“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

animación dia-noche.pptx