international journal of hydrogen energy 34 (2009) 12531259

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Application of desirability function based on neural network

for optimizing biohydrogen production process

Jianlong Wang*, Wei Wan

Laboratory of Environmental Technology, INET, Tsinghua University, Tsinghua Yuan, Beijing 100084, PR China

article info abstract

Article history:
Received 6 October 2008
Received in revised form
20 November 2008
Accepted 20 November 2008
Available online 23 December 2008
Keywords:
Neural network
Genetic algorithm
Method of desirability function
Biohydrogen production
Modified logistic model
A fractional factorial design was carried out to investigate the effects of temperature, initial
pH and glucose concentration on fermentative hydrogen production by mixed cultures in
batch tests and then the experimental data of substrate degradation efficiency, hydrogen
yield and average hydrogen production rate were described by a neural network, based on
which the simultaneous optimization of the three responses was performed by the method
of desirability function. The analysis showed that the neural network could successfully
describe the effects of temperature, initial pH and glucose concentration on the substrate
degradation efficiency, hydrogen yield and average hydrogen production rate of this study.
The maximum substrate degradation efficiency of 95.3%, hydrogen yield of 305.3 mL/g
glucose and average hydrogen production rate of 23.9 mL/h were all obtained at the
optimal temperature of 39.0  C, initial pH of 7.0 and glucose concentration of 24.6 g/L
identified by the method of desirability function based on a neural network. In sum, the
method of desirability function based on a neural network was a useful tool to optimize
several responses for fermentative hydrogen production processes simultaneously.
2008 International Association for Hydrogen Energy. Published by Elsevier Ltd. All rights
reserved.

1. Introduction
Environmental pollution due to the use of fossil fuels as well
as their shortfall makes it necessary to find alternative energy
sources that are environmentally friendly and renewable.
Hydrogen satisfies the above requirements because it
produces only water, when it is combusted as a fuel or con-
verted to electricity [1,2]. Among various hydrogen production
processes, the biological method is known to be less energy
intensive, for it can be carried out at ambient temperature and
pressure. The biological method mainly includes photosyn-
thetic hydrogen production and fermentative hydrogen
production. The efficiency of photosynthetic hydrogen
production is low and it cannot be operated in the absence of
light, while fermentative hydrogen production can produce
hydrogen all day long without light using various kinds of
substrates such as organic wastes [3,4]. Thus fermentative
hydrogen production is more feasible and widely used. In
addition, it is of great significance to produce hydrogen from
organic wastes by fermentative hydrogen production, because
it plays the dual role of waste reduction and energy produc-
tion. Therefore fermentative hydrogen production has been
receiving increasing attention in recent years [5].
Many factors such as temperature, initial pH and substrate
concentration can influence the fermentative hydrogen
production, because they can affect the activity of hydrogen-
producing bacteria by influencing the activity of some essen-
tial enzymes such as hydrogenases for fermentative hydrogen
production [1]. It has been demonstrated that in an appro-
priate range, increasing temperature, initial pH and substrate

* Corresponding author. Tel.: 86 10 6278 4843; fax: 86 10 6277 1150.

E-mail address: wangjl@tsinghua.edu.cn (J. Wang).
0360-3199/$ see front matter 2008 International Association for Hydrogen Energy. Published by Elsevier Ltd. All rights reserved.
doi:10.1016/j.ijhydene.2008.11.055
1254 international journal of hydrogen energy 34 (2009) 12531259
concentration could increase the ability of hydrogen-
producing bacteria to produce hydrogen during fermentative
hydrogen production, but temperature, initial pH and
substrate concentration at much higher levels could decrease
it with increasing levels [510]. Thus, appropriate levels of the
above three factors can be chosen to optimize the fermenta-
tive hydrogen production process to increase the ability of
hydrogen-producing bacteria to produce hydrogen.
There have been a great number of studies reporting the
optimization of fermentative hydrogen production process
with one-factor-at-a-time experimental design, but such
design does not depict the interactive effects among different
factors. On the contrary, response surface methodology based
on a factorial design can depict the interactive effects among
different factors and allows the effects of many factors with
two or more levels on a response to be studied in a relatively
small number of runs [1114]. In recent years, a neural
network which is able to depict the interactive effects among
different factors in a complicated bioprocess has been applied
successfully in multivariate non-linear bioprocesses as
a useful tool to construct models. It has been shown that
a neural network is a superior and a more accurate modeling
technique compared with the response surface methodology,
as it represents the non-linearities in a much better way
[1517]. A neural network can be considered as the objective
function for optimization by genetic algorithm, which has
established itself as a powerful search and optimization
technique to solve problems with objective functions that are
not continuous or differentiable. It has been reported that the
genetic algorithm based on a neural network had a higher
optimizing ability than response surface methodology [1517].
Moreover, the method of desirability function can be used to
optimize several responses simultaneously when there are
many responses to be optimized for a bioprocess. Otherwise,
without multiple-response optimization, several responses
would have to be optimized separately [1820]. Simultaneous
optimization of multiple responses by the method of desir-
ability function involves first building an appropriate model
for each response and then trying to find a set of operating
conditions that in some sense optimizes all responses or at
least keeps them within desired ranges.
To the best of our knowledge, however, there is little
literature reporting the optimization of fermentative
hydrogen production processes by the method of desirability
function based on a neural network using a genetic algorithm.
Thus, in this study the effects of temperature, initial pH and
glucose concentration on fermentative hydrogen production
by mixed cultures were investigated in batch tests and
described by a neural network, and then the process was
optimized by the method of desirability function based on the
neural network, with the purpose of obtaining the optimal
conditions for the fermentative hydrogen production process.
2. Materials and methods
2.1. Seed sludge
The digested sludge collected from a primary anaerobic
digester at Beijing Gaobeidian Sewage Treatment Plant (China)
was used as the seed sludge. Heat-shock was used in this
study to enrich hydrogen-producing bacteria by heating the
seed sludge at 100  C for 15 min [1,2].
2.2. Experimental design and procedures
In this study, a three-factor fractional factorial design was
used to design the experiment for constructing the neural
network. Temperature, initial pH and glucose concentration
were chosen as the factors, while substrate degradation effi-
ciency, hydrogen yield and average hydrogen production rate
were chosen as the responses. The experimental design is
shown in Table 1.
According to the experimental design in Table 1, batch
tests were conducted in 150 mL glass bottles. One liter of the
nutrient solution contained NaHCO3, 40,000 mg; NH4Cl,
5000 mg; NaH2PO4 $ 2H2O, 5000 mg; K2HPO4 $ 3H2O, 5000 mg;
FeSO4 $ 7H2O, 15,000 mg; MgCl2 $ 6H2O, 85 mg; NiCl2 $ 6H2O,
4 mg. Fifteen milliliters of the pretreated seed sludge, 10 mL of
nutrient solution and certain amount of glucose were added to
each glass bottle. Then the total working volume of the bottles
was filled to 100 mL by de-ionized water. The initial pH of the
mixed solution in each bottle was adjusted by 1 mol/L HCl or
1 mol/L NaOH. Each bottle was flushed with argon for 3 min to
provide anaerobic condition, capped with a rubber stopper,
and placed in a reciprocal shaker (reciprocation: 150 strokes/
min) at different temperature. Each batch test was done three-
fold.
2.3. Analytical methods
The water displacement method was used to collect and
measure the biogas produced. The fraction of hydrogen in the
biogas was determined by a gas chromatograph (Model 122,
Shanghai, China) equipped with a thermal conductivity
detector (TCD) and a 2 m column packed with 5 A molecular
sieves. Helium was used as the carrying gas at the flow rate of
12 mL/min. The operating temperature of the column,
detector and injector were 40  C, 80  C and 50  C, respectively.
All gas production data reported were standardized to the
standard temperature (0  C) and pressure (760 mmHg). The pH
in the solution was measured by a pH meter (Model 526,
Germany). The concentration of glucose after reaction was
determined by the DNS colorimetric method [21]. The volatile
suspended solid (VSS) of the seed sludge was determined
according to the standard methods [22].
In this study, using non-linear regression with the software
Origin 7.5, the modified Logistic model (Eq. (1)) was used to fit
the cumulative hydrogen production data obtained from each
batch test to obtain H, Rm and l [5,23].
P
H (1)
1 exp4Rm ,l  t=P 2
where H (mL) is the cumulative hydrogen production at the
reaction time t (h), P (mL) is the hydrogen production poten-
tial, Rm (mL/h) is the maximum hydrogen production rate and
l (h) is the lag time.
Once the three parameters were obtained, Eq. (2) was used
to calculate the average hydrogen production rate in each
batch test.
 international journal of hydrogen energy 34 (2009) 12531259 1255

Table 1 Experimental design for constructing neural network.

Temperature ( C) Initial pH Glucose Substrate degradation Hydrogen yield Average hydrogen
concentration (g/L) efficiency (%) (mL/g glucose) production rate (mL/h)

30.0 6.0 10.0 70.5 236.8 8.4

30.0 7.0 10.0 80.7 262.0 9.8
30.0 8.0 10.0 75.8 224.7 9.1
30.0 9.0 10.0 52.5 136.5 6.3
31.6 7.0 25.0 79.0 131.9 12.3
35.0 6.0 10.0 82.6 278.8 9.7
35.0 6.0 20.0 74.8 123.1 12.1
35.0 6.0 30.0 83.0 71.6 12.2
35.0 8.0 10.0 83.6 264.6 10.5
35.0 8.0 20.0 75.0 200.9 15.6
35.0 8.0 30.0 83.0 169.3 18.9
35.0 9.0 10.0 64.6 160.7 7.3
40.0 5.3 25.0 79.6 49.0 8.3
40.0 6.0 10.0 80.4 277.9 9.8
40.0 7.0 16.6 70.5 195.0 9.6
40.0 7.0 25.0 95.2 282.3 23.6
40.0 7.0 33.4 84.4 158.6 16.5
40.0 8.0 10.0 87.5 263.7 10.7
40.0 8.7 25.0 80.5 145.4 9.5
40.0 9.0 10.0 60.2 160.2 7.4
45.0 6.0 10.0 71.0 228.2 8.3
45.0 6.0 20.0 75.7 75.1 5.8
45.0 6.0 30.0 81.7 37.7 4.5
45.0 7.0 10.0 80.0 252.5 9.7
45.0 8.0 10.0 72.2 216.5 9.0
45.0 8.0 20.0 73.6 94.8 5.7
45.0 8.0 30.0 78.2 40.5 5.3
45.0 9.0 10.0 52.3 131.5 6.3
48.4 7.0 25.0 79.8 30.1 3.5

P Xmax  Xmin X  0:1

R (2) X Xmin (4)
l P=Rm 0:8

The hydrogen yield was calculated by dividing the hydrogen
production potential by the amount of glucose consumed in
each batch test. The substrate degradation efficiency was
estimated by dividing the amount of glucose consumed by the
amount of initial glucose.
2.4. Neural network
In general, a feed-forward neural network with one hidden
layer containing certain hidden neurons has been shown to be
capable of providing accurate approximations to many non-
linear functions [17]. Thus, a feed-forward neural network
with back propagation algorithm was used in this study. The
inputs chosen in this study were temperature, initial pH and
glucose concentration, respectively, while the outputs were
substrate degradation efficiency, hydrogen yield and average
hydrogen production rate, respectively. In this study, the
transfer function in the hidden layer and the output layer
were logistic function and linear function, respectively. All the
inputs and outputs were normalized within a uniform range
of (0.1, 0.9) to ensure that they received equal attention during
the training process [16,24]. The new scaled variables were
calculated by Eq. (3). These values were rescaled by Eq. (4).
X  Xmin
X 0:8  0:1 (3)
Xmax  Xmin
where X is the variable in a group to be scaled, Xmin is the
minimum variable in a group to be scaled, Xmax is the maximum
variable in a group to be scaled, and X* is the scaled variable.
The first step in training the neural network is to design the
topology of the neural network. The number of neurons in
the input layer is fixed by the number of inputs, whereas the
number of neurons in the output layer equals the number of
outputs. The critical aspect is the choice of the number of
neurons in the hidden layers [24]. To obtain the optimal
number of neurons in the hidden layer of the neural network,
the number of neurons in the hidden layer was investigated. In
the training process, the mean square error between the
experimental data and the corresponding predicted data is
calculated and propagated backward through the network.
The back propagation algorithm adjusts the weights in each
successive layer to reduce the error. This procedure is repeated
until the error between the experimental data and the corre-
sponding predicted data satisfies certain error criteria.
Eq. (5) was used to calculate the coefficient of determina-
tion (R2) of the neural network.
Pk  0 2
2 i1 yi  y
R Pk  2 (5)
i1 yi  y
where yi is the experimental data, y0i is the corresponding
predicted data by the neural network, and y is the mean of all
the experimental data.
1256 international journal of hydrogen energy 34 (2009) 12531259

R2 can be interpreted as the proportion of variability that 3000

can be accounted for by the neural network around the mean

Mean square error for hydrogen yield

for the responses. R2 equals 0 when the values of the factors do
not allow any prediction of the responses, and equals 1 when
the neural network can perfectly predict the responses from
2000
the factors studied. In other words, R2 is the measure of how
well the neural network describes the experimental data. The
closer R2 approaches 1, the higher the modeling ability the
neural network has.
In addition, the data predicted by neural network were 1000
plotted against the corresponding experimental data to visu-
alize the modeling ability of the neural network. How much
closer to the line of perfect prediction (the line on which
the data predicted by the neural network are all equal to the 0
corresponding experimental data) of the data points for the 1 2 3 4 5 6 7 8 9 10
experimental and predicted data, the higher the modeling Number of neurons in the hidden layer of neural network
ability of the neural network.
Fig. 2 Effects of number of neurons in the hidden layer of
the neural network for hydrogen yield on the mean square
2.5. Method of desirability function error.

The method of desirability function was used to obtain the

maximum substrate degradation efficiency, hydrogen yield 1
Y
k 1k
and average hydrogen production rate simultaneously in this D d1  d2  ,,,  dk k di (7)
i1
study. For the application of the method of desirability func-
tion, each response yi was converted to an individual desir- In other words, the simultaneous optimization of several
ability function di that ranges from 0 to 1, according to Eq. (6). responses can be achieved by determining the maximum of
the overall desirability.
8 0 if yi  L
<  w
yi L
di if L < yi < U (6) 2.6. Genetic algorithm
: UL
1 if yi  U

where yi is the response to be optimized, L is the minimum The genetic algorithm is based on the principles of evolution
acceptable value of yi, U is the maximum value beyond which through natural selection, that is, the survival of the fittest
improvements would serve no further benefit, and w is strategy. It explores all regions of the solution space using
a weight factor. a population of individuals. Each individual represents a set of
Then the optimal conditions for several responses were factors. The individual chosen in this study was a set of
obtained by maximizing the overall desirability D, which is the temperature, initial pH and glucose concentration. Initially,
geometric mean of all the individual desirability functions a population of individuals is formed randomly. The fitness of
(Eq. (7)). each individual is evaluated using a fitness function. Upon

9
Mean square error for average hydrogen

40
Mean square error for substrate degradation

30 6
production rate
efficiency

20

10

0
0 1 2 3 4 5 6 7 8 9 10
1 2 3 4 5 6 7 8 9 10 Number of neurons in the hidden layer of neural
Number of neurons in the hidden layer of neural network network

Fig. 1 Effects of number of neurons in the hidden layer of Fig. 3 Effects of number of neurons in the hidden layer of
the neural network for substrate degradation efficiency on the neural network for average hydrogen production rate
the mean square error. on the mean square error.
 international journal of hydrogen energy 34 (2009) 12531259 1257

Predicted substrate degradation efficiency ( )

Predicted average hydrogen production rate (mL/h)

100
25
Line of perfect prediction
Data points Line of perfect prediction
90 Data points
20

80
15

70
10

60
5

50
50 60 70 80 90 100 0
Experimental substrate degradation efficiency ( ) 0 5 10 15 20 25
Experimental average hydrogen production rate (mL/h)
Fig. 4 Experimental and predicted data of substrate
degradation efficiency. Fig. 6 Experimental and predicted data of average
hydrogen production rate.

completion of the fitness evaluation, genetic operations such

as mutation and crossover are applied to individuals selected, degradation efficiency, hydrogen yield and average hydrogen
according to their fitness, to produce the next generation of production rate are shown in Table 1.
individuals for fitness evaluation. This process continues until
an optimal solution is found [17]. 3.1. Effects of temperature, initial pH and glucose
In this study, the desirability function based on the neural concentration on substrate degradation efficiency, hydrogen
network was used as the fitness function for the genetic yield and average hydrogen production rate
algorithm to optimize the fermentative hydrogen production
process. The experimental data of substrate degradation efficiency,
hydrogen yield and average hydrogen production rate in Table 1
were used to train the neural network for them. To obtain the
3. Results and discussion optimal number of neurons in the hidden layer of the neural
network, the number of neurons in the hidden layer was
In this study, the modified Logistic model was used to fit the investigated. Figs. 13 show the mean square error between
cumulative hydrogen production data obtained from each the experimental data and the corresponding predicted data
batch test. The coefficients of determination (R2) of all the at different numbers of neurons in the hidden layer of the
fittings were close to 1.000, which indicated that the modified neural network for substrate degradation efficiency, hydrogen
Logistic model could successfully describe the progress of yield and average hydrogen production rate, respectively.
cumulative hydrogen production in the batch tests of this The mean square error between the experimental data and
study. The calculated experimental results of substrate the corresponding predicted data for substrate degradation
efficiency and hydrogen yield decreased with increasing
number of neurons in the hidden layer of the neural network
350 from 1 to 5, while it changed little with further increasing the
Predicted hydrogen yield (mL/g glucose)

Line of perfect prediction neuron number from 5 to 10. This indicated that when
300 Data points the number of neurons in the hidden layer was from 5 to 10, the
neural network was better able to model the experimental data
250
of substrate degradation efficiency and hydrogen yield data. In
addition, when the number of the neurons in the hidden layer
200
of the neural network is larger, the neural network is very
150 complex and will take a longer time to train the neural

100

Table 2 Parameters for optimization by method of

50
desirability function.
Response yi L U w
0
0 50 100 150 200 250 300 350 Substrate degradation efficiency (%) y1 90.0 96.0 1
Experimental hydrogen yield (mL/g glucose) Hydrogen yield (mL/g glucose) y2 230.0 310.0 1
Average hydrogen production rate (mL/h) y3 20.0 25.0 1
Fig. 5 Experimental and predicted data of hydrogen yield.
1258 international journal of hydrogen energy 34 (2009) 12531259
Table 3 Experimental design for validating the optimal conditions identified by the method of desirability function based
on neural network.
Response Temperature ( C) Initial pH
Substrate degradation efficiency (%) 39.0
Hydrogen yield (mL/g glucose) 39.0
Average hydrogen production rate (mL/h) 39.0
network. Thus, 5 neurons were chosen in the hidden layer of
the neural network for substrate degradation efficiency and
hydrogen yield, respectively. The neural network architecture
for substrate degradation efficiency in this study consisted of 3
neurons (temperature, initial pH and glucose concentration) in
the input layer, 5 neurons in the hidden layer, and 1 neuron
(substrate degradation efficiency and hydrogen yield, respec-
tively) in the output layer (topology 3-5-1). In a similar way, 6
neurons were chosen in the hidden layer of the neural network
for average hydrogen production rate. Training a neural
network is accomplished by adjusting the weight coefficients
in each successive layer to minimize the mean square error
between experimental data and the corresponding predicted
data of substrate degradation efficiency, hydrogen yield and
average hydrogen production rate, respectively.
Coefficients of determination of the neural network for
substrate degradation efficiency, hydrogen yield and average
hydrogen production rate were calculated as 0.984, 0.994 and
0.984, respectively. In addition, as shown in Figs. 46, the
experimental and predicted data points of them were very
close to the line of perfect prediction. All of this indicated that
the neural network for substrate degradation efficiency,
hydrogen yield and average hydrogen production rate could
successfully describe the effects of temperature, initial pH and
glucose concentration on them in this study.
3.2. Optimization by the method of desirability function
The method of desirability function was used to obtain the
maximum substrate degradation efficiency, hydrogen yield
and average hydrogen production rate simultaneously. The
parameters for optimization by method of desirability func-
tion are shown in Table 2.
When yi is between L and U, the overall desirability can be
expressed by Eq. (8).
 1
y1  90 y2  230 y3  20 3
D , , (8)
96  90 310  230 25  20
Subsequently, the overall desirability was optimized by
genetic algorithm and the maximum overall desirability of
0.819 was estimated at the temperature of 39.0  C, the initial
pH of 7.0 and the glucose concentration of 24.6 g/L. Accord-
ingly, the maximum substrate degradation efficiency of 94.5%,
hydrogen yield of 307.0 mL/g glucose and average hydrogen
production rate of 23.8 mL/h were predicted at the tempera-
ture of 39.0  C, the initial pH of 7.0 and the glucose concen-
tration of 24.6 g/L.
The optimal temperature for fermentative hydrogen
production obtained in this study was close to those (3540  C)
reported by Wang and Wan [5], Lin et al. [25] and Zhang and
Shen [26]. However, it was much lower than those (5060  C)
Glucose concentration (g/L) Identified Experimental
7.0 24.6 94.5 95.3
7.0 24.6 307.0 305.3
7.0 24.6 23.8 23.9
reported by Lin et al. [6], Yokoyama et al. [27] and Valdez-
Vazquez et al. [28]. The optimal initial pH for fermentative
hydrogen production obtained in this study was close to that
(6.5) reported by Lin et al. [29], but was much lower than that
(9.0) reported by Lee et al. [7], and much higher than that (4.5)
reported by Khanal et al. [8]. The optimal glucose concentra-
tion for fermentative hydrogen production obtained in this
study was close to that (23.8 g/L) reported by Pan et al. [13], but
was much higher than that (1 g/L) reported by Zheng et al. [30].
Several other experiments (Table 3) were carried out to
validate the optimal conditions identified by the method of
desirability function based on the neural network. As shown
in Table 3, under the optimal conditions identified the
experimental data were very close to those identified by the
method of desirability function based on the neural network,
indicating that the method of desirability function based on
the neural network was a useful tool to optimize the
fermentative hydrogen production process.
4. Conclusions
A fractional factorial design was carried out to investigate the
effects of temperature, initial pH and glucose concentration
on fermentative hydrogen production by mixed cultures in
batch tests and then the experimental data of substrate
degradation efficiency, hydrogen yield and average hydrogen
production rate were described by a neural network, based on
which the simultaneous optimization of the three responses
was performed by the method of desirability function. The
following conclusions could be drawn.
The neural network was able to successfully describe the
effects of temperature, initial pH and glucose concentration
on the substrate degradation efficiency, hydrogen yield and
average hydrogen production rate of this study. The
maximum substrate degradation efficiency of 95.3%,
hydrogen yield of 305.3 mL/g glucose and average hydrogen
production rate of 23.9 mL/h were all obtained at the optimal
temperature of 39.0  C, initial pH of 7.0 and glucose concen-
tration of 24.6 g/L identified by the method of desirability
function based on the neural network. In sum, the method of
desirability function based on a neural network was a useful
tool to optimize several responses for fermentative hydrogen
production processes simultaneously.
Acknowledgement
The authors would also like to thank the National Natural
Science Foundation of China for financially supporting this
research under Contract No. 50325824.
 international journal of hydrogen energy 34 (2009) 12531259 1259

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