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Membranes Are Decisive for Maximum Freezing Efficiency of Bacterial Ice Nucleators.

ABSTRACT: Ice-nucleating proteins (INPs) from Pseudomonas syringae are among the most active ice nucleators known, enabling ice formation at temperatures close to the melting point of water. The working mechanisms of INPs remain elusive, but their ice nucleation activity has been proposed to depend on the ability to form large INP aggregates. Here, we provide experimental evidence that INPs alone are not sufficient to achieve maximum freezing efficiency and that intact membranes are critical. Ice nucleation measurements of phospholipids and lipopolysaccharides show that these membrane components are not part of the active nucleation site but rather enable INP assembly. Substantially improved ice nucleation by INP assemblies is observed for deuterated water, indicating stabilization of assemblies by the stronger hydrogen bonds of D₂O. Together, these results show that the degree of order/disorder and the assembly size are critically important in determining the extent to which bacterial INPs can facilitate ice nucleation.

SUBMITTER: Schwidetzky R

PROVIDER: S-EPMC8591660 | biostudies-literature | 2021 Nov

REPOSITORIES: biostudies-literature

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Membranes Are Decisive for Maximum Freezing Efficiency of Bacterial Ice Nucleators.

Schwidetzky R R Sudera P P Backes A T AT Pöschl U U Bonn M M Fröhlich-Nowoisky J J Meister K K

The journal of physical chemistry letters 20211101 44

Ice-nucleating proteins (INPs) from <i>Pseudomonas syringae</i> are among the most active ice nucleators known, enabling ice formation at temperatures close to the melting point of water. The working mechanisms of INPs remain elusive, but their ice nucleation activity has been proposed to depend on the ability to form large INP aggregates. Here, we provide experimental evidence that INPs alone are not sufficient to achieve maximum freezing efficiency and that intact membranes are critical. Ice n ...[more]

PMID: 34723523

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Project description:This paper introduces hands-on curricular modules integrated with research in atmospheric ice nucleation, which is an important phenomenon potentially influencing global climate change. The primary goal of this work is to promote meaningful laboratory exercises to enhance the competence of students in the fields of science, technology, engineering, and math (STEM) by applying an appropriate methodology to laboratory ice nucleation measurements. To achieve this goal, three laboratory modules were developed with 18 STEM interns and tested by 28 students in a classroom setting. Students were trained to experimentally simulate atmospheric ice nucleation and cloud droplet freezing. For practical training, this work utilized a simple freezing assay device called the West Texas Cryogenic Refrigerator Applied to Freezing Test (WT-CRAFT) system. More specifically, students were provided with hands-on lessons to calibrate WT-CRAFT with deionized water and apply analytical techniques to understand the physicochemical properties of bulk water and droplet freezing. All procedures to implement the developed modules were typewritten during this process, and shareable read-ahead exploration materials were developed and compiled as a curricular product. Additionally, students conducted complementary analyses to identify possible catalysts of heterogeneous freezing in the water. The water analyses included: pH, conductivity, surface tension, and electron microscopy-energy-dispersive X-ray spectroscopy. During the data and image analysis process, students learned how to analyze droplet freezing spectra as a function of temperature, screen and interpret the data, perform uncertainty analyses, and estimate ice nucleation efficiency using computer programs. Based on the formal program assessment of learning outcomes and direct (yet deidentified) student feedback, we broadly achieved our goals to (1) improve their problem-solving skills by combining multidisciplinary science and math skills and (2) disseminate data and results with variability and uncertainty. The developed modules can be applied at any institute to advance undergraduate and graduate curricula in environmental science.

Dataset Information

Membranes Are Decisive for Maximum Freezing Efficiency of Bacterial Ice Nucleators.

Publications

Membranes Are Decisive for Maximum Freezing Efficiency of Bacterial Ice Nucleators.

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