Studies on the Assisted Actions of Furfuryl Alcohol for the Renaturation Efficiency of α-Chymotrysin by Liquid Chromatography
Shen Yehua, Wang Haibo, Wei Yinmo, Geng Xindu * (Institute of Modern Separation Science, Key Laboratory of Modern Separation Science in Shaanxi Province, Northwest University, Xi′an 710069)
Using high performance hydrophobic interaction chromatography (HPHIC), the assisted actions of furfuryl alcohol in a guanidine hydrochloride (GuHCl) solution or on silica surface bonding ligands with the end group of furfuryl alcohol to the renaturation efficiency of α-chymotrysin (α-Chy) were investigated. The magnitudes of bioactive recovery of α-Chy were employed to characterize the contributions of furfuryl alcohol to α-Chy refolding in the two circumstances. By comparison of the results from size exclusion chromatogrpahy (SEC) HPHIC it may understand the different magnitudes in renaturation efficiency of furfuryl alcohol for both of solution and stationary phase, and the mechanism of the assisted actions of furfuryl alcohol may be attributed to form a complex between α-Chy and furfuryl alcohol either in solution, or on the surface of the HPHIC stationary phase. Furthermore, the optimazation of the amount of furfuryl alcohol in the GuHCl solution was also carried out.