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th

European Symposium on Computer Aided Process Engineering ESCAPE20

S. Pierucci and G. Buzzi Ferraris (Editors)

2010 Elsevier B.V. All rights reserved.

A hybrid neural approach to model batch

fermentation of dairy industry wastes

Alessandra Saraceno, Sascha Sansonetti, Stefano Curcio, Vincenza Calabr,

Gabriele Iorio

Department of Enginering Modeling-University of Calabria, via P. Bucci-cubo 42/A,

Arcavacata di Rende 87036, Italy, alessandra.saraceno@unical.it

Abstract

In this work, the fermentation of Ricotta cheese whey for the production of ethanol

was simulated by means of a Hybrid Neural Model (HNM), obtained by coupling neural

network approach to mass balance equations describing the time evolution of lactose

(substrate), ethanol (product) and biomass concentrations. The realized HNM was

compared with a pure neural network model (NM) and the advantages gained from the

hybrid approach were emphasized. The experimental data, necessary to develop the

model, were collected during batch fermentation runs. For all the proposed networks,

the inputs were chosen as the operating variables exhibiting the highest influence on the

reaction rate. The simulation results showed that the HNM was capable of an accurate

representation of system behavior by predicting biomass, lactose and ethanol

concentration profiles with an average error percentage lower than 10%. Moreover,

especially if compared with the NM, the HNM showed good forecasting capability even

with fermentation run never seen during the training phase.

Keywords: grey-box models, artificial neural networks, batch fermentation, modeling

1. Introduction

Modeling of biotechnological processes represents a key issue to achieve proper design

and control aimed at process optimization (Lubbert and Jorgensen, 2001). Biochemical

reactions actually involve many parallel-serial reaction steps and depend on several

transport phenomena that may limit the observed reaction rates. Usually, the

fundamental approach, largely considered as the most rigorous, cannot be applied in

reaction kinetic modeling due to inherent non-linearity, lack of information,

experimental inaccuracy, deviations from ideal conditions (Feyo de Azevedo et al.,

1997). On the contrary, black-box models, suggested by the empirical approach, suffer

from a restricted validity domain depending on the range of data collected during the

experiments. A reasonable trade-off between theoretical and empirical approach is

represented by hybrid modeling, leading to a grey-box model capable of good

performance in terms of data interpolation and extrapolation. The main advantage of

hybrid modeling regards the possibility of describing some well-assessed phenomena by

means of a fundamental theoretical approach, leaving the analysis of other aspects, very

difficult to interpret and describe in a traditional way, to rather simple cause-effect

models. Among these, Artificial Neural Networks (ANNs) were successfully used in

bioreactor modeling (James at al., 2002; Laursen et al., 2007; Simutis et al., 1995)

ANNs consist of interconnected computational elements called neurons or nodes: each

A.Saraceno et al.

neuron receives input signals from the related units, elaborates these stimuli by an

activation or transfer function and generates an output signal that can be transferred to

other neurons. ANN can be used both in a pure empirical model structure and in a

hybrid model formulation. When ANNs are utilized in a hybrid model, a very important

step is identifying the respective domain of theoretical and empirical parts in such a way

as to decide which aspects of the process are to be described by a black-box model.

Ricotta cheese whey (RCW),scotta, is a dairy industry waste characterized by a

lactose content of about 45-50 g/l and a very low amount of proteins ranging from 0.22

0.24 % in weight. BOD reduction of scotta can be achieved, for instance, by

alcoholic fermentation of lactose carried out by Kluyveromyces Marxianus. Even if

several authors dealt with biotechnological utilization of cheese whey (Gonzles Siso,

1996; Ozmihci and Kargi, 2006, 2007), as far as our knowledge is concerned, a single

work exists about Ricotta Cheese whey(Sansonetti et al., 2009).

The aim of this paper was to propose a model that could describe batch fermentation of

lactose in ethanol investigating two different strategies. At first, the concentration

profiles of the reacting species were described by a pure neural model; afterwards, with

the aim of introducing theoretical information (mass balance) into the model, a multiple

hybrid neural model was realized.

2. Development of the models

The proposed models, i.e. NM and HNM, are aimed at the description of the batch

fermentation of RCW predicting biomass, lactose (substrate) and ethanol (product)

concentration profiles. A preliminary phase of the model development was performed.

It consisted on the identification of the operating variables that, among all the

parameters that could affect reaction progress, exhibited the highest influence on

process performance. These variables, i.e.: temperature (T), pH (pH), reactor stirring

rate (rpm), initial lactose concentration (C

0

lat

) and reaction time (t), represented the

inputs for both the pure neural and the hybrid neural model.

2.1 Neural model structure

A multiple neural model was proposed to predict concentration profiles. It consisted of

three neural networks, i.e. NM1, NM2 and NM3, respectively capable of predicting

biomass, lactose and ethanol concentration profiles as outputs. The structure of the three

neural networks was very similar: they were characterized by the same number of input

nodes, corresponding to the pre-identified input variables and by a single output node

that had to produce, as output, the corresponding concentration profile. Between input

and output nodes two hidden layers were put and the number of nodes in each layer was

determined according to an iterative procedure.

2.2 Hybrid model structure

The realized hybrid neural model consisted of three models characterized by a common

structure. A mass balance equation was written for each of the compounds and an ANN

was set up to evaluate the kinetic parameters actually necessary to solve the balance

equations. Some logic conditions, expressing the fulfilment of physical constraints,

were also introduced into the model.

The three mass balance equations, written with reference to biomass, lactose and

ethanol assumed the following form:

A hybrid neural approach to model batch fermentation of dairy industry

=

=

=

X

dt

dP

qX

dt

dS

X

dt

dX

P

(1)

where X, S, P were, respectively, biomass, lactose and ethanol concentration profiles,

was biomass specific growth rate, q was the lactose consumption rate function,

p

was

the product growth rate and t was the reaction time. Eqs. (1) were approximated by

Eulers discretization and the discretized form was used recursively to determine the

biomass, lactose and ethanol concentration values at the time instant t+t, knowing all

the information concerning time instant t. The values of kinetic parameters at time

instant t, strictly necessary to solve the discretized form of the equations, were provided

by three neural networks named HNM1, HNM2 and HNM3, respectively predicting ,

q and

p.

The three neural networks were set up according to an iterative procedure. The

kinetic parameters, evaluated using HNM1, HNM2 and HNM3, were subsequently

processed by some logic conditions that verified the agreement between the net outputs

and some well-assessed process information. The logic condition assumed the following

form:

= <

= <

= <

0 0

0 0

0 0

p p

if

q q if

if

(2)

Eqs. (2) faced the fact that, during the fermentation runs, biomass and ethanol

concentrations always grew, whereas lactose concentration always decreased.

3. Materials and methods

3.1. Experimental design

The experimental data necessary to develop the models were collected from a set of

anaerobic fermentations performed on RCW. According to the factorial design method

(Box et al., 1978), the following variables and operating condition were chosen: 1)

Temperature in the range 32- 40 C; 2) pH in the range 4-6; 3) Stirring rate in the range

100-300 rpm; 4) The lactose concentration was varied between 50-90 g/l since this is

the expected range in which the process could be operated. The lowest value of the

range was set to 50 g/l due to the typical lactose concentration of RCW, i.e. 45-50 g/l.

The highest value of the range, i.e. 90 g/l, derived from the consideration that it could be

useful to concentrate RCW before the fermentation process so to achieve a more

concentrated ethanol solution flowing out the reactor. Nevertheless this value cannot be

increased as much as one likes since a limit imposed by the osmotic pressure at the cell

wall does actually exist. The value of 90 g/l value was chosen according to several

preliminary experiments (not reported in the paper), which showed the technical and

economical feasibility of RCW pre-concentration (up to 90 g/l) by nanofiltration and,

therefore, by a well-assessed procedure.

A total of 16 batch runs, having a duration of 18 hours with a sampling time of one

hour, were therefore performed and a total of 912 experimental points, expressing the

A.Saraceno et al.

time evolutions of biomass, lactose and ethanol concentration, were collected during

fermentation.

3.2. Neural network development

To realize both the NMs and the HNMs, six different neural networks were developed

following a common iterative procedure implemented according to Matlab Neural

Network Toolbox Ver. 4.0.1. For the neural model only the concentration values

relative to the batch fermentation runs were sufficient to train the networks. For the

hybrid model development it was necessary to determine the experimental values of ,

q,

p

by interpolating the collected experimental data by cubic splines. Subsequently,

the experimental points relative to 15 of the 16 available runs were used to train/test the

developed networks: the data points were randomly split into two groups, reserving 2/3

of data (570 points) to the training phase and remaining 1/3 (285 points) to test the

neural networks. Finally, the six realized neural models were validated using two

complete experiments never exploited during both the previous training/test phases. The

neuron transfer function was always the hyperbolic tangent, except for the output layer

where a linear transfer function was used. To develop the ANNs, an iterative procedure

was implemented. The performance index, chosen to evaluate simulation results

reliability, was the percentage error between each predicted (C

p,k

) and measured (C

m,k

)

concentration value: in a whole batch run the average value of the percentage error had

to be lower than 10%. The iterative procedure consisted in placing a layer with a single

neuron between the input and output layers, thus verifying the fulfillment of

convergence criterion; if at the end of the training procedure the performance objective

was not achieved, a neuron was added into the hidden layer, repeating the procedure

until the performance objective was achieved. If the neurons of the 1

st

hidden layer

reached the number of 10, an additional hidden layer with a single neuron was put into

the network and, the procedure described for the first hidden layer iterated.

4. Results and discussion

On the basis of the previous discussion, a multiple neural model and a multiple hybrid

neural model were realized. Table 1 summarizes the architecture of all the developed

networks.

Table 1. ANNs architecture

Neural Network

Number of neurons

1

st

hidden layer

Number of neurons

2

nd

hidden layer

Number of neurons

output layer

NM1 8 5 1

NM2 10 10 1

NM3 10 4 1

HNM1 6 5 1

HNM2 10 3 1

HNM3 10 2 1

A comparison between models predictions and experimental data is reported in Fig.1.

Fig. 1 reports the simulation results relative to a batch run used during training/test

phase of the models. A remarkable agreement between model predictions and the

corresponding experimental data was found, both with the pure neural and the hybrid-

neural approach so that an average percentage error much lower than 10% was found.

A hybrid neural approach to model batch fermentation of dairy industry

Figure 1. Predictions during training/test. Run conditions: T=32C, pH=6, rpm=100, C

0

lat

=90 g/l

0.00

30.00

60.00

90.00

0 2 4 6 8 10 12 14 16 18

time [h]

c

o

n

c

e

n

t

r

a

t

i

o

n

[

g

/

l

]

biomass (experimental) lactose (experimental) ethanol (experimental)

biomass (NM1) lactose (NM2) ethanol (NM3)

biomass (HNM1) lactose (HNM2) ethanol (HNM3)

Moreover, it should be observed that even if models performance is very similar, NMs

were more accurate in predicting the system dynamics with the experimental point

reserved to training/test phase. This result emphasizes how neural networks, being data-

based models, are able to perform very well on training data. Indeed, the training phase

of a neural network consists in structuring it so as to predict the training points

extremely well. Fig. 2. shows two different validation tests aimed at comparing hybrid

neural model and pure neural model predictions under a combination of operating

conditions never exploited during the training and the test phases, even if included in

the same range chosen to perform the experimental design. From Fig. 2 it is possible to

appreciate how NMs and HNMs validation results were quite different. The hybrid

model was capable of giving good simulation results even during validation: the model

performance, measured in terms of %, is comparable to that obtained with the

experimental data used to perform the training and the test phases.

Figure 2. Models validation. Run conditions: a) T=32C, pH=4, rpm=100, C

0

lat

=50 g/l; b)

T=37C, pH=5, rpm=300, C

0

lat

=50 g/l

0

30

60

0 4 8 12 16

a) time [h]

c

o

n

c

e

n

t

r

a

t

i

o

n

[

g

/

l

]

0

30

60

0 2 4 6 8

b) time [h]

c

o

n

c

e

n

t

r

a

t

i

o

n

[

g

/

l

]

This result shows how hybrid models do not just learn in order to recognize the training

points; instead, the models are able to predict the system behaviour even when they are

operated in conditions unexploited during training phase. On the contrary, neural

A.Saraceno et al.

models, during the validation, give worse performance that those obtained during the

training phase. Indeed, even if the neural models are able to predict smoothly the system

behaviour, the error percentage is significantly higher than 10% thus confirming how

pure black-box models are very reliable into their definition domain whereas they

become less accurate when used in interpolation and extrapolation. Therefore, the

results obtained in this paper, indicate how the introduction of theoretical information

into black-box models can lead to a more robust modeling approach.

5. Conclusions

The purpose of this work was to make a comparison between a pure neural and a hybrid

neural approach in modeling the batch fermentation of RCW. The hybrid modeling

approach showed better forecasting capability than the pure neural model, being capable

to predict the system behavior even in operating conditions never exploited during the

training phase. On the other hand, NMs did not show the same reliability during the

model validation thus stating how the introduction of simple theoretical information into

black-box models can lead to a strong improvement in model performance. As a matter

of fact, HNMs turned out to be a very efficient tool for the simulation of the RCW

fermentation process and, more generally, it could be used in modeling several complex

biotechnological processes. Overcoming the difficulties to describe very complex

reaction mechanism, HNMs are able to exploit all the available theoretical knowledge

all the same. This modeling effort can lead to more robust models than empirical

models and this feature make the hybrid neural approach a promising alternative in

process engineering because most of the advanced control and optimization strategies

are based on mathematical models.

References

G. Box, W. Hunter and S.Hunter , 1978, Statistics for Experimenters, An Introduction to Design,

Data Analysis and Model Building, John Wiley and Sons, New York.

S. Feyo de Azevedo, B. Dahm and F.R. Oliveira, 1997, Hybrid modelling of Biochemical

Processes: A comparison with the conventional approach., Comput. Chem. Eng., 21,751.

M. I. Gonzles Siso, 1996, The biotechnological utilization of cheese whey: a review, Bioresour.

Technol., 57, 1.

S. James, R. Legge and H.Budman, 2002, Comparative study of black-box and hybrid estimation

methods in fed-batch fermentation, J. Process Control, 12, 113.

S. O.Laursen, ,D. Webb and W. F. Ramirez, 2007, Dynamic HNM of an industrial fed-batch

fermentation process to produce foreign protein, Comput. Chem. Eng., 31, 163.

A. Lubbert and S. B. Jorgensen, 2001, Bioreactor performance: a more scientific approach for

practice, J. Biotechnol., 85, 187.

S. Ozmihci and F. Kargi, 2006,Utilization of cheese whey powder for ethanol fermentation:

effects of operating parameters, Enzyme and Microbial Technol., 38, 711.

S. Ozmihci and F. Kargi, 2007, Kinetics of batch ethanol fermentation of (CWP) solution as a

function of substrate and yeast concentrations, Bioresour. Technol., 98, 2978.

S. Sansonetti, S. Curcio, V. Calabr and G. Iorio, 2009, Bio-ethanol production by fermentation

of Ricotta Cheese Whey as an effective alternative non-vegetable source, Biomass and

Bioenerg., 12, 1692.

R. Simutis, M. Dors and A. Lubbert, 1995, Bioprocess optimization and control: Application of

hybrid modelling, J. Biotechnol., 42, 285.

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