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Mechanistic insights into the adaptive evolvability of spore heat resistance in Bacillus cereus sensu lato

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Abstract
Wet heat treatment is a commonly applied method in the food and medical industries for the inactivation of microorganisms, and bacterial spores in particular. While many studies have delved into the mechanisms underlying wet heat killing and spore resistance, little attention has so far been dedicated to the capacity of sporeforming bacteria to tune their resistance through adaptive evolution. Nevertheless, a recent study from our group revealed that a psychrotrophic strain of the Bacillus cereus sensu lato group (i.e. Bacillus weihenstephanensis LMG 18989) could readily and reproducibly evolve to acquire enhanced spore wet heat resistance without compromising its vegetative cell growth ability at low temperatures. In the current study, we demonstrate that another B. cereus strain (i.e. the mesophilic B. cereus sensu stricto ATCC 14579) can acquire significantly increased spore wet heat resistance as well, and we subjected both the previously and currently obtained mutants to whole genome sequencing. This revealed that five out of six mutants were affected in genes encoding regulators of the spore coat and exosporium pathway (i.e. spoIVFB, sigK and gerE), with three of them being affected in gerE. A synthetically constructed ATCC 14579 Delta gerE mutant likewise yielded spores with increased wet heat resistance, and incurred a compromised spore coat and exosporium. Further investigation revealed significantly increased spore DPA levels and core dehydration as the likely causes for the observed enhanced spore wet heat resistance. Interestingly, deletion of gerE in Bacillus subtilis 168 did not impose increased spore wet heat resistance, underscoring potentially different adaptive evolutionary paths in B. cereus and B. subtilis.
Keywords
THERMAL INACTIVATION, SUBTILIS SPORES, SPORULATION, SEQUENCE, GERE, Endospore, Directed evolution, gerE, Spore coat, Lysozyme, DPA

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MLA
Khanal, Sadhana, et al. “Mechanistic Insights into the Adaptive Evolvability of Spore Heat Resistance in Bacillus Cereus Sensu Lato.” INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY, vol. 418, 2024, doi:10.1016/j.ijfoodmicro.2024.110709.
APA
Khanal, S., Kim, T. D., Begyn, K., Duverger, W., Kramer, G., Brul, S., … Aertsen, A. (2024). Mechanistic insights into the adaptive evolvability of spore heat resistance in Bacillus cereus sensu lato. INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY, 418. https://doi.org/10.1016/j.ijfoodmicro.2024.110709
Chicago author-date
Khanal, Sadhana, Tom Dongmin Kim, Katrien Begyn, Wouter Duverger, Gertjan Kramer, Stanley Brul, Andreja Rajkovic, et al. 2024. “Mechanistic Insights into the Adaptive Evolvability of Spore Heat Resistance in Bacillus Cereus Sensu Lato.” INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY 418. https://doi.org/10.1016/j.ijfoodmicro.2024.110709.
Chicago author-date (all authors)
Khanal, Sadhana, Tom Dongmin Kim, Katrien Begyn, Wouter Duverger, Gertjan Kramer, Stanley Brul, Andreja Rajkovic, Frank Devlieghere, Marc Heyndrickx, Joost Schymkowitz, Frederic Rousseau, Veronique Broussolle, Chris Michiels, and Abram Aertsen. 2024. “Mechanistic Insights into the Adaptive Evolvability of Spore Heat Resistance in Bacillus Cereus Sensu Lato.” INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY 418. doi:10.1016/j.ijfoodmicro.2024.110709.
Vancouver
1.
Khanal S, Kim TD, Begyn K, Duverger W, Kramer G, Brul S, et al. Mechanistic insights into the adaptive evolvability of spore heat resistance in Bacillus cereus sensu lato. INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY. 2024;418.
IEEE
[1]
S. Khanal et al., “Mechanistic insights into the adaptive evolvability of spore heat resistance in Bacillus cereus sensu lato,” INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY, vol. 418, 2024.
@article{01JSM3K1CQ4P93ZAH0Q9YJCZW5,
  abstract     = {{Wet heat treatment is a commonly applied method in the food and medical industries for the inactivation of microorganisms, and bacterial spores in particular. While many studies have delved into the mechanisms underlying wet heat killing and spore resistance, little attention has so far been dedicated to the capacity of sporeforming bacteria to tune their resistance through adaptive evolution. Nevertheless, a recent study from our group revealed that a psychrotrophic strain of the Bacillus cereus sensu lato group (i.e. Bacillus weihenstephanensis LMG 18989) could readily and reproducibly evolve to acquire enhanced spore wet heat resistance without compromising its vegetative cell growth ability at low temperatures. In the current study, we demonstrate that another B. cereus strain (i.e. the mesophilic B. cereus sensu stricto ATCC 14579) can acquire significantly increased spore wet heat resistance as well, and we subjected both the previously and currently obtained mutants to whole genome sequencing. This revealed that five out of six mutants were affected in genes encoding regulators of the spore coat and exosporium pathway (i.e. spoIVFB, sigK and gerE), with three of them being affected in gerE. A synthetically constructed ATCC 14579 Delta gerE mutant likewise yielded spores with increased wet heat resistance, and incurred a compromised spore coat and exosporium. Further investigation revealed significantly increased spore DPA levels and core dehydration as the likely causes for the observed enhanced spore wet heat resistance. Interestingly, deletion of gerE in Bacillus subtilis 168 did not impose increased spore wet heat resistance, underscoring potentially different adaptive evolutionary paths in B. cereus and B. subtilis.}},
  articleno    = {{110709}},
  author       = {{Khanal, Sadhana and Kim, Tom Dongmin and Begyn, Katrien and Duverger, Wouter and Kramer, Gertjan and Brul, Stanley and Rajkovic, Andreja and Devlieghere, Frank and Heyndrickx, Marc and Schymkowitz, Joost and Rousseau, Frederic and Broussolle, Veronique and Michiels, Chris and Aertsen, Abram}},
  issn         = {{0168-1605}},
  journal      = {{INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY}},
  keywords     = {{THERMAL INACTIVATION,SUBTILIS SPORES,SPORULATION,SEQUENCE,GERE,Endospore,Directed evolution,gerE,Spore coat,Lysozyme,DPA}},
  language     = {{eng}},
  pages        = {{11}},
  title        = {{Mechanistic insights into the adaptive evolvability of spore heat resistance in Bacillus cereus sensu lato}},
  url          = {{http://doi.org/10.1016/j.ijfoodmicro.2024.110709}},
  volume       = {{418}},
  year         = {{2024}},
}

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