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Purification of human milk fucosyltransferase

par Vandenbroucke, Anne Thérèse;Dolmans, Marcel ;Petiau-de Vries, Ghislaine;Leonis, José
Référence Archives internationales de physiologie et de biochimie, 84, 5, page (1111-1112)
Publication Publié, 1976

Article révisé par les pairs

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Résumé :

The enzyme has been purified from skimmed milk concentrated on an Amicon PM 10 membrane and desalted on a Sephadex G 25 column equilibrated with 0.02 M MOPS buffer pH 7.4 contaning 10 mM MgCl2. This 2.5 fold concentrated preparation has been purified by affinity chromatography on a GDP Sepharose 4B column according to Barker et al. The fucosyltransferase active fractions were eluted by a 0.02 M MOPS buffer pH 7.4 containing 0.3 M lactose and 0.02 M EDTA. More than 99.4% of total proteins were eliminated in one step by this procedure. Two activity peaks were detected. This suggests the existence of different lactose: fucosyltransferases. In a second method, the concentrated human milk serum has been submitted to chromatography on DEAE cellulose suspended in 0.025 M phosphate buffer pH 6.9 and eluted with the same buffer. The first fractions contained maximal fucosyltransferase activity and were submitted to affinity chromatography. In this case, only one peak of fucosyltransferase activity has been detected. Acrylamide gel electrophoresis in the presence of sodium dodecylsulphate has been performed at each step of purification.