Application of taste modifying protein, neoculin and miraculin for food preperation

2009 Fiscal Year Final Research Report

• Project/Area Number: 19300248
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Eating habits, studies on eating habits
• Research Institution: The University of Tokyo
• Principal Investigator: ASAKURA Tomiko The University of Tokyo, 大学院・農学生命科学研究科, 特任准教授 (20259013)
• Co-Investigator(Renkei-kenkyūsha): MINAMI Michiko 東京学芸大学, 教育学部, 教授 (70272432) ; FUNAKI Junko 福岡女子大学, 人間環境学部, 准教授 (60219029) ; OKADA Shinji 東京大学, 大学院・農学生命科学研究科, 助教 (50376563)
• Project Period (FY): 2007 – 2009
• Keywords: ネオクリン / ミラクリン / 甘味受容体

Research Abstract

Neoculin and miraculin are taste modifying proteins which convert sourness to sweetness. They are occurring in tropical fruits, Curculigo latiforia and Synsepalum dulcificum, respectively. Each native protein is difficult to obtain constantly. We succeeded to produce recombinant neoculin and miraculin with Aspergillus oryzae that was designated to GRAS. To verify the safety of neoculin, viability of Caco-2 cells exposed to neoculin was investigated. The viability of cells was not changed with neoculin exposure. DNA microarray analysis also revealed that neoculin did not affect the gene expression of Caco-2 cells. Neoculin is received by human sweet taste receptor, T1R2-T1R3. The interaction sites of neoculin to T1R2-T1R3 were analyzed using HEK293T cells that express T1R2-T1R3. As a result, neoculin was received at the N-terminal extra cellar domain in T1R3.

Research Products

• [Journal Article] pH-dependent Structural Change of Neoculin in Special Reference to Its Taste-modifying (2009)
  • Author(s): Morita, Y., Nakajima, K., Iizuka, K., Terada, T., Shimizu-Ibuka, A., Ito, K., Koizumi, A., Asakura, T., Misaka, T., Abe, K.
  • Journal Title: Activity. Biosci. Biotech. Biochem. 73 Pages: 2552-2555
• [Journal Article] pH-Dependent Inhibition of the Human Bitter Taste Receptor hTAS2R16 by a Variety of Acidic Substances (2009)
  • Author(s): Sakurai, T., Misaka,T., Nagai, T., Ishimaru, Y., Matsuo, S., Asakura, T., Abe, K.
  • Journal Title: J.Agric.Food Chem. 57 Pages: 2508-25141
• [Journal Article] Neoculin a taste-modifying sweet protein, accumulates in ripening fruits of cultivated Curculigo latifolia (2008)
  • Author(s): Okubo, S., Asakura, T., Okubo, K., Abe, K., Misaka, T., Akita, T., Abe K.
  • Journal Title: J.Plant Physiol. 165(18) Pages: 1964-9
• [Journal Article] Biochemical and Genomic Analysis of Neoculin Compared to Monocot Mannose-Binding Lectins (2008)
  • Author(s): Shimizu-Ibuka, A., Nakai, Y., Nakamori, K., Morita, Y., Nakajima, K., Kadota, K., Watanabe, H., Okubo, S., Terada, T., Asakura, T., Misaka, T., Abe, K.
  • Journal Title: J.Agric.Food Chem. 56 Pages: 5338-5344
• [Journal Article] Acid-induced sweetness of neoculin is ascribed to its pH-dependent agonistic-antagonistic interaction with human sweet taste receptor (2008)
  • Author(s): Nakajima, K., Morita, Y., Koizumi, A., Asakura, T., Terada, T., Ito, K., Shimizu-Ibuka, A., Maruyama, J., Kitamoto, K., Misaka, T., Abe K.
  • Journal Title: FASEB J. 22 Pages: 2323-2330
• [Journal Article] Signal peptide peptidase and its homologs in Arabidopsis thaliana-plant tissue-specific expression and distinct subcellular localization (2008)
  • Author(s): Tamura, T., Asakura, T., Uemura, T., Ueda, T., Terauchi, K., Misaka, T., Abe, K.
  • Journal Title: FEBS J. 275 Pages: 34-43
• [Journal Article] Microbial production of sensory-active miraculin (2007)
  • Author(s): Ito, K., Asakura, T., Morita, Y., Nakajima, K., Koizumi, A., Shimizu-Ibuka, A., Masuda, K., Ishiguro, M., Terada, T., Maruyama, J., Kitamoto, K., Misaka, T., Abe, K.
  • Journal Title: Biochem. Biophys. Res. Commun. 360 Pages: 407-411
• [Journal Article] Taste-modifying sweet protein, neoculin, is received at human T1R3 amino terminal domain (2007)
  • Author(s): Koizumi, A., Nakajima, K., Asakura, T., Morita, Y., Ito, K., Shmizu-Ibuka, A., Misaka, T., Abe, K.
  • Journal Title: Biochem. Biophys. Res. Commun. 358 Pages: 585-589
• [Journal Article] Gliadain, a gibberellin-inducible cysteine proteinase occurring in germinating seeds of wheat, Triticum aestivum L., specifically digests gliadin and is regulated by intrinsic cystatins (2007)
  • Author(s): Kiyosaki, T., Matsumoto, I., Asakura, T., Funaki, J., Kuroda, M., Misaka, T., Arai, S., Abe, K.
  • Journal Title: FEBS J. 274 Pages: 1908-1917
• [Journal Article] Differential functions of wheat aspartic proteinases, WAP1 and WAP2, in germinating and maturing seeds (2007)
  • Author(s): Tamura, T., Terauchi, K., Kiyosaki, T., Asakura, T., Funaki, J., Matsumoto, I., Misaka, T., Abe, K.
  • Journal Title: J.Plant Physiol 164 Pages: 470-477
• [Book] 味覚修飾蛋白質ネオクリンの活性化機構と新規甘味蛋白質の開発, 蛋白質核酸酵素 (2009)
  • Author(s): 中島健一郎, 古泉文子, 朝倉富子, 三坂巧
  • Publisher: 共立出版
• [Book] 植物のアスパラギン酸プロテアーゼ, 化学と生物 (2009)
  • Author(s): 朝倉富子, 田村倫子, 寺内かえで, 阿部啓子
  • Publisher: 日本農芸化学会
• [Book] Neoculin as a novel sweet protein with taste-modifying activity Current Topics in Biotechnology (2009)
  • Author(s): Nakajima, K., Asakura, T., Misaka, T., Abe, K.
• [Book] 食と味覚(ネスレ栄養科学会議監修) (2008)
  • Author(s): 朝倉富子, 阿部啓子, 山本隆, 的場輝佳, ジェーン シュミレット, 共著
  • Publisher: 建帛社
• [Book] Neoculin as a new sweet protein with taste-modyfying activity : purification, characterization, and X-ray crystallography. Sweetness and Sweeteners, Biology, Chemistry and Psychophysics. ACS symposium series (2008)
  • Author(s): Shimizu-Ibuka, A., Morita, Y., Nakajima, K., Asakura, T., Terada, T., Misaka, T., Abe, K.
• [Book] 機能性食品の事典 (2007)
  • Author(s): 朝倉富子, 荒井綜一, 阿部啓子, 吉川敏一, 金沢和樹, 渡辺昌, 編集
  • Publisher: 朝倉書店
• [Book] 体験活動に必要な作物ーグリーンアドベンチャー作物編ー (2005)
  • Author(s): 朝倉富子
  • Publisher: 国立信州高遠少年自然の家
• [Book] 調理学実習テキスト (2005)
  • Author(s): 朝倉富子, 久木野睦子
  • Publisher: 京大出版センター
• [Book] 調理科学実験 (2004)
  • Author(s): 朝倉富子, 久木野睦子
  • Publisher: 京大出版センター
• [Remarks] ホームページ等
• [Patent(Industrial Property Rights)] 阿部啓子, 中島健一朗, 朝倉富子, 三坂巧, 石井翔 (2007)
  • Inventor(s): 甘味を有するタンパク質
  • Industrial Property Number: 2008-228690
  • Filing Date: 20070300