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Study of analysis methods to monitor sporulation during fermentation of Bacillus subtilis to produce endospores

Hannemann, F. and Albin, A. and Bader, J. and Prowe, S. and Senz, M. and Dounia, S. and Moeller, R. (2017) Study of analysis methods to monitor sporulation during fermentation of Bacillus subtilis to produce endospores. In: 19th INTERNATIONAL CONFERENCE on Bacilli & Gram-Positive Bacteria - Abstractbook. 19th INTERNATIONAL CONFERENCE on Bacilli & Gram-Positive Bacteria, 11–15 June 2017, Berlin, Germany.

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Abstract

Bacterial endospores, mainly of Bacillus subtilis, B. atrophaeus or Geobacillus stearothermophilus, are used as biological indicator for the validation of sterilization processes in various industrial processes and applications. Commonly used spores for sterilization studies are spores of B. subtilis, due to their high degree of resistance to chemical and physical treatments, reproducible inactivation response, and ease of use. To prevent falsified results in the validation of sterilization processes, theses spores must have high and comparable D-values as well as a homogeneous distribution of the resistance within every charge. The main influence on the resistance properties is the fermentation process, which lead to the necessity of well-defined methods and analytics of the spore formation for the production process. To produce suspensions containing homogeneously equipped spores, a reproducible fed batch fermentation strategy with optimized concentrations of media components has been developed (VLB; data not yet published). To increase spore resistance, different influence factors such as fermentation time, temperature and composition of sporulation media were investigated. A systematic screening of fermentation capabilities towards optimal spore formation, production and efficiency was performed. A collection of commonly used sterilization-recommended B. subtilis strains was evaluated by applying a set of different molecular biological techniques and classical microbiological assays. Quantitative real time PCR was used to study transcriptomic changes during the sporulation; here major sporulation events (e.g., sigma factors; from Stuelke, Joerg, SubtiWiki) were studied. MALDI-TOF-TOF analyses were conducted to follow changes in the protein spectra (Momo et al, 2013) from during initiation of sporulation until maturation of the dormant spore. The EloTrace® System (Junne et al, 2010) allowed us to characterize the morphological and physiological of the transition of the sporulating cultures using an electro-optical approach. Established and reproducible methods for measuring spore resistance were used to determine batch-specific resistance properties to selected sterilization methods (hydrogen peroxide or UV-C radiation treatments; Raguse et al, 2016).

Item URL in elib:https://elib.dlr.de/118088/
Document Type:Conference or Workshop Item (Poster)
Title:Study of analysis methods to monitor sporulation during fermentation of Bacillus subtilis to produce endospores
Authors:
AuthorsInstitution or Email of AuthorsAuthors ORCID iD
Hannemann, F.Beuth University of Applied Sciences, Dept. V, Biotechnology, Lab for Microbiology, Berlin, GermanyUNSPECIFIED
Albin, A.Beuth University of Applied Sciences Berlin, Dept. V, Lab for Bioprocess technology, Berlin, GermanyUNSPECIFIED
Bader, J.Beuth University of Applied Sciences Berlin, Dept. V, Lab for Bioprocess technology, Berlin, GermanyUNSPECIFIED
Prowe, S.Beuth University of Applied Sciences, Dept. V, Biotechnology, Lab for Microbiology, Berlin, GermanyUNSPECIFIED
Senz, M.Research and Teaching Institute for Brewing in Berlin (VLB), Berlin, Germany 4German Aerospace Center (DLR e.V., Institute of Aerospace Medicine, Radiation Biology Department, Space Microbiology Research Group, Cologne, GermanyUNSPECIFIED
Dounia, S.Research and Teaching Institute for Brewing in Berlin (VLB), Berlin, Germany 4German Aerospace Center (DLR e.V., Institute of Aerospace Medicine, Radiation Biology Department, Space Microbiology Research Group, Cologne, GermanyUNSPECIFIED
Moeller, R.Radiation Biology Department, Institute of Aerospace Medicine, German Aerospace Center (dlr), Cologne, Germany; ralf.moeller (at) dlr.dehttps://orcid.org/0000-0002-2371-0676
Date:2017
Journal or Publication Title:19th INTERNATIONAL CONFERENCE on Bacilli & Gram-Positive Bacteria - Abstractbook
Refereed publication:Yes
Open Access:Yes
Gold Open Access:No
In SCOPUS:No
In ISI Web of Science:No
Status:Published
Keywords:Bacillus subtilis, sporulation
Event Title:19th INTERNATIONAL CONFERENCE on Bacilli & Gram-Positive Bacteria
Event Location:Berlin, Germany
Event Type:international Conference
Event Dates:11–15 June 2017
HGF - Research field:Aeronautics, Space and Transport
HGF - Program:Space
HGF - Program Themes:Research under Space Conditions
DLR - Research area:Raumfahrt
DLR - Program:R FR - Forschung unter Weltraumbedingungen
DLR - Research theme (Project):R - Vorhaben Strahlenbiologie
Location: Köln-Porz
Institutes and Institutions:Institute of Aerospace Medicine > Radiation Biology
Deposited By: Kopp, Kerstin
Deposited On:11 Jan 2018 13:43
Last Modified:31 Jul 2019 20:15

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