Single-Cell Analysis of Microbial Production Strains in Microfluidic Bioreactors
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Beschreibung
Industrial biotechnology is concerned with the sustainable production of, for
example, fine and bulk chemicals, pharmaceuticals and proteins by utilizing
microorganisms for the conversion of renewable carbon sources. Well known examples
include the production of amino acids by Corynebacterium glutamicum at a million ton
scale per year worldwide, or the recombinant production of insulin by Escherichia coli.
Growth and productivity of the underlying host microorganisms are two key
performance indicators in biotechnological production processes. Assuming isogenic
starting populations, optimal reactor control and mixing, a uniform cell behavior during
growth might be expected. However, as confirmed in recent years, isogenic bacterial
populations can be physiologically heterogeneous. Obviously, there is a strong demand
to unravel microbial population heterogeneity, understand its origin and gain knowledge
on its impact on large scale biotechnological production. Therefore, new analytical
techniques addressing single-cell behavior are the key for further optimization.
In particular, state-of-the-art microfluidic cultivation systems facilitating single-cell
resolution and accurate environmental control over long time periods at the same time,
offer completely new experimental assays on microbial populations. In contrast to
conventional systems, for example, fluorescence activated cell sorting, microfluidic
cultivations enable the analysis of cell dynamics by automated time-lapse microscopy
with full spatio-temporal resolution.
The aim of the present thesis was to develop and establish a new microfluidic
platform technology for microbial single-cell analysis in order to address key concerns
on population heterogeneity and reactor inhomogeneity in industrial biotechnology.
Several unique single-cell cultivation chips were successfully fabricated and validated
with a variety of industrially applied microorganisms. Each device contained up to several
thousand micrometer sized cultivation structures in parallel intended for high-throughput
analysis of single cells and isogenic microcolonies
Autor*in
Alexander Manuel Grünberger
Themen in »Single-Cell Analysis of Microbial Production Strains in Microfluidic Bioreactors«
Bioprocess Microbial Microfluidic