Glycoprotein from Ginkgo biloba kernel may be allergic. In this paper, the allergic proteins were identified with Western blotting, and the 32 kDa glycoprotein was purified with ion exchange chromatography and gel chr...Glycoprotein from Ginkgo biloba kernel may be allergic. In this paper, the allergic proteins were identified with Western blotting, and the 32 kDa glycoprotein was purified with ion exchange chromatography and gel chromatography. With Western blotting, there were 3 allergic proteins with molecular weight 21, 32, and 36 kDa. With SDS-PAGE analysis and measurements of the protein and sugar contents, the isolation and purification technology of 32 kDa was confirmed. Ginkgo crude protein extract was precipitated with ammonium sulphate （saturation gradient： 40-80%）. The precipitate was purified by chromatography with DEAE-cellulose 52, then chromatography with Sephadex G-200 and the target glycoprotein was finally obtained. The analysis results showed the molecule of the glycoprotein was 32.12 kDa and the ratio of protein to sugar was 20.56：1. In conclusion, the purification method could be used in preparation of the glycoprotein, and the study could provide a basis for the further research of the glycoprotein.展开更多
基金supported by Research Fund for the Doctoral Program of Higher Education of China(200802980004 and 20070298008)the National Natural Science Fundation of China (30872055)
文摘Glycoprotein from Ginkgo biloba kernel may be allergic. In this paper, the allergic proteins were identified with Western blotting, and the 32 kDa glycoprotein was purified with ion exchange chromatography and gel chromatography. With Western blotting, there were 3 allergic proteins with molecular weight 21, 32, and 36 kDa. With SDS-PAGE analysis and measurements of the protein and sugar contents, the isolation and purification technology of 32 kDa was confirmed. Ginkgo crude protein extract was precipitated with ammonium sulphate （saturation gradient： 40-80%）. The precipitate was purified by chromatography with DEAE-cellulose 52, then chromatography with Sephadex G-200 and the target glycoprotein was finally obtained. The analysis results showed the molecule of the glycoprotein was 32.12 kDa and the ratio of protein to sugar was 20.56：1. In conclusion, the purification method could be used in preparation of the glycoprotein, and the study could provide a basis for the further research of the glycoprotein.
