Ethanol_21novx - aos-hci-2012-research-program

messengerrushBiotechnology

Feb 22, 2013 (4 years and 8 months ago)

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Aman

Mangalmurti


Kara Newman

Leong Qi Dong

Soh Han
Wei

Depletion of non
-
renewable fossil
fuels

Heavy metal ions
accumulate inside
organisms and affect
the ecosystem

Heavy metal water
contamination of
water is rampant in
many countries

Various diseases
afflicting bananas
leads to organic
waste

Conversion of
renewable sources to
fuel ensures continual
energy supply

Biosorption

in
removal of heavy
metal ions by fruit
peel wastes

Utilizing organic
waste

Problems faced
previously

With this
project

Use of chemical water filtration
methods


Expensive, unaffordable to those
who need it most


Low efficiency at low metal ion
concentrations

Affordable, accessible
water filter


Helps more people in many
countries

Renewable source of
energy


Less pollution

To prepare extracts of fruit peel for ethanol
fermentation

To determine which fruit peel gives highest ethanol
yield

To determine which fruit peel waste adsorbs heavy
metal ions best

To determine a protocol which maximizes efficiency of
fruit waste


Ethanol yield from fermentation differs for
both peels


The efficiency of heavy metal ion adsorption
differs
for both peels


The order of adsorption and fermentation has
an effect on the ethanol yield and the
efficiency of adsorption

Preparation of
fruit peel extract,
microbe, heavy
metal solution

Adsorption
of
Ions

Extraction of
sugars

Ethanol
Fermentation

Extraction of
sugars

Ethanol
Fermentation

Residue for
Adsorption
of
Ions


Banana
Peels

AOS


Mango
Peels

HCI

Independent


Fruit peels used
(AOS: banana,
HCI: mango)


Heavy metal ions


Order of
Procedures

Dependent


Initial
concentration of
reducing sugars in
fruit peel extracts


Ratio of ethanol
yield to initial
sugar
concentration


Final ethanol yield


Final
concentration of
heavy metal ions

Constant


Mass of fruit peel
used


Type of
microorganism
used


Immobilisation of
microorganism


Fermentation
conditions


Adsorption
conditions


Procedures

APPARATUS


Centrifuge


Centrifuge tube


Spectrophotometer


Spectrophotometer cuvettes


Glass rod


Dropper


Sieve


Blender


Boiling water bath


Shaking incubator


Fractional
distillatory


Quincy Lab Model 30 GC hot
-
air
oven


Rotary Mill


Sieve: 0.25mm (60 Mesh)


MATERIALS


Zymomonas

mobilis


Glucose
-
yeast medium


Sodium
alginate medium


Calcium
chloride solution


Sodium
Chloride
solution


Fruit
peel


Deionised
water


Dinitrosalicylic

acid


Acidified
potassium chromate
solution


Lead
(II
), Copper
(II
), Zinc
(II) ion
solutions


Lead (II), Copper (II), Zinc (II)
r
eagent kits


ETHANOL FERMENTATION

Growth of
Z.
mobilis

Immobilisation of cells

Extraction of sugars from fruit peels

Determination of sugars in extracts

Ethanol fermentation by immobilized
Z. mobilis

cells

Determination of ethanol yield with the
dichromate test

ADSORPTION OF HEAVY
METAL IONS

Pre
-
treatment of peel

Creation of heavy metal mixture

Adsorption

Determination of final ion
concentration

Preparation of
Z.
mobilis
, Extraction of Sugars, Fermentation,
Determination of Yield

Z.
mobilis

cells are inoculated in 20 ml GY
medium (2% glucose, 0.5% yeast extract) and
incubated at 30
°
C for 2 days with shaking to
obtain the
preculture
.

The Z.
mobilis

preculture

is
centrifuged at 7000
rpm for 10 minutes

The
cell pellets are
resuspended

in 7.5 ml of
fresh GY medium.

The absorbance of the
cultures are taken at 600
nm.

7.5 ml of 2% sodium
alginate is added to the
cell suspension and
mixed well.

The mixture is dropped
into 0.1 mol dm‐3
calcium chloride solution
to form Z.
mobilis

alginate beads.

The beads are rinsed
with 0.85% sodium
chloride solution.

30 g of fruit peels are
blended in 300 ml of
deionised water
using a blender.

The liquid is passed
through a sieve to
remove the residue.

To 0.5 ml of extract,
0.5 ml of DNS
(
dinitrosalicylic

acid)
is added.

The mixture is left in
a boiling water bath
for 5 minutes.

4 ml of water is then
added.

The samples are placed in
spectrophotometer cuvettes and
the absorbance is taken at 530 nm
using a spectrophotometer.

The concentration of reducing
sugars in
μmol
/ml is read from
a maltose standard curve.

200 beads are
added to 50 ml
waste extract.

A control is
prepared in which
200 empty alginate
beads are added to
the same volume of
waste extract
instead.

All the set‐ups are
incubated with
shaking at 30
°
C for
2 days for ethanol
fermentation to
occur.

The beads are then
removed and the
extracts are distilled
to obtain ethanol.

2.5 ml of acidified
potassium
dichromate solution
is added to 0.5 ml of
distillate in a ratio
of 5:1.

The samples are
placed in a boiling
water bath for 15
minutes.

The absorbance is
measured at 590
nm using a
spectrophotometer,
and the
concentration of
ethanol is read from
an ethanol standard
curve.

Pre
-
treatment of peel, Creation of heavy metal mixture, Adsorption,
Determination of final ion concentration

Desiccate fruit peel
residue, (put the residue
in the hot air oven and
dry them at 60 degrees
for 23 hours)

Using a rotary mill to
grind desiccated residue

Sieve to 0.25 mm particle
size.

A mixture is made of
0.5mols of each metal:
Pb
2+
, Zn
2+
, Cu
2+
in 1L of
distilled water

Add powder to mixture

Allow solution to set for
20 min at 100rpm to
increase contact time

Fruit product is removed
by centrifuging

Using respective
reagent kits, the
remaining concentration
of lead(II),copper (II) and
zinc(II) ions will be
found.


The ratio of ethanol yield to
amount of initial reducing sugar


µmol of ethanol per g of fruit peel

Ethanol yield
would be
evaluate by
comparing


The ratio of the final
concentrations of metal ions to the
initial concentrations


% of heavy metal ions adsorbed

Heavy metal ion
adsorption
efficiency would
be evaluated by
comparing

Cost
-
effective
method of
producing ethanol

Reduces reliance on
non
-
renewable fossil
fuels

Using by
-
product
waste

Viable method in
wastewater
treatment


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, T. J.
Ugye
, T.D.
Nyiaatagher

(2009). Chemical composition of Musa
sapientum

(Banana) peels.
Electronic Journal of Environmental, Agricultural and
Food Chemistry, 8,
437
-
442.

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Koffi
, L. & Han, Y.W. (1990). Alcohol production from pineapple waste. World
Journal of Microbiology and Biotechnology, 6(3), 281‐284.


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, G. Burke, J. Foster, S.
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, A.B.M.S. &
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, A.R. (2010). Creation of alternative energy by
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, C.C. &
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, I.N. (2010). Novel method of wine production from banana
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saccharification

and
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Saccharomyces

cerevisiae

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Biochemistry,
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Mishra
, V.,
Balomajumder
, C. &
Agarwal
, V.K. (2010).
Biosorption

of Zn(II) onto the surface of
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