Large Scale Purification of a-D-Galactosidase from Taro

• Main Authors: Chau-Shan Lii, 李釗杉
• Other Authors: Su-Fang Chien
• Format: Others
• Language: zh-TW
• Published: 2000
• Online Access: http://ndltd.ncl.edu.tw/handle/78245282098653363554

碩士 === 淡江大學 === 化學學系 === 88 === Taro is rich in a-D-galactosidase . It can modify B red blood cells into O red blood cells. In order to have large quantity of enzyme, the purpose of this experiment is designed a faster method to purify a-D-galactosidase from taro to a form of 3600-fold of purification fold. The yield is 25% and the specific activity is 53 unit/mg protein. The a-D-galactosidases was purified by extraction, gel filtration on Sephadex G-100 column, and Sephadex G-75 column, ion exchange chromatography on DEAE-Sephadex A-50 column, and CM- Sepharose fast flow column. The molecular weight of the enzyme was estimated to be 40 kDa by SDS-PAGE. This method is rather fast, efficient and requires just one week. About 3.5 mg of pure a-D-galactosidase protein was obtained from 7.5 kg of taro. It results in a 90% pure protein.