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“Light to live, live to light” Valencia 2012
PHOTOSYNTHETIC
BIOLAMP
The project:
The project:
AUTOSUFFICIENT
The project:
SOLAR POWER
The project:
AUTOREGULATED
The project:
ENERGY EFFICIENT
The project:
COLD LIGHT
The project:
NON ELECTRIC
The project:
ENVIRONMENTALLY FRIENDLY
The project:
NO FUEL INPUT
The project:
CO2 SINK
The project:
SYNTHETIC ECOLOGY
The project:
ARTIFICIAL TROPHIC
SYMBIOSIS
INTERSPECIES CELL-TALK
The project:
Synechococcus elongatus Aliivibrio fischeri
Synergy can be the
greatest power:
BATTERY AND SWITCH
BIOLAMP
Energy Signal
molecule
The
idea:
Aliivibrio fischeri
Yummy!
suc: sucrose
PpsbAI λ P LuxI
cscB
LuxR LuxICDABE
S. elongatus cscB
LuxR LuxR
LuxR
cI repressor
Lac P
BATTERY AND SWITCH
BIOLAMP
Aliivibrio fischeri
AHL
Yummy!
suc: sucrose
psbAI cI repressor λ P
cscB
LuxR LuxICDABE
S. elongatus cscB
AHL
AHL
LuxR
AHL
LuxR
LuxR
LuxI
Lac P
BATTERY AND SWITCH
BIOLAMP
Integrate both organisms with maximum
efficiency
•Continuous culture •Separate modules
Photosynthesis and Export
Synechococcus elongatus
Biolamp Aliivibrio
fischeri
Designing the common
medium Meet requirements of both organisms:
Synechococcus elongatus Aliivibrio fischeri
High pH High pH
Saline medium Saline medium
Micronutrients C, N organic sources
Synechococcus strain growing in BG-11M enriched with 250mM of salt.
Aliivibrio strain growing in BG-11M enriched with salt and organic sources.
Submitted parts
Overview
Human Practices
Achievements
Modeling
Modelling the
parameters Find the optimal bioreactor design parameters:
• Volumes of the cultures
• Cell densities
In order to optimize:
• Enough light emission
• Stable day/night switch
• Sucrose balance to keep our system autonomous
Metabolic network with 898 reactions + 803 metabolites.
Synechococcus metabolic model + 2 extra reactions: - sucrose export - AHL
LIGHT CO2
+ BIOLOGICAL CONSTRAINTS
Sucrose AHL Growth
Synechococcus
model
Assay production of AHL and sucrose
0.0221mmol/ gDCW·h
Synechococcus
model Growth: exponential rate
(mmol/ gDCW·h)
(mmol/ gDCW·h)
Belta et al, 2001
9 differential equations to predict light emission:
x7´= (Vc[Ai] + Vv.X7 )/Vt
Aliivibrio QS
model
where:
Solar Light CO2
Synechococcus Metabolic network
Aliivibrio Bioluminescence model
AHL AHL SUC SUC
AHL
AHL
Luminescence
SUC
Synergic
model
Submitted parts
Overview
Modeling
Achievements
Human Practices
YES 56% NO
14%
N/A 25%
Population Poll
Human practices:
Synth(ethic) Biology Blog http://igemvalencia2012.wordpress.com/
Kids lectures
Seminars
Indifferent 5%
Overview
Modeling
Human Practices
Achievements
Submitted parts
Submitted parts:
Our submitted part in the pSB1C3 standard BioBrick
Submitted parts:
Testing the part:
Dark (h)
Light (h)
Measure starts at
PpsbAI::luxAB 1 1 Light cycle Dark cycle
PpsbAI::luxAB 8 16 Light cycle Dark cycle
PpsbAI::luxAB 16 8 Light cycle Dark cycle
PpsbAI::luxAB 12 12 Light cycle Dark cycle
PpsbAI::luxAB 0 24 Light cycle Dark cycle
Negative control (WT)
Same Same Same Same At three different light intensities
PpsbAI::luxCDABE fusion vector
Submitted parts
Overview
Modeling
Human Practices
Achievements
Achievements:
Designed a self-sufficient photosynthetically-powered biolamp.
Studied an artificial consortium between two marine organisms.
Built an effective model for the different biolamp processes.
Transformed Synechococcus elongatus PCC7942 to characterize the psbAI promoter.
Submitted a well characterized part to the Registry of Standard Biological Parts.
Opened a bilingual weblog about synthetic biology, focusing on divulgation.
Established collaborations with others iGEM teams: Uppsala, UC London, Copenhagen, Chile.
Tested and characterized new culture medium for growing cyanobacteria in our lab conditions.
Acknowledgments
• Daniel Ducat
• Susan Golden
• Javier Espinosa, from Fisiology, Genetic and Microbiology departments of University of Alicante
• Dr. Carratalá
• Marisa Salvador, from the Molecular Biology Department of University of Valencia
• Joaquín Moreno, from Molecular Biology Department of University of Valencia
• Diego Orzáez, from IBMCP(UPV-CSIC)
• Borja Martinez-Clavel Vallés
• Marta Chillerón Rueda
• Raul Gancedo Sabater
