Rastädter, K., Tramontano, A., Wurm, D. J., Spadiut, O., & Quehenberger, J. (2022, March 29). Dead or alive: a novel PAT tool for the extremophile Sulfolobus acidocaldarius [Conference Presentation]. 7th BioProScale Symposium 2022, Germany. http://hdl.handle.net/20.500.12708/152493
Viability of Sulfolobus acidocaldarius, an extremophilic Archaeon thriving at 75 °C and pH 3.0, is still determined via laborious, time- and material-intensive plating assays. Sulfolobus harbor great potential as still untapped production hosts and biocatalysts in biotechnological applications, nevertheless no industrial processes have been developed yet. During development and scaling of industrial bioprocesses it is crucial to monitor the impact of process parameters on the cultivated organism. Flow cytometry (FCM) combines the ability to analyze single-cell properties in a cell population with viability assessment via the use of fluorescent-active dyes. In this study, commercially available fluorescent- active dyes applicable in S. acidocaldarius were identified. The dyes, Fluorescein diacetate and Concanvalin A conjugated with rhodamin, are suitable for viability determination via FCM. For showing the applicability of the developed at-line PAT tool for bioprocess monitoring a chemostat cultivation at 75°C and pH 3.0 was conducted. Over 800 hours, this novel PAT tool successfully allowed monitoring of viability. In my talk, I will introduce our exciting PAT tool paving the way for an untapped Archaeon into industrial application.
