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Insights in Enzyme Research
ISSN: 2573-4466
E u r o S c i C o n C o n g r e s s o n
Enzymology and
Molecular Biology
A u g u s t 1 3 - 1 4 , 2 0 1 8
P a r i s , F r a n c e
Enzymology 2018
T
he present project presents a combination of methodologies that manages to turn around the design-construction-test
cycle of bacterial strains of metabolic engineering production. We started with an in silico design generated by the genomic
scale model of last generation Escherichia coli (ME-iOL1554). From this, the strains were generated using molecular biology
tools. The strains generated were characterized in a simple experimental system but with strict micro aerobic conditions and
underwent a process of adaptive evolution in the same experimental system, managing to generate strains with fermentative
pathways interrupted but that manage to grow under strict micro aerobic conditions. The strains generated produced L-alanine
(although not in titles close to that predicted by the metabolic model at genomic scale), the exo-metabolomic analyzes of one
of the strains show that it is igniting latent fermentation pathways not previously described. This is why this work constitutes a
conceptual advance for several reasons. 1) Test the use of computer models as a design tool, a combination of systems biology
and synthetic biology is achieved. Both sciences are of great importance and relevance today. 2) The concept of growth-coupled
(Growth-Coupled), a fundamental quality in a production strain, is experimentally validated. 3) A combination of methodologies
was implemented: computational design, molecular biology, fermentations, adaptive evolution and exo-metabolomics by H1-
NMR. 4) An advance was achieved in the generation of L-Alanine producing strains, however the most important result of the
project was the use of computational models as a design tool and the discovery of latent fermentation pathways (ethylene glycol
and methanol) in Escherichia coli, which could reinforce what has been said and proposed by other researchers. At the moment,
there are two strains whose characteristics make them candidates for strains ¨Chasis¨
.
biolex.corp@gmail.comGeneration of bacterial strains of production,
with a growth-coupled focus for its application
in synthetic biology
Cruz Rodriguez Francisco
1
, Feist Adam
2
and Utrilla
Carreri Jose
3
1
Metropolitan Autonomous University, Mexico
2
University of California San Diego (UCSD), California, USA
3
Genomics Sciences Center-UNAM, Systems and Synthetic Biology Lab, Mexico
Insights Enzyme Res 2018, Volume 2
DOI: 10.21767/2573-4466-C1-003