This is my slide presentation for candidature defence in Master of Biotechnology at University of Malaya.

1. Bioethanol Production from
Leucaena leucocephala Seeds
atikzufar@siswa.um.edu.my
Candidature Defence
29th December 2016
University Malaya
Name : Atik Zufar bin Haji Mohd Razaki
Matrix No : SGF150008
Supervisor : Dr. Zul Ilham bin Zulkiflee Lubes

2. Bioethanol
Renewable fuel
Produced from edible biomass (Sugar cane etc.)
Conversion of sugar into ethanol by Saccharomyces
cerevisiae (Yeast)
Shrubs
Fermentation
Figure 1: Chemical equation for fermentation
+ CO2

3. Problem Statement
Fuel vs food demand (Edible biomass).
High demand for fuel ethanol could be
unsustainable.
(Hahn-Hägerdal et al, 2006)

4. - Non-edible biomass.
- Lead; ipil ipil; lamtoro;
Petai Belalang
- Quick growing, Short rotation,
no fertilization needed
- Limited information on
bioconversion of monosugars
from seed of Leucaena
leucocephala into bioethanol.
Leucaena leucocephala
Tree
Seeds
Pod
Bushy- 2 years

5. Leucaena leucocephala
Peru Type – 1 year Plantation Giant - 8 months
Wood

6. Objectives
To determine the optimum parameters for the
fermentation of anhydrous glucose with yeast.
To convert carbohydrate, cellulose and glucose from
Leucaena leucocephala seeds to bioethanol using
optimized conditions.
To detect the presence of bioethanol from Leucaena
leucocephala seeds by using GCMS headspace.
To calculate the percentage yield of bioethanol.

7. Overall Procedure
Autoclave
Optimization
Fermentation
Filtration
Distillation
Analysis by GCMS Headspace
Days
Amount of
Glucose
Amount of
Yeast
pH
Cellulose obtained from
Leucaena leucocephala
seeds, (CLLS)
Leucaena leucocephala
seeds, (LLS)
Glucose obtained from
Leucaena leucocephala
seeds, (GLLS)

8. GCMS Headspace
Detect the presence of the ethanol in the term of
qualitative.
Involved two separating column in order to change
retention of ethanol and other volatiles after injection
of sample.
(Tiscione et al, 2011)

9. Data Analysis
Figure 2: Retention time of ethanol for the optimized day

10. Calculation
Optimization of Day 3
Mass of glucose = 6.02 g
Molar mass of glucose = 180.18 g mol-1
n = 0.03 mol
Molar mass of ethanol = 46.08 g mol-1
Theoretical mass = 2.76 g
Actual mass = 0.13 g
Yield of Ethanol = Actual mass x 100
Theoretical mass
= 4.71 %
C6H12O6 2 CH3CH2OH + CO2
Fermentation

11. Data Collection
Optimization of Parameters Yield of Ethanol, %
Days
1 1.45
3 4.71
8 4.35
10 2.54
Amount of
Glucose, g
10 0.36
15 0.95
20 3.25
30 2.36

12. Data Collection
Yield of
Ethanol,
%
Period of Time Taken, day
Figure 3: Yield of ethanol for optimization of days

13. Data Collection
Optimization of Parameters Yield of Ethanol, %
Amount of
Yeast, g
1 5.92
2 4.14
3 6.80
4 0.89
5 0.79
pH Range
4.0 – 4.6 1.87
6.0 – 6.6 1.58
7.0 – 7.6 1.78

14. Data Collection
Sample Optimized
Day
Optimized
Amount of
Glucose, g
Optimized
Amount of
Yeast, g
Optimized
pH Range
Yield of
Ethanol, %
CLLS
3 20 3 4.0 – 4.6
0.07
LLS 0.08
GLLS

15. Data Analysis
Figure 4: Yield of ethanol from cellulose and seeds
of Leucaena leucocephala

16. Discussion
Pure ethanol had successfully obtained from distillation
process at 78°C – 80°C.
GCMS headspace had successfully identified ethanol in
each parameter.
Problem in controlling temperature during distillation.

17. Plan for Future Works
To convert cellulose obtained from Leucaena
leucocephala seeds into glucose.
To convert glucose obtained from Leucaena
leucocephala seeds into bioethanol by using optimized
conditions.
To detect presence of ethanol from GLLS by using
GCMS headspace.
To calculate yield of ethanol from GLLS.

18. Conclusion
Leucaena leucocephala seeds could be potentially
replace edible biomass in production of ethanol

19. References
Hahn-Hägerdal, B., Galbe, M., Gorwa-Grauslund, M. F., Lidén, G., & Zacchi, G.
(2006). Bio-ethanol–the fuel of tomorrow from the residues of today.Trends in
biotechnology, 24(12), 549-556.
Tiscione, N. B., Alford, I., Yeatman, D. T., & Shan, X. (2011). Ethanol analysis by
headspace gas chromatography with simultaneous flame-ionization and mass
spectrometry detection. Journal of analytical toxicology,35(7), 501-511.

20. Thank you