ENGINEERING AN EDIBLE PROBIOTIC CONSORTIUM TO REGULATE APPETITE AND DIGESTION COLUMBIA NYC Jacky Cheung, Samuel Magaziner, Suppawat Kongthong, Hudson Lee, Kenya Velez
ENGINEERING AN EDIBLE PROBIOTIC CONSORTIUM TO REGULATE APPETITE
AND DIGESTION
COLUMBIA NYC
Jacky Cheung, Samuel Magaziner, Suppawat Kongthong, Hudson Lee, Kenya Velez
THE RISE OF METABOLIC DISEASES
CHANGES IN CALORIC INTAKE PER CAPITA PER
DAY OVER TIME IN THE U.S
COMPARISON OF AVERAGE CALORIC INTAKE PER
CAPITA AROUND THE WORLD
OBESITY & DIABETES MEDICAL EXPENSES QUALITY OF LIFE
FOOD AS THE SOLUTION – NOT THE PROBLEM
HISTORICAL AND PROJECT SALES OF YOGURT IN THE UNITED STATES WHAT ARE PROBIOTICS?
Live Bacteria & Yeasts
That Improve Health
DIGESTIVE FUNCTIONS
IMMUNE SYSTEM
FUNCTIONS
SUPPORT FRIENDLY
BACTERIA
“Let food be thy medicine and medicine be they food” - Hippocrates
GOAL: ENGINEER A PROBIOTIC FOOD THAT COUNTERS OBESITY AND DIABETES
METHOD TO COUNTER OBESITY AND DIABETES
GLUCAGON-LIKE PEPTIDE I (GLP-I)
INSULIN LEVELS
SATIETY LEVELS
PEPTIDE YY (PYY)
APPETITE LEVELS
GI MOVEMENT
GHRELIN
APPETITE LEVELS
CARDIOVASCULAR HEALTH
ENGINEERING AN EDIBLE PROBIOTIC TO SECRETE GUT PEPTIDES TO REGULATE APPETITE AND DIGESTION
E. C
oli N
issle
Lact
obaci
llus
Reu
teri
phoA
pelB
Lp_3050
M6
GLP-1
Ghrelin PYY
Signal Peptides
Sig
nal Pe
ptid
es
(SPs)
x
Gut Pep
tid
es
(GPs)
EXPERIMENTAL SETUP: SIGNAL PEPTIDE SECRETION
Anhydro-Tetracycline (or
Nisin in Lactobacillus model)
pTet HormoneSP His-TagLysate
1X101
1X10-1
1X10-2
1X102
1X101
1X10-1
Supernatant
His-tagged
Protein
Concentrate
supernatant +
lyse cells
Grow-up, spin
down cell pellet,
extract
supernatant
Blot
RESULTS: HORMONES (NO SIGNAL PEPTIDE)
Ghrelin PYYGlp-1
Note: We expect to see no secretion
Lysate (Left) 1x101 Supernatant (Right) 1x102
Lysate (Left) 1x10-1 Supernatant (Right) 1x101
Lysate (Left) 1x10-2 Supernatant (Right) 1x10-1
RESULTS: SUCCESS WITH PHOA-GLP1 (IMPORTANCE OF INDUCTION TIME)
Lysate (Left) 1x101 Supernatant (Right) 1x102
Lysate (Left) 1x10-1 Supernatant (Right) 1x101
Lysate (Left) 1x10-2 Supernatant (Right) 1x10-1
PhoA-Glp1(36hr induction)PhoA-Glp1 (12hr Induction)
Lysate 1x101
Supernatant 1x101
Supernatant 1x10-1
Note: Same construct, Wildly different Results; also
improvement of previous biobrick BBa_K817000
RESULTS: SUCCESS WITH PELB
Lysate 1x101
Supernatant 1x101
Supernatant 1x102
PelB-Glp1 (36hr) PelB-PYY (36hr)
Here we demonstrate the generalizable
nature of signal peptide secretion
IMPLEMENTING A CONSORTIUM TO PROVIDE REGULATIONS AND SAFETY MECHANISM FEATURES
COMMUNICATION
REGULATION
SAFETY
OUR SYSTEM IS TO BE
TIGHTLY REGULATED AS A
SAFETY PRECAUTION
THE SYSTEM: SAFETY, REGULATION, COMMUNICATION, AND SECRETION MEET
THE SYSTEM BREAKDOWN: THE AHL FACTORY
THE SYSTEM BREAKDOWN: TARGET SECRETION
THE SYSTEM BREAKDOWN: TIME-DELAYED CELL LYSIS (BBA_K1848006)
RESULTS
• Timer-Lysis Cassette system submitted as
biobrick (BBa_K1848006)
• System successfully cloned in
• Time constraints prevented its testing
• However…
1 32
Heat whole milk to
185ºF
Cool to 115ºF and
add engineered L.
reuteri + yogurt
starter culture
Incubate at 37ºC for
8 hr and refrigerate
IN PRACTICUM: YOGURT
Note: Yogurt was not intended for human consumption, nor did it leave the lab
SURVEYING THE PUBLIC• Online survey
• Polled public on GMOs and probiotics, to inform design of safety measures and yogurt
products
• Informed survey takers about probiotics and GMOs
• Twitter and blog
Comfort level in consuming probiotics (On scale of 1-5) before
informing the survey takers about probiotics
Comfort level in consuming probiotics (On scale of 1-5)
after informing the survey takers about probiotics
THE FUTURE: YOGHURT (OR LOOKING WHEY INTO THE FUTURE)
• Going Forward:
1. Finish screen of submitted quorum sensing system
2. Move system into L. reuteri
3. Test active system’s ability to function within yogurt
culture
4. Screen more hormones
• The Foreseeable future:
• Catalog of peptide secreting yogurt
• Biotech branch based around personalized consumable
medicines
IN SUMMATION…
• We managed to:
1. Submit 6 bio-bricks (5 of which are well-
characterized) (BBa_K1848001-006)
2. Successfully secrete 3 gut hormones and 2 signal
peptides in E. coli
3. Develop a promising system quorum sensing based
secretion and lysis
4. Move an engineered L. reuteri system in yogurt
5. Poll on public opinion of probiotics and willingness to
consume GMOs
6. Have an amazing summer and lay the foundation for
future iGEM teams from our University
Student Team MembersThe Whole Team
ATTRIBUTIONS AND ACKNOWLEDGMENT
• Undergraduate Team Members: Suppawat Kongthong, Jacky Cheung, Kenya
Velez, Hudson Lee, Samuel Magaziner
• Mentors: Nathan Johns, Sway Chen, and Sonja Billerbeck
• PIs: Harris Wang, Virginia Cornish, Ken Shepard, Dana Pe’er
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