Difference between revisions of "Team:Toulouse/Modeling"
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− | Formate is | + | Formate is naturally produced by <i>E.coli</i> but at a level that is quite low. Our project requires that Apicoli produces more. |
− | to | + | Hence we had to optimize its biosynthesis by studying the genes coding for the enzymes involved in the pathway. We decided to focus our efforts on the Pyruvate Formate Lyase (PFL), the enzyme that causes degradation of pyruvate, thus yielding formate. |
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Revision as of 16:06, 7 September 2015
Modeling
Content
Metabolic networks
As said before, the aim of our project is to create a biological system able to produce two molecules:
butyric acid and formic acid.
To achieve this, we need to modify the existing balance between the metabolic pathways present in E.Coli.
Indeed, we want to optimize butyrate and formate productions in our bacterium by adjusting environmental conditions in order to obtain the desired concentrations of the associated acids.
The following metabolic network represents all of the known metabolites and metabolic pathways for Escherichia coli K12 MG1655(best known model). It was obtained from KEGG database [1].
Our first step was to identify the pathways in which our molecules take part, in order to have a clear understanding of their role and effect.
Figure 1: Kegg Metabolic pathways - Escherichia coli K-12 MG1655
Formate network
Formate is naturally produced by E.coli but at a level that is quite low. Our project requires that Apicoli produces more. Hence we had to optimize its biosynthesis by studying the genes coding for the enzymes involved in the pathway. We decided to focus our efforts on the Pyruvate Formate Lyase (PFL), the enzyme that causes degradation of pyruvate, thus yielding formate.
Figure 2: Reaction catalyzed by PFL
The subnetwork presented below is the result of this mapping, it was realised with the MetExplore platform [2] and present all known enzymes from the Kegg and ByoCyc databases involved with the production or consumption of formate. This map will help us predict the consequences of formate overproduction in Apicoli. Formate, harmful if to much is produced, is metabolized into others products most of the time. We thus have to find the balance between producing enough formate without killing Apicoli.
Figure 3: Metabolic network of all reactions involving formate happening in E.coli
Butyrate network
Flux Balance Analysis (FBA)
Presentation
To help ourselves creating Apicoli we modelised our system by using Flux Balance Analysis (FBA) and Flux Variability Analysis (FVA). We used the most recent described model for E.coli K12 MG1655, describing all metabolic pathways known and up to date. This XML file
Annexes
References
- [1] KEGG Metabolic pathways - Escherichia coli K-12 MG1655
- [2] Le Conte Y, Arnold G, Trouiller J, Masson C, Chappe B, Ourisson G. 1989. Attraction of the parasitic mite varroa to the drone larvae of honey bees by simple aliphatic esters. Science 245:638–639.
- [3] Methods for attracting honey bee parasitic mites. [accessed 2015 Jul 24].
- [4] Louis P, Flint HJ. 2009. Diversity, metabolism and microbial ecology of butyrate-producing bacteria from the human large intestine. FEMS Microbiol. Lett. 294:1–8. [5] Atsumi S, Cann AF, Connor MR, Shen CR, Smith KM, Brynildsen MP, Chou KJY, Hanai T, Liao JC. 2008. Metabolic engineering of Escherichia coli for 1-butanol production. Metabolic Engineering 10:305–311.
- [6] Wallace KK, Bao Z-Y, Dai H, Digate R, Schuler G, Speedie MK, Reynolds KA. 1995. Purification of Crotonyl-CoA Reductase from Streptomyces collinus and Cloning, Sequencing and Expression of the Corresponding Gene in Escherichia coli. European Journal of Biochemistry 233:954–962.