k.u.leuven essencia coli: the fragrance factory essencia...
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K.U.LeuvenEssencia coliEssencia coli : The Fragrance Factory
The multidisciplinary team from the University of Leuven Belgium
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University of Leuven, Belgium
K.U.LeuvenEssencia coli
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K.U.LeuvenEssencia coliThe Fragrance Factory
Aim:Aim:Design and build a bacterium that sets and controls the vanillin concentration in the environment
Results:
vanillin concentration in the environment
Results:
• fully integrated modelfully integrated model• 19 parts and devices made• 7 new parts!7 new parts!
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K.U.LeuvenEssencia coliSubsystems of Essencia coli
Input = blue light receptor
Production = tyrosine to vanillin conversion
Sensing = vanillin receptorSensing = vanillin receptor
Controlling = key/antisense key/lock4
Controlling key/antisense key/lock
K.U.LeuvenEssencia coliIntegrated system of Essencia coli
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K.U.LeuvenEssencia coliInput: the Blue light receptor
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K.U.LeuvenEssencia coliBlue light drives the vanillin production
7N. Tschowri, S. Busse and R. Hengge. Genes Dev 23: 522-534 (2009)
K.U.LeuvenEssencia coliBlue light drives the vanillin production
8N. Tschowri, S. Busse and R. Hengge. Genes Dev 23: 522-534 (2009)
K.U.LeuvenEssencia coliThe Blue light responsive promoter is weak
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K.U.LeuvenEssencia coliBlue light increases expression
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K.U.LeuvenEssencia coliProduction: converting tyrosine to vanillin
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K.U.LeuvenEssencia coliFive step conversion to vanillin
12Follow-up on previous work done by Prof. C. French and the iGEM team 2007 of the University of Edinburgh
K.U.LeuvenEssencia coliSensing: the Vanillin receptor
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K.U.LeuvenEssencia coliSensing drives antisense key expression
PvirB
14Natural BioBricks made from Agrobacterium tumefaciens
K.U.LeuvenEssencia coliControlling: the key/antisense key system
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K.U.LeuvenEssencia coliControl by key/antisense key balance
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Vanillin sensing
Blue light100
molecules
Production of vanillin
Vanillin sensing
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16Follow-up on previous work done by the 2006 iGEM team from the University of Berkeley
0 2000 4000 6000 8000 time (s)
K.U.LeuvenEssencia coliSummary animation
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K.U.LeuvenEssencia coliUsing a model to realise the concept
Solution
ConceptSimplified
Conceptmodel
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K.U.LeuvenEssencia coliSimplified model
Disturbance
+‐
[Vanillin][Key] [Key] ++‐ +
[Antikey][Antikey]
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K.U.LeuvenEssencia coliFully integrated kinetic model
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K.U.LeuvenEssencia coliTwo operational modes
Deficiency Abundance Deficiency Abundance
+ + +[L k]+‐
++ ‐
++
[Leak]
Speed of feedback ∆t Speed of feedback
Effect of copy number
∆t
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Effect of copy number
K.U.LeuvenEssencia coliTagging reduces feedback delay
Half life determines delay
KEY
tration
Production enzymes
T i V illi tial con
cen
τ=10τ=22
Tyrosine Vanillin
% init
key
τ=2τ=4
τ 10
Time [au]Use tag to reduce the feedback delay
Half lifeTAG
No TAG
COMT
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0 500 1000Half life [min]
K.U.LeuvenEssencia coliThe effect of copy number
45 copies
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15
25
0
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K.U.LeuvenEssencia coliThe effect of copy number
45 copies
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15
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0
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K.U.LeuvenEssencia coliWhat in the case of abundance?
‐+[Leak]
+
Evaporation rate determines feedback!
But vanillin is not very volatile
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K.U.LeuvenEssencia coliVirA: internal sensing is not an issue
Phenol sensor senses intracellular vanillin
OM
Cout Dif
PP
ffu
τ ≈ 5 [ms]k = 1/ τ ≈ 200 [1/s]
VirA IM
Cin
sioCin
nVanillin
But fast diffusion over cell membrane!
(C C ) 1/k S26
(Cin – Cout) = 1/k Svanillin
K.U.LeuvenEssencia coliAccomplishments
made sequenced
parts
Fully integrated model
made sequenced
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Pitfalls in system tackled2 2
Characterized new natural BioBrick: Blue light receptor11 7
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OtherEthics & human practice Helped other teams
Wiki Had a great summer
27Poster follow our team members
K.U.LeuvenEssencia coliThanks to
Gold Sponsors
BioSCENTer l f
CMPG
Faculty of Bioscience EngineeringFaculty of EngineeringFaculty of ScienceFaculty of ScienceFaculty of Biomedical Sciences
Silver Sponsors
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K.U.Leuven sympathizer
K.U.LeuvenEssencia coliThanks for your attention!
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