Monitoring of population dynamics of Corynebacterium glutamicum by multiparameter flow cytometry.

TitleMonitoring of population dynamics of Corynebacterium glutamicum by multiparameter flow cytometry.
Publication TypeJournal Article
Year of Publication2013
AuthorsNeumeyer, A, Hübschmann, T, Müller, S, Frunzke, J
JournalMicrob Biotechnol
Volume6
Issue2
Pagination157-67
Date Published2013 Mar
ISSN1751-7915
KeywordsBiotechnology, Cell Division, Corynebacterium glutamicum, DNA, Bacterial, Flow Cytometry, Fluorescent Dyes, Indoles, Membrane Potentials, Population Dynamics, Staining and Labeling
Abstract

Phenotypic variation of microbial populations is a well-known phenomenon and may have significant impact on the success of industrial bioprocesses. Flow cytometry (FC) and the large repertoire of fluorescent dyes bring the high-throughput analysis of multiple parameters in single bacterial cells into reach. In this study, we evaluated a set of different fluorescent dyes for suitability in FC single cell analysis of the biotechnological platform organism Corynebacterium glutamicum. Already simple scattering properties of C. glutamicum cells in the flow cytometer were shown to provide valuable information on the growth activity of analysed cells. Furthermore, we used DAPI staining for a FC-based determination of the DNA content of C. glutamicum cells grown on standard minimal or complex media. Characteristic DNA patterns were observed mirroring the typical uncoupled DNA synthesis in the logarithmic (log) growth phase and are in agreement with a symmetric type of cell division of C. glutamicum. Application of the fluorescent dyes Syto 9, propidium iodide, and DiOC(2)(3) allowed the identification of subpopulations with reduced viability and membrane potential within early log and stationary phase populations. The presented data highlight the potential of FC-based analyses for online monitoring of C. glutamicum bioprocesses and provide a first reference for future applications and protocols.

DOI10.1111/1751-7915.12018
Alternate JournalMicrob Biotechnol