a comparative study of the yield of bioethanol in algae, corn and newspaper group: 01-36 team...
TRANSCRIPT
![Page 1: A COMPARATIVE STUDY OF THE YIELD OF BIOETHANOL IN ALGAE, CORN AND NEWSPAPER Group: 01-36 Team Members Tang Kwan Hou (L) (4S123) Robin Ho (4S116) Jerroy](https://reader038.vdocuments.site/reader038/viewer/2022102808/56649e875503460f94b8b4e6/html5/thumbnails/1.jpg)
A COMPARATIVE STUDY OF THE YIELD OF BIOETHANOL IN ALGAE, CORN AND NEWSPAPER
Group: 01-36
Team MembersTang Kwan Hou (L) (4S123)
Robin Ho (4S116)Jerroy Chang (4S203)
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Content
• Aim
• Hypothesis
• Variables
• Materials and Method
• Results and Analysis
• Conclusions
• Extensions
• References
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Problem to be addressed
• Methods of extracting bioethanol too expensive or energy-consuming
• Find out an effective and cheap way to produce bioethanol
oBioethanol is rising in demand across the world
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Aim
• To investigate and compare yield of bioethanol per unit mass of different substrates at optimum conditions
• To investigate the optimum concentration of cellulase and amylase to use for each substrate
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• Ulva• Macroalgae contain significant amount of sugars (at least 50%) that could
be used in fermentation for bioethanol production (Wi et al., 2009)
• Most green algae can have a cellulose content of up to 70% of dry mass (B. Baldan, P. Andolfo, L. Navazio, C. Tolomio, P. Mariani, 2002)
• Corn• An increase in the ethanol production means an increase in the demand
of corn (Pimental D., 2009)
• Corn kernels contain 75.2% starch and 30% cellulose. (Yong T., Zhao D., Cristhian C., Jiang J., 2011)
Literature Review
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• Paper• The presence of 70% cellulose & hemicellulose, α-cellulose
(60%) and lignin (16%) makes it a prospective and renewable biomass for bioethanol production (Alok K.D. et. al, 2012)
• Husk• Corn husks contain 42% cellulose and 13% lignin. (Y.
Mahalaxmi, et. al, 2009)
• Often discarded when people prepare corn
Literature Review
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• Sargassum• The brown seaweed Sargassum sp. is a promising feedstock
for ethanol production because of its relatively high content (41.6% dry basis) of holocellulose. It also contains 22.0% of alpha-cellulose and 19.6% of hemicellulose. (Jeylnne P. et. al, 2014)
Literature Review
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• Commercial Production• Acid Hydrolysis
• Algae species were hydrolysed in dilute 1.0ml of 0.70% H2SO4 and were heated at
105°C for 6h. (Gupta R. et al, 2012)
• Required 95.103 kWh power which costs $24.42 according to Singapore’s electrical tariff of $0.2568 between 1 July 2014 to 30 Sep 2014
• Wet Milling
• Corn kernel is steeped in water, with or without sulphur dioxide, to soften the seed kernel in order to help separate the kernel’s various components.
• For example, it can separate a 56-pound bushel of corn into more than 31 pounds of corn starch, which in turn can be converted into corn ethanol (J. Womach et al, 2005)
Literature Review
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• Cellulase has an optimum pH between 4 to 5 and an optimum temperature between 40 to 50ºC (Carl B. Z., n.d.)
• The optimum temperature of the α-amylase is 50ºC and optimum pH value is 6 (Atiyeh M., Reza H. S., Mehdi R., Vahab J. , 2010)
• Optimum temperature for fermentation by Saccharomyces cerevisiae is at 45ºC but will ethanol yield will drop above that (Lin Y. et al, 2012)
Literature Review
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Optimal pH and Temperature
Optimum pH
Cellulase
4.0-5.0
Alpha-Amylas
e
6.0
Optimum Temperature
Cellulase
40-50℃
Alpha-Amylas
e
50℃
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Hypothesis
• Paper produces the greatest yield of bioethanol (cm3/g), after enzymatic action and fermentation.
• The usage of pH 5.0 acetate buffer and enzymatic action at 45°C will increase yield of bioethanol (cm3/g).
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Variables• Independent:
• Type of starting product
• Concentration of cellulase added (%)
• Concentration of amylase added (%)
• Dependent:
• Yield of bioethanol after a fixed period of time
• Controlled:
• Mass of starting material used (6.0g)
• Temperature of surroundings (Room temperature or 45°C)
• Duration of fermentation (1 day)
• pH value of solution (7.0 or 5.0)
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MATERIALS AND METHODS
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MATERIALS TO BE TESTED ON
• AlgaeUlva sp. (green algae)Sargassum sp. (brown algae)
• Zea mays (maize)• Kernel
• Husk
• Newspaper
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• Potato Dextrose Broth
• Cultured Yeast (Saccharomyces cerevisiae)
• Cellulase
• Alpha-Amylase
• Deionised Water
OTHER MATERIALS USED
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APPARATUS
• Rack Shaker
• Weighing Scale
• Centrifuge machine
• Centrifuge tubes
• Incubator
• Water Bath
• Blender
• Ethanol Probe
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60mlamylase
60ml cellulase
6g materi
al
60ml DI
water
24:00:00
37°C
• Independent variable – Starting materials (Paper, Ulva sp. , Kernel, Husk)
Methodology
Homogenisation
Enzymatic action
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Supernatant
25°C90°C
00:10:00
5000 rpm
Methodology
• Heated at 90 degrees Celsius to halt enzyme catalysis reaction by inactivating it (Nam S. W., n.d.)
Denaturing
CentrifugationDecanting
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60ml pH5.0
Acetate buffer
60ml DI
water
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1L DI Water
24g PDB(Potato Dextrose
Broth)
1L PDB
00:15:00
121°C
Methodology
Preparing yeast broth
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Yeast
Yeast
30mL PDB
30mL PDB1L PDB
37°C
24:00:00
Methodology
Preparing yeast broth
Inoculation
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6.7mL
yeast
3.3mL
extract
30mL PDBSupernata
nt
24:00:00
37°C
Methodology
Inoculation
Fermentation
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Methodology
Reading Results
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RESULTS AND ANALYSIS
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Results - Husk
0.25 0.5 1 20
0.05
0.1
0.15
0.2
0.25
0.3
0.2000.245
0.180 0.190
Bar chart showing the effect of concentration of cellulase
on ethanol yield/%
Cellulase concentration/%
Eth
an
ol Y
ield
/%
0.25 0.5 1 20.320.330.340.350.360.370.38
0.343 0.350
0.373 0.370
Bar chart showing the effect of concentration of amylase
on ethanol yield/%
Amylase concentration/%
Eth
an
ol Y
ield
/%From the graph we can see that:Best Cellulase Concentration: 0.50%Best Amylase Concentration: 1.00%
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Results - Kernel
0.25 0.5 1 20
0.02
0.04
0.06
0.08
0.030
0.0600.070
0.030
Bar chart showing the effect of concentration of cellulase on
ethanol yield/%
Cellulase concentration/%
Eth
an
ol Y
ield
/%
0.25 0.5 1 20
0.1
0.2
0.3
0.4
0.237 0.2700.323 0.370
Bar chart showing the effect of concentration of amylase on
ethanol yield/%
Amylase concentration/%
Eth
an
ol Y
ield
/%From the graph we can see that:Best Cellulase Concentration: 1.00%Best Amylase Concentration: 2.00%
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Results - Paper
0.25 0.5 1 20
0.05
0.1
0.15
0.2
0.1200.150
0.180 0.160
Bar chart showing the effect of concentration of cellulase
on ethanol yield/%
Cellulase concentration/%
Eth
an
ol Y
ield
/%
0.25 0.5 1 20
0.10.20.30.40.50.6
0.480 0.407 0.390 0.387
Bar chart showing the effect of concentration of amylase
on ethanol yield/%
Amylase concentration/%
Eth
an
ol Y
ield
/%From the graph we can see that:Best Cellulase Concentration: 1.00%Best Amylase Concentration: 0.25%
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Results – Ulva sp.
0.25 0.5 1 20.22
0.24
0.26
0.28
0.3
0.2750.293
0.273
0.247
Bar chart showing the effect of concentration of amylase on
ethanol yield/%
Amylase concentration/%
Eth
an
ol Y
ield
/%
1 2 3 40.000
0.020
0.040
0.060
0.080
0.100
0.057 0.0600.077 0.077
Bar chart showing the effect of concentration of cellulase on
ethanol yield/%
Cellulase concentration/%
Eth
an
ol Y
ield
/%
From the graph we can see that:Best Cellulase Concentration: 1.00%Best Amylase Concentration: 0.50%
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• Best amylase concentration varies with each extract.
• However, Mann-Whitney U and Kruskal-Wallis Test shows that the difference in results are insignificant.
• Best cellulase concentration for All Starting Materials: 1.00%
• Except husk (0.50%)
Data Analysis
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Best Cellulase Concentration/%
Best Amylase Concentration/%
Husk 0.50 1.00
Kernel 1.00 2.00
Paper 1.00 0.25
Ulva sp. 1.00 0.50
Summary
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Results – Sargassum sp.
0.25 0.50 1.00 2.000.00
0.05
0.10
0.15
0.20
0.25
0.30
0.35
0.140.18
0.22
0.29
Graph showing effect of varying concentration of cellulase/% on
ethanol yield/%
Eth
an
ol Y
ield
/%
0.25 0.50 1.00 2.000
0.05
0.1
0.15
0.2
0.25
0.3
0.35
0.08
0.18 0.19
0.3
Graph showing effect of varying concentration of cellulase/% on
ethanol yield/%
Eth
an
ol Y
ield
/%
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Results - Paper
0.25 0.50 1.00 2.000
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
0.12 0.15 0.18 0.160.25 0.25
0.59
0.95
Graph showing effect of varying concentration of cellulase/% on ethanol
yield/%
BeforeAfter
Eth
anol Yie
ld/%
0.25 0.50 1.00 2.000
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.480.407 0.39 0.387
0.24 0.25
0.53
0.71
Graph showing effect of varying concentration of amylase/% on ethanol
yield/%
BeforeAfter
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Results - Kernel
0.25 0.50 1.00 2.000
0.05
0.1
0.15
0.2
0.25
0.030.06 0.07
0.03
0.13
0.170.19
0.22
Graph showing effect of varying concentration of cellulase/% on
ethanol yield/%
BeforeAfter
Eth
anol Yie
ld/%
0.25 0.50 1.00 2.000
0.05
0.1
0.15
0.2
0.25
0.3
0.35
0.4
0.2370.27
0.3230.37
0.07
0.17 0.16
0.28
Graph showing effect of varying concentration of amylase/% on ethanol
yield/%
BeforeAfter
Eth
anol Yie
ld/%
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Results - Husk
0.25 0.50 1.00 2.000
0.05
0.1
0.15
0.2
0.25
0.3
0.35
0.4
0.45
0.343 0.35 0.373 0.37
0.110.07
0.3
0.42
Graph showing effect of varying concentration of amylase/% on ethanol
yield/%
BeforeAfter
Eth
anol Yie
ld/%
0.25 0.50 1.00 2.000
0.05
0.1
0.15
0.2
0.25
0.3
0.35
0.4
0.45
0.20.245
0.307
0.19
0.020.07
0.380.43
Graph showing effect of varying concentration of cellulase/% on
ethanol yield/%
BeforeAfter
Eth
anol Yie
ld/%
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Results – Ulva sp.
0.25 0.50 1.00 2.000
0.1
0.2
0.3
0.4
0.5
0.6
0.0650000000000001 0.06 0.065 0.04
0.440.5
0.420.346666666666667
Graph showing effect of varying concentration of cellulase/% on
ethanol yield/%
BeforeAfter
Eth
anol Yie
ld/%
0.25 0.50 1.00 2.000
0.1
0.2
0.3
0.4
0.5
0.6
0.275 0.293 0.273 0.247
0.1
0.5
0.120.19
Graph showing effect of varying concentration of cellulase/% on
ethanol yield/%
BeforeAfter
Eth
anol Yie
ld/%
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Best Cellulase Concentration/%
Best Amylase Concentration/%
Sargassum sp. 2.00 2.00
Paper 2.00 2.00
Kernel 2.00 2.00
Husk 2.00 2.00
Ulva sp. 0.50 0.50
Summary
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Mann-Whitney U Test + Kruskal-Wallis Test (pH 7.0 and room temperature)
-
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Kruskal Wallis Test (pH 5.0 and 45 degrees Celsius)
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• Best amylase concentration varies with each extract.
• However, Mann-Whitney U and Kruskal-Wallis Test shows that the difference in results are insignificant.
• Best cellulase concentration for All Starting Materials: 1.00%
• Except husk (0.50%)
Data Analysis
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• Most graphs were positive functions
• More enzyme, more ethanol produced
• Best concentration of both cellulase and amylase were 2.00%
• In all cases (except for Ulva sp.)
Data Analysis
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COMPARISON OF RESULTS•Paper – 0.480%
•Husk – 0.373%
•Kernel – 0.370%
•Ulva sp. – 0.293%
•Paper – 0.950%
•Husk – 0.430%
•Kernel – 0.280%
•Ulva sp. – 0.500%
•Sargassum sp. – 0.300%
Before
After
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• Converting ethanol yield/% into cm3/g:
• “It takes about 20 lb (9.1kg) of corn … to produce a gallon (3.9L) of ethanol” (The Energy Collective, 2013)
• 0.417cm3/g
Conclusion
Material Ethanol yield/%
Ethanol/cm3 per setup
Ethanol per gram (cm3/g)
Paper 0.950 0.0950 0.855
Husk 0.430 0.0430 0.387
Kernel 0.280 0.0280 0.252
Ulva sp. 0.500 0.0500 0.450
Sargassum sp.
0.300 0.0300 0.270
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• Paper produces the greatest yield of bioethanol (cm3/g), after enzymatic action and fermentation.
• The usage of pH 5.0 acetate buffer and enzymatic action at 45°C increased yield of bioethanol (cm3/g).
Conclusion
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EXTENSIONS
• Create a bioreactor using calcium chloride beads of immobilized enzymes and yeast
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Sources of error and how to overcome them
• Ethanol probe was wetClean the probe and calibrate each time before reading
results
• Amount of yeast in each set-up was differentUse spectrometer to check turbidity of each PDB for
consistency
• Contamination of starting material (Bacteria entering solution)Micro-filter and do it in sterile environment
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• Ethanol Probe may not be accurate in reading the ethanol yield due to the low yield
• KMnO4 can be added to the ethanol produced and titrated to
get a more accurate concentration
• However, it required a few weeks to prepare the KMnO4 at
the specific concentration required
• Less time-consuming if ethanol probe is used
Sources of error and how to overcome them
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Shetty P. R. (2009) Corn husk as a novel substrate for the production of rifamycin B by isolated amycolatopsis sp. RSP 3 under SSF. Retrieved from: http://www.Academia.Edu/201890/corn_husk_as_a_novel_substrate_for_the_production_of_rifamycin_b_by_isolated_amycolatopsis_sp._Rsp_3_under_ssf
The Energy Collective. (2013) The US Corn-to-Ethanol Program. Retrieved from: http://theenergycollective.com/willem-post/287061/us-corn-ethanol-program
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References
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THANK YOU!ANY QUESTIONS?