PRODUCTION OF SWEETNESS-INDUCING SUBSTANCE

摘要

PURPOSE:To obtain purified miraculin having stable sweetnessinducing activity and free from coloring impurities on an industrial scale at a low cost by extracting the compound from a miraculin-containing raw material such as miracle fruit with an aqueous solution of an acidic buffer salt. CONSTITUTION:An aqueous solution of an acidic buffer salt of pH3.5-5.6 (e.g. 0.1M acetic acid-sodium acetate buffer solution) is added at about 0 deg.C (in ice water) to a miraculin-containing raw material such as freeze-dried pulp of miracle fruit, homogenized with ultrasonic wave (20kHz) and centrifuged at 10,000rpm for 45min to extract the substance. The supernatant is collected and the obtained extract liquid is desalted by gel-filtration chromatography and salted out with ammonium sulfate. The precipitate is dissolved in a neutral phosphate buffer solution and purified by ion exchange chromatography and affinity chromatography to obtain the objective sweetness-inducing miraculin having stable sweetnessinducing activity and free from coloring impurities.