| Summary: | 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.
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