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Discussion papers
https://doi.org/10.5194/acp-2019-23
© Author(s) 2019. This work is distributed under
the Creative Commons Attribution 4.0 License.
https://doi.org/10.5194/acp-2019-23
© Author(s) 2019. This work is distributed under
the Creative Commons Attribution 4.0 License.

Submitted as: research article 15 Jan 2019

Submitted as: research article | 15 Jan 2019

Review status
This discussion paper is a preprint. It has been under review for the journal Atmospheric Chemistry and Physics (ACP). A final paper in ACP is not foreseen.

Freezing from the inside. Ice nucleation in Escherichia coli and Escherichia coli ghosts by inner membrane bound ice nucleation protein InaZ

Johannes Kassmannhuber1,2, Sergio Mauri3, Mascha Rauscher1, Nadja Brait4, Lea Schöner1, Angela Witte4, Tobias Weidner5, and Werner Lubitz1 Johannes Kassmannhuber et al.
  • 1BIRD-C GmbH, Vienna, Austria
  • 2Centre of Molecular Biology, University of Vienna, Vienna, Austria
  • 3Max Planck Institute for Polymer Research, Mainz, Germany
  • 4Department of Microbiology, Immunobiology and Genetics, Max F. Perutz Laboratories, University of Vienna, Vienna, Austria
  • 5Department of Chemistry, Aarhus University, Aarhus C, Denmark

Abstract. An N-terminal truncated form of the ice nucleation protein (INP) of Pseudomonas syringae lacking the transport sequence for the localization of InaZ in the outer membrane was fused to N- and C- terminal inner membrane (IM) anchors and expressed in Escherichia coli C41. The ice nucleation (IN) activity of the corresponding living recombinant E. coli catalyzing heterogeneous ice formation of super-cooled water at high subzero temperatures was tested by droplet freezing assay. Median freezing temperature (T50) of the parental living E. coli C41 cells without INP was detected at −20.1 °C and with inner membrane anchored INPs at T50 value between −7 °C and −9 °C demonstrating that IM anchored INPs facing the luminal IM site are able to induce IN from the inside of the bacterium almost similar to bacterial INPs located at the outer membrane. Bacterial Ghosts (BGs) derived from the different constructs showed first droplet freezing values between −6 °C and −8 °C whereas C41 BGs alone without carrying IM anchored INPs exhibit a T50 of −18.9 °C. The more efficient IN of INP-BGs compared to their living parental strains can be explained by the free access of IM anchored INP constructs to ultrapure water filling the inner space of the BGs. The cell killing rate of -NINP carrying E. coli at subzero temperatures is higher when compared to survival rates of the parental C41 strain.

Johannes Kassmannhuber et al.
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Interactive discussion
Status: closed
Status: closed
AC: Author comment | RC: Referee comment | SC: Short comment | EC: Editor comment
Printer-friendly Version - Printer-friendly version Supplement - Supplement
Johannes Kassmannhuber et al.
Johannes Kassmannhuber et al.
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Short summary
Empty cell envelopes of E. coli, called Bacterial Ghosts (BGs), carrying the ice nucleation protein of P. syringae on their inner membrane were successfully tested for their ability to act as ice nuclei at low subzero temperature (−6 °C) using a freezing droplet assay. Influence of airborne ice-active bacteria on cloud condensation and rain promotion within troposphere is well known. Illustrated by our results, BGs prove themselves as promising and environmental-friendly tool for cloud seeding.
Empty cell envelopes of E. coli, called Bacterial Ghosts (BGs), carrying the ice nucleation...
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