Complete purification and characterization of the taste-modifying protein, miraculin, from miracle fruit.

Open AccessPublished:August 15, 1988DOI:
      This paper is only available as a PDF. To read, Please Download here.
      The taste-modifying protein, miraculin, has the unusual property of modifying a sour taste into a sweet taste. Previous attempts to isolate miraculin from deeply colored alkaline extracts of the miracle fruit were unsuccessful. We found that miraculin is extracted with 0.5 M NaCl solution. The extracted solution is colorless and shows the strong sweet-inducing activity. Miraculin was purified from the extracted solution by ammonium sulfate fractionation, CM-Sepharose ion-exchange chromatography, and concanavalin A-Sepharose affinity chromatography. The purified miraculin thus obtained gave a single sharp peak in reverse phase high performance liquid chromatography, indicating that it is highly pure. The sample also gave a single band having molecular weight 28,000 in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This value was much lower than the values reported previously (40,000-48,000). The amino acid composition of the purified miraculin was determined. Sequence analysis of the purified miraculin indicated that it is composed of a pure single polypeptide and identified 20 amino-terminal amino acids. The purified miraculin contained as much as 13.9% of sugars, which consisted of glucosamine, mannose, galactose, xylose, and fucose in a molar ratio of 3.03:3.00:0.69:0.96:2.12.


        • Kurihara K.
        • Beidler L.M.
        Science. 1968; 161: 1241-1243
        • Brouwer J.N.
        • Van der Wel H.
        • Francke A.
        • Henning G.J.
        Nature. 1968; 220: 373-374
        • Giroux E.L.
        • Henkin R.I.
        J. Agric. Food Chem. 1974; 22: 594-601
        • Kurihara Y.
        • Terasaki S.
        Biochim. Biophys. Acta. 1982; 719: 444-449
        • Laemmli U.K.
        Nature. 1970; 227: 680-685
        • Heinrickson R.L.
        • Meridith S.C.
        Anal. Biochem. 1984; 136: 65-74
        • Scholze H.
        J. Chromatogr. 1985; 350: 453-460
        • Edelhoch H.
        Biochemistry. 1967; 6: 1948-1954
        • Mikami H.
        • Ishida Y.
        Bunseki Kagaku. 1983; 32: E207
        • Kurihara K.
        • Beidler L.M.
        Nature. 1969; 222: 1176-1179
        • Kurihara K.
        • Kurihara Y.
        • Beidler L.M.
        Olfaction and Taste HI.
        Rockefeller University Press, New York1969: 450-469
        • Lowry O.H.
        • Rosebrough N.J.
        • Farr A.L.
        • Randall R.J.
        J. Biol. Chem. 1951; 193: 265-275
        • Segrest J.P.
        • Jackson R.L.
        Methods Enzymol. 1972; 28: 54-63