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TLC‐based assay of ganglioside synthesis
Author(s) -
Czarnowski Megan N,
Ettinger Anna C,
Moore Kathleen H
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.800.1
Subject(s) - lactosylceramide , ganglioside , chemistry , skimmed milk , food science , capric acid , incubation , biochemistry , chromatography , enzyme , fatty acid , lauric acid
Ganglioside content in cultured dairy products is significant and may contribute to antimicrobial effects associated with such foods ( J Food Comp Anal 13 : 783, 2000). The source of gangliosides is unclear as dairy products are generated from ganglioside‐free components (skim milk and culture bacteria). By a terminal synthetic reaction, sialyltransferase (ST) transfers N‐acetylneuraminic acid (NANA) to lactosylceramide (LC): CMP‐NANA + HO‐ lactosylceramide → NANA‐lactosylceramide + CMP. Formation of gangliosides (eg NANA‐LC) is visualized by thin‐layer chromatography (TLC) via resorcinol staining. Incubation conditions have been optimized. Analysis with multiple NANA acceptors (LC solution, whey, dialyzed yogurt, and milk) indicates that milk contains an appropriate acceptor, yielding a NANA containing product (putative ganglioside) with an R f (0.71) that is distinct from the R f (0.63) of free NANA. Now that it has been determined that appropriate NANA acceptors are inherent in milk, the presence of ST in dairy components can be assessed. Rich ganglioside content may be a synergistic by‐product among dairy components. (Supported by OU Provost's Undergraduate Research Award)