Accepted Manuscript

Potentiometric of bioethanol production from cantaloupe waste (Magassi Neish-

abouri Cultivar)

Rouhollah Salehi, Ahmad Taghizadeh-Alisaraei, Fakhri Shahidi, Ahmad

Jahanbakhshi

PII: S1658-077X(18)30025-0
DOI: https://doi.org/10.1016/j.jssas.2018.05.006
Reference: JSSAS 322

To appear in: Journal of the Saudi Society of Agricultural Sciences

Received Date: 12 January 2018

Revised Date: 9 May 2018
Accepted Date: 28 May 2018

Please cite this article as: Salehi, R., Taghizadeh-Alisaraei, A., Shahidi, F., Jahanbakhshi, A., Potentiometric of
bioethanol production from cantaloupe waste (Magassi Neishabouri Cultivar), Journal of the Saudi Society of
Agricultural Sciences (2018), doi: https://doi.org/10.1016/j.jssas.2018.05.006

This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers
we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and
review of the resulting proof before it is published in its final form. Please note that during the production process
errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
 Potentiometric of bioethanol production from cantaloupe waste (Magassi
Neishabouri Cultivar)
Rouhollah Salehi1*, Ahmad Taghizadeh-Alisaraei1, Fakhri Shahidi2, Ahmad
Jahanbakhshi3

1
Department of Biosystems Engineering, University of Gorgan, Gorgan, Iran.
2
Department of Food Science and Technology, Ferdowsi University of Mashad, Mashhad, Iran.
3
Department of Biosystems Engineering, University of Mohaghegh Ardabili, Ardabil, Iran.
*
Corresponding Author Email:salehirouhollah@yahoo.com

Abstract

The ever-increasing need of the human beings to energy has brought about concerns
regarding the decrease in the amount of energy resources. So, renewable energies have
become very important and have obtained a special status in today's world. Using bioethanol
as a renewable resource to be blended with other fuels in internal combustion engines has
attracted many researchers to itself. Thus, in this study, we assess the amount of bioethanol
extracted from cantaloupe waste (Magassi Neishabouri Cultivar) to be used as a fuel. To do
so, considering the 15 kg capacity of the device's fermenter, 12 kg of refined cantaloupe
sugar syrup and three kilograms of water (with the standard ratio of 4 to 1), i.e. a total of 15
kg of substrate with the brix degree of 20, as well as 75 g of saccharomyces cervisiae yeast,
cultured in standard conditions (5 g of yeast for each kilogram of substrate) were transferred
into the fermenter. The tests were conducted in 35 hours with three replications and at three
different rotation speeds of the fermenter's mixer. Sampling took place every five hours.
Fermentation temperature was set as 30οC and distillation temperature was set to be 78.5οC
which is the standard temperature of bioethanol's boiling point. The results showed that about
49.5 g of bioethanol were extracted from each kilogram of cantaloupe syrup. In comparison
to the results obtained by other researchers, it became clear that due to its high sugar level,
cantaloupe waste is one of the appropriate options for the production of bioethanol. In
addition, considering the rate of the alcohol produced and in order to optimize energy
consumption, it was seen that the best speed for the fermenter's mixer, was 120 rpm.

Keywords: Bioethanol, Renewable Energy, Cantaloupe Waste.

1. Introduction

1
Nowadays, preparation of energy resources has become one of the most important concerns
of countries (Ghobadian et al., 2009; Taghizadeh-Alisaraei et al., 2017). Fossil fuels have
long been used as the source of energy in internal combustion engines, power plants, etc. The
ever-increasing population and increase in the use of fossil fuels, increase in the cost of oil
products and decrease in the reserves of such resources, have encouraged researchers to
search for new alternative resources. Besides, increase in environmental pollution due to
increase in the amount of pollutants such as unburned hydrocarbons (UHC), nitrogen oxides
(NOx), carbon monoxides (CO) and carbon dioxide (CO2) produced by the use of fossil fuels,
is the main shortcoming for such fuels. In order to preserve, save and make a more optimized
use of the existing resources and reduce environmental pollutions, it seems self-evident and
inevitable to be inclined to renewable energies (Kalam et al., 2003). Ethanol is considered as
renewable fuel since it has a plant base. Having one oxygen molecule in its molecular
structure, ethanol needs less oxygen for combustion compared to diesel fuels and gasoline.
For the same reason it produces less pollution (Li et al., 2005; Hansen et al., 2005). In
addition, using waste and producing fuels by them would reduce the expenses made to
transport and dump them. Through fermentation of some cucurbits some alcohol can be
produced which is flammable. Considering the high amount of waste from cucurbits such as
melon, cantaloupe, watermelon, etc., research studies conducted on the processing of the
above products in the world are very few (Rahimi, 2005).
Cantaloupe or Cucumis melo L. is highly popular among people for its high sugar level.
According to global statistics provided by the FAO, the area annually used to cultivate
cantaloupe in Iran and in the world is 79992 and 1308018 hectares, respectively and the mean
production levels for Iran and the world are 15.4 and 21.6 ton/ha, respectively (FAO, 2006).
In their study and modeling of sugar reduction in producing bioethanol and carbon dioxide in
the process of date fermentation, Vaezi et al. (2010) reported that during the fermentation
process, changes in the amount of carbon dioxide dropped significantly by the passage of
time and then stopped. Moreover, at the end of the process, the rate of the soluble sugar
decreased from a brix degree of 20 to a brix degree of around 8 and the total amount of
bioethanol production from each kilogram of syrup with a brix degree of 20 was about 72 g.
Louhichi et al. (2013) studied the rate of bioethanol extraction from three date cultivars. They
reported that for all the cultivars, the amount of the bioethanol obtained was almost 25
volume percent and about 316 g of bioethanol was produced for each kilogram of substrate.
In a study, Ghorbani et al. (2009) reported that sugar concentration is a very important factor
in the fermentation process. They stated that a high concentration of substrate prevents the
growth of the ferment, stops the production of alcohol and disturbs the metabolism of the
microorganism. Moreover, a sugar concentration within the 5-25 range had a significant
inhibitory effect and in 40 percent of the cases led to the complete stop of the microorganism
activity. Corbin et al. (2015) studied ethanol production from grape sugar syrup. They carried
out a test one two different varieties of grapes, namely red and white grapes. They reported
that if the grape sugar syrup is fermented without hydrolysis, 270 g of bioethanol is can be
extracted from each liter of syrup and if the hydrolysis takes place first and then fermentation
happens, 400 g of bioethanol can be extracted from each liter of sugar syrup. Khalil et al.
(2015) conducted a study on the production of bioethanol from sweet cluster corn waste.
First, they hydrolyzed the product under acidity conditions for 60 minutes at the temperature

2
of 120οC. In the end, they reported 50 g of alcohol production for each kilogram of corn
sugar syrup. Moreover, they estimated that the SS-301 species had the potential to produce
160 g of bioethanol from each liter of sugar syrup.
Iran is one of the important poles of cantaloupe production and the production of other
cucurbits. Unfortunately, more than 30 percent of these products are destroyed in the
production chain between the farm and the consumption areas. This is a very high percentage
and thus we are required to conduct deep analyses about the processing of different parts of
the products and the production of essential products and think about ways to optimally use
the wastes. In this regard, the present study aims at assessing the amount of bioethanol
extraction from cantaloupe waste (Magassi Neishabouri Cultivar) which can significantly
contribute to the optimization of cantaloupe waste use.

2. Materials and Methods

Ethanol can be obtained from the cellulose, sugar and starch sources in agricultural products
(Chintagunta et al., 2016; Smuga-Kogut et al., 2017). Thus, in this study the rate of the sugars
in cantaloupe waste that could be transformed using an HPLC Waters 600 Controller was
measured. Bioethanol extraction device used in this study consisted of two different parts: 1)
hydrolysis reactor (which precisely controls temperature parameters and the rotation speed of
the mixer so that pressure cooking is made possible to hydrolyze starch and turn it into simple
transformable sugars. 2) fermentation reactor (which controls temperature and rotation speed
of the mixer, makes sampling possible, has a system for drying bioethanol and adjusting the
aeration rate for better and faster growth of the yeast) (Fig. 1).

Figure 1 Bioethanol extraction device: 1) Control box, 2) Electromotor, 3)

Hydrolysis tank, 4) Fermenter tank, 5) Refrigerant and 6) Chassis.

3
The stages of the research are reported in Fig. 2.

Figure 2 Different stages of bioethanol extraction from cantaloupe waste.

In this study, cantaloupe (Magassi Neishabouri Cultivar) waste was used to extract
bioethanol. To do so, cantaloupe waste was turned into a strong pulpy liquid by an electric
fruit juicer. Syrup was made with a 70 percent concentration after refining the liquid. Then,
considering the 15 kg capacity of the device's fermenter, 12 kg of refined cantaloupe sugar
syrup and three kilograms of water (with the standard ratio of 4 to 1), i.e. a total of 15 kg of
substrate with the brix degree of 20 was transferred into the fermenter. In the next step, 75 g
of the saccharomyces cervisiae (5 g of yeast for every kilogram of substrate) which had been
cultured under standard conditions was added to the 15 kg of substrate in the tank (Sarkar et
al., 2012; Duarte et al., 2009). After that, the rate of sugar that existed in the substrate with
the brix degree of 20 was measured by the Hand Held Refractometer (PAL-3) which was
made in Japan.

In order to assess the device's work and specify the standard rotation speed for the
fermentation operation in the fermenter, the device was tested under anaerobic fermentation
conditions for 35 hours which included the time needed to make a suspension at the constant
fermentation temperature of 30 οC (the temperature 28-30οC is the appropriate and optimal
temperature for the growth of the saccharomyces yeast), the rotation speeds of 120, 130 and
140 rpms and the distillation temperature of 78.5οC.

During the time the samples were being fermented (35 hours), sampling took place seven
times (every five hours). After each sampling, the rate of the ferment growth and production

4
of alcohol in the substrate was measured using a Digital Alcohol Tester with the brand name
of Milwaukeeinst-MA884. The little differences in the data obtained after measuring the
alcohol in the substrate (the 5th, the 6th and the 7th sampling) indicated that almost all of the
convertible sugar that existed in the substrate could be turned into alcohol through the
saccharomyces yeast. Thus, the continuation of that trend could affect the device's
performance regarding energy consumption. Measuring the amount of alcohol produced by
fermentation can be a better determiner of fermentation rate. The alcohol tester used in this
study, reported the amount of the alcohol produced in the substrate based on the volume
percentage of alcohol. Thus, to obtain the weight percentage of the alcohol produced, the
values obtained were divided by alcohol's density (0.79). In similar studies, Zeng et al. (2012)
reported that for all the substrates applied, the amount of the sugar remained after 36 hours of
fermentation could be ignored.

According to relation (1), measuring the rate of glucose to ethanol transformation in the
process of fermentation, one mole glucose is turned into two moles of carbon dioxide and two
moles of ethanol:

CH O
6 12 6
 2C2 H 5 O H  2C O
2
(1)

This phenomenon results in the reduction of weight and can give an ascending trend to
ethanol production. Theoretically, each gram of glucose can produce 0.51 g of ethanol. So,
50% of glucose turns into ethanol and 50% of it changes to carbon dioxide. In the next step,
the alcohol produced must turn into vapor. For this, the distillation temperature of 78.5οC
which is the standard temperature of ethanol's boiling point was used. High purity ethanol
cannot be produced through usual distillation methods. Although ethanol's boiling point is
close to 79οC and water's boiling point is 100οC, when the solution of ethanol and water is
heated up to ethanol's boiling temperature, only 95% alcohol can be obtained. After the
distillation of ethanol and water, ethanol-water azeotrope (95% ethanol and 5% water) can be
obtained. Therefore, for drying bioethanol, the output path for the bioethanol to exit the
fermenter where bioethanol is at the vapor phase, was designed in a way that it passed
through a zeolite column and the water was absorbed by the zeolite on the way (Tosheva,
2001). At the last stage, after passing through the zeolite column, the vaporized bioethanol
enters a double-shelled cooling path where water is used as the liquid for cooling bioethanol
vapor. Later, the bioethanol vapor is turned into bioethanol liquid and enters the tank where
the produced ethanol is stored.

Each of the tests was replicated three times. In the end, the means of the data obtained were
analyzed using Excel 2013 and the results were reported in the form of graphs.

3. Results and Conclusion

The rate of the transformable sugars that exist in cantaloupe is reported in Fig. 3.

5
 80

The amount of sugar per kilogram

69
70

60

50

40

30

20
9.1
10

0
Glucose Sucrose
Convertible sugars

Figure 3 Convertible sugars existing in cantaloupe.

As can be seen, due to the high amount of glucose and sucrose in cantaloupe waste, it can be
used as one of the main resources for bioethanol production in the country. Changes in the
amounts of alcohol produced during the fermentation process at different mixer speeds are
reported in Fig. 4.

6
Amount of alcohol produced (%)

120 rpm
3
130 rpm
2 140 rpm

0
5 10 15 20 25 30 35

Time (h)

Figure 4 Changes in the rate of alcohol produced in the process of fermenting cantaloupe
waste at different rotation speeds of the fermenter's mixer.

The results indicate that at the mixer speed of 120 rpm, ethanol production is more favorable
compared to other speeds and this might be due to better blending and lower turbulence of the
flow. It can also be seen that by increasing the speed of the fermenter's mixer, the rate of the
alcohol produced decreases and this is while there is not much difference in the amount of the
alcohol produced at different speeds. This might be due to improper comparisons that lead to
inaccurate judgments stating there is not much difference among different mixer speeds in the

6
fermenter. Therefore, it is suggested that in order to prevent energy waste, it is better not to
use speeds above 120 rpm in the fermenter. These results are consistent with the results
obtained by Apar and Özbek (2004).

Changes in the amount of alcohol produced in the process of cantaloupe waste fermentation
show that within 30 hours from the beginning of the fermentation process (the sixth stage of
measuring alcohol rate), the rate of the alcohol produced at all the mixer speeds, is about five
percent. In other words, for each kilogram of substrate, around 50 g of alcohol produced. In
the times higher than 30 hours, it can be seen that the alcohol production rate is very
imperceptible and unstable. To save energy and time in the process of producing bioethanol
from cantaloupe waste, it is better to reduce the time from 35 hours to 30 hours. These results
are consistent with the findings of Vaezizadeh et al. (2010) in investigating and modeling
sugar reduction and bioethanol and carbon dioxide production in the process of date
fermentation.

The mean rate of bioethanol produced from each kilogram of cantaloupe syrup at different
mixer speeds in the fermenter is reported in Fig. 5.

60
[VALUE]a
50 [VALUE]b
Bioethanol produced (g)

[VALUE]c

40

30

20

10

0
120 130 140
Speeds of the fermenter's mixer (rpm)

Figure 5 The rate of the bioethanol produced for each kilogram of cantaloupe syrup at
different speeds of the fermenter's mixer.

After the complete distillation of the substrate at of 78.5οC, which is the standard temperature
of the bioethanol boiling point and weighing the amount of the extracted bioethanol, the
results showed that the mean amounts of the extracted bioethanol from the wastes of the
Mgassi Neishabouri cultivar of cantaloupe at different speeds of the fermenter mixer (120,
130 and 140 rpm) were respectively 49.5, 47 and 44.5 g of bioethanol from each kilogram of
cantaloupe syrup with the brix degree of around 20. Therefore, considering the results of the
present study, it can be stated that the best performance of the device takes place at the mixer
speed of 120 rpm not only with respect to the amount of the alcohol produced in the
fermenter but also regarding the rate of the bioethanol produced compared to other speeds. In
addition, in similar studies, the rates of the bioethanol obtained from apples were 43 g and

7
from grapes were 53 g (Hang et al., 1981; Hang et al., 1986). So, considering the rate of the
fermented sugar in the fermenter chamber, the amount of the ethanol produced in this study is
considerable compared to similar studies mentioned above. This, in fact, can be considered as
a good performance for the fermenter device too. Also, in the statistical analysis because of
the absence of common alphabets on mean of treatments, the results showed a significant
difference at the probability level of 1%.

4. Conclusion

Based on the accurate data obtained in the present study, the following general findings can
be stated:

1- Although bioethanol production has improved due to the use of advanced technologies,
there is still room for more studies on this topic. These studies include preserving yeasts
in industrial production, more improvement of the lignocellulose material preparation
technologies and investigation and optimization of the necessary parameters for
economizing ethanol production systems.
2- Considering the results obtained regarding bioethanol production from cantaloupe waste
(Magassi Neishabouri culrivar) which showed that about 49.5 g of bioethanol was
extracted from each kilogram of cantaloupe waste. Comparing these results with those
obtained by other researchers, it became known that cantaloupe waste is one of the best
alternatives for bioethanol production due to its high level of sugar.

References

Apar, D. K., Özbek, B., 2004. σ-Amylase inactivation during corn starch hydrolysis process.
Process Biochemistry. 39, 1877-1892.
Chintagunta, A. D., Jacob, S., Banerjee, R., 2016. Integrated bioethanol and biomanure
production from potato waste. Waste Management. 49, 320-325.
Corbin, K. R., Hsieh, Y. S., Betts, N. S., Byrt, C. S., Henderson, M., Stork, J., Burton, R. A.,
2015. Grape marc as a source of carbohydrates for bioethanol: chemical composition,
pre-treatment and saccharification. Bioresource technology. 193, 76-83.
Duarte, J. C., Sàágua, M. C., Baeta-Hall, L., Correia, A., Ribeiro, B., Lourenço, V., Paixão,
S. M., 2009. Bioethanol production from agricultural wastes. ECOMONDO: 13th
International Trade Fair of Material & Energy Recovery and Sustainability.
Faostat. 2006. FAO Statistical Databases. Available from http: //faostat.fao.org.
Ghobadian, B., Najafi, G., Rahimi, H., Yusaf, T. F., 2009. Future of renewable energies in
Iran. Renewable and sustainable energy reviews. 13, 689-695.
Ghorbani, F., Yunesi, H. A., Esmaeili Sari, A., Ghasempouri, S. M., Amini, M., Daneshi, A.,
2009. Production of ethanol in a batch fermentation system using the Saccharomyces
cervisiae from molasses waste of a sugar factory. Journal of Enviromental Science and
Technology. 11, 139-148. (In Persian).
Hang, Y. D., Lee, C. Y., Woodams, E. E., 1986. Solid-state fermentation of grape pomace for
ethanol production. Biotechnology letters. 8, 53-56.

8
Hang, Y. D., Lee, C. Y., Woodams, E. E., Cooley, H. J., 1981. Production of alcohol from
apple pomace. Applied and environmental microbiology. 42, 1128-1129.
Hansen, A. C., Zhang, Q., Lyne, P. W., 2005. Ethanol–diesel fuel blends––a review.
Bioresource technology. 96, 277-285.
Kalam, M. A., Husnawan, M., Masjuki, H. H., 2003. Exhaust emission and combustion
evaluation of coconut oil-powered indirect injection diesel engine. Renewable Energy.
28, 2405-2415.
Khalil, S. R., Abdelhafez, A. A., Amer, E. A. M., 2015. Evaluation of bioethanol production
from juice and bagasse of some sweet sorghum varieties. Annals of Agricultural
Sciences. 60, 317-324.
Li, D. G., Zhen, H., Xingcai, L., Wu-gao, Z., Jian-Guang, Y., 2005. Physico-chemical
properties of ethanol–diesel blend fuel and its effect on performance and emissions of
diesel engines. Renewable energy. 30, 967-976.
Louhichi, B., Belgaib, J., Hajji, N., 2013. Production of bio-ethanol from three varieties of
dates. Renewable energy. 51, 170-174.
Rahimi, H., 2005. A project presented in diesel engine performance using bioethanol
derived from potato waste for the degree master of science in mechanics of farm
machinery. Thesis, Department mechanics of farm machinery school of
agriculture, Tarbiat Modares University, Tehran, Iran. (In Persian).
Sarkar, N., Ghosh, S. K., Bannerjee, S., Aikat, K., 2012. Bioethanol production from
agricultural wastes: an overview. Renewable energy. 37, 19-27.
Smuga-Kogut, M., Zgórska, K., Kogut, T., Kukiełka, K., Wojdalski, J., Kupczyk, A.,
Wielewska, I., 2017. The use of ionic liquid pretreatment of rye straw for bioethanol
production. Fuel. 191, 266-274.
Taghizadeh-Alisaraei, A., Assar, H. A., Ghobadian, B., Motevali, A., 2017. Potential of
biofuel production from pistachio waste in Iran. Renewable and Sustainable Energy
Reviews. 72, 510-522.
Tosheva, L., 2001. Molecular sieve macrostructures prepared by resin templating (Doctoral
dissertation, Lulea University of Technology).
Vaezizadeh, M., Ghazanfari Moghaddam, A., Fouladi, M. H., 2010. Examining and modeling
sugar reduction and bioethanol and carbon dioxide production in the process of date
fermentation, Iranian Journal of Biosystem Engineering. 2, 121-126. (In Persian).
Zheng, Y., Yu, C., Cheng, Y. S., Lee, C., Simmons, C. W., Dooley, T. M., VanderGheynst, J.
S., 2012. Integrating sugar beet pulp storage, hydrolysis and fermentation for fuel
ethanol production. Applied Energy. 93, 168-175.