title: Quantifying biomass composition by gas chromatography/mass spectrometry.
abstract: We developed a set of methods for the quantification of four major components of microbial biomass using gas chromatography/mass spectrometry (GC/MS). Specifically, methods are described to quantify amino acids, RNA, fatty acids, and glycogen, which comprise an estimated 88% of the dry weight of Escherichia coli. Quantification is performed by isotope ratio analysis with fully (13)C-labeled biomass as internal standard, which is generated by growing E. coli on [U-(13)C]glucose. This convenient, reliable, and accurate single-platform (GC/MS) workflow for measuring biomass composition offers significant advantages over existing methods. We demonstrate the consistency, accuracy, precision, and utility of this procedure by applying it to three metabolically unique E. coli strains. The presented methods will have widespread applicability in systems microbiology and bioengineering.
authors.affiliation: Department of Chemical and Biomolecular Engineering, Metabolic Engineering and Systems Biology Laboratory, University of Delaware , Newark, Delaware 19716, United States.
authors.forename: Christopher P
authors.initials: CP
authors.lastname: Long
authors.forename: Maciek R
authors.initials: MR
authors.lastname: Antoniewicz
mesh_terms: D000596:Amino Acids / Q000032:analysis*; D018533:Biomass; D002247:Carbon Isotopes; D004926:Escherichia coli / Q000737:chemistry* / Q000378:metabolism; D005227:Fatty Acids / Q000032:analysis*; D005285:Fermentation; D008401:Gas Chromatography-Mass Spectrometry; D005947:Glucose / Q000378:metabolism; D006003:Glycogen / Q000032:analysis*; D012329:RNA, Bacterial / Q000032:analysis*
chemical_list: D000596:Amino Acids; D002247:Carbon Isotopes; D005227:Fatty Acids; D012329:RNA, Bacterial; D006003:Glycogen; D005947:Glucose