Group: 1
-
124
Leong Qi Dong
4S216
Soh Han Wei
4I324
Aman
Mangalmurti
AOS
Kara Newman
AOS
Depletion of non
-
renewable fossil
fuels
Heavy metal water
contamination of
water is rampant in
many countries
Heavy metal ions
accumulate inside
organisms and affect
the ecosystem
Conversion to
biofuel, ensuring
continual energy
supply
Biosorption
in
removal of heavy
metal ions by fruit
peel wastes
Fruit peel waste
Ethanol
Fermentation
Shorter fermentation
time (300%
-
400%
faster than yeast)
higher ethanol yield
(92%
-
94% versus
88%
-
90% for yeast)
Microorganism
used:
Zymomonas
mobilis
Nguyen, T., and
Glassner
, D. (2001 )
To investigate the production of ethanol from
fruit peels
To investigate the efficiency of adsorption of
heavy metal ions by fruit peels
To determine the procedure which
maximises
ethanol yield and efficiency of heavy metal
ion adsorption
Mango peels contain reducing sugars that
can be fermented to ethanol.
Mango peels show different efficiency levels
in the adsorption of copper, zinc and lead
ions.
Preparation of
fruit peel extract,
microbe, heavy
metal solution
Extraction of
sugars
Ethanol
Fermentation
Residue for
Adsorption
of
Ions
Adsorption
of
Ions
Extraction of
sugars
Ethanol
Fermentation
•
Mango
Peels
Fruit
tested
•
Pb
2+
•
Cu
2+
•
Zn
2+
Ions
tested
Independent
•
Heavy metal ion
(Pb
2+
, Cu
2+
, Zn
2+
)
•
Sequence of
procedures
Dependent
•
Initial
concentration of
reducing sugars in
fruit peel extracts
•
Ratio of ethanol
yield to sugar
concentration
•
Amount of ethanol
per g of fruit peel
•
Final concentration
of heavy metal
ions
Controlled
•
Mass of fruit peel
used
•
Type of
microorganism
used (
Z.
mobilis
)
•
Duration and
temperature of
fermentation
Centrifuge
Centrifuge tube
Spectrophotometer
Glass
rod
Sieve
Blender
Dry blender
Shaking incubator
Oven
Incubator
Weighing Balance
Zymomonas
mobilis
Glucose
-
yeast medium
Sodium alginate
Calcium chloride solution
Sodium chloride solution
Fruit peel
Cuvettes
Deionised water
Dinitrosalicylic
acid
Acidified potassium chromate solution
Lead (II), Copper (II), Zinc (II) ion solutions
Copper (II) and Zinc (II) reagent kits
Preparation of
Z.
mobilis
, Extraction of Sugars, Fermentation,
Determination of Yield
Growth of
Z.
mobilis
Immobilisation
of cells
Extraction of sugars from fruit peels
Ethanol fermentation by immobilized
Z. mobilis
cells
Determination of ethanol yield with the dichromate test
Ethanol fermentation
Z.
mobilis
cells were inoculated in 20 ml GY
medium (2% glucose, 0.5% yeast extract)
Incubated at 30
°
C for 2 days with shaking for
growth to occur
Culture was
centrifuged at
7000 rpm for 10
min
Cell pellets were
resuspended
in
7.5 ml GY
medium.
Absorbance of
the cultures
were taken at
600 nm.
7.5 ml of 2%
sodium alginate
is added to the
cells.
Added
dropwise
to 0.1 M calcium
chloride solution
to form beads.
Beads were
rinsed in 0.85%
sodium chloride
solution.
40 g of fruit
peels were
blended in 400
ml of deionised
water using a
blender.
The extract was
passed through
a sieve to
remove the
residue.
Suspension was
centrifuged at
7000rpm.
Supernatant
and residue
were collected.
200
Z.
mobilis
beads were
added to 50 ml
mango peel
extract.
Control : 200
empty beads
were added to
the same volume
of mango peel
extract.
Set
-
ups were
incubated with
shaking at 30
°
C
for 2 days for
ethanol
fermentation.
Beads were
removed and the
extracts are
distilled to obtain
ethanol.
2.5 ml of acidified potassium dichromate
solution was added to 0.5 ml of distillate.
Samples were placed in a boiling water
bath for 15 min.
Absorbance was measured at 590 nm
using a spectrophotometer
Concentration of ethanol was read from
an ethanol standard curve.
Dessication
of peel, Preparation of ion solution, Adsorption, Determination
of final ion concentration
Fruit peel
residue
was dried
in an oven.
Dried peel was
blended to
powder using the
dry blender.
0.2g fruit peel powder
was added to 10ml 50ppm
of each ion solution (test).
No peel was added in the
control set
-
up.
Mixtures were
placed on a
rocker for 24 h
at room
temperature.
Peel powder
was removed
by
centrifugation
.
Using reagent kits for copper
(II) and zinc(II) and AAS for
lead (II) the final
concentration of ions in
solution was determined.
•
µmol of ethanol per g of fruit peel
Ethanol yield
•
% of heavy metal ions adsorbed
•
(Final
-
Initial)/Initial x 100%
•
t
-
test to determine if difference between
the control and the test is significant
Heavy metal ion
adsorption
efficiency
y = 0.8101x
-
0.0691
R² = 0.9969
0.0
0.5
1.0
1.5
2.0
2.5
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
Absorbance at 530 nm
Maltose concentration / µmol/ml
Maltose standard curve
y = 0.6392x
-
0.0034
R² = 0.9996
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
0
0.5
1
1.5
Absorbance at 590 nm
Concentration of ethanol / %
Ethanol standard curve
Sugar Extraction First, Ethanol Fermentation, Ion Adsorption
0.196
0.205
0.00
0.05
0.10
0.15
0.20
0.25
0.30
1
2
Concentration of ethanol / %
Ethanol yield from mango peels
46.5
63.2
36.2
9.4
26.3
1.9
0
10
20
30
40
50
60
70
Copper
Zinc
Lead
Final concentration of ions / ppm
Adsorption of ions by mango peels (1st
round)
Control
without
peels
Test with
mango
peels
45.9
53.8
32.5
4.4
15.8
0.5
0
10
20
30
40
50
60
Copper
Zinc
Lead
Final concentration of ions / ppm
Adsorption of ions by mango peels (2nd
round)
Control
without
peels
Test with
mango
peels
Ion
t
-
test p value to show difference between
control and test
Round 1
Round 2
Copper
0.000399
0.000229
Zinc
0.00037
0.00101
Lead
0.00001
0.000405
All differences were significant as p < 0.05
85.1
64.5
96.6
0
20
40
60
80
100
120
Copper
Zinc
Lead
Ion adsorbed by mango peels / %
A comparison of the efficiency of adsorption of
ions by mango peels
Ion
Adsorption First, Sugar Extraction, Ethanol Fermentation
48.6
51.2
55.3
15.0
33.3
4.3
0
10
20
30
40
50
60
Copper
Zinc
Lead
Final concentration of ions / ppm
Adsorption of ions by mango peels (1st
round)
Control
without
peels
Test with
mango
peels
50.4
51.6
32.4
19.8
31.7
5.8
0
10
20
30
40
50
60
Copper
Zinc
Lead
Final concentration of ions / ppm
Adsorption of ions by mango peels (2nd
round)
Control
without
peels
Test with
mango
peels
49.0
51.2
31.9
15.1
24.4
7.2
0
10
20
30
40
50
60
Copper
Zinc
Lead
Final concentration of ions / ppm
Adsorption of ions by mango peels (3rd
round)
Control
without
peels
Test with
mango
peels
Ion
t
-
test p value to show difference between
control and test
Round 1
Round 2
Round
3
Copper
0.0000000723
0.000485
0.000000139
Zinc
0.000542
0.002072
0.000619
Lead
0.00000471
0.004109
0.00000225
All differences were significant as p < 0.05
64.9
36.7
87.2
0
10
20
30
40
50
60
70
80
90
100
Copper
Zinc
Lead
Ion adsorbed by mango peels / %
A comparison of the efficiency of adsorption of ions
by mango peels
Ethanol yield
Extraction of sugars
first
Adsorption of
ions first
Round 1
Round 2
Round 1
Mean ethanol
concentration / %
0.196
0.205
0.223
Amount of ethanol / µmol
334.91
351.88
382.25
Amount of ethanol / µmol
per g of fruit peel
66.98
70.38
148.02
Sequence 2 (Adsorption of ions followed by extraction of
sugars) resulted in a higher yield of ethanol
Metal ion
Mean % of ions adsorbed
Extraction of
sugars first
Adsorption
of ions first
Copper ion adsorbed / %
85.1
64.9
Zinc ion adsorbed / %
64.5
36.7
Lead ions adsorbed / %
96.6
87.2
Sequence 1 (Extraction of sugars followed by adsorption
of ions) resulted in higher efficiency of adsorption of ions
C
-
H
stretch
Some changes in the
1000
-
1800cm
-
1
wavenumbers
weaker
C
-
H
stretch
Adsorption of ions has resulted in changes in
FTIR
spectra
Weaker C
-
H stretch after lead ion adsorption
Stretching of more bonds in between 1800
-
1000 cm
-
1
after all three ion adsorption
We
believe that the carboxylic
acid, ester and
lactone
(1700cm
-
1
) and
alkene groups
(1600cm
-
1
)
are responsible for adsorption.
Difficulty in
standardising
batch of mango
peels for all tests performed
May yield inconsistent results for each repeat
Cost
-
effective
method of
producing ethanol
Reduces reliance
on non
-
renewable
fossil fuels
Using by
-
product
waste
Low cost, viable
method in
wastewater
treatment
Investigate the effect of pH of ion solution on
adsorption
Investigate the production of ethanol and
adsorption of ions on peels of other locally
available fruits such as pineapple
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, T. J.
Ugye
, T.D.
Nyiaatagher
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, A.B.M.S. &
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, A.R. (2010). Creation of alternative energy by bio‐ethanol production
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