Ice binding proteins (IBPs) have been attracting significant interest on account 
of their characteristic of inhibiting ice growth and recrystallization. Owing to 
their unique characteristics, IBPs have been studied for applications in food, 
pharmaceuticals, and medicine, as well as from a general scientific point of 
view. In this study, we have used differential scanning calorimetry (DSC) and 
Raman spectroscopy as tools to understand the ice binding activity of the 
Arctic-yeast-originating extracellular ice binding glycoprotein (LeIBP) isolated 
from Leucosporidium sp. AY30. From the DSC results, an increase in the specific 
heat capacity was confirmed for 1 mg/mL LeIBP, which suggested that additional 
heat flow was required for the change in temperature. In addition, the 
temperature corresponding to the phase change of the solution was measured, and 
Raman spectroscopy was carried out on the frozen and molten phases, 
respectively. From the results of Raman analysis, we confirmed that the helical 
vibrations related to the ice binding sites on LeIBP were dramatically 
suppressed when the LeIBP solution was frozen. Furthermore, principal component 
analysis (PCA) of the Raman spectra yielded the contrast factor between the 
freezing and melting states. Both DSC and Raman spectroscopy are widely used to 
study the ice binding activity and the structural changes associated with 
molecular vibrations in cryobiology.