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16 SESJA A VI SYMPOZJUM GLIKOKONIUGATÓW 13 PURIFICATION AND CHARACTERIZATION OF GDP-L-FUC:N-ACETYL p-d-glucosaminide <xl->6fucosyltrans- FERASE FROM HUMAN BLOOD PLATELETS A-07 Joanna Kamińska'. Mary Catherine Glick 2, and Jerzy Kościelak 1 'Department of Biochemistry, Institute of Hematology and Blood Transfusion, Warsaw, Poland; department of Pediatrics, School of Modicine, University of Pennsylvania and The Joseph Stokes Jr. Research Institute, The Children's Hospital of Philadelphia, Pennsylvania a-6-l-fucosyltransferase (a1,6fuct; EC ) from human platelets is an enzyme that is raleased into serum during coagulation of blood. Total activity of al,6fuct in the serum is proportional to blood platelet concentration. In platelets, however, the enzyme activity is higher, the lower is platelet concentration in blood. Molecular mechanism by which platelets acquire high activity of al,6fuct is unknown. It's been hypothesized that this activity is a measure of ploidy level of megakaryocytes. In the present report al,6fuct was purified 100,000-fold. The purification required three sequential chromatographic steps: chromatofocusing, affinity column on GnGn-Gp-CH-Sepharose, and gel filtration on Sephadex G-200. The final preparation contained a protein that migrated as a single discrete band M r of 58,000 in SDS-PAGE under non-reducing conditions, and as a single enzymatically active peak M r of 58,000 in gel filtration., Although the purified enzyme utilized the biantennary GnGn-Gp as substrate, it was twice as active with the triantennary oligosaccharide when the Man a1,3 antenna was substituted with GlcNAcbl,4. On the other hand the tetraantennary oligosaccharide was not a preferred Km values for the substrate asialoagalactransferrin-glycopeptide, and GDP-L-fucose were 29 and 28 mm, respectively. The optimum ph of the enzyme was . The activity of al,6fuct was abolished in the presence of p-mercaptoethanol. Divalent cations such as Wig 2 * and Ca 2ł activated, but Cu 2ł, Zn 2t and Ni 2 * strongly inhibited the activity.

Abstract: With the global implementation of the “Framework Convention on Tobacco Control”, tobacco industry in different countries would meet more pressure in future development to a certain extent. This essay is based on Kunming flue-cured acquisition data 2006-2010 combined with quality control theory, non- constant process of quality control and logistic non-linear regression analysis to examine annual acquisition trend in history multi-levelly and multi-dimensionally and to excavate current situation deeply so as to discover natural discipline in flue-cured production, then establish an effective quality control index system of tobacco acquisition course assisted by current demands, which could lay the foundation for ensuring the acquisition process being accomplished efficiently and enhancing the ability of supplying high grade tobacco leaves, as well as strengthen the core competencies of Yunnan tobacco industry.

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