To illuminate the impacts of sugar concentration and stereochemistry on protein protection we used attenuated-total-reflectance Fourier-transform infrared-spectroscopy (ATR-FTIR) to study the effects of four aldohexoses on poly-N-isopropylacrylamide (PNIPA) phase transition. Protein stability in aqueous solutions is essential in numerous fields, predominantly biotechnology and food science. Saccharides protect proteins against thermal denaturation, but the mechanisms are still debatable. We therefore studied the effect of sugar concentration and stereochemistry on the LCST phase transition of PNIPA as a model for protein cold renaturation, using ATR-FTIR. The transition temperature of PNIPA, as observed by both the shift in amide II peak and its area, revealed the following order: galactose > glucose > mannose > talose, i.e., galactose is the most kosmotropic of the four isomers and talose is the least. We concluded that the soluting-out effect exerted on the polymer by these sugars positively correlates with the sugar hydration number governed by sugar stereochemistry.
