Reducing Recon 2 for steady-state flux analysis of HEK cell culture.

Abstract:

:A representative stoichiometric model is essential to perform metabolic flux analysis (MFA) using experimentally measured consumption (or production) rates as constraints. For Human Embryonic Kidney (HEK) cell culture, there is the opportunity to use an extremely well-curated and annotated human genome-scale model Recon 2 for MFA. Performing MFA using Recon 2 without any modification would have implied that cells have access to all functionality encoded by the genome, which is not realistic. The majority of intracellular fluxes are poorly determined as only extracellular exchange rates are measured. This is compounded by the fact that there is no suitable metabolic objective function to suppress non-specific fluxes. We devised a heuristic to systematically reduce Recon 2 to emphasize flux through core metabolic reactions. This implies that cells would engage these dominant metabolic pathways to grow, and any significant changes in gross metabolic phenotypes would have invoked changes in these pathways. The reduced metabolic model becomes a functionalized version of Recon 2 used for identifying significant metabolic changes in cells by flux analysis.

journal_name

J Biotechnol

journal_title

Journal of biotechnology

authors

Quek LE,Dietmair S,Hanscho M,Martínez VS,Borth N,Nielsen LK

doi

10.1016/j.jbiotec.2014.05.021

subject

Has Abstract

pub_date

2014-08-20 00:00:00

pages

172-8

eissn

0168-1656

issn

1873-4863

pii

S0168-1656(14)00267-3

journal_volume

184

pub_type

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