scholarly journals Protection of Alcohol Dehydrogenase against Freeze–Thaw Stress by Ice-Binding Proteins Is Proportional to Their Ice Recrystallization Inhibition Property

Marine Drugs ◽  
2020 ◽  
Vol 18 (12) ◽  
pp. 638
Author(s):  
Young Hoon Lee ◽  
Kitae Kim ◽  
Jun Hyuck Lee ◽  
Hak Jun Kim

Ice-binding proteins (IBPs) have ice recrystallization inhibition (IRI) activity. IRI property has been extensively utilized for the cryopreservation of different types of cells and tissues. Recent reports demonstrated that IRI can also play a significant role in protecting proteins from freezing damage during freeze–thaw cycles. In this study, we hypothesized that the protective capability of IBPs on proteins against freeze–thaw damage is proportional to their IRI activity. Hence we used two IBPs: one with higher IRI activity (LeIBP) and the other with lower activity (FfIBP). Yeast alcohol dehydrogenase (ADH) was used as a freeze-labile model protein. IBPs and ADH were mixed, frozen at −20 °C, and thawed repeatedly. The structure of ADH was assessed using fluorescence emission spectra probed by 1-anilinonaphthalene-8-sulfonate over the repeated freeze–thaw cycles. The activity was monitored at 340 nm spectrophotometrically. Fluorescence data and activity clearly indicated that ADH without IBP was freeze-labile. However, ADH maintained about 70% residual activity after five repeated cycles at a minimal concentration of 0.1 mg mL-1 of high IRI-active LeIBP, but only 50% activity at 4 mg mL−1 of low active FfIBP. These results showed that the protection of proteins from freeze–thaw stress by IBPs is proportional to their IRI activity.

Biomolecules ◽  
2020 ◽  
Vol 10 (2) ◽  
pp. 274 ◽  
Author(s):  
Aneta Białkowska ◽  
Edyta Majewska ◽  
Aleksandra Olczak ◽  
Aleksandra Twarda-Clapa

More than 80% of Earth’s surface is exposed periodically or continuously to temperatures below 5 °C. Organisms that can live in these areas are called psychrophilic or psychrotolerant. They have evolved many adaptations that allow them to survive low temperatures. One of the most interesting modifications is production of specific substances that prevent living organisms from freezing. Psychrophiles can synthesize special peptides and proteins that modulate the growth of ice crystals and are generally called ice binding proteins (IBPs). Among them, antifreeze proteins (AFPs) inhibit the formation of large ice grains inside the cells that may damage cellular organelles or cause cell death. AFPs, with their unique properties of thermal hysteresis (TH) and ice recrystallization inhibition (IRI), have become one of the promising tools in industrial applications like cryobiology, food storage, and others. Attention of the industry was also caught by another group of IBPs exhibiting a different activity—ice-nucleating proteins (INPs). This review summarizes the current state of art and possible utilizations of the large group of IBPs.


2019 ◽  
Author(s):  
Zhengyao Qu ◽  
Shuaiqi Guo ◽  
Christian C. M. Sproncken ◽  
Romà Surís-Valls ◽  
qingliang yu ◽  
...  

Frost weathering of porous materials caused by seasonal temperature changes is a major source of damage to the world’s infrastructure and cultural heritage. Here we investigate poly(vinyl alcohol) (PVA) addition as a means to enhance the freeze-thaw durability of concrete without compromising its structural or mechanical integrity. We evaluate the ice recrystallization inhibition activity of PVA in a cementitious environment and the impact of PVA on key structural and mechanical properties, such as cement hydration (products), microstructure, strength, as well as freeze‑thaw resistance. We find that a low amount of PVA significantly reduces the surface scaling of concrete and displays excellent ice recrystallization inhibition in the saturated Ca(OH)<sub>2 </sub>solution which has a similar pH value as cement pore solution, while it does not affect cement hydration, microstructure, nor its mechanical properties. These findings contribute to new insights on freeze-thaw damage mechanism and more importantly we disclose a new direction for the design of concrete with excellent freeze‑thaw resistance.


Biomolecules ◽  
2019 ◽  
Vol 9 (8) ◽  
pp. 347 ◽  
Author(s):  
Romà Surís-Valls ◽  
Ilja Voets

Antifreeze (glyco)proteins (AF(G)Ps) have received increasing attention as potential cryopreservation agents since their discovery in the 1970s. While cryopreservation strategies for specific cells (such as red blood cells) are successful and widely implemented, preservation of other cell types, tissues and whole organs remains challenging. This is due to the multifactorial nature of the freeze-thaw damage, the complexity of preserving biological matter and the (country-to-country) variability of the employed procedures and regulations. AF(G)Ps are well-known for their ability to modulate ice crystal growth morphology and ice recrystallization inhibition (IRI), both of which are considered key contributors to freeze-thaw damage. To date, however, the impact of AF(G)Ps on cell survival remains at best partially understood as conflicting results on the benefits or disadvantages of including AF(G)P in cryopreservation strategies remain unelucidated. We hypothesize that variability in the additives in the cryopreservation media contributes to the observed discrepancies. To critically examine this idea, we monitored the inhibition of ice recrystallization by AF(G)P in the presence of various salts using a quantitative analysis of optical microscopy images via the Lifshitz-Slyozov-Wagner (LSW) theory for Oswald ripening. We found that the addition of salts, which are used in culture and cryopreservation media, enhances the IRI activity of AF(G)Ps, and that the magnitude of the enhancement was in line with the Hofmeister series. The size of ice crystals grown in AFGP1–5 and type III AFP samples containing chloride, phosphate and citrate ions were statistically smaller after 90 min of incubation than crystals grown in the absence of these salts. The ice recrystallization rates (kd) of AFGP1–5 and type III AFP samples prepared at a fixed overall ionic strength of 100 mM progressively decreased following the Hofmeister series for anions. Our results demonstrate that the performance of AF(G)Ps is significantly influenced by additives present in common cryopreservation media. It is thus important to conduct excipient compatibility experiments to identify potential incompatibilities between additives and AF(G)Ps in cryopreservation formulations.


2012 ◽  
Vol 58 (4) ◽  
pp. 402-412 ◽  
Author(s):  
Sandra L. Wilson ◽  
Paul Grogan ◽  
Virginia K. Walker

Freeze–thaw stress has previously been shown to alter soil community structure and function. We sought to further investigate this stress on enriched microbial consortia with the aim of identifying microbes with ice-associating adaptations that facilitate survival. Enrichments were established to obtain culturable psychrotolerant microbes from soil samples from the latitudinal extremes of the Canadian Shield plateau. The resulting consortia were subjected to consecutive freeze–thaw cycles, and survivors were putatively identified by their 16S rRNA gene sequences. Even though the northerly site was exposed to longer, colder winters and large spring-time temperature fluctuations, the selective regime similarly affected both enriched consortia. Quantitative PCR and metagenomic sequencing were used to determine the frequency of a subset of the resistant microbes in the original enrichments. The metagenomes showed 22 initial genera, only 6 survived and these were not dominant prior to selection. When survivors were assayed for ice recrystallization inhibition and ice nucleation activities, over 60% had at least one of these properties. These phenotypes were not more prevalent in the northern enrichment, indicating that regarding these adaptations, the enrichment strategy yielded seemingly functionally similar consortia from each site.


ACS Omega ◽  
2020 ◽  
Vol 5 (22) ◽  
pp. 12825-12831
Author(s):  
Zhengyao Qu ◽  
Shuaiqi Guo ◽  
Christian C. M. Sproncken ◽  
Romà Surís-Valls ◽  
Qingliang Yu ◽  
...  

Author(s):  
Zhengyao Qu ◽  
Shuaiqi Guo ◽  
Christian C. M. Sproncken ◽  
Romà Surís-Valls ◽  
qingliang yu ◽  
...  

Frost weathering of porous materials caused by seasonal temperature changes is a major source of damage to the world’s infrastructure and cultural heritage. Here we investigate poly(vinyl alcohol) (PVA) addition as a means to enhance the freeze-thaw durability of concrete without compromising its structural or mechanical integrity. We evaluate the ice recrystallization inhibition activity of PVA in a cementitious environment and the impact of PVA on key structural and mechanical properties, such as cement hydration (products), microstructure, strength, as well as freeze‑thaw resistance. We find that a low amount of PVA significantly reduces the surface scaling of concrete and displays excellent ice recrystallization inhibition in the saturated Ca(OH)<sub>2 </sub>solution which has a similar pH value as cement pore solution, while it does not affect cement hydration, microstructure, nor its mechanical properties. These findings contribute to new insights on freeze-thaw damage mechanism and more importantly we disclose a new direction for the design of concrete with excellent freeze‑thaw resistance.


Author(s):  
Lindsay Smith

At subzero temperatures extracellular ice growth can kill plants by dehydrating cells and rupturing their membranes. Some grasses can protect themselves from this damage by producing antifreeze proteins (AFPs). These AFPs irreversibly adsorb to growing ice crystals and prevent further gowth. This is measured by ice-recrystallization inhibition (IRI), whereby ice crystals remain small at high sub-zero temperatures. An AFP from Brachypodium distachyon, a temperate grass, has been structurally modelled as a left-handed beta helix with two flat ‘faces’ on either side of the molecule. I am trying to determine which ‘face’ is important for ice adsorption. I have made mutations in the sequence encoding the AFP so that a small, flat amino acid is replaced by a bulky residue, which will likely interfere with the “fit” of the protein to ice. A mutation on one of the flat ‘faces’ of the protein seems to retain all AFP activity, whereas mutations on the opposite ‘face’ appear to cause a loss in activity. Therefore, I believe that this latter ‘face’ is the one important for ice-binding. By understanding how proteins interact with ice, it may be possible to develop new technologies such as environmentally-friendly de-icing agents.


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