NUCLEIC ACID SEQUENCES ENCODING TYPE III TENEBRIO ANTIFREEZE PROTEINS AND METHOD FOR ASSAYING ACTIVITY

摘要

Thermal hysteresis proteins and their nucleotide sequences derived from the Tenebrionoidea Superfamily which lower the freezing point of a solution without effecting the melting point. Related methods for preparing said proteins and for providing antifreeze or recrystallization inhibition properties to a subject formulation. Also provided is a recrystallisation inibition method for determining the presence, relative concentration, and/or activity of thermal hysteresis proteins comprising: providing a proteinaceous composition in a solvent to form a test solution; flash freezing said solution; raising the temperature of the frozen solution to an appropriate annealing temperature that allows for a partial melt, while limiting heterogeneity in ice grain sizes within said solution; maintaining said frozen solution at the annealing temperature for a length of time sufficient to allow for recrystallization; monitoring the ice crystal grain size changes over time; and determining the presence of functional thermal hysteresis proteins in said solution given the retention of significantly smaller ice crystal grain sizes relative to at least one control solution.