Enzyme purification and immobilization with bifunction regenerated cellulose membrane

• Main Authors: Tzu-Ning Lin, 林子寧
• Other Authors: Sung-Chyr Lin
• Format: Others
• Language: zh-TW
• Published: 2018
• Online Access: http://ndltd.ncl.edu.tw/handle/26aa82

碩士 === 國立中興大學 === 化學工程學系所 === 106 === In this study, the regenerated cellulose membrane was used as a substrate, and it was modified into a bifunctional adsorption substrate by chemical synthesis. It can adsorb the His-tagged at the end of the protein with IMAC functional group and the protein can form a covalent bond with the epoxy group on the membrane. By using this method, the regenerated cellulose membrane can simultaneously purify and immobilze enzyme. Through integrating purification and immobilization steps, we can save a lot of time and cost. In the results of this study, it was found that when the metal chelator has a density of about 30%, the enzyme retains the highest activity. During the purification process, a large amount of protein can be obtained by adsorption for 3 hours, and excessive impurity proteins are not adsorbed. After physical adsorption, it takes about 16 hours to form covalent bond. The immobilized enzyme formed by covalent bond can improve its thermal stability, and can maintain a relative activity of more than 90% in the case of repeated operations 20 times, which is sufficient to confirm the covalent bonding is more stable than physical adsorption.