Fermentation Analysis of Cellulolytic Microorganism Thermobifida Fusca
1. FERMENTATION ANALYSIS OF
CELLULOLYTIC MICROORGANISM
THERMOBIFIDA FUSCA
Engineering Researcher: Karan Sharma
Supervisor: Dr. Stephen S. Fong, PhD
Date: 09/26/2011
Graduation Date: May 2012
2. SYSTEMS BIOLOGICAL ENGINEERING LAB
Number of PhD candidates: 3
Number of Undergraduates: 5
Research Topics:
Whole Cell Modeling
Genetic Programming
Metabolic Pathway Engineering
Laboratory Adaptive Evolution
Organisms: Thermobifida fusca, E.Coli
3. LOCATION
Fermentation Research Engineer
Principle Investigator: Dr. Stephen S. Fong
Systems Biological Lab
Rm. 409 West Engineering Hall
4. BIOFUELS
What is a Biofuel?
A fuel that is derived from natural and biological raw
masses.
Kind of biofuels:
Ethanol
Biodiesel
Bio-butanol, propanol
Other higher alcohols
5. AN ILLUSTRATION OF
HOW BIOFUELS ARE
PRODUCED TODAY!
Lawrence Berkeley National Lab.
6. WHAT IS MOST IMPORTANT IN THE PROCESS!
Cellulose:
is an organic compound. A polysaccharide consisting of
a linear chain of several hundred to over ten thousand
β(14) linked by D-glucose units.
is the structural component of the primary cell
wall of green plants.
7. THERMOBIFIDA FUSCA
An aerobic filamentous bacterium.
Rod shaped.
Major degrader of plant cell wall.
Extremely high thermo-stability of Cellulases (up to
70°Celcius)
Cellulases produced by T.fusca can degrade the
cellulose to sugars which are then used to make
biofuels.
8. THERMOBIFIDA FUSCA B6
B6 is an engineered strain by T.fusca made Dr. Yu
Deng and
Dr. S.S. Fong.
Systems
Biological
Lab
10. INTEREST – THE PROJECT
Fermentation Engineering of T.fusca & B6
To improve overall production and yield of Butyrate and
1-proponal.
Parameters to improve
Aeration Rate
Agitation (Stir) Rate
pH
Different carbon sources (c.s.) used for growth
(glucose, glycerol & cellbiose)
Temperature
Instruments used
Bioreactor
Flask Test
11. DATA ANALYSIS METHODS
Data was analyzed by two different methods.
HPLC – High Performance Liquid Chromatography
is a chromatographic technique that can separate a mixture of
compounds to identify, quantify and purify the individual
components of the mixture.
Key Peaks in the HPLC
0.5% Butyrate 23.095
0.5% Glycerol 13.868
1% Propanol 25.328
Acetate 16.412 -- 16.147
Acetic Acid 15.927
Butyric Acid 18.227
Cellibiose 8.366
Propionic Acid 18.607 (Ri) -- 18.834
Sucrose 8.415
12. DATA ANALYSIS METHODS (CONTI.)
Bradford Protein Assay
is an analytical spectroscopic procedure used to measure the
concentration of a protein in a solution. It is dependent on the
amino acid composition of the measured protein.
4.000
3.500
3.000
2.500
2.000 Carbon Source
Glycerol
1.500 Butyrate (g/L)
O.D.
1.000
0.500
0.000
0 2 4 6 10 13 17 31.5 33 34 35 37.5
Time (hours)
15. FUTURE WORK
Explore future aspects of T.Fusca fermentation
engineering.
More specific & different parameters
Explore future aspects of metabolic engineering of
T.Fusca
16. ACKNOWLEDGMENTS!
Dr. Stephen S. Fong PhD
Systems Biological Engineering Lab
Colleagues: Cindy Lovelace, Dr. Yu Deng, Dr. Chris
Gowen & Niti Vanee