Basidiomycetous Yeast, Glaciozyma antarctica, Forming Frost-Columnar Colonies on Frozen Medium

The basidiomycetous yeast, Glaciozyma antarctica, was isolated from various terrestrial materials collected from the Sôya coast, East Antarctica, and formed frost-columnar colonies on agar plates frozen at −1 °C. Thawed colonies were highly viscous, indicating that the yeast produced a large number of extracellular polysaccharides (EPS). G. antarctica was then cultured on frozen media containing red food coloring to observe the dynamics of solutes in unfrozen water; pigments accumulated in frozen yeast colonies, indicating that solutes were concentrated in unfrozen water of yeast colonies. Moreover, the yeast produced a small quantity of ice-binding proteins (IBPs) which inhibited ice crystal growth. Solutes in unfrozen water were considered to accumulate in the pore of frozen colonies. The extracellular IBPs may have held an unfrozen state of medium water after accumulation in the frost-columnar colony.

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Growth suppression of ice crystal basal face in the presence of a moderate ice-binding protein does not confer hyperactivity

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1807461115 ◽

2018 ◽

Vol 115 (29) ◽

pp. 7479-7484 ◽

Cited By ~ 10

Author(s):

Maddalena Bayer-Giraldi ◽

Gen Sazaki ◽

Ken Nagashima ◽

Sepp Kipfstuhl ◽

Dmitry A. Vorontsov ◽

...

Keyword(s):

Crystal Growth ◽

Heat Dissipation ◽

Freezing Point ◽

Binding Protein ◽

Ice Crystals ◽

Ice Crystal ◽

Freezing Point Depression ◽

Basal Face ◽

Ice Binding ◽

Ice Binding Protein

Ice-binding proteins (IBPs) affect ice crystal growth by attaching to crystal faces. We present the effects on the growth of an ice single crystal caused by an ice-binding protein from the sea ice microalga Fragilariopsis cylindrus (fcIBP) that is characterized by the widespread domain of unknown function 3494 (DUF3494) and known to cause a moderate freezing point depression (below 1 °C). By the application of interferometry, bright-field microscopy, and fluorescence microscopy, we observed that the fcIBP attaches to the basal faces of ice crystals, thereby inhibiting their growth in the c direction and resulting in an increase in the effective supercooling with increasing fcIBP concentration. In addition, we observed that the fcIBP attaches to prism faces and inhibits their growth. In the event that the effective supercooling is small and crystals are faceted, this process causes an emergence of prism faces and suppresses crystal growth in the a direction. When the effective supercooling is large and ice crystals have developed into a dendritic shape, the suppression of prism face growth results in thinner dendrite branches, and growth in the a direction is accelerated due to enhanced latent heat dissipation. Our observations clearly indicate that the fcIBP occupies a separate position in the classification of IBPs due to the fact that it suppresses the growth of basal faces, despite its moderate freezing point depression.

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Systematic Affiliation and Genome Analysis of Subtercola vilae DB165T with Particular Emphasis on Cold Adaptation of an Isolate from a High-Altitude Cold Volcano Lake

Microorganisms ◽

10.3390/microorganisms7040107 ◽

2019 ◽

Vol 7 (4) ◽

pp. 107 ◽

Cited By ~ 1

Author(s):

Alvaro S. Villalobos ◽

Jutta Wiese ◽

Johannes F. Imhoff ◽

Cristina Dorador ◽

Alexander Keller ◽

...

Keyword(s):

Binding Proteins ◽

Cold Shock ◽

Extreme Environment ◽

Crystal Formation ◽

Ice Crystal ◽

Cold Environments ◽

Ice Binding ◽

Shock Proteins ◽

Harsh Conditions ◽

Insight Into

Among the Microbacteriaceae the species of Subtercola and Agreia form closely associated clusters. Phylogenetic analysis demonstrated three major phylogenetic branches of these species. One of these branches contains the two psychrophilic species Subtercola frigoramans and Subtercola vilae, together with a larger number of isolates from various cold environments. Genomic evidence supports the separation of Agreia and Subtercola species. In order to gain insight into the ability of S. vilae to adapt to life in this extreme environment, we analyzed the genome with a particular focus on properties related to possible adaptation to a cold environment. General properties of the genome are presented, including carbon and energy metabolism, as well as secondary metabolite production. The repertoire of genes in the genome of S. vilae DB165T linked to adaptations to the harsh conditions found in Llullaillaco Volcano Lake includes several mechanisms to transcribe proteins under low temperatures, such as a high number of tRNAs and cold shock proteins. In addition, S. vilae DB165T is capable of producing a number of proteins to cope with oxidative stress, which is of particular relevance at low temperature environments, in which reactive oxygen species are more abundant. Most important, it obtains capacities to produce cryo-protectants, and to combat against ice crystal formation, it produces ice-binding proteins. Two new ice-binding proteins were identified which are unique to S. vilae DB165T. These results indicate that S. vilae has the capacity to employ different mechanisms to live under the extreme and cold conditions prevalent in Llullaillaco Volcano Lake.

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Three-dimensional simulation of ice growth in the presence of antifreeze proteins

Canadian Journal of Physics ◽

10.1139/p03-015 ◽

2003 ◽

Vol 81 (1-2) ◽

pp. 39-45 ◽

Cited By ~ 2

Author(s):

B Wathen ◽

M J Kuiper ◽

V K Walker ◽

Z Jia

Keyword(s):

Crystal Growth ◽

Growth Inhibition ◽

Antifreeze Proteins ◽

Three Dimensional ◽

Computational Method ◽

Inhibition Mechanism ◽

Ice Crystal ◽

Ice Growth ◽

Ice Binding ◽

Dimensional Simulation

A Monte Carlo computational method for simulating the growth of entire ice crystals from the liquid phase has been developed specifically to study the inhibition of ice-crystal growth by antifreeze proteins (AFPs). AFPs are found in the fluids of certain organisms that inhabit freezing environments and constrain ice-crystal growth by adsorbtion to the ice surface, but their inhibition mechanism is still poorly understood. Thus, it was of interest to incorporate these molecules into the dynamic ice simulations to examine the inhibition phenomenon on a whole-crystal basis. We have undertaken simulations with AFPs from two different organisms that differ in activity; the insect AFP has up to 100 times the activity of the fish AFP on a molar basis. Simulations involving insect and fish AFPs have achieved ice-growth inhibition at simulation temperatures within reported activity ranges for both fish and insect AFPs, accompanied by resulting ice morphologies similar to those observed experimentally. These results, as well as other studies on ice-etching patterns and ice burst growth at temperatures below known AFP ice-growth inhibition abilities suggest that AFP activity is dominated by the AFP ice-binding position rather than AFP ice-binding strength. PACS No.: 07.05T

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Ice-binding proteins that accumulate on different ice crystal planes produce distinct thermal hysteresis dynamics

Journal of The Royal Society Interface ◽

10.1098/rsif.2014.0526 ◽

2014 ◽

Vol 11 (98) ◽

pp. 20140526 ◽

Cited By ~ 44

Author(s):

Ran Drori ◽

Yeliz Celik ◽

Peter L. Davies ◽

Ido Braslavsky

Keyword(s):

Basal Plane ◽

Binding Proteins ◽

Thermal Hysteresis ◽

Freezing Point ◽

Ice Crystal ◽

Ice Binding ◽

Time Sensitivity ◽

Custom Made ◽

Order Of Magnitude ◽

Accumulation Kinetics

Ice-binding proteins that aid the survival of freeze-avoiding, cold-adapted organisms by inhibiting the growth of endogenous ice crystals are called antifreeze proteins (AFPs). The binding of AFPs to ice causes a separation between the melting point and the freezing point of the ice crystal (thermal hysteresis, TH). TH produced by hyperactive AFPs is an order of magnitude higher than that produced by a typical fish AFP. The basis for this difference in activity remains unclear. Here, we have compared the time dependence of TH activity for both hyperactive and moderately active AFPs using a custom-made nanolitre osmometer and a novel microfluidics system. We found that the TH activities of hyperactive AFPs were time-dependent, and that the TH activity of a moderate AFP was almost insensitive to time. Fluorescence microscopy measurement revealed that despite their higher TH activity, hyperactive AFPs from two insects (moth and beetle) took far longer to accumulate on the ice surface than did a moderately active fish AFP. An ice-binding protein from a bacterium that functions as an ice adhesin rather than as an antifreeze had intermediate TH properties. Nevertheless, the accumulation of this ice adhesion protein and the two hyperactive AFPs on the basal plane of ice is distinct and extensive, but not detectable for moderately active AFPs. Basal ice plane binding is the distinguishing feature of antifreeze hyperactivity, which is not strictly needed in fish that require only approximately 1°C of TH. Here, we found a correlation between the accumulation kinetics of the hyperactive AFP at the basal plane and the time sensitivity of the measured TH.

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Ice-binding proteins: a remarkable ice crystal regulator for frozen foods

Critical Reviews in Food Science and Nutrition ◽

10.1080/10408398.2020.1798354 ◽

2020 ◽

pp. 1-14

Author(s):

Xu Chen ◽

Xiaodan Shi ◽

Xixi Cai ◽

Fujia Yang ◽

Ling Li ◽

...

Keyword(s):

Binding Proteins ◽

Ice Crystal ◽

Frozen Foods ◽

Ice Binding

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Ice-binding proteins and the applicability and limitations of the kinetic pinning model

Philosophical Transactions of The Royal Society A Mathematical Physical and Engineering Sciences ◽

10.1098/rsta.2018.0391 ◽

2019 ◽

Vol 377 (2146) ◽

pp. 20180391 ◽

Cited By ~ 2

Author(s):

Michael Chasnitsky ◽

Ido Braslavsky

Keyword(s):

Binding Proteins ◽

Thermal Hysteresis ◽

Freezing Point ◽

Ice Crystal ◽

Water Ice ◽

Ice Binding ◽

Production Technologies ◽

Seeding Time ◽

Two Phases ◽

Key Parameter

Ice-binding proteins (IBPs) are unique molecules that bind to and are active on the interface between two phases of water: ice and liquid water. This property allows them to affect ice growth in multiple ways: shaping ice crystals, suppressing the freezing point, inhibiting recrystallization and promoting nucleation. Advances in the protein's production technologies make these proteins promising agents for medical applications among others. Here, we focus on a special class of IBPs that suppress freezing by causing thermal hysteresis (TH): antifreeze proteins (AFPs). The kinetic pinning model describes the dynamics of a growing ice face with proteins binding to it, which eventually slow it down to a halt. We use the kinetic pinning model, with some adjustments made, to study the TH dependence on the solution's concentration of AFPs by fitting the model to published experimental data. We find this model describes the activity of (moderate) type III AFPs well, but is inadequate for the (hyperactive) Tenebrio molitor AFPs. We also find the engulfment resistance to be a key parameter, which depends on the protein's size. Finally, we explain intuitively how TH depends on the seeding time of the ice crystal in the protein solution. Using this insight, we explain the discrepancy in TH measurements between different assays. This article is part of the theme issue ‘The physics and chemistry of ice: scaffolding across scales, from the viability of life to the formation of planets’.

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Ice-Binding Protein from Shewanella frigidimarinas Inhibits Ice Crystal Growth in Highly Alkaline Solutions

Polymers ◽

10.3390/polym11020299 ◽

2019 ◽

Vol 11 (2) ◽

pp. 299 ◽

Cited By ~ 2

Author(s):

Elizabeth Delesky ◽

Shane Frazier ◽

Jaqueline Wallat ◽

Kendra Bannister ◽

Chelsea Heveran ◽

...

Keyword(s):

Crystal Growth ◽

Ionic Strength ◽

Binding Protein ◽

Sodium Dodecyl ◽

Size Exclusion ◽

Alkaline Solutions ◽

Ice Crystal ◽

Ice Binding ◽

Sds Page ◽

Ice Binding Protein

The ability of a natural ice-binding protein from Shewanella frigidimarina (SfIBP) to inhibit ice crystal growth in highly alkaline solutions with increasing pH and ionic strength was investigated in this work. The purity of isolated SfIBP was first confirmed via sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and size-exclusion chromatography with an ultraviolet detector (SEC-UV). Protein stability was evaluated in the alkaline solutions using circular dichroism spectroscopy, SEC-UV, and SDS-PAGE. SfIBP ice recrystallization inhibition (IRI) activity, a measure of ice crystal growth inhibition, was assessed using a modified splat assay. Statistical analysis of results substantiated that, despite partial denaturation and misfolding, SfIBP limited ice crystal growth in alkaline solutions (pH ≤ 12.7) with ionic strength I ≤ 0.05 mol/L, but did not exhibit IRI activity in alkaline solutions where pH ≥ 13.2 and I ≥ 0.16 mol/L. IRI activity of SfIBP in solutions with pH ≤ 12.7 and I ≤ 0.05 mol/L demonstrated up to ≈ 66% reduction in ice crystal size compared to neat solutions.

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