Abstract
The goal of this work was to alter the composition of amide-linked FA of bovine buttermilk gangliosides, particularly the disialoganglioside GD3, to adjust lipid sources to special food specifications and pharamacological or cosmetic applications. The chemical transacylation of amide-linked FA of buttermilk gangliosides with free arachidic acid (20∶0) by a combination of basic hydrolysis and diethylphosphorylcyanide/triethylamine-catalyzed reacylation was compared to an enzymatic sphingolipid ceramide N-deacylase (EC 3.5.1.23)-catalyzed FA exchange by GC analysis and nano electrospray ionization-MS. The buttermilk predominantly contained the disialoganlioside GD3 and the monosialoganglioside GM3. The heterogeneity of FA that are incorporated into gangliosides, mainly palmitic acid (29.4 wt%), stearic acid (16.9 wt%), oleic acid (17.8 wt%), and myristic acid (8.5 wt%), was effectively altered by both transes-terification techniques. Arachidic acid, which was not integrated into the initial buttermilk gangliosides, was transacylated to total gangliosides with 23.2 wt% (GD3, 6.7 wt%) by the chemical process and with 8.7 wt% (GD3, 13.8 wt%) when catalyzed enzymatically. Mainly behenic acid and lignoceric acid of GD3 were exchanged chemically, and stearic acid was exchanged by the enzymatic process. This observation might depend on hydrolytic sensitivities of amide-linked very long chain saturated FA or specific enzyme subtrate affinities, respectively. Results of chemical hydrolysis indicated there was a risk of sialic acid decomposition and unspecific degradations. Regarding specificity and avoidance of critical agents, the enzymatic transesterification is recommended for industrial-scale production of consumer goods.
Similar content being viewed by others
Abbreviations
- GD:
-
disialoganglioside
- GM:
-
monosialoganglioside
- HPTLC:
-
high performance TLC
- nano ESI-MSn :
-
nano electrospray ionization-MS
- RT:
-
retention time
References
Lloyd, K.O., and Furukawa, K. (1998) Biosynthesis and Functions of Gangliosides: Recent Advances, Glycoconj. J. 15, 627–636.
Ebel, F., Schmitt, E., Peter-Katalinic, J., Kniep, B., and Mühlradt, P.F. (1992) Gangliosides: Differentiation Markers for Murine T Helper Lymphocyte Subpopulations TH1 and TH2, Biochemistry 31, 12190–12197.
Natomi, H., Saitoh, T., Iwamori, M., Fukayama, M., and Nagai, Y. (1993) Systemic Analysis of Glycosphingolipids in the Human Gastrointestinal Tract: Enriched Sulfatides with Hydroxylated Longer-Chain Fatty Acids in the Gastric and Duodenal Mucosa, Lipids 28, 737–742.
Merrill, A.H., Jr. (2002) De novo Sphingolipid Biosynthesis: A Necessary but Dangerous Pathway, J. Biol. Chem. 277, 25843–25846.
Vesper, H., Schmelz, E.M., Nikolova-Karakashian, M.N., Dillehay, D.L., Lynch, D.V., and Merrill, A.H., Jr. (1999) Sphingolipids in Food and the Emerging Importance of Sphingolipids to Nutrition, J. Nutr. 129, 1239–1250.
Elias, P.M., and Menon, G.K. (1991) Structural and Lipid Biochemical Correlates of the Epidermal Permeability Barrier, Adv. Lipid Res. 24, 1–26.
Kolstø Otnæss, A.B., Lægreid, A., and Ertresvåg, K. (1983) Inhibition of Enterotoxin from Escherichia coli and Vibrio cholerae by Gangliosides from Human Milk, Infect. Immun. 40, 563–569.
Hakomori, S. (1981) Glycosphingolipids in Cellular Interaction, Differentiation, and Oncogenesis, Annu. Rev. Biochem. 50, 733–764.
Tsui, Z.C., Hou, W.H., Yang, L., and Zhu, Z.M. (1990) Effect of a Cell Differentiation Inducer, Ganglioside GM3, on the Neutral Glycosphingolipid Composition and Cell Membrane Fluidity of a Human Promyelocytic Leukemia Cell Line HL-60, In Vivo 4, 205–208.
Kappel, T., Anken, R.H., Hanke, W., and Rahmann, H. (2000) Gangliosides Affect Membrane-Channel Activities Dependent on Ambient Temperature, Cell. Mol. Neurobiol. 20, 579–590.
Brown, D.A., and London, E. (2000) Structure and Function of Sphingolipid- and Cholesterol-Rich Membrane Rafts J. Biol. Chem. 275, 17221–17224.
Van Meer, G., and Lisman, Q. (2002) Sphingolipid Transport: Rafts and Translocators, J. Biol. Chem. 277, 25855–25858.
Perry, D.K., and Hannun, Y.A. (1998) The Role of Ceramide in Cell Signaling, Biochim. Biophys. Acta 1436, 233–243.
Hannun, Y.A., and Obeid, L.M. (2002) The Ceramide-Centric Universe of Lipid-Mediated Cell Regulation: Stress Encounters of the Lipid Kind, J. Biol. Chem. 277, 25847–25850.
Ladisch, S., Hasegawa, A., Li, R., and Kiso, M. (1995) Immuno-suppressive Activity of Chemically Synthesized Gangliosides, Biochemistry 34, 1197–1202.
Bouhours, J.-F., and Bouhours D. (1981) Ceramide Structure of Sphingomyelin from Human Milk Fat Globule Membrane, Lipids 16, 726–731.
Nakano, T., Sugawara, M., and Kawakami, H. (2001) Sialic Acid in Human Milk: Composition and Functions, Acta Paediatr. Taiwan 42, 11–17.
Huang, R.T.C. (1973) Isolation and Characterization of the Gangliosides of Buttermilk, Biochim. Biophys. 306, 82–84.
Hauttecoeur, B., Sonnino, S., and Ghidoni, R. (1985) Characterization of Two Molecular Species GD3 Ganglioside from Bovine Buttermilk, Biochim. Biophys. Acta 833, 303–307.
Rueda, R., Maldonado, J., Narbona, E., and Gil, A. (1998) Neonatal Dietary Gangliosides, Early Hum. Dev. 53, S135-S147.
Martin, M.J., Martin-Sosa, S., and Hueso, P. (2001) Bovine Milk Gangliosides: Changes in Ceramide Moiety with Stage of Lactation, Lipids 36, 291–298.
Jensen, R.G., Bitman, J., Carslon, S.E., Couch, S.C., Hamosh, M., and Newburg, D.S. (1995) Human Milk Lipids, in Handbook of Milk Composition (Jensen R.G., ed.), pp. 495–542, Academic Press, New York.
Hirabayashi, Y., Kimura, M., Matsumoto, M., Yamamoto, K., Kadowaki, S., and Tochikura, T. (1988) A Novel Glycosphingolipid-Hydrolyzing Enzyme, Glycosphingolipid Ceramide Deacylase, Which Cleaves the Linkage Between the Fatty Acid and Sphingosine Base in Glycosphingolipids, J. Biochem. 103, 1–4.
Kurita, T., Izu, H., Sano, M., Ito, M., and Kato, I. (2000) Enhancement of Hydrolytic Activity of Sphingolipid Ceramide N-Deacylase in the Aqueous-Organic Biphasic System, J. Lipid Res. 41, 846–851.
Ito, M., Kurita, T., and Kita, K. (1995) A Novel Enzyme That Cleaves the N-Acyl Linkage of Ceramides in Various Glycosphingolipids as Well as Sphingomyelin to Produce Their Lyso Forms, J. Biol. Chem. 270, 24370–24374.
Kita, K., Kurita, T., and Ito, M. (2001) Characterization of the Reversible Nature of the Reaction Catalyzed by Sphingolipid Ceramide N-Deacylase. A Novel Form of Reverse Hydrolysis Reaction, Eur. J. Biochem. 268, 592–602.
Anand, J.K., Sadozai, K.K., and Hakomori, S. (1996) A Simple Method for the Synthesis of Ceramides and Radiolabeled Analogues, Lipids 31, 995–998.
Ladisch, S., and Gillard, B. (1985) A Solvent Partition Method for Microscale Ganglioside Purification, Anal. Biochem. 146, 220–231.
Sonnino, S., Kirschner, G., Ghidoni, R., Acquotti, D., and Tettamani, G. (1985) Preparation of GM1 Ganglioside Molecular Species Having Homogeneous Fatty Acid and Long Chain Base Moities, J. Lipid Res. 26, 248–257.
Kadowaki H., Bremer, E.G., Evans, J.E., Jungalwala F.B., and McCluer, R.H. (1983) Acetonitrile-Hydrochloric Acid Hydrolysis of Gangliosides for High Performance Liquid Chromatographic Analysis of Their Long Chain Bases, J. Lipid. Res. 24, 1389–1397.
Williams, M.A., and McCluer, R.H. (1980) The Use of Sep-Pak™ C18 Cartridges During the Isolation of Gangliosides, J. Neurochem. 35, 266–269.
Müthing, J. (1996) High-Resolution Thin-Layer Chromatography of Gangliosides, J. Chromatogr A. 720, 3–25.
Svennerholm, L. (1957) Quantitative Estimation of Sialic Acids, II. A Colorimetric Resorcinol-Hydrochloric Acid Method, Biochim. Biophys. Acta 24, 604–611.
Gazzotti, G., Sonnino, S., and Ghidoni, R. (1985) Normal-Phase High-Performance Liquid Chromatographic Separation of Nonderivatized Ganglioside Mixtures, J. Chromatogr. 348, 371–378.
Lepage, G., and Roy, C.C. (1986) Direct Transesterification of All Classes of Lipids in a One-Step Reaction, J. Lipid Res. 27, 114–120.
Kohn, G., Van Der Ploeg, P., Möbius, M., and Sawatzki, G. (1996) Influence of the Derivatization Procedure on the Results of the Gas Chromatographic Fatty Acid Analysis of Human Milk and Infant Formulae, Z. Ernährungswiss. 35, 226–234.
Hsu, F.F., and Turk, J. (2001) Structural Determination of Glycosphingolipids as Lithiated Adducts by Electrospray Ionization Mass Spectrometry Using Energy Collisional-Activated Dissociation on a Triple Stage Quadrupole Instrument, J. Am. Soc. Mass Spectrom. 12, 61–79.
Ii, T., Ohashi, Y., and Nagai, Y. (1995) Structural Elucidation of Underivatized Gangliosides by Electrospray-Ionization Tandem Mass Spectrometry (ESIMS/MS), Carbohydr. Res. 273, 27–40.
Wilm, M., and Mann, M. (1996) Analytical Properties of the Nanoelectrospray Ion Source, Anal. Chem. 68, 1–8.
Creaser, C.S., and Stygall, J.W. (1998) Recent Developments in Analytical Ion Trap Mass Spectrometry, Trends Anal. Chem. 17, 583–593.
Author information
Authors and Affiliations
Corresponding author
About this article
Cite this article
Beermann, C., Röhrig, AK. & Boehm, G. Chemical and enzymatic transacylation of amide-linked FA of buttermilk gangliosides. Lipids 38, 855–864 (2003). https://doi.org/10.1007/s11745-003-1136-3
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s11745-003-1136-3