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PMID: 15667247 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

High-throughput metabolic state analysis: the missing link in integrated functional genomics of yeasts.

The Biochemical journal ·Vol. 388 ·No. Pt 2 ·2005-06-01 ·Pages 669-77

Villas-Bôas SG, Moxley JF, Akesson M, Stephanopoulos G, Nielsen J

Abstract

The lack of comparable metabolic state assays severely limits understanding the metabolic changes caused by genetic or environmental perturbations. The present study reports the application of a novel derivatization method for metabolome analysis of yeast, coupled to data-mining software that achieve comparable throughput, effort and cost compared with DNA arrays. Our sample workup method enables simultaneous metabolite measurements throughout central carbon metabolism and amino acid biosynthesis, using a standard GC-MS platform that was optimized for this purpose. As an implementation proof-of-concept, we assayed metabolite levels in two yeast strains and two different environmental conditions in the context of metabolic pathway reconstruction. We demonstrate that these differential metabolite level data distinguish among sample types, such as typical metabolic fingerprinting or footprinting. More importantly, we demonstrate that this differential metabolite level data provides insight into specific metabolic pathways and lays the groundwork for integrated transcription-metabolism studies of yeasts.

MeSH Terms
Aerobiosis Amino Acids/biosynthesis Anaerobiosis Carboxylic Acids/metabolism Gene Expression Profiling/methods Genomics/methods Mutation Saccharomyces cerevisiae/genetics,metabolism Sensitivity and Specificity
Chemicals
Amino Acids Carboxylic Acids
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Villas-Bôas Silas G
Centre for Microbial Biotechnology, Technical University of Denmark, BioCentrum-DTU, Building 223, DK-2800 Kongens Lyngby, Denmark.
Moxley Joel F
Akesson Mats
Stephanopoulos Gregory
Nielsen Jens
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
1470-8728
Published
2005-06-01
Pages
669-77
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1138975
Subset
IM
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