A nonprotein thermal hysteresis-producing xylomannan antifreeze in the freeze-tolerant Alaskan beetle Upis ceramboides

Abstract

Thermal hysteresis (TH), a difference between the melting and freezing points of a solution that is indicative of the presence of large-molecular-mass antifreezes (e.g., antifreeze proteins), has been described in animals, plants, bacteria, and fungi. Although all previously described TH-producing biomolecules are proteins, most thermal hysteresis factors (THFs) have not yet been structurally characterized, and none have been characterized from a freeze-tolerant animal. We isolated a highly active THF from the freeze-tolerant beetle, Upis ceramboides, by means of ice affinity. Amino acid chromatographic analysis, polyacrylamide gel electrophoresis, UV-Vis spectrophotometry, and NMR spectroscopy indicated that the THF contained little or no protein, yet it produced 3.7 ؎ 0.3°C of TH at 5 mg/ml, comparable to that of the most active insect antifreeze proteins. Compositional and structural analyses indicated that this antifreeze contains a ␤-mannopyranosyl-(134) ␤-xylopyranose backbone and a fatty acid component, although the lipid may not be covalently linked to the saccharide. Consistent with the proposed structure, treatment with endo-␤-(134)xylanase ablated TH activity. This xylomannan is the first TH-producing antifreeze isolated from a freeze-tolerant animal and the first in a new class of highly active THFs that contain little or no protein.antifreeze protein ͉ insect cold tolerance ͉ glycolipid ͉ membrane associated antifreeze

A Unique Capsular Polysaccharide Structure from the Psychrophilic Marine Bacterium Colwellia psychrerythraea 34H That Mimics Antifreeze (Glyco)proteins

“…66 The fact that the polysaccharide examined also contained lipid leaves unresolved whether the polysaccharide or the lipid component was responsible for the activity. 66 This study has demonstrated for the first time the existence of a capsular polysaccharide that in its purified form is endowed with ice recrystallization inhibition activity. Its unique structure, in contrast to that isolated from the beetle U. ceramboides, is strongly related to that of AFGPs, both for the presence of a Thr residue and for the Gal-Gal disaccharide motif.…”

Conversion of the chill susceptible fruit fly larva ( Drosophila melanogaster ) to a freeze tolerant organism

“…Such lineages had to overcome selection pressures of shorter growing season, exaggerated daily temperature fluctuations, and seasonal drop of temperatures below the physiological thresholds for activity, growth, and development (7). It is now widely accepted that cold adaptation in insects is highly complex and requires adjustments at all levels of biological organization (8)(9)(10)(11)(12)(13)(14)(15)(16). Supercooling and freeze tolerance are two main strategies that help insects to cope with the risk of water freezing in a cryothermic state.…”

It's Not a Bug, It's a Feature: Functional Materials in Insects

“…During the extreme winter months, the spruce budworm, Choristoneura hebenstreitella, resists freezing at temperatures approaching −30 °C, while the Alaskan beetle Upis ceramboides can survive in a temperature of −60 °C. [423] …”

“…[420][421][422] AFPs adsorb to the surface of ice and prevent water from joining the crystal lattice, thereby preventing freezing of a solution in the presence of ice until a new, lower freezing point is reached. [423,424] AFPs create a difference between the melting point and freezing point; this phenomenon is known as thermal hysteresis, and it allows insects to survive while their body temperature is below the melting point.…”

“…Pigmentary coloration Swallowtail butterflies, [368] butterflies [298,299] Thermoregulation Cooling Moths, [408] bumblebees [658] Freeze resistance Spruce budworm, [423,659] Alaskan beetles [423,659] Thermosensing Fruit flies [660] with this behavior are reduviid nymphs and chrysopoid larvae (Figure 3). [47] Both of these insect subsets rely on adhesive properties of physical microstructures for their camouflaging abilities.…”