International Journal of Scientific Research Engineering & Technology (IJSRET)

Volume 2 Issue 6 pp 362-365 September 2013 www.ijsret.org ISSN 2278 – 0882

Production of Bioethanol From Fruit Rinds by Saccharification and

Fermentation
Suhas V Bhandari*, Arun Panchapakesan, Naveen Shankar, H G Ashok Kumar
Department of Biotechnology, RV College of Engineering, Mysore Road, Bangalore - 560059

ABSTRACT
The generation of bio-fuels from wastes forms an
attractive solution towards both waste management and
energy generation. The most important roadblock faced
in large scale application is the aeration during
saccharification. The utility of fruit rinds as a possible
source of cellulosic ethanol in a process without aeration
was investigated by using rinds of four fruits namely
Pineapple, Jackfruit, Watermelon and Muskmelon. The
powdered rinds were subjected to saccharification by
Trichoderma viride followed by fermentation with
Saccharomyces cerevisiae. Significant amounts of
reducing sugars were obtained at the end of the
saccharification process, with jackfruit and pineapple
rinds being the most effective at 10.28 mg.ml-1 and
10.18 mg.ml-1 respectively. The amount of ethanol
produced after fermentation was analyzed by gas
chromatography and found to be highest for the same
fruits with yields of 4.64 g.l-1 and 4.38 g.l-1 respectively.
The results indicate the promising future for generation
of ethanol from cellulosic wastes on a large scale.
Keywords – Bioethanol, Fermentation, Fruit rinds,
Trichoderma, Saccharification.
I. INTRODUCTION
In a world where the dumping of wastes is causing
serious harm to the flora and fauna of the areas
surrounding the dumping sites, the concept of using the
wastes for production of energy forms a solution which
is easily adoptable, cheap and efficient.
One of the most abundant sources of energy in the
world is the bio-polymer cellulose, which forms a major
component of most plant and algal cell walls. The ability
of organisms such as species of Trichoderma,
Aspergillus, Clostridium, etc. to produce the enzyme
cellulase enables them to hydrolyze this cellulose into its
constituent glucose units [1-3]. The glucose can then be
utilized by organisms of the genera Saccharomyces
which can ferment the glucose into fuels such as ethanol
[4].
India being an agro based economy generates nearly
350 million tonnes of wastefrom the vegetables, fruits
and other organic materials [5]. Organic matter including
fruit rinds is a major part of wastes generated daily by
households, agricultural sector and food processing
industries. Fruits are used on a small and large scale for
household consumption and by food processing
industries like pulp and jam manufacturers. In urban
areas, a considerable portion of solid waste includes fruit
waste generated by fruit juice vendors and restaurants.
These industries and establishments usually discard the
inedible parts of the fruits which include the exocarp
commonly referred to as ‘rind’ or ‘peel’. In most cases,
these waste materials are dumped in landfills which lead
unhygienic conditions. However, utilization of these
waste materials in production of bio-fuels would be of
great environmental and economic benefit as it could
reduce the burden on conventional sources of energy and
also get rid of the wastes.
Ethanol an important biofuel, having high calorific
value has the added advantage of being less polluting
than most sources of energy that are in use today.Reports
available suggest that previous natural substrates for
ethanol production via saccharification have included
sugarcane bagasse, wheat straw, corn, softwood etc. [6-
9]. Fruit rinds are excellent sources of cellulose which
can be used for the production of ethanol via
saccharification followed by fermentation [10].
Most of the procedures followed in the laboratory are
not feasible when the process is scaled up for application
on a scale that would be useful to a large community, the
most important parameter being the amount of power
consumed just for aeration [11]. This study is thus aimed
at processes which can be scaled up without great
investments or tedious procedures. The work showcases
a comparative study on utilization of the rinds of
Pineapple, Watermelon, Jackfruit and Muskmelon by T.
viride and fermentation of these sugars by S. cerevisiae.

IJSRET @ 2013
 International Journal of Scientific Research Engineering & Technology (IJSRET)
Volume 2 Issue 6 pp 362-365 September 2013
II. MATERIALS AND METHODS
1. Microorganism and Culture media:
T. viride and S. cerevisiae were procured from MTCC
(Microbial Type Culture Collection, Chandigarh)
Accession number 9699 and 170 respectively. T. viride
was cultured on Potato Dextrose Agar (PDA) and S.
cerevisiae on Yeast extract Peptone Dextrose (YPD) at
30˚C. The cultures were stored at 4˚C and subcultured
every 30 days.
2. Processing of the substrates:
The fruit rinds of Ananas comosus (Pineapple),
Artocarpus heterophyllus (Jackfruit), Citrullus lanatus
(Watermelon) and Cucumis melo (Muskmelon) were
obtained from local shops and washed twice with
distilled water before being prepared for use. The rinds
were dried in a hot air oven at 65˚C for 24 hours and
powdered using a grinder. 50 g of each rind was
weighed and utilized as the substrate.
3. Saccharification:
The processed fruit rinds were added to a broth
containing yeast extract (5g/l) and peptone (10g/l). The
media was autoclaved at 1210C and 15 psi pressure for
20 min and T. viride was inoculated under aseptic
conditions. The flasks were then incubated at room
temperature for a period of 144 hrs.
Figure 1 Amount of reducing sugars produced with different fruit rinds as substrates.
IJSRET @ 2013
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4. Reducing sugars assay:
Reducing sugars assay was carried out according to the
Dinitrosalicylic method [12]. Un-inoculated media was
used as the control for the assay. The optical densities of
the samples were measured against the blank at 540nm.
The glucose concentration was then calculated using
standard glucose curve.
5. Fermentation:
After saccharification, the cultures were autoclaved and
filtered using Whatman No 1 Filter Paper. The filtrate
was then transferred into 250ml Erlenmeyer flasks, made
airtight with cork and autoclaved. The flasks were then
aseptically inoculated with 15mlovernight grown S.
cerevisiae and incubated at room temperature for 96hrs.
The ethanol produced was determined by Gas
Chromatography using a NUCON Gas Chromatograph
(5765 EPC) with a flame ionization detector. The carrier
gas used was Nitrogen.
III. RESULTS
Jackfruit and Pineapple rinds displayed similar reducing
sugar concentrations over the period of the assay,
terminating at 10.28 mg.ml-1 and 10.18mg.ml-
1
respectively after 144 hours, as can be observed in Fig.
1.
 International Journal of Scientific Research Engineering & Technology (IJSRET)
Volume 2 Issue 6 pp 362-365 September 2013
Figure 2. Ethanol yield after fermentation of sugars produced using various substrates.
Watermelon and Muskmelon produced
comparatively lower concentrations of reducing sugars
with 4.16 mg.ml-1 and 7.43 mg.ml-1 respectively at 144
hours when the process was terminated by autoclaving.
The amount of ethanol produced after 96 hours of
fermentation with S. cerevisiae in various flasks is as
shown in Fig. 2.
Ethanol yield of jackfruit rinds was the maximum at
4.64 g.L-1 of the media followed by Pineapple,
Muskmelon and Watermelon rinds with yields of 4.38
g.L-1, 3.08 g.L-1 and 1.89 g.L-1 respectively. The
percentage conversion was calculated to be 84.2 %,
89%, 88.2% and 81.1% for Pineapple, Watermelon,
Jackfruit and Muskmelon rinds respectively.
IV. DISCUSSION
The amount of reducing sugars produced at the end of
saccharification is significantly higher than that reported
by Omojasola et al [9], which can be attributed to the
different rinds and organisms used. Reducing sugar
concentrations can be further increased by pre-treatment
of the cellulosic wastes as has been reported by Tewari
et al and Gomathi et al [13,14]. Nevertheless, on a large
scale pre-treatment and agitation are tedious procedures
and usage of a saccharification process without aeration
forms an ideal solution. However, the comparative yield
IJSRET @ 2013
www.ijsret.org ISSN 2278 – 0882
by use of this kind of a process is less. Hence, a balance
between the two needs to be established for an economic
process with sufficient yield.
The amount of ethanol produced is slightly below
that of Itelima et al, which can be explained by the same
reasons, cited above. The ethanol yields can be further
increased by using mutant strains of S. cerevisiae, as has
been reported by Manikandan et al [15,16].
V. CONCLUSION
This study has explored the possibility of using
Pineapple, Jackfruit, Muskmelon and Watermelon fruit
rinds for the purpose of bioethanol production. These
fruit rinds usually end up in garbage dumps or biogas
reactors.
This study shows that there is potential for use of
these fruit rinds in bioethanol production with minimal
energy consumption to provide aeration.
These cellulosic substrates usually have a lot of
lignin content which prevents easy access for the
microorganisms for saccharification. Thus some
pretreatment methods like Acid or Base treatment is
necessary to delignify these wastes and to obtain higher
reducing sugar yields and hence higher ethanol yields
also.
 International Journal of Scientific Research Engineering & Technology (IJSRET)
Volume 2 Issue 6 pp 362-365 September 2013 www.ijsret.org ISSN 2278 – 0882

Also the use of engineered strains of T. viride [12]Miller GL (1959). . Use of dinitrosalicylic acid
capable of high cellulase activities will also help in reagent for determination of reducing sugar. Anal
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[13]Tewari HK, Marwaha SS, Rupal K (1986). Ethanol
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