Computational Platform for Flux Analysis Using 13C-Label Tracing- Phase I SBIR Final Report

Van Dien, Stephen J.

Genomatica, Inc.

关键词: Isotopomer Label Tracing;    Strains Flux Analysis;    10 Synthetic Fuels;    Amino Acids;    Methylobacterium Extorquens;   

DOI  :  10.2172/859298
RP-ID  :  DOE ER83965-1
RP-ID  :  FG02-04ER83965
RP-ID  :  859298

美国|英语

来源: UNT Digital Library

【 摘 要 】

Isotopic label tracing is a powerful experimental technique that can be combined with metabolic models to quantify metabolic fluxes in an organism under a particular set of growth conditions. In this work we constructed a genome-scale metabolic model of Methylobacterium extorquens, a facultative methylotroph with potential application in the production of useful chemicals from methanol. A series of labeling experiments were performed using 13C-methanol, and the resulting distribution of labeled carbon in the proteinogenic amino acids was determined by mass spectrometry. Algorithms were developed to analyze this data in context of the metabolic model, yielding flux distributions for wild-type and several engineered strains of M. extorquens. These fluxes were compared to those predicted by model simulation alone, and also integrated with microarray data to give an improved understanding of the metabolic physiology of this organism.