, Pilar EscalanteMinakata, Arturo Moiss ChvezRodrguez, Isabel Alicia ComparanDueas, et al.
Food and Bioprocess Technology
An International Journal
ISSN 1935-5130
Food Bioprocess Technol
DOI 10.1007/s11947-013-1136-2

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Food Bioprocess Technol
DOI 10.1007/s11947-013-1136-2

ORIGINAL PAPER

Optimization, Modeling, and Online Monitoring

of the Enzymatic Extraction of Banana Juice
Vrani Ibarra-Junquera & Pilar Escalante-Minakata & Arturo Moiss Chvez-Rodrguez &
Isabel Alicia Comparan-Dueas & Juan Alberto Osuna-Castro & Jos de Jess OrnelasPaz & Jaime David Prez-Martnez & Cristbal No Aguilar

Received: 25 May 2012 / Accepted: 20 May 2013

# Springer Science+Business Media New York 2013

Abstract This article focuses on the optimization, modeling, and online monitoring of banana juice production
through an enzymatic method. In order to perform this
task, a batch reactor was designed with automatic control
over the temperature and the agitation speed as well as
online monitoring of torque. The experiments were carried
out with the Musa AAA Cavendish banana variety (Enano
gigante), the main variety planted in Mexico. Three different
ripening stages were evaluated. Optimization of juice extraction was evaluated as a function of the pulp/water relationship
and the concentration of the enzyme complex. The results
showed that the adding of water had no influence on the
extraction of banana juice, and the optimal enzyme concentration per kilogram of banana pulp was found. Based on a
fuzzy logic approach, it was possible to relate the initial torque
V. Ibarra-Junquera (*) : P. Escalante-Minakata :
A. M. Chvez-Rodrguez : I. A. Comparan-Dueas
Bioengineering Laboratory, University of Colima, Carretera
Colima-Coquimatln, Km 9,
Coquimatln, Colima State, Mexico
e-mail: vij@ucol.mx
J. J. Ornelas-Paz
Centro de Investigacin en Alimentacin y Desarrollo A.C.
(CIAD), Unidad Cuauhtmoc, Chihuahua State, Mexico
J. D. Prez-Martnez
Faculty of Chemical Sciences, Autonomous
University of San Luis Potos,
San Luis Potos, S.L.P. State, Mexico
J. A. Osuna-Castro
Faculty of Biological and Agricultural Sciences,
University of Colima, km 40 Autopista Colima-Manzanillo,
28100, Tecomn, Colima, Mexico
C. N. Aguilar
Department of Food Science and Technology, School of
Chemistry, Autonomous University of Coahuila, Saltillo
Coahuila State, Mexico

with the ripeness stage. Furthermore, an observable dynamical

model based on ordinary differential equations and fuzzy logic
is presented. With this model, the relationship between the
torque dynamic and the instant juice yield was found to depend
on the amount of enzyme, the temperature, and the maturity
stage of the banana used. In addition, a principal components
analysis was used to classify and to relate the final juice
characteristics (e.g., L, a, and b colorimetric components) to
the processing conditions and the final appreciation of a group
of sensorial panelists. Additionally, a robust observer was
designed and implemented to filter the noise present in the
torque signal and to predict the instant juice yield.
Keywords Banana . Juice . Modeling . Observer .
Optimization . Processing

Introduction
Bananas are one of the worlds most important food crops,
consumed by millions as part of a daily diet and for nutrient
enrichment (Mohapatra et al. 2011) since they are an important
source of polyphenols, minerals, and carbohydrates (Kiyoshi and
Wahachiro 2003; Wall 2006; Escalante-Minakata et al. 2013).
Furthermore, the banana is widely appreciated for its flavor and
aroma (Boudhrioua et al. 2003; Mohapatra et al. 2011). Though
commonly consumed as fresh fruit, bananas have an unfortunately short shelf-life due to softening and thus are often used in
banana juice processing as an alternative (Kyamuhangire et al.
2002; Lee et al. 2006a, b; Lpez-Nicols et al. 2007; Mohapatra
et al. 2011; Chvez-Rodrguez et al. 2013).
A relevant issue in banana juice extraction is the retention
of the juice in the pulp due to the great amount of polysaccharides, which prevent the release of intra-cell components,
affecting the extraction yield and the clarification. The juice
can be extracted by a mechanical press and/or through the

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action of pectinolytic enzymes, such as pectinase and

polygalactouronase (Casimir and Jayaraman 1971; Viquez et
al. 1981; Kyamuhangire et al. 2002; Lee et al. 2006a). For that
reason, banana juice extraction can be considered a bioprocess
when an enzymatic bioreactor is at its core. Such a process
demands high levels of quality and safety in food production, which calls for high standards in quality assurance and
process control; satisfying this demand, in turn, requires
appropriate analytical tools for food analysis both during
and after production (Zhong 2010). Desirable features of
such tools include speed, ease of use, minimal or no
sample preparation, and the avoidance of sample destruction. It is also necessary to control and optimize the bioreactor environment via operating variables in order to favor
the desired functions and achieve cost-effective, large-scale
manufacture (Zhong 2010). Traditionally, a method to
achieve this goal utilizes laboratory instruments to treat
samples taken from the processing plant and analyzed offline (Ibarra-Junquera et al. 2010). This allows process
investigation, checking of the specifications of raw materials and final products, which aids in management and
decision making (Cozzolino et al. 2011). However, recent
developments in on-line measurements offer the possibility
of following process dynamics, achieving better and more
rapid control, and eliminating questions about the statistical
significance of quality control based on sampling (Zhang
2009; Ibarra-Junquera et al. 2010; Cozzolino et al. 2011).
Several studies have described on-line instrumentation for
determining the characteristics of particle systems in situ in
the processing plant and that describe how on-line measurements from such instruments lead to process improvements. In particular, better monitoring and control of bioreactors requires reliable on-line estimation of process variables and parameters that often cannot be measured directly
(Zhang 2009; Ibarra-Junquera et al. 2010).
Therefore, this study has three main goals. The first goal
is to optimize the enzymatic method for banana juice extraction by finding the minimum amount of water and
enzymatic complex needed to obtain the maximum juice
yield in a given span of time. Second, this study aims to
develop a minimalist model to describe, in general terms,
banana juice extraction dynamics, allowing its usage for online monitoring and automatic control. Thus, such a model
should include on-line measurable variables and the juice
yield as one of its states, and then construct a model-based
observer algorithm to infer in real time the juice yield from
the on-line measurable output. Under the hypothesis that the
torque evolution can be associated with the juice yield
dynamic, a batch reactor was designed with automatic control over the temperature and the agitation speed and with
on-line monitoring of torque. The studys third goal is to
relate the visual characteristics of the final banana juice to
the processing conditions.

Methodology
Bioreactor
The enzymatic juice extractions were performed in a 4-L
stainless steel batch-jacketed reactor (i.d. 12.5 cm),
equipped with automatic control over the temperature, based
on a cooling thermostat with a precision of 0.02 C (RE
630S; Lauda Eco Silver, Germany). The bioreactor agitation
speed control is composed of the Compact cRio-9074 data
acquisition system (DAS) (National Instruments, TX, USA),
a PC for the control of the DAS, a stepper motor (NEMA 23
Stepper Motor; National Instruments) connected to a
Microstepping Drive (National Instruments), and two independent power supplies. The stepper motor movement is
directed by using an application developed within the
Labview platform (National Instruments). An open-paddle
impeller (width 9.6 cm, height 12.3 cm) is connected to the
motor through a drill chuck. The torque was monitored
online using a rotary torque sensor with a maximum capacity of 2 Nm (FSH01979; FUTEK, CA, USA) connected to a
PC through a data acquisition system from National Instruments (Cryo-9074) and Labview-based interface.
Biological Material
Bananas of the cultivar Enano gigante (Musa AAA, subgroup Cavendish) were acquired in the local market when
they were green (Fig. 1) with a minimum length of 13 cm.
The banana bunches were ripened in laboratory conditions
at 20 C without atmospheric control for different storage
times. The color of the peel was monitored by comparison
with a scale ranging from 1 to 3, with values representing
the following stages of ripeness: Green (1), ranging from
more yellow than green to yellow with green endings;
Yellow (2), ranging from completely yellow to yellow with
slight brown specks; and Brown (3), which included yellow
with many brown specks. Additionally, the parts of individual samples were weighed in an analytical balance to determine the pulp-to-peel ratio. The results were expressed as
the percent weight of pulp relative to peel.

Fig. 1 Peel color changes in different stages of banana ripeness, the

numbers 1, 2, and 3 corresponds to green, yellow, and brown,
respectively

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Juice Extraction Procedure

Colorimetric Analysis

The banana fruits were washed in 0.2 % (v/v) sodium

hypochlorite aqueous solutions for 5 min, peeled, and cut
into pieces. In order to homogenize the banana pulp, appropriate volumes of water were added to achieve a banana
pulp/water ratio of 0.8, 0.9, and 1.0. Then, the mixture was
homogenized to puree in an electrical blender. The enzymatic extractions were performed using a commercial mixture of three plant-cell-wall-degrading enzymespectinase,
cellulase, and hemicellulase (Macerex PM, ENMEX, S.A.
de C.V. (2003)), which hydrolyze the polysaccharide substrates from banana cell wallranging from 100 to 600 L
of enzyme mixture per kilogram of puree. The enzymatic
extractions took place in a batch reactor (filled with 3 kg of
banana puree) and continuously agitated (100 rpm) at room
temperature (25 C) for 2 h. Subsequently, the juice was
centrifuged at 12,000g using a centrifuge (RC6+; Sorvall,
Newtown, CT, USA) in an SLA-3000 rotor for 15 min, and
the supernatant was collected and filtered. Finally, the total
soluble solids content was determined for each banana juice
obtained, using a digital refractometer (RP-101; Atago, Tokyo, Japan) with a scale ranging between 0 and 45 degrees
Brix (Bx). The soluble solids content were reported as
degrees Brix.

Analysis of color variation was performed with a total

volume of 20 mL from each banana juice sample and
dispensed into separate Petri dishes. The color values of
each sample were obtained with a LABSCAN XE chroma
meter (Hunterlab, VA, USA) at room temperature. Equipment was set up for illuminant D65 and 10 observer angle.
CIE-Lab values of L (lightness), a (redness), and b
(yellowness) were determined to describe the precise location of a color inside a three-dimensional visible color
space.

Juice Yield
The banana juice yield was calculated based on the weight
of the resulting juice, the added water, and the banana pulp,
according to the following equation:
Juice yield%

jwgwwg
 100
bwg

where jw is the amount of juice recovered, ww is the weight

of the added water, and bw is the weight of the total banana
puree used. All weight measurements were expressed in
grams (g). The juice was recovered by centrifugation at
the same conditions mentioned in the juice extraction methodology. The samples weights were obtained through an
analytical balance (OHAUS; Explorer Pro, Pine Brook,
NJ, USA; accuracy 0.1 mg).

Statistical Analysis
All statistical analyses were carried out using MatLab software (MathWorks Inc., USA). The analysis of variances
(ANOVA) was applied to compare the mean values of
samples according to the different factors. In addition, principal component analysis (PCA) was used as a data multivariate technique. The purpose of this method is to decompose the data matrix and concentrate the source of variability in the data into the first few principal components. Here,
PCA was applied to two different groups of values: the
matrix of maturity mean attribute ratings across banana
samples (initial torque and the peel-to-pulp ratio) and the
matrix of mean color measurements (L, a, and b). The two
cases were also analyzed by cluster analysis (average linkage method) in an attempt to classify the samples in terms of
their maturity and processing method.
Visual Evaluation
The juice data, coming from the colorimetric analysis, were
evaluated using PCA and cluster analysis. As an additional
reference, the resulting groups were classified in terms of
visual acceptance by a five-judge panel belonging to the
faculty of Chemical Sciences and research scholars of the
Bioengineering Laboratory (both of the University Colima
in Mexico). This general appearance of the juice was measured subjectively on a three-point hedonic scale: poor
(dislike), good (like), and excellent (like very much).
Optimization Procedure

Protein Quantification
The protein concentration of the enzymatic complex was
determined by the Bradford method (1976) as modified by
BioRad (using bovine serum albumin protein as standard).
The samples were diluted to a ratio of 1:20 by adding
distilled water. The absorbance of the samples was determined at 595 nm using a lambda 25 UVVis spectrophotometer (Perkin-Elmer Instrument, USA).

The optimization methodology used in this work is based on

a three-step process: (1) generating a mesh of experimental
data within the design space, (2) modeling the data sets
through nonlinear fitting, and (3) using the fitted model
and ANOVA analysis to obtain the optimal condition.
The dependent variables selected for these studies were
X, enzyme concentration (L/kg), taking values in the set
{0, 25, 50, 100, 150, 300, 600 L/kg}; Y, ratio of

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water/banana pulp taking values in the set {1, 0.9, 0.8}; and
Z, response variable. Given are a total of 63 experiments, to
be performed in triplicate. The juice yield, as evaluated in
Eq. 1, was the response variable. Based on the preliminary
data, the following generalized MichaelisMenten response
surface (Hirst et al. 1996) is used to model the data behavior:

Z X ; Y

bXY
c dYZ

2
Fig. 2 Memberships function with triangular membership distribution

Exclusively for the optimization process, the previously

mentioned banana juice extraction methodology was slightly modified since all experiments were carried out at 50 C
for 30 min, and the yield was calculated at the end of the
process.

Fuzzy Logic Approach

The complexity of biological processes often renders impractical the development of detailed, structured phenomenological models. Fuzzy modeling is based on sets of fuzzy
ifthen rules derived from expert domain knowledge to
handle uncertainty. The concept of fuzzy logic provides a
natural way of dealing with problems where the absence of
well-defined criteria could be a source of imprecision. Since
fuzzy systems can simultaneously handle numerical data
and linguistic knowledge, they provide opportunities for
modeling of conditions that are inherently imprecisely defined, like fruit ripeness.
The quality of banana juice is defined by its physical appearance and taste, both of which are directly
related to its stage of ripeness. Thus, the ripeness
stage directly influences the quality of the final juice.
Therefore, the purpose of the fuzzy modeling is to
relate the banana maturity as perceived by human eyes
based on color and general physical characteristics
(e.g., appearance of spots) to standard measurements
that can allow the modeling and monitoring of the
juice extraction process.
Triangular membership functions have been used for
similar problems (Sinija and Mishra 2011). For example,
triangle a1b1c1 represents the membership function for
ripeness stage 1, triangle a2b2c2 represents the distribution function for ripeness stage 2, etc. Figure 2 represents
the triplets associated with three ripeness stages. The second
number of the triplet denotes the coordinate of the abscissa
at which the value of the membership function is 1. The first
and third numbers of the triplet designate the distance to the
left and right, respectively, of the second number where the
membership function is 0.

Enzymatic Juice Extraction Model

The main goal of this paper is to develop a minimalist model
to describe, in general terms, banana juice extraction dynamics to be used for monitoring and control. This unstructured and nonsegregated model is given by Eqs. 3 and 4.






 
dx1
m T ; x2;0 xmax T ; x2;0 x1
dt




 
m T ; x2;0

dx2
 xmax T ; x2;0 x1


dt
y T ; x2;0

where the state variables are as follows: x1 stands for juice

yield and x2 stand for torque. The parameters are as follows:
T stands for temperature (C) and M for the maturity stage,
i.e., M (1,2,3). The rest of the functions are given in
Table 1. The mathematical expressions given in Eqs. 3 and
4 were selected since the preliminary data showed an exponential growth of the juice yield and an exponential decay of
the torque signal as well as a temperature and maturity stage
dependence of the kinetic rate, m, the maximum juice yield,
xmax, and the torque initial condition x2,0. The function
y(T,x2,0) allows to relate the juice yield x1(t) with the torque
Table 1 Functions and units used in dynamical model given by Eqs. 3
and 4
Variable

Dependence

Units

xmax(T,x2,0)

f1(T,M)

y(T,x2,0)
m(T,x2,0)

f2(T,M)

1/N m

km
max

max S
k m S

Be

M X 2;0

1
Ae T
( M(X2,0))2 +w M(X2,0)+

e Y

M X 2;0

L/kg

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measurements x2(t). Thus, this methodology can be considered as a data-driven approach to the modeling of the
enzymatic extraction of banana juice in which its structure
and the parameter identification is based on off-line and online time series obtained from a rigorous experimental set.
Software Sensor Design
Taking into account the fact that rarely can one have a
sensor on every state variable, and some form of reconstruction from the available measured output data is needed,
software can be constructed using the mathematical model
of the process to obtain an estimate of the true state x. Ever
since the original work by Luenberger (1971), the use of
state observers has proven useful in process monitoring and
for many other tasks. In the sense of control theory, an
algorithm capable of giving a reasonable estimation of the
unmeasured variables of a process will be called an
observer.
Numerous attempts have been made to develop observer
design methods for bioprocess applications (EscalanteMinakata et al. 2009; Zhang 2009; Fernndez-Fernndez
and Prez-Correa 2010). The first systematic approach for
the development of a theory of observers was proposed
some time ago by Krener and Isidori (1983). Nevertheless,
it is well known that classical proportional observers tend to
amplify the noise of on-line measurements, which can lead
to the degradation of the observer performance. In order to
avoid this drawback, in this paper, the observer algorithm is
based on the work of Ibarra-Junquera et al. (2005) because
the proposed integral observer provides robustness against
noisy measurements and uncertainties.
In this section, the design of the software sensor is
presented in which xj, for j(1n), is the naturally measured state (i.e., the variable easiest to measure). Therefore,
it seems logical to take xj as the output of the system, y=h(x)
=xj. Now, considering that the output function h(x) is contaminated with a Gaussian noise, the model given by the
aforementioned Eqs. 3 and 4 acquires the form:

X f X
y cX

where f(X) is a matrix containing the left-hand side of Eqs. 3

and 4, y represents the noisy on-line measurable output, is
the additive bounded measurement noise, X=[x1,x2] is the
vector of states, and the vector cX=[0,1]T[x1,x2] defines the
on-line measurable output, x2 in this particular case. Then,
the task of designing an observer for the system Eqs. 3 and 4
is to estimate the vector of states X, despite the noise but
considering that y is measured on-line and that the system is
observable.

Results and Discussion

Optimization Procedure
Many recent works show that the use of polysaccharidases
facilitates the release of juice and increases the extraction
yield (Kyamuhangire et al. 2002; Sreenath et al. 1994;
Cheirsilp and Umsakul 2008; Buenrostro-Figueroa et al.
2010; Bahramian et al. 2011). In addition, in the case of
banana and mango juice extraction, many protocols include
dilution of pulp in water, ranging from undiluted extraction
processes (Khalil et al. 1989; Reddy and Reddy 2005;
Buenrostro-Figueroa et al. 2010) to water/pulp ratios of
1:1 (Cheirsilp and Umsakul 2008; Kyamuhangire and
Pehrson 1999; Onwuka and Awam 2001), 1:2 (Lee et al.
2006a, b; Lee et al. 2007), 1:4 (Falade and Babalola 2004),
and 1:5 (Akubor 1996). However, none of these works
present a justification for the selected dilution ratio. While
there is clear evidence that the application of enzyme complexes in the extraction of banana juice is efficient, no study
has reported on the variables of ripeness stage and
water/pulp ratio.
The results of the Bradford assay show that the commercial enzymatic complex contains 12.06 1.20 mg
protein/mL. The juice yield was affected by the enzymatic
complex concentration and water/pulp ratio within the studied conditions (P<0.05). Nevertheless, the water/pulp ratio
had a significant effect only when extractions were carried
out without addition of the enzymatic complex (P<0.05).
In juice extractions with enzymatic complex, independently of the water/pulp ratio, the juice yield increased with
the enzyme complex concentration from 25 L/kg
(65.47 %) to 150 L/kg (71.11 %), while higher enzyme
concentrations (i.e., 300 L/kg and 600 L/kg) did not
produce significant increase in the juice yield (P>0.05).
Hence, the optimal (i.e., more sustainable) conditions for
juice extraction were 150 L/kg enzyme complex concentration without water addition.
As shown in Fig. 3, the complete experimental data set were
properly represented by Eq. 2 (R2 =0.9808). As previously
mentioned, a slight effect of dilution was observed only in
extraction without enzymatic complex, while a high increment
in juice yield was observed with the addition of the enzymatic
complex to reach a plateau at concentrations ranging from 150
to 600 L/kg independently of the water/pulp ratio.
Modeling the Enzymatic Juice Extraction
It is well known that modeling involves the process of
interpreting a problem-solving situation where a system of
interest needs to be represented by a mathematical system,
which will simplify, delete, maintain, and distort various features (Zawojewski 2010). In particular, this work aims to

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juice yield (%)

75
70
65
60
55
50
45
1
0.95
0.9
600

0.85

400

water/pulp ratio
0.8

200
0

enzyme concentration (L/kg)

Fig. 3 Graphic shows the nonlinear fitting of the experimental data

(blue circles) and the model given by Eq. (2). The model estimated
parameters are a=49.5, b=225.4, c=100, and d=10, with R2 =0.9808

develop a simplified model that relates on-line measurable

variables with off-line ones. In addition, the resulting model
should allow the construction of an observer scheme to rebuild, in real time, the non-measurable on-line variables.
It has been previously reported that the use of
polysaccharidases improves the juice yield and modifies
the physicochemical and rheological properties of the mixture during the process (Aguilar et al. 2008; Bahramian et al.
2011). Formerly, off-line viscosity measurements of pectin
solution were used to determine the activity of
polysaccharidases because a decrement of this parameter is
related to this endohydrolase activity (Combo et al. 2012).
Since torque is the measurement of the force that rotates an
object around its axis (e.g., an open-paddle impeller), this
force represents the resistance that a fluid offers to rotational
motion. Based on that fact, Virgen-Ortz et al. (2012) measured the pectinolytic activity throughout the slope of the
-3

x 10

-3

a)

x 10

It is important to note that the kinetic rate m decreases at

50 C in all of the enzymatic complex concentrations and
ripeness stages studied. Thus, the data obtained at 50 C
were not taken into account for the temperature dependence
studies (although this phenomenon is briefly studied at the
end of this subsection).
The calculated values of km reveal no change as a function
of the temperature, while the dependence of max on the
-3

b)

2
2
4 x 10
Concentration (L/kg)

0
0

x 10

c)

1
0
0

3
m

decrement of the on-line torque in a pectin solution when

the pectinase was added. Although this procedure involves
the on-line measure of the torque, it is in fact an off-line
procedure since the analysis is performed after the enzymatic
process has ended. Thus, the relation between juice yield and
the torque on-line monitoring is investigated here.
In order to effectively model the juice extraction process,
96 experiments were carried out (only for dynamical modeling purposes), at various enzymatic complex concentrations,
fruit ripeness stages, and the process temperatures but with
no additional water, since the optimization has shown that
this is not necessary. From these experiments, the maximum
juice yield was identified as a function of ripeness stage and
temperature. Then, for each temperature, ripeness stage, and
enzymatic complex concentration, the right-hand side of
Eq. 3 can be integrated as follows: ln(x1max x1)=mt. Then,
the juice yield rate m was determined by plotting the natural
logarithm of maximum juice yield minus the instant juice
yield (obtained from the off-line monitoring of the process)
versus time. The slope of the line is the kinetic rate m. Thus,
a specific value of m was obtained for each temperature,
enzymatic complex concentration, and ripeness stage.
Next, the equation mE kmmaxEE was used to model the
relation between the value of m and the enzymatic complex
concentration (E) at each temperature and ripeness stage. The
nonlinear fitting was performed in MatLab (R 2010b). The
observed deviations when plotting the predicted m values versus
the experimental data are graphically presented in Fig. 4, where it
is possible to appreciate the good accuracy of the prediction.

x 10
2
4
Concentration (L/kg)

Fig. 4 Plots show the nonlinear regression plots for kinetic of juice
yield by the enzymatic complex, the lines (. . .), (__), (-+-+), and (- - -)
stand for the model prediction at 20, 30, 40, and 50 C, respectively.

0
0

x 10
2
4
Concentration (L/kg)

The symbols (), (*), (), and () stand for experimental data at 20, 30,
40, and 50 C, respectively. Plots (a), (b), and (c) correspond to
ripeness stages 1, 2, and 3, respectively

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Table 2 Identified model parameters and functions

temperature was modeled using the following equation

1
Ae T . To perform the identification of A and , a linear fitting

using the natural logarithm of max versus the inverse of

temperature was performed for each of the three ripeness
stages. Figure 5 graphically demonstrates the accuracy of the
aforementioned procedure.
Then, the dependence of km, A, and values with respect
to the ripeness stages were calculated. To perform the identification of these parameters, a linear fitting using the
natural logarithm of A and versus the inverse of the
ripeness stages (expressed as 1, 2, and 3) was performed,
while a nonlinear correlation was needed for the case of the
and ripeness stage relation. Figure 5 graphically demonstrates the accuracy of these results.
Data presented in Table 2 summarizes the identified
model parameters values and their units. It is worth mentioning that the mathematical model has been formulated in
such a way as to allow the description of the juice production rate and its relation with the torque over the work range
of ripeness stages and temperatures.
At this point, two more correlations were needed in order to
fully model the processthe relation xmax(T,M) and y(T,M). In
both cases, the relations given in Fig. 6 were found.
Since bananas are a climacteric fruit, when harvested at the
preclimacteric matured green stage, the fruit undergoes
various physicochemical changes in terms of composition,
color, texture, aroma, and taste, pertaining to changes in
metabolic rates and biochemical reactions like respiration,
ripening, and senescence in the climacteric phase (Mohapatra
et al. 2011). In fact, such changes in the physicochemical
properties are the manifestation of various complex biochemical reactions (Zolfaghari et al. 2010). Thus, bananas can have
different uses in function of its maturity stage.
In order to correlate the fruits physicochemical characteristics (pulp-to-peel ratio, average of the data corresponding to the first 10 min of torque values and the Brix degree

-0.5

1/N m

L/kg

C
C
C

-5.5

1/T

Ripeness stage

-6
-6.5

0.03

0.04

c)

-6
-6.5
-7

0.05

1/T

-5

d)

-5.5

b)

-7

0.05

0.03

0.04

-10

f)

-20

-5.5

-6

0.05

1/T

e)

-1

-1.5
1

0.14T+1.5M(X2,0)+62.63
46T1,400M(X2,0)373.33
0.008325768
0.3308
0.234898
0.25541
9.1405
10.4698
38.422

Ln max

-6.5

0.04

xmax(T,x2,0)
y(T,x2,0)

Ln max

a)

0.03

Units

measured in the obtained juice) with the visual banana

ripeness stage (Fig. 1), a PCA analysis was performed.
The scatter plot for the first two principal components is
presented in the Fig. 7, where it is possible to appreciate an
overlap between the groups corresponding to the fruit ripeness stages visually classified according to Fig. 1. Similarly,
the ANOVA analysis indicates that there are no significant
differences between the nearest neighbors corresponding to
the mean values of first 10 min of the torque signal of each
ripeness stages and processing temperature, except in the
ripeness extremes; however, it is possible to appreciate a
clear gradient of change in the torque signal as the ripeness
stages advance.
Thus, to correlate the initial torque measurements with
the ripeness stages, a fuzzy logic approach is proposed that
makes it possible to correlate the visual criteria used to
classify the bananas with the initial online torque measurements. The fuzzy logic algorithm is based on the knowledge
acquired with the previous results. To infer the maturity
stage from on-line torque measurements M(X2,0), it is first
necessary to create the membership functions for the torque
measurements (input of the fuzzy logic model) and the

-6

-7

Value

Ln A

Ln max

-5.5

Ln Km

Fig. 5 The plots (a), (b), and

(c) graphically show the
correlation between the natural
logarithm of the experimental
values and inverse of
temperature (C) for each
ripeness stage. From left to
right, the results for ripeness
stages 1, 2, and 3 are shown.
Plots (d) and (e) correspond to
the correlation between the
natural logarithm of the
experimental and values and the
ripeness stage, while the plot (f)
corresponds to the second-order
polynomial used to model the
relation between and the
ripeness stage

Parameter

Ripeness stage

-30

Ripeness stage

Author's personal copy

Food Bioprocess Technol

75

0
y (T,M)

Xmax (T,M)

Fig. 6 From left to right, the

plots correspond to the linear
correlation between the
experimental values obtained at
different temperatures and
ripeness stages

70

65
50

40

30
Temperature (C)

20 3

Fig. 7 The plot corresponds to

the scatter plot for the first two
principal components of the
fruit physicochemical
characteristics matrix; green,
blue, and brown stand for
ripeness stages 1, 2, and 3,
respectively. The percentage of
the total variance explained by
each principal component is
indicated in brackets

Second Principal Component (18.2%)

membership functions for the maturity stage (output of the

fuzzy logic model) and name them. Each membership function is described in the form of a triplet (a, b, and c) as
represented in Fig. 2. The triplets corresponding to the input
and output sets as well as the group names are shown in
Table 3.
The fuzzy rules were set according to the understanding
of the behavior of the system. Since the relation between
torque and ripeness stage is a single-input single-output
relation, the following rules were used: If (input is Brown)
then (output is 3); If (input is Yellow) then (output is 2); If
(input is Green) then (output is 1).
In order to infer the juice yield initial condition from
online torque and temperature measurements, the membership functions for the error (input of the fuzzy logic controller) and the gains (output of the fuzzy logic controller) are
needed. The group names are shown in Table 3 and an
illustration of the functions is shown in Fig. 2.
Since the relation between torque and ripeness stage is a
multiple-input single-output relation, the following rules
were used: If (input is Brown), then (output is 3); If (input
is Yellow), then (output is 2); If (input is Green), then
(output is 1). The rules were stated as follows: If (Torque
is Brown) and (Temperature is 20 C), then (Output is 60); If
(Torque is Brown) and (Temperature is 30 C), then (Output
is 60); If (Torque is Brown) and (Temperature is 40 C),

2
Ripeness stage

-5000
-10000
50
40

Temperature (C)

30
20 3

1
2
Ripeness stage

then (Output is 65); If (Torque is Brown) and (Temperature

is 50 C), then (Output is 65); If (Torque is Yellow) and
(Temperature is 20 C), then (Output is 50); If (Torque is
Yellow) and (Temperature is 30 C), then (Output is 55); If
(Torque is Yellow) and (Temperature is 40 C), then (Output
is 60); If (Torque is Yellow) and (Temperature is 50 C),
then (Output is 60); If (Torque is Green) and (Temperature is
20 C), then (Output is 40); If (Torque is Green) and (Temperature is 30 C), then (Output is 40); If (Torque is Green)
and (Temperature is 40 C), then (Output is 45); If (Torque
is Green) and (Temperature is 50 C), then (Output is 45).
Remarks on Enzyme Activity and Temperature
As shown Fig. 8, the reaction rate decreases at 50 C in all
three stages of maturity. This apparently contradicts the
enzymatic complex data sheet, regarding the optimal temperature. Therefore, we sought to measure the protein concentration by Bradford both as banana juice in distilled
water at the same pH 5 (pH featuring banana juice) and
pH 3.5, which is recommended as optimal in the technical
sheet. The one-way ANOVA results of the Bradford
performed to the protein solutions have shown a significant
decrement in the concentration when the solution has the
same pH as the juice and 50 C. This can be due to a lack of
solubility, and this can explain decrement in the enzyme

2
1
0
-1
-2
-3

-2

-1
0
1
First Principal Component (71.1%)

Author's personal copy

Food Bioprocess Technol
Table 3 Values of the different membership functions used in the
fuzzy logic algorithm that relates the on-line torque measurements to
banana ripeness stage, i.e., M(X2,0), as well as the values of the
different membership functions used in the fuzzy logic algorithm that
infer the initial juice yield condition [x1,0 = x1(0)] from the on-line
measurements of temperature and torque
Set name

Torque
Brown
(0.008,0)
Yellow
(0.0135,0)
Green
(0.0195,0)
Ripeness stage
1
(0.5,0)
2
3
Temperature
20 C
30 C
40 C
50 C
Initial yield
40
45
50
55
60
65

Set type

(0.0113,1)
(0.0149,1)
(0.024,1)

(0.0145,0)
(0.0205,0)
(0.0285,0)

Input
Input
Input

(1,1)

(1.5,0)

Output

(1.4,0)
(2.4,0)

(2,1)
(3,1)

(2.45,0)
(3.6,0)

Output
Output

(14,0)
(24,0)
(34,0)
(44,0)

(20,1)
(30,1)
(40,1)
(50,1)

(26,0)
(36,0)
(46,0)
(56,0)

Input
Input
Input
Input

(36,0)
(41,0)
(46,0)
(51,0)
(56,0)
(61,0)

(40,1)
(45,1)
(50,1)
(55,1)
(60,1)
(65,1)

(44,0)
(49,0)
(54,0)
(59,0)
(64,0)
(69,0)

Output
Output
Output
Output
Output
Output

Colorimetric Analysis
The color of a product is one of the most important
properties that influence the consumers response to it.
Therefore, one of the objectives of this study was to
determine how the characteristic of the raw material and
the process conditions influence the final color attributes
of the banana juice. In order to allow the interpretation
the color results from the L.a.b. analysis, a PCA technique plus a clustering analysis were performed. The
results appear in Fig. 9.
Cluster analysis groups data objects with similar characteristics that were also made then different from objects of
other group. Therefore, the three clusters formed in Fig. 9
correspond to the banana juice with similar colorimetric components. The sensorial panelists were asked to categorize the
samples corresponding to each attribute of all samples. From
the PCA score, the cluster analysis, and opinions of the
panelists, it can be concluded that the clusters of green diamonds obtained in Fig. 9 are the ones with the best visual
attributes. This group corresponds to ripeness stage 1 (green)
and with no influence of the temperature or the enzyme
concentration; however, it is important to mention that the
closest neighbors were samples of the blue star group corresponding to ripeness stage 2 (yellow) and produced at temperatures of 40 and 50 C.
The Software Sensor

Set type refers to the nature of the set that is an input and/or output to
the fuzzy logic algorithm

activity. It is known that the tridimensional structure of proteins

is sensitive to minor changes in factors in the environment,
including pH, temperature, and medium composition, and such
structure changes could alter the catalysis activity of the proteins (Srivatsa 1996).

On-line availability of the juice yield measurement is very

important for the control and particularly for the process of
supervision and fault detection; however, there exists no
device that provides an on-line juice yield measurement.
One way to overcome this problem is to use software
sensors to estimate missing state variables on-line. As it
is described in the Methodology section, a robust observer
was constructed. In order to provide the observer with
robust properties, the following representation of the system
(Eqs. 3 and 4) is proposed:


x0 x2


x1 mxmax x1

m
xmax x1
Y

x2

Fig. 8 The plot corresponds to the correlation between the kinetic rate
and temperature in the banana juice extraction process. Green squares,
blue triangles, and red circles correspond to ripeness stages 1, 2, and 3,
with and R2 of 0.885, 0.998, and 0.992, respectively

y0 x0

where x0 is the dynamical extension that allows us to

integrate the noisy signal in order to recover a filtered

Author's personal copy

Second Principal Component (21.4%)

Food Bioprocess Technol

2
G-30-100L

-1

G-40-100L

G-50-100L

G-50-300L
G-30-500L
G-20-300L
G-40-300L

B-20-500L
Y-30-300L
Y-50-300L
Y-20-500L
Y-30-500L
B-20-300L
B-40-100L
G-30-300L
Y-20-500L
B-20-100L
Y-50-500L
Y-20-300L
Y-30-100L
G-50-500L
Y-20-100L
Y-40-500L
B-30-500L
B-30-100L
Y-40-100L
B-30-300L
B-40-500L
Y-40-300L
B-50-300L
B-50-500L
B-50-100L
B-40-300L
Y-50-100L

-2
-3

-2

-1

First Principal Component (74.2%)

Fig. 9 Scatter plot for the first two principal components with colored
clusters corresponding to the matrix of juice color (L, a, and b) at
different process conditions: temperatures, enzymatic complex concentration, and ripening stages (the three generated clusters are marked as
red triangles, blue five-pointed stars, and green diamonds). The letters

B, Y, and G stand for brown, yellow, and green, corresponding to

ripeness stages 1, 2, and 3, respectively. The percentage of the total
variance explained by each principal component is indicated in
brackets

signal. Thus, the task becomes the estimation of this

new state Xbi (a standard task for an observer).

definite matrix that do not depend on . Thus, St,j are

such that S is a positive solution of the algebraic
Riccati equation,

 


S A I A I S C T C:
2
2

X AX B
y CX
Where
2
3
2
3
0
0 1

A 4 0 m
0 5; B 4 m xmax 5and C 1 0 0 :
0 m=Y 0
m xmax =Y
An asymptotic-type observer of the system is given as
follows:



c
b 2 k 1 y0 X
b0
X0 X






b
b 1 k 2 y0 X
b0
X 1 m xmax X





m
b
b 1 k 3 y0 X
b0
X max X
X2
Y

10

11

where the gain vector of the observer is given by:

T
K S1
C 
S i; j
S i; j
i j1

Each entry of the matrix S is given by the above

equation, where S is an n n matrix (i and j run from
1 to n) and St,j are entries of a symmetric positive

In particular, for our case, the resulting vector K is given

by:
2 3 "
#
32m
k1
2

2m
Y
K 4 k2 5
m
2m3m
k3
It is worth mentioning that we can think of this observer
as a slave system that follows the master system, which
is precisely the real experimental system. In addition, S, as
functional components of the gain vector, guarantee the
accurate estimation of the observer through the convergence
to zero of the error dynamics, i.e., the dynamics of the
difference between the measured state and its corresponding
estimated state. One can see that generates an extra degree
of freedom that can be tuned by the user such that the
performance of the software sensor becomes satisfactory
for him (here, we use = 0.01).
The reason for the filtering effect is that the dynamic
extension acts at the level of the observer as an integration of the output of the original system (see the first
equation of the system given by Eqs. 58 and the error
part in the equations of system given by Eqs. 911).
The integration has averaging effects upon the noisy
measured states. More exactly, the difference between
the integral of the output of the slave part of system
and the integral of the output of the original system

Author's personal copy

a)

0.025

Torque (Nm)

Torque (Nm)

Food Bioprocess Technol

0.02
0.015
0.01
0

500

1000

1500

2000

2500

3000

3500

4000

0.025

b)

0.02
0.015
0.01
0

4500

500

1000

1500

2000

Juice Yield (%)

Juice Yield (%)

3000

3500

4000

4500

3000

3500

4000

4500

80

80
60
40
20
0

2500

Time(s)

Time(s)

500

1000

1500

2000

2500

3000

3500

4000

4500

60
40
20
0

500

1000

1500

2000

2500

Time(s)

Time(s)

Torque (Nm)

Torque (Nm)

0.02

c)

0.02
0.015
0.01
0

500

1000

1500

2000

2500

3000

3500

4000

d)
0.015

0.01
0

4500

500

1000

1500

Juice Yield (%)

Juice Yield (%)

80
60
40
0

500

1000

1500

2000

2500

3000

3500

4000

4500

1000

1500

2000

2500

4000

4500

3000

3500

4000

4500

60
40
0

500

1000

1500

3000

3500

4000

2000

2500

f)

0.014
0.012
0.01
0

4500

500

1000

1500

2000

2500

3000

3500

4000

4500

3000

3500

4000

4500

Time (s)

80

80
Juice Yield (%)

Juice Yield (%)

3500

0.016

Time (s)

60
40
0

3000

Time (s)

Torque (Nm)

Torque (Nm)

e)
500

2500

80

Time (s)

0.016
0.014
0.012
0.01
0

2000

Time (s)

Time (s)

500

1000

1500

2000

2500

3000

3500

4000

4500

Time (s)

60
40
0

500

1000

1500

2000

2500

Time (s)

Fig. 10 The plots show the application of the software sensor to

different experimental situations: in all cases, red solid lines represent
the filtered states and juice yield prediction, while blue lines represent
the noisy measured torque signal, and blue points represent offline
juice yield quantification. In all cases, the enzymatic complex was

added to the 1,320 s (22 min). Ripeness stage 1 at 20 C (a) and at

30 C (b) both with 100 L of enzyme complex. Ripeness stage 2 at
30 C (c) with 500 L of enzyme complex and at 50 C (d) with
300 L of enzyme complex. Ripeness stage 3 at 30 C (e) with 300 l
of enzyme complex and at 20 C (f) with 300 l of enzyme complex

gives the error, and the observer is planned in such a

way that the error dynamics go asymptotically to zero,
which results in the recovering of both the filtered state
and the unmeasured states.
From Fig. 10, it is possible to appreciate that the
algorithm developed here is able to infer very accurately
the instant juice yield only based on the online measures of torque and temperature. Since the algorithm
does not need to be fed with the ripeness stage, it can
be also used to confirm the adequate selection of banana ripeness. Additionally, from Fig. 10, it is possible
to see that the torque measurements by themselves are
not enough to detect with precision the end of the
process, while the prediction of juice yield could be
the criterion to stop the enzymatic process and continue
the next downstream step. Thus, the software sensor
developed here effectively rebuilds the unmeasured online juice yield and also is able to filter the noisy torque
signal under different experimental situations.

Conclusions
The optimization of the banana juice extraction process
shows that the addition of water did not significantly affect
the juice yield, while the enzymatic complex concentration
exhibits a positive impact on the juice yield. As such, the
optimal conditions for juice extraction (i.e., more sustainable) were 150 L/kg enzyme complex concentration without water addition. Moreover, from this study, it can be
concluded that the model here developed could be a valuable instrument for monitoring and controlling enzymatic
banana juice extraction. Furthermore, enzyme reactors can
operate fed-batch-wise or continuously, and the model is a
helpful tool for optimization. Moreover, the software sensor
developed here effectively rebuilds the unmeasured on-line
juice yield and also is able to filter the noisy torque signal.
In addition, the computational scheme provides a very appropriate tool for fast and reliable quality control and can be
used to ensure the homogeneity of the final product. The

Author's personal copy

Food Bioprocess Technol

methodology presented in this paper is general and can be

used also in automatic control applications. This supervision
scheme provides instantaneous access to batch records, and
provides audit tracking and traceability. This tool presents
valuable characteristics like real-time usage, ease of use, no
need for sample preparation, and no sample destruction. In
addition, PCA proves to be a valuable tool to relate the best
visual attributes of the banana juice with the maturity fruit
ripeness stage selection and the operation conditions, showing that the juice processing temperature presents positive
effects on juice color attributes.
Acknowledgments This work was supported by the CONACyT,
Mxico (No. 169048) and to PROMEP-SEP by funding the project.
The authors would like to thank the anonymous reviewers for their
valuable comments and suggestions to improve the document.

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