PMID: 11607651Apr 16, 1996Paper

Analysis of metabolic pathways by the growth of cells in the presence of organic solvents

Proceedings of the National Academy of Sciences of the United States of America
H E SpinnlerE N Vulfson

Abstract

A new approach to the analysis of metabolic pathways involving poorly water-soluble intermediates is proposed. It relies upon the ability of the hydrophobic intermediates formed by a sequence of intracellular reactions to cross the membrane(s) and partition between aqueous and organic phases, when cells are incubated in the presence of a nonpolar and nontoxic organic solvent. As a result of this thermodynamically driven efflux of the formed intermediates from the cell, they accumulate in the organic medium in sufficient quantities for GC-MS analysis and identification. This enables direct determination of the sequence of chemical reactions involved with no requirement for the isolation of each individual metabolite from a cell-free extract. The feasibility of the proposed methodology has been demonstrated by the elucidation of the biosynthesis of (R)-gamma-decalactone from (R)-ricinoleic acid catalyzed by the yeast Sporidiobolus ruinenii grown in the presence of decane. The corresponding 4-hydroxy-acid intermediates, formed in the course of beta-oxidation of (R)-ricinoleic acid, were simultaneously observed in a single experiment on the same chromatogram. Potential applications of this proposed methodology are briefly discussed.

References

Jun 21, 1991·Science·G Stephanopoulos, J J Vallino
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Mar 1, 1993·Antimicrobial Agents and Chemotherapy·M N PrichardC Shipman

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Citations

May 24, 2003·Applied Microbiology and Biotechnology·Y WachéJ-M Belin
Sep 19, 2015·Critical Reviews in Biotechnology·Carrie PerkinsArnold L Demain
Oct 30, 2003·Journal of Proteome Research·Tomoyoshi SogaTakaaki Nishioka

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