Purification Of Trypsin By Affinity Chromatography With Chitosan As Carrier

Trypsin catalyzes the hydrolysis of peptide bonds in the interior of a polypeptide chain or protein molecule. Trypsin is an enzyme specific for peptides and esters of the amino acids lysine and arginine. It is useful and valuable. The goals of this study is to purify trypsin with a cheap affinity adsorbent, research the technique for the affinity adsorbent and the purification of trypsin, characterize the purified trypsin.In this study, we put forward a preparation technique for a new affinity adsorbent. The affinity adsorbent uses chitosan as affinity adsorbent carrier, chicken ovomucoid as affinity ligand. Chicken ovomucoid was salted out from egg white, and then purified by ion exchange. Chitosan, cheap and resourceful, was dissolved by hydrochloric solution. The chitosan beads were prepared by coagulating technique. Chicken ovomucoid as the affinity ligand was immobilized on the chitosan beads via a cross-linker, glutaraldehyde. This method of preparing affinity adsorbent using chitosan as carrier is more simple and safe than that using sepharose as carrier.The optimum conditions for immobilization were studied. The results were as follows: We get most chicken ovomucoid when we salted it out at 4℃,pH3.5. For preparing the chitosan beads, the composing of coagulating solution contains 4%...