In-Silico - Comparative Growth Analysis of Wild type vs PR Transformed cells

(Metabolic Modeling using COnstraint Based Reconstruction and Analysis)

Hypothesis

1. Increase in growth rate due to Proteorhodopsin proton efflux in minimal carbon media
2. Proton efflux generated by Proteorhodopsin increases ATP production

Model Design

1. Genome scale metabolic model thermodynamic data for genome scale E.coli K-12 MG1655 was derived. This was done by alignment with genomic annotation and the metabolic content of EcoCyc, characterization and quantification of biomass components and maintenance requirements of cell required for growth of the cell and thermodynamic data for reactions[1].


2. Reconstruction of the pathway was carried out to suit our project, hence involving the effects due to Proteorhodpsin pumping activity. Data for pH gradient [2], the delta [H+] [3] was taken from literature and hence flux was calculated to formulate a comprehensive model.

Validation of Model

Simulations for Proof of Concept

Reference

1. "A genome-scale metabolic reconstruction for Escherichia coli K-12 MG1655 that accounts for 1260 ORFs and thermodynamic information" Adam M Feist[1], Christopher S Henry[2], Jennifer L Reed[1], Markus Krummenacker[3], Andrew R Joyce[1], Peter D Karp[3],Linda J Broadbelt[2], Vassily Hatzimanikatis[4] and Bernhard Ø Palsson[1],*
2. "Proteorhodopsin photosystem gene expression enables photophosphorylation in a heterologous host" A. Martinez*, A. S. Bradley†, J. R. Waldbauer‡, R. E. Summons†, and E. F. DeLong*§
3. "Light-powering Escherichia coli with proteorhodopsin" Jessica M. Walter*†, Derek Greenfield*‡, Carlos Bustamante*†‡§¶_, and Jan Liphardt*†‡**






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