Complete purification and characterization of the taste-modifying protein, miraculin, from miracle fruit.
Complete purification and characterization of the taste-modifying protein, miraculin, from miracle fruit.
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从神秘果中完全纯化和表征味道修饰蛋白 Miraculin。
DOI：
10.1016/s0021-9258(18)37991-2
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发表时间：
1988
期刊：
影响因子：
--
通讯作者：
Y. Kurihara
中科院分区：
文献类型：
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作者：
S. Theerasilp;Y. Kurihara
味道改变蛋白，神秘果蛋白，具有将酸味改变为甜味的不寻常特性。以前试图从神秘果的深色碱性提取物中分离神秘果素是不成功的。我们发现用0.5M NaCl溶液提取神秘果素。提取液无色，具有较强的甜味诱导活性。通过硫酸铵分级分离、CM-Sepharose离子交换层析和伴刀豆球蛋白A-Sepharose亲和层析从提取液中纯化神秘果蛋白。由此获得的纯化的神秘果素在反相高效液相色谱中给出单一尖锐峰，表明其是高纯度的。该样品在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳中也得到分子量为28，000的单一条带。这一数值远低于以前报告的数值（40，000 - 48，000）。对纯化的神秘果素的氨基酸组成进行了测定。序列分析表明，它是由一个纯的单一多肽，并确定了20个氨基末端氨基酸。纯化的神秘果素含有高达13.9%的糖，其由葡萄糖胺、甘露糖、半乳糖、木糖和岩藻糖组成，摩尔比为3.03：3.00：0.69：0.96：2.12。
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.