BIOETHANOL PRODUCTION FROM GREEN COCONUT SHELL

Project Reference No.: 42S_B_MSC_008

COLLEGE : GARDEN CITY UNIVERSITY, BENGALURU

BRANCH : DEPARTMENT OF BIOTECHNOLOGY
GUIDE : Dr. PREMA KULKARNI
Dr. NAGAMANI J E
STUDENTS : Ms. CHAHAT SHARMA
Ms. SHALMALI CHATTERJEE

Introduction: Modern life demands high mobility and, as a result, transport is one of the largest and fastest growing
energy demanding sectors. Also, increase in competitive agribusiness automatization leads to a high energy
demand. However, due to concern on the negative impact of fossil fuels on the environment, the use of biofuels
emerges as a promising alternative that is gradually becoming technically and economically feasible.
Ethanol is the leading liquid biofuel used for transportation. First-generation ethanol has a simple production process
using sugar or grain as raw material (sugarcane juice in Brazil and corn in the USA and EU, for example), while 2G
ethanol (bioethanol) has more complex steps of production and uses lignocellulosic material as a substrate. Among
the major byproducts generated by agroindustry’s, lignocellulosic biomass is one of the most abundant, conflict-free
with food production and is available throughout the year at low prices. All of these characteristics show that
lignocellulose waste might be considered the most feasible option for fossil fuel replacement, having a significant
potential for bioethanol productivity while giving a destination for an environmental liability.
There are many projects around the world focusing on the use of lignocellulosic residues for biofuel production.
These residues can come from homes or city dumps. Nowadays, biofuels have an important part in the global liquid
fuel market and over a hundred companies in different countries base their production on various types of 2G
biofuels. Coconut husk and shell is a very promising substrate that can be used as raw material for 2G ethanol
production, since coconut palm plays an important role in the economy of several tropical countries. Coconut palm
tree is a perennial crop grown in tropical climate countries. Due to the coconut structure, many valuable products can
be obtained from it, such as meat (copra), oil, water, milk, and fibers; therefore, this fruit is of great economic
importance. Coconut harvesting time is determined by its purpose and is usually carried out in two stages of ripening.
The green fruits are destined to the coconut water market, while mature fruits are destined to the dry coconut market
(for meat, milk and oil). Therefore, depending on the plantation site, the residue is made of green or mature coconut
husks, which have different compositions.
The estimated annual worldwide coconut production in 2015 was around 55 million tons and the main producing
countries are Brazil, India, Indonesia, the Philippines, and Sri Lanka. The food industry uses coconuts to obtain
various products leaving the husk and shell as waste. Moreover, coconut husk encompasses 80 to 85% of the weight
of the fruit, which is left as waste. It is important to note that coconut husk has a high lignin content that during husk
decomposition penetrates the soil and can reach the water table imposing a great environmental risk. Since it is
discarded in high volumes it is mandatory to find a safe destination for this waste. Therefore, the use of coconut husk
for 2G ethanol production may be a solution to reduce the environmental impact. Moreover, if the technology is
cheap and simple enough it can be used by small producers. As mentioned earlier, coconut producing countries are
part of the third-world economy; in this way, turning trash into jobs and income, which also an extra advantage of
bioethanol production.
OBJECTIVES
The major objectives of this project are:
 Delignification of the husk fibres using alkaline pre-treatment
 Enzymatic hydrolysis of cellulose and hemi-cellulose present in the husk fibres.
 Isolation of saccharomyces sp. from fruit peels.
 Fermentation of the husk fibres using saccharomyces sp. for production of ethanol.
METHODOLOGY:
The husk was cut to smaller sizes, sanitized with 100ppm sodium hypochlorite solution for 15min and dried at 50°C
until constant weight. Subsequently, the coconut husk pieces were crushed mechanically and finally blended in
grinder. The fibers were separated from the powder by sieving. After sieving four major steps are being followed ie.
1. Alkaline pre-treatment,
2. enzymatic hydrolysis,
3. isolation of saccharomyces sp. from fruit peels, and
4. Fermentation

RESULTS AND CONCLUSIONS

alkaline pre-treatment: Alkaline pretreatment was carried out by adding 5g of coconut fiber to 100mL of 5%
NaOH solution in an Erlenmeyer flask with capacity of 500mL. Mixture was then autoclaved at 121°C and 1atm for
40min. Afterwards, the reacted mixture was filtered to separate the liquid phase from the solid fraction. total reducing
sugars were measured by hydrolysis with 1.5M H2SO4 solution and neutralization with 2N NaOH. The solid fraction
was washed with distilled water at room temperature until pH becomes neutral and dried in an oven at 50°C until
constant weight.

The solid phase recovered after alkaline pre-treatment (husk fibres).

The liquid phase recovered after alkaline pre-treatment (lignin).

Isolation of saccharomyces sp. from fruit peels.

Fruit samples (pineapple) were collected from local market, and their peels were extracted. One gram of
sample was soaked in 250 ml yeast maintenance media (YMM) broth at 30 °C for 3 days. After 3-day
incubation, each 100 μl of suspension was spread on a plate containing YMM, which consisted of 3 g yeast
extract, 3 g malt extract, 5 g peptone, 10 g glucose, and 15 g agar, in 1 l water, initial pH 5.5, and was
incubated aerobically at 30 °C for 3 days. Single colony formed was picked, and the cells were observed
under a microscope. the morphology of cells of the selected isolates and their appearance on yeast extract
peptone dextrose (YPD) agar media was examined. the plates were inoculated with 48-hour-old yeast
strain and incubated at 30 °C for 48 h.

Two major steps ie. Enzymatic hydrolysis and fermentation is still need to be done to conclude this
project.
SCOPE FOR FUTURE WORK:
Nowadays, the most economical way to produce 2G bioethanol is the biorefinery scheme, which is important for
strengthening and supporting the growing biobased economy. The world is entering a new scenario where many
countries are taking substantial steps towards a biobased economy. New bioproducts are beginning to replace fossil-
based products, greenhouse gas emissions are decreasing and innovative policies are emerging to support these
changes. To establish the foundation of a biobased economy, the use of biomass resources must be efficient and
sustainable. That goal can be achieved by biorefinery systems.
In an energy driven biorefinery system, the biomass is primarily used to produce energy (biofuel, power and/or heat),
and other byproducts are upgraded to more added-value products to optimize the economic and ecological
performance of the whole production process.
Advances in biorefineries allow the development of alternative products to avoid the accumulation of different
residues. As an example, 1,3-propanediol obtained from maize residues is important in the formation of polymers.
Also, succinic acid removed from various lignocellulosic residues is used in the chemical and pharmaceutical
industries. An important alternative to polyethylene is the use of ether amylose derived from various wastes such as
sugarcane, potato, and corn.
Other applications for coconut husks different from ethanol production and possible byproducts were found, such as
polymer composites and adsorbents to remove a wide range of water pollutants. As the focus of this work is ethanol
production, further studies should be made to see if the biomass remaining after the chosen process to obtain
ethanol can still be used for these purposes and analyze if this strategy is economically viable.

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