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Multi-capillary column ion mobility spectrometry of volatile metabolites for phenotyping of microorganisms

  • Rational strain engineering requires solid testing of phenotypes including productivity and ideally contributes thereby directly to our understanding of the genotype-phenotype relationship. Actually, the test step of the strain engineering cycle becomes the limiting step, as ever advancing tools for generating genetic diversity exist. Here, we briefly define the challenge one faces in quantifiying phenotypes and summarize existing analytical techniques that partially overcome this challenge. We argue that the evolution of volatile metabolites can be used as proxy for cellular metabolism. In the simplest case, the product of interest is a volatile (e.g., from bulk alcohols to special fragrances) that is directly quantified over time. But also nonvolatile products (e.g., from bulk long-chain fatty acids to natural products) require major flux rerouting that result potentially in altered volatile production. While alternative techniques for volatile determination exist, rather few can be envisaged for medium to high-throughput analysis required for phenotype testing. Here, we contribute a detailed protocol for an ion mobility spectrometry (IMS) analysis that allows volatile metabolite quantification down to the ppb range. The sensivity can be exploited for small-scale fermentation monitoring. The insights shared might contribute to a more frequent use of IMS in biotechnology, while the experimented aspects are of general use for researchers interested in volatile monitoring.

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Author of HS ReutlingenBaumbach, Jörg Ingo
Erschienen in:Synthetic metabolic pathways : methods and protocols. - (Methods in molecular biology ; 1671)
Place of publication:New York
Editor:Michael Krogh Jensen
Document Type:Book chapter
Publication year:2018
Tag:ion mobility spectrometry; multi-capillary column; on-site analysis; online analysis; phenotype screening; volatile metabolites; volatile organic compounds
Page Number:30
First Page:229
Last Page:258
PPN:Im Katalog der Hochschule Reutlingen ansehen
DDC classes:660 Technische Chemie
Open access?:Nein