Site-specific Immobilization of α-amylase Protein

• Main Authors: Kuan-Wei Tseng, 曾冠維
• Other Authors: Ruoh-Chyu Ruaan
• Format: Others
• Language: zh-TW
• Published: 2008
• Online Access: http://ndltd.ncl.edu.tw/handle/55525489566686551462

碩士 === 國立中央大學 === 化學工程與材料工程研究所 === 96 === Random protein immobilization usually suffers from serious loss of the specific bioactivity of the immobilized protein. Oriented protein immobilization of histidine tagged protein on metal chelating resin does not guarantee 100% exposure of the active site. In this study, we develop a new method for oriented immobilization. Design an affinity ligand according to the characteristic and distribution of amino acids at the opposite to the active site. The target protein is α-amylase from Aspergillus oryzae, and the searched ligand is 3,3’,4,4’ - Biphenyltetracarboxylic dianhydride (BPDA). We predict the possible binding sites by using molecular dockikng. And at the experiments, we attach BPDA to the surface of silica gel and via isotherm adsorb and bioactivity assay show oriented immobilization owns superior specific activity than random immobilization.