Eau d’e coli igem 2006: mit. meet the team The Undergrads: Kate Broadbent Biological Engineering Andre Green Biology / Physics Stephen Payne Biological.

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Presentation transcript:

eau d’e coli igem 2006: mit

meet the team 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 Austin Che Jason Kelly Reshma Shetty Samantha Sutton The Faculty: Drew Endy Tom Knight

motivations eau d’e coli mit igem 2006

background A. thaliana JMT jasmine O. basilicum CCMT cinnamon A. majus BAMT fresh / floral BSMT wintergreen C. breweriS. cerevisiae ATF1 banana

background O. basilicum CCMT cinnamon A. majus BAMT fresh / floral BSMT wintergreen C. breweriS. cerevisiae ATF1 banana

background A. majus BAMT fresh / floral BSMT wintergreen C. breweriS. cerevisiae ATF1 banana

background BSMT wintergreen C. breweriS. cerevisiae ATF1 banana

timeline: initial planning initial planning scent production self-generating scent production regulation eliminating indole

timeline: scent production initial 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 elimination initial planning scent production self-generating scent production regulation eliminating indole

wintergreen generating device

device generates wintergreen methyl salicylate MW = 152 no methyl salicylate [no peak at 9.1 minutes] E. coli E. coli + precursor (SA) + WGD

banana generating device

precursor biosynthesis

goal: the regulated system

growth dependent regulation osmY: active in stationary phase & under high osmotic pressure conditions osm Y osmY + inverter

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] Indole Peak at 10 minutes MW = 117 wild type E. colichassis: tnaA5 - E. coli

final system

smell test results

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

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.

questions and answers

growth curves