Study On Immobilizing Papain On Aminated Resin By Flexible Connection

Based on the traditional enzyme immobilization methods this paper poses the model of enzyme flexible immobilization, which means connecting the carrier to the enzyme with a high molecular mass, hydrophilic flexible chain. Thus the carrier with flexible chain can make buffering , maintain the conformation of enzyme and reduce the loss of enzyme activity that is brought from the direct collision between carriers and enzymes. And the object of present work is the preparation of flexible carrier with amino group which is used to immobilize papain flexibly.Firstly, we generalize the developments of enzyme immobilization home and abroad, and summarize the methods and the principles of immobilization and function of carriers.Secondly we mainly studies the preparation of aminated polystyrene which is used as the flexible immobilization carrier. The aminated resin could be obtained by chloroacetyl polystyrene which is synthesized by Friedel-Crafts Reaction, thus avoid the classic chloromethylation process. And the effects of solvent, dispersing time, temperature and catalyst as well as diameter of the resin and other related factors are investigated in attempt to achieve excellent results, which are analyzed by IR and element analysis. The results show: the optimal solvent is DMF; the dispersing time is at least 8.5h; the optimal temperature is 80℃ the weight increment percent of aminated resin is maximal when the volume of tetraethylene pentamine exceed 4ml; and the weight increment percent is more close to the ideal weight increment percent when using resin with small diameter and low substitution percent. Based on the optimization, we could prepare the aminated resin with following characters: weight increment percent is 21.8%; N% is 8.7% and the mol of amino per aminated resin is 6.2mmol/g0 At the same time by analysis of rudimental chlorine content we could know that the reaction is involved in both primary amine and secondary amine.Thirdly we immobilize papain on the aminated resin It shows that if immobilizing papain by direct connection of glytaraldehyde, optimal concentration ofglytaraldehyde is 4%; while if immobilizing papain on the special resin, the product of glytaraldehyde and aminated resin, the effect of immobilization is better than the first method.Finally following discussion above we put forward some suggestion to future work on model of flexible immobilization.