Variability in subpopulation formation propagates into biocatalytic variability of engineered Pseudomonas putida strains Article Swipe
Martin Lindmeyer
,
Michael Jahn
,
Carsten Vorpahl
,
Susann Müller
,
Andreas Schmid
,
Bruno Bühler
·
YOU?
·
· 2015
· Open Access
·
· DOI: https://doi.org/10.3389/fmicb.2015.01042
YOU?
·
· 2015
· Open Access
·
· DOI: https://doi.org/10.3389/fmicb.2015.01042
Pivotal challenges in industrial biotechnology are the identification and overcoming of cell-to-cell heterogeneity in microbial processes. While the development of subpopulations of isogenic cells in bioprocesses is well described (intra-population variability), a possible variability between genetically identical cultures growing under macroscopically identical conditions (clonal variability) is not. A high such clonal variability has been found for the recombinant expression of the styrene monooxygenase genes styAB from Pseudomonas taiwanensis VLB120 in solvent-tolerant Pseudomonas putida DOT-T1E using the alk-regulatory system from P. putida GPo1. In this study, the oxygenase subunit StyA fused to eGFP was used as readout tool to characterize the population structure in P. putida DOT-T1E regarding recombinant protein content. Flow cytometric analyses revealed that in individual cultures, at least two subpopulations with highly differing recombinant StyA-eGFP protein contents appeared (intra-population variability). Interestingly, subpopulation sizes varied from culture-to-culture correlating with the specific styrene epoxidation activity of cells derived from respective cultures (clonal variability). In addition, flow cytometric cell sorting coupled to plasmid copy number (PCN) determination revealed that detected clonal variations cannot be correlated to the PCN, but depend on the combination of the regulatory system and the host strain employed. This is, to the best of our knowledge, the first work reporting that intra-population variability (with differing protein contents in the presented case study) causes clonal variability of genetically identical cultures. Respective impacts on bioprocess reliability and performance and strategies to overcome respective reliability issues are discussed.
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- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.3389/fmicb.2015.01042
- https://www.frontiersin.org/articles/10.3389/fmicb.2015.01042/pdf
- OA Status
- gold
- Cited By
- 17
- References
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- Related Works
- 10
- OpenAlex ID
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All OpenAlex metadata
Raw OpenAlex JSON
- OpenAlex ID
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https://openalex.org/W1816510943Canonical identifier for this work in OpenAlex
- DOI
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https://doi.org/10.3389/fmicb.2015.01042Digital Object Identifier
- Title
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Variability in subpopulation formation propagates into biocatalytic variability of engineered Pseudomonas putida strainsWork title
- Type
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articleOpenAlex work type
- Language
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enPrimary language
- Publication year
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2015Year of publication
- Publication date
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2015-10-01Full publication date if available
- Authors
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Martin Lindmeyer, Michael Jahn, Carsten Vorpahl, Susann Müller, Andreas Schmid, Bruno BühlerList of authors in order
- Landing page
-
https://doi.org/10.3389/fmicb.2015.01042Publisher landing page
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https://www.frontiersin.org/articles/10.3389/fmicb.2015.01042/pdfDirect link to full text PDF
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YesWhether a free full text is available
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goldOpen access status per OpenAlex
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https://www.frontiersin.org/articles/10.3389/fmicb.2015.01042/pdfDirect OA link when available
- Concepts
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Pseudomonas putida, Biology, Population, Bioprocess, Recombinant DNA, Genetics, Cell sorting, Computational biology, Gene, Flow cytometry, Demography, Sociology, PaleontologyTop concepts (fields/topics) attached by OpenAlex
- Cited by
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17Total citation count in OpenAlex
- Citations by year (recent)
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2024: 2, 2022: 1, 2021: 1, 2020: 3, 2019: 4Per-year citation counts (last 5 years)
- References (count)
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87Number of works referenced by this work
- Related works (count)
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10Other works algorithmically related by OpenAlex
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