title: Comparative study on human milk and serum biotinidase.
abstract: Biotinidase was purified from human breast milk (4,000-fold), and was compared with human serum biotinidase (enriched 30,000-fold). The molecular weight of milk enzyme was 68,000 Da as determined by SDS-PAGE. It was definitely smaller than that of serum biotinidase (Mr = 76,000). Isoelectric point of milk biotinidase was 4.6, whereas that of serum biotinidase was 4.3. Sialic acid content in milk biotinidase was less than that found in serum enzyme. N-Acetyl-galactosamine was present in milk enzyme, whereas it was absent in serum enzyme. Milk biotinidase is O-glycosylated, whereas serum biotinidase is N-glycosylated. These differences in glycosylation suggest the existence of different types of excretion mechanisms between milk and serum biotinidase. Both biotinidases were found to be thiol-type enzyme, however, the extent of activation of the enzyme by 2-mercaptoethanol was 13-fold in milk, whilst the serum enzyme was activated only 1.5-fold. Km for biotinyl-4-amino-benzoate was 22 microM in milk enzyme and 50 microM in serum enzyme. Competitive inhibition by biotin (Ki) of milk enzyme was 43 microM and 1.3 mM for serum enzyme. These results suggest the structural differences at or near the active site of the each enzyme.
authors.affiliation: National Children's Medical Research Center, Division of Metabolism, Tokyo, Japan.
authors.forename: J
authors.initials: J
authors.lastname: Oizumi
authors.forename: K
authors.initials: K
authors.lastname: Hayakawa
authors.forename: M
authors.initials: M
authors.lastname: Hosoya
mesh_terms: D000581:Amidohydrolases / Q000097:blood / Q000378:metabolism*; D000596:Amino Acids / Q000032:analysis; D043523:Biotinidase; D002241:Carbohydrates / Q000032:analysis; D004789:Enzyme Activation / Q000187:drug effects; D005260:Female; D006358:Hot Temperature; D006801:Humans; D007526:Isoelectric Point; D007700:Kinetics; D008623:Mercaptoethanol / Q000494:pharmacology; D008895:Milk, Human / Q000201:enzymology*; D008970:Molecular Weight
chemical_list: D000596:Amino Acids; D002241:Carbohydrates; D008623:Mercaptoethanol; D000581:Amidohydrolases; D043523:Biotinidase