Thermal glycation of proteins by D-glucose and D-fructose.

The dry thermal glycation method was used to conjugate D-glucose and D-fructose with bovine serum albumin. Reactions were conducted at 50-104 degrees C for 30 min. Depending on temperature, different levels of substitution were achieved. In the case of D-glucose, average substitution levels of up to 53 mol glucose/mol bovine serum albumin were obtained. D-fructose turned out to be less reactive than D-glucose. The pH of the reaction mixture was also found to affect the efficiency of the glycation reaction. The levels of substitution were estimated using matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry and SDS-polyacrylamide gel electrophoresis. Glycated molecules, and a process of glycation itself are implicated in diabetes complications. Hypothetically bovine spongiform encephelopathy (BSE) was also derived from glycation.