STUDIES ON PROTEIN DENATURATION BY SIZE-E XCLUSION CHROMATOGRAPHY

A size-exclusion chromatography able to study conformational change du ring protein denaturation was proposed by comparing the correlations b etween protein biophysical properties and chromatographic behaviors. T he relative volume of denatured protein may be compared by size-exclus ion chromatography in term of changes in retention time. The number of forming denatured species can be determined by chromatographic peak n umber. The expansion extent of protein may be described using changes in peak shape and peak number. The exposure of aromatic amino acid res idues in denatured proteins can also be deduced from peak height under different wave lengths. We utilized the established size-exclusion ch romatography to examine the denatured aspects for liquid and solid a-a mylase upon long term storage under lower temperature, to discuss the influence of denaturation time and denaturation temperature on unfoldi ng behavior of proteins.