Abstract
The use of mixed microbial cultures (MMCs) is seen as an attractive strategy for polyhydroxyalkanoate (PHA) production. In order to optimize the MMC-PHA production process, tools are required to improve our understanding of the physiological state of the PHA-storing microorganisms within the MMC. In the present study, we explored the use of flow cytometry to analyse the metabolic state and polyhydroxybutyrate (PHB) content of the microorganisms from an MMC-PHA production process. A sequencing batch reactor under a feast and famine regime was used to enrich an MMC with PHB-storing microorganisms. Interestingly, once the PHB-storing microorganisms are selected, the level of PHB accumulation depends largely on the metabolic state of these microorganisms and not exclusively on the consortium composition. These results demonstrate that flow cytometry is a powerful tool to help to understand the PHA storage response of an MMC-PHA production process.
Key points
• Flow cytometry allows to measure PHB content and metabolic activity over time.
• Microorganisms showing high PHB content also have high metabolic activity.
• PHB producers with low metabolic activity show low PHB content.
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Funding
This work was financially supported by the Agencia Nacional de Investigación y Desarrollo (ANID) – Chile, with the Science Project for doctoral studies in Chile #21191476, and Proyecto Anillo ACT 172128: Genomics and Applied Microbiology for Biodegradation and Bioproducts (GAMBIO). AS was supported by FONDECYT Postdoctoral Grant #3200748.
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KG: investigation, conceptualisation, analysis, writing—review and editing, visualisation, writing—original draft. AS: writing—review and editing, visualisation. FP: writing—review and editing, visualisation. RN: funding acquisition, project administration. SL: writing—review and editing. MC: conceptualisation, funding acquisition, supervision, writing—review and editing. All authors read and approved the manuscript.
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González, K., Salinas, A., Pinto, F. et al. Flow cytometry: a tool for understanding the behaviour of polyhydroxyalkanoate accumulators. Appl Microbiol Biotechnol 107, 581–590 (2023). https://doi.org/10.1007/s00253-022-12318-x
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DOI: https://doi.org/10.1007/s00253-022-12318-x