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Euro Biotechnology 2016
November 07-09, 2016
Volume 6, Issue 7(Suppl) J Biotechnol Biomater
ISSN: 2155-952X JBTBM, an open access journal
conferenceseries .com
November 07-09, 2016 Alicante, Spain
12th Euro Biotechnology Congress
Daniela Ehgartner, J Biotechnol Biomater 2016, 6:7(Suppl)
Methods and applications concerning spore inoculum quality and spore germination in filamentous
bioprocesses
Daniela Ehgartner
Vienna University of Technology, Austria
S pore inoculum quality in filamentous bioprocesses is a critical parameter associated with viable spore concentration and
spore germination. It influences pellet morphology and consequently, process performance. The state-of-the-art method to
investigate this quality attribute is colony forming unit (CFU), being tedious, associated with significant inherent bias and not
applicable in real time. The approach presented here is based on combining viability staining and large-particle flow cytometry
enabling measurements in real-time. It is compatible with complex medium background and allows the quantification of
metabolically active spores and monitoring of spore germination. In an industrial bioprocess with filamentous fungi, a good
correlation to CFU was found. Spore swelling and spore germination were followed over the initial process phase with close
temporal resolution. The validation of the method showed an error of spore classification of less than 5%. In this contribution
bioprocesses with various spore inoculum qualities were monitored at-line. Thereby, critical parameters concerning the amount
of germinating spores in spore inoculum were found. Related to these critical parameters, differences of spore germination on
agar plates vs. liquid environment were observed. These findings challenge the CFU as appropriate method for spore inoculum
quality determination. Two applications of the method are proposed: First, the determination of spore inoculum quality before
starting the cultivation to guarantee a common starting point for the batches. Secondly, monitoring of spore swelling to adjust
the process in real-time to prevent batch failure. These common starting points ensure homogeneity of morphology between
cultivations and consequently, better process performance.
Biography
Daniela Ehgartner is currently pursuing PhD at the University of Technology in Vienna, Austria. She has completed her undergraduate studies in Nutritional Science
and her Masters in Biotechnology and Bioanalytics.