eau d’e coli igem 2006: mit
DESCRIPTION
eau d’e coli igem 2006: mit. The Undergrads: Kate Broadbent Biological Engineering Andre Green Biology / Physics Stephen Payne Biological Engineering Veena Venkatachalam Chemistry / Physics Boyuan Zhu Electrical Engineering and Computer Science The Grad Students: Barry Canton - PowerPoint PPT PresentationTRANSCRIPT
eau d’e coliigem 2006: mit
meet the teamThe Undergrads:Kate Broadbent Biological Engineering
Andre Green Biology / Physics
Stephen Payne Biological Engineering
Veena Venkatachalam Chemistry / Physics
Boyuan Zhu Electrical Engineering and Computer Science
The Grad Students:Barry CantonAustin CheJason KellyReshma ShettySamantha Sutton
The Faculty:Drew EndyTom Knight
motivations
eau d’e coli
mit igem 2006
background
A. thaliana
JMTjasmine
O. basilicu
m
CCMTcinnamon
A. majus
BAMTfresh / floral
BSMTwintergreen
C. breweri S. cerevisia
e
ATF1banana
background
O. basilicu
m
CCMTcinnamon
A. majus
BAMTfresh / floral
BSMTwintergreen
C. breweri S. cerevisia
e
ATF1banana
background
A. majus
BAMTfresh / floral
BSMTwintergreen
C. breweri S. cerevisia
e
ATF1banana
background
BSMTwintergreen
C. breweri S. cerevisia
e
ATF1banana
timeline:initial planning
initial planning • scent production • self-generating scent • production regulation • eliminating indole
timeline:scent productioninitial planning • scent production • self-generating scent • production regulation •
eliminating indole
timeline:self-generating scent
initial planning • scent production • self-generating scent • production regulation • eliminating indole
timeline:production regulation
initial planning • scent production • self-generating scent • production regulation • eliminating indole
timeline:indole eliminationinitial planning • scent production • self-generating scent • production regulation •
eliminating indole
wintergreen generating device
device generates wintergreen
O
O
OH
methyl salicylateMW = 152no methyl
salicylate [no peak at 9.1 minutes]
E. coli E. coli + precursor (SA) + WGD
banana generating device
precursor biosynthesis
precursor biosynthesis
goal: the regulated system
growth dependent regulation
osmY: active in stationary phase & under high osmotic pressure conditions
osmY
osmY + inverter
regulation is growth dependent
0E+0
2E+4
4E+4
6E+4
8E+4
1E+5
1E+5
1E+5
2E+5
2E+5
2E+5
0.0 0.1 0.2 0.3 0.4 0.5
Absorbance 600nm (relative units)
GFP Signal (relative units)
BBa_J45995 (pOsmY-GFP)
BBa_J45017 (Negative control)
BBa_I7100 (Constitutive promoter-GFP)
regulation is growth dependent
regulated system: exponential
regulated system: stationary
regulated system
chassis (tnaA5-) does not produce indole
No indole [no peak at 10
minutes]
IndolePeak at 10 minutes
MW = 117
wild type E. coli chassis: tnaA5- E. coli
final system
smell test results
0
10
20
30
40
50
60
70
80
90
100
A (Mint) B (Stinky) C (Banana)
Percentage
Banana
Mint
Stinky
future applications
Improve the workplace environment for microbiologists working with E. coli
Port our system to bacterial species involved in bioremediation.
Implement our system in bacteria responsible for bad human odor in the mouth, armpits and feet.
Implement our system in yeast used in bread and beer production
conclusion
Designed, built, and tested a synthetic biological system over the course of summer
Engineered synthetic scent systems in E. coli
Regulated via transcription control devices
acknowledgmentsNatalia Dudareva, Purdue University: gifts of expression vectors encoding BAMT, SAMT and BSMT enzymes.
Eran Pichersky, University of Michigan: suggestion of eliminating indole from Escherichia coli to mitigate the natural "bad" smell.
Mary Berlyn, CGSC, The Coli Genetic Stock Center: Esherichia coli strain YYC912.
Brian Cook, MIT: valuable discussions.
Cornelia Reimmann, University of Lausanne: gift of an expression vector carrying the pchBA coding region.
Dieter Haas, University of Lausanne: gift of the Pseudomonas fluorescens strain CHA0.
Peter Bakker, Utrecht University, The Netherlands: gift of an expression vector carrying the pmsCEAB coding region, as well as, gift of the Pseudomonas fluorescens strain WCS374.
Herbert P. Schweizer, Colorado State University: gift of the pUCP22 Esherichia coli to Pseudomonas shuttle vector.
Pamela Silver, Harvard Medical School: gift of yeast BioBricks vector.
questions and answers
growth curves
0.00
0.10
0.20
0.30
0.40
0.50
0.60
0.70
0 10000 20000 30000 40000 50000 60000 70000
Time (min)
Absorbance 600nm (relative units)
Bba_J45995 (pOsmY-GFP)
Bba_J45017 (Negative control)
BBa_I7100 (Constitutive promoter-GFP)