eau d e coli igem 2006: mit
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1 eau d e coli igem 2006: mit
2 The Undergrads: meet the team 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 Austin Che Jason Kelly Reshma Shetty Samantha Sutton The Faculty: Drew Endy Tom Knight
3 motivations eau d e coli mit igem 2006
4 background A. thaliana O. basilicum A. majus C. breweri S. cerevisiae JMT CCMT BAMT BSMT ATF1 jasmine cinnamon fresh / floral wintergreen banana
5 background O. basilicum A. majus C. breweri S. cerevisiae CCMT BAMT BSMT ATF1 cinnamon fresh / floral wintergreen banana
6 background A. majus C. breweri S. cerevisiae BAMT BSMT ATF1 fresh / floral wintergreen banana
7 background C. breweri S. cerevisiae BSMT wintergreen ATF1 banana
8 timeline: initial planning initial planning scent production self-generating scent production regulation eliminating indole
9 timeline: scent production initial planning scent production self-generating scent production regulation eliminating indole
10 timeline: self-generating scent initial planning scent production self-generating scent production regulation eliminating indole
11 timeline: production regulation initial planning scent production self-generating scent production regulation eliminating indole
12 timeline: indole elimination initial planning scent production self-generating scent production regulation eliminating indole
13 wintergreen generating device
14 device generates wintergreen E. coli E. coli + precursor (SA) + WGD no methyl salicylate [no peak at 9.1 minutes] methyl salicylate MW = 152 O O OH
15 banana generating device
16 precursor biosynthesis
17 precursor biosynthesis
18 goal: the regulated system
19 growth dependent regulation osmy: active in stationary phase & under high osmotic pressure conditions osmy osmy + inverter
20 regulation is growth dependent BBa_J45995 (posmy-gfp) BBa_J45017 (Negative control) BBa_I7100 (Constitutive promoter-gfp)
21 regulation is growth dependent
22 regulated system: exponential
23 regulated system: stationary
24 regulated system
25 chassis (tnaa5 - ) does not produce indole wild type E. coli chassis: tnaa5 - E. coli Indole Peak at 10 minutes MW = 117 No indole [no peak at 10 minutes]
26 final system
27 smell test results Banana Mint Stinky A (Mint) B (Stinky) C (Banana)
28 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
29 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
30 acknowledgments Natalia 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.
31 questions and answers
32 growth curves Bba_J45995 (posmy-gfp) Bba_J45017 (Negative control) BBa_I7100 (Constitutive promoter-gfp) 0 20,000 40,000 60,000 80,000
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