AN INNOVATIVE TECHNIQUE CONVERTS WINERY WASTE INTO BIOFUEL

1. AN INNOVATIVE TECHNIQUE
CONVERTS WINERY WASTE INTO BIOFUEL
By
Dr. Thirunahari Ugandhar
Asst. Prof of Botany
Govt Degree College Mahabubabad

3. Abstract
• The chemical components that power biofuel may be found
in every part of the grape plant, including the vines, the
seeds, the peels, the pulp, and even the stems. laboratory
analysis to determine their potential application in
producing bio-ethanol.
• A recently developed method for breaking down waste
products from the process of making wine could potentially
be used to produce biogas and biofuel.
• a new approach to the generation of biofuel that makes use
of the components of grapes and vines that are discarded
during the winemaking process. found a way to convert
waste from the wine industry, which is mostly made up of
cellulose, pectins, and lignins, into simpler molecules that
may be utilized to make ethanol or other types of biofuels.
• These enzymes convert the waste to soluble sugars, which
may subsequently be transformed into other products

4. • Grape fruit peels had the highest polyphenol concentration of
54.45%.
• In the mixed fruit pulps, the dilute acid (H2SO4) pretreatment (DAP)
followed by Various fungi is known to decompose this waste by
producing an assortment of enzymes.
• This fermentation process can take anywhere from one to three
weeks, and it results in the production of alcohols, acids, and
simple sugars that have potential use in industry and medicine.
• The method of biodegradation through the use of fungus is not
particularly novel; nevertheless, the implementation of symbiotic
fungi to enhance the process of biomass degradation is relatively
recent.
• In order for this method to be a financial success, chemists,
engineers, and economists will need to contribute significantly.
• We are working on developing a method that any nearby
vineyard would be able to utilize to convert its waste materials
into either biofuel like ethanol or other compounds that are
significant from a business perspective.

5. • At 42 hours of incubation, the hydrolysates derived from
dilute H2SO4 pretreated Grapefruit peels generated a
high of 13.84 percent ethanol with a fermentation
efficiency of 27.13 percent
• The fermentation of hydrolysates obtained from dilute
acid pretreatment followed by enzymatic saccharification
of mixed fruit pulps (Grape ) was determined to be the
best for higher ethanol production under the optimum
condition in the current study.

7. Introduction
• Since it is produced from biomass, biofuel is considered
a sustainable fuel source.
• Ethanol is clear, colorless alcohol made from a variety of
biomass materials called feedstocks (the raw materials
used to make a product).
• U.S. producers mostly use food grains and crops with
high starch and sugar content as feedstocks for making
ethanol such as corn, sorghum, barley, sugar cane, and
sugar beets.
• Ethanol can also be made from grasses, trees, and
agricultural and forestry residues such as corn cobs and
stocks, rice straw, sawdust, and wood chips. Ethanol is
made from these feedstocks in several ways.

9. Descriptionof the ResearchProblem:
• Urbanization and rapid population growth are
responsible for increasing the consumption of energy in
the world.
• The worldwide consumption of energy has increased 17-
20 % in the last century".
• On the other hand, conventional energy sources (non-
renewable) such as fossil fuels are limited and thus
cannot meet the demand in the long term.
• Moreover, the use of fossil fuels also has a negative
environmental impact, e.g. increased greenhouse gas
emissions.

11. Biofuel push: India is set to be the third-
largest ethanol market by Prime Minister Narendra
Modi had said in June that the government has
resolved to meet the target of 20% ethanol
blending in petrol by 2025. Earlier the target
was set for 2030.
 India is set to become the third-largest market for ethanol in the
world after the US and Brazil by 2026, a recent report by the
International Energy Agency (IEA) said, adding that the country has
tripled ethanol demand to an estimated 3 billion litres between
2017 and 2021.
 Buoyed by India’s growing appetite, Asia is set to overtake Europe
in terms of biofuels production by 2026, the agency said. While

13. •Objectives
• The major objectives of large-scale production of biofuel
are: diversification of the sugarcane Ripen fruits, industry,
reduction of energy imports, better resource use, and,
indirectly, better environmental management.
• Develop a method to convert cellulose and hemicellulose
from amaranth lignocellulose Into fermentable sugars.
• For bioethanol to become more sustainable to replace petrol,
the production process has to be more efficient.
• Reducing cost of conversion
• Increasing yields Increase the diversity of crops used.
• Enhance participant understanding of fermentation processes
and chemistry.
• Enhance participant understanding of microbial growth
processes.
• Provide an opportunity for participants to gain hands-on
experience with state-of-the-art fermentors and fermentation
monitoring equipment.

14. • Fermentation is the most common method for producing
fuel ethanol
• The most common ethanol production processes today
use yeast to ferment the starch and sugars in corn, sugar cane,
and sugar beets.
• Corn is the main feedstock for fuel ethanol in the United
States because of its abundance and relatively low price
historically.
• The starch in corn kernels is fermented into sugar, which is
then fermented into alcohol.
• Sugar cane and sugar beets are the most common feedstocks
used to make fuel ethanol in other parts of the world.
Because alcohol is made by fermenting sugar, sugar crops are
the easiest ingredients to convert into alcohol.
• Brazil, the world's second-largest fuel ethanol producer after
the United States, makes most of its fuel ethanol from sugar
cane. Most of the cars in Brazil can run on pure ethanol or on
a blend of gasoline and ethanol.

15. •Review of Literature
• Fuel ethanol production has been fascinating
now because many countries look for reducing oil
imports, boosting rural economies, and improving
air quality.
• The world ethanol production has reached about
51,000 million liters (RFA (2007), being the USA
and Brazil being the first producers and India
standing fourth among the top fuel ethanol
producers.
• Main feedstocks for bioethanol production are
sugarcane (in Brazil) and corn grains (in the USA),
while many other agricultural raw materials are
also used worldwide.

16. • Among the three major types of raw materials, the
production of ethanol from sugary and starchy materials
are easier as compared to lingo cellulosic materials since
it requires additional technical challenges such as
pretreatment (Petersson et. al., 2007).
• Furthermore, the use of high technology and complicated
instrumentation methods with high operating costs may
in turn limit their commercialization and industrial
application in the developing countries (Isarankura et al.,
2007).
• Ethanol was first prepared synthetically in 1826, through
the independent effort of Henry Hennel in Britain and S.G
in France. Michael Faraday prepared ethanol by the acid-
catalysed hydration of ethylene in 1828, in a process
similar to that used for industrial synthesis of ethanol
today.

17. • Materials and methods:
• Grape vines, grape seeds, skins, pulp, and stalks were collected
from the local wholesale fruit market.
• The fresh plant materials were brought to the laboratory,
washed, and separated into pulp and peels.
• The fruit pulps were homogenized by using a simple wet
milling technique without adding water and the pulverized
puree was taken for further experimental studies.
• A mixed fruit pulp sample was also prepared by mixing both
fruits in an equal ratio (B1:O1 w/w).
• The fresh Grape vines, grape seeds, skins, and pulp were
chopped into small pieces (2-3 cm) and dried in a hot air oven
at 65ºC for 24 h.
• The dried materials were ground in a Wiley mill to pass
through a 1 mm screen and the powdered materials were
stored at room temperature for further analysis.
• Composite samples of both fruit peels were also prepared in
the same manner.

19. Mechanism of the acid-catalyzed transesterification of vegetable oils

20. • Preparation of Growth Medium
• The growth medium prepared for ethanol production
consists of 20 g substrate (Winery pulp)
• in 250 ml of the conical flask containing 100 ml of distilled
water (pH-5.5). The flasks were autoclaved at 121°C for
20 minutes. This medium is poured into Petri plates and
set aside to solidify.
• Preparation of Inoculum:
• The cells of S.cereviacae were aseptically cultured in
Yeast Extract Peptone Dextrose (YEPD) broth and
incubated at 30°C for 24hrs. 3.5 Saccharification of the
Fermentation Medium with Aspergillus
• The substrate medium as prepared above was inoculated
with spores of Aspergillus. The culture was incubated at
28C for 7 days under rigorous aerobic culture.

21. • The samples were taken at regular intervals every 24 hr for
analysis. After 7 days in culture maximum total sugar was
obtained. 3.6 Ethanol Production Medium (270 ml) was
prepared and transferred to a Duran wide-mouth bottle.
• The media was autoclaved at 121C for 20 minutes and
cooled. The culture broth (30 ml) from the Saccharification
step was added to the medium. This broth contains the
cellulolytic enzyme complex elaborated by Aspergillus.
• The bottle was inoculated with 5% v/v of previous activate
Saccharomyces cerevisiae.
• The bottles were cultured in static conditions. The samples
were withdrawn at regular time intervals every 24 h. 3.7
• Primary Product Isolation:
• The samples were centrifuged at 5000 rpm for 5
minutes and stored at -20C for further analysis.
• The raw ethanol yield was measured by ethanol assay
using the potassium dichromate method. 3.8

22. • Estimation of Biofuel:
• 10 ml of sample was drawn in a 250 ml volumetric flask.
The water was added to make the volume up to 250 ml.
From this diluted sample, a 20 ml aliquot was drawn and
drawn in a conical flask.
• 10 ml of 40% sulphuric acid was added to each flask using
a measuring cylinder. Stopper each flask loosely and heat
for 10 minutes in a water bath at 45–50°C. After 10
minutes the flasks were removed from the bath and 2 g of
potassium iodide was added to each flask.
• A burette was filled with standard thiosulphate solution.
• Titrate the contents of the flask with the thiosulphate
solution. 1– 2 ml of starch was added to the solution
when the brown colour of the solution becomes green.
The equivalence point of the titration is reached when
blue colour of the starch iodine complex just disappears,
leaving a clear green colour.

23. Results & Observations
• Pre-treatment of Fruit peels and pulp: The
media consisting of the oven-dried substrate
(20gms) was used and volume was made up to 250
ml and pH was adjusted to 5.5.
• The media was inoculated with spores of
Aspergillus niger. The culture was incubated at
28°C for 7 days under aerobic culture. The enzyme
activity was qualitatively determined by Starch
Agar plate assay and Cellulose Agar plate.
• The maximum residual sugar was obtained during
192 hr- 216 hrs. During the fermentation
concentration of reducing sugars was determined
by finding out the refractive index of the solution.

24. • The culture was replenished with fresh media and
inoculated with an activated culture of Sacchromyces
cerevisiae.
• The culture was transferred to Duran wide mouthed
bottles. The bottles were sealed and kept static in an
incubator at 28°C.
• The samples were withdrawn in every 24hrs and the
changes in pH, RI and ethanol concentration are
displayed graphically as follows’. Similar procedure was
carried out with different substrates. The profile of
change in pH, residual sugar and ethanol production was
monitored regularly at 24hr intervals

25. • Conclusion
• The massive utilization of fuel ethanol in the
world requires that its production technology be
cost-effective and environmentally sustainable.
• The current research tendencies for improving fuel
ethanol production are linked to the nature of
employed raw materials, processing steps, and
related process engineering issues.
• Agro-processing techno-economic activities are to
be generated for the conservation and handling of
agricultural produce and to make it usable as food,
feed, fiber, fuel, or industrial raw material.

26. • In this present study, efforts were made to identify the
fruit wastes as potential raw material for bioethanol
production and the results showed that mixed ripened
fruit biomass of banana and Orange can yield 36% of
ethanol and similarly the banana fruit peels treated with
dilute acid and microbial enzymes showed a potential
production of 14% ethanol.
• The high nonstructural carbohydrates, reserve starch
content and low fiber contents showed the potentiality of
bananas as a good feedstock for ethanol production.
• The major costs for a bio refinerys are feedstock
collection, storage, transportation, and pretreatment,
hence, optimizing these unit operations through using
fruit peels and un-marketed fruit wastages should prove
beneficial

27. tugandharbiotech@gmail.comin
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9/21/2022