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Studies on the structure-function relationships in sweet-tasting proteins and the effects of the masking of bitterness by sweet-tasting proteins

Research Project

Project/Area Number 25450167
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Food science
Research InstitutionKyoto University

Principal Investigator

masuda tetsuya  京都大学, (連合)農学研究科(研究院), 助教 (80311744)

Project Period (FY) 2013-04-01 – 2016-03-31
Project Status Completed (Fiscal Year 2015)
Budget Amount *help
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2015: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2014: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2013: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
Keywords甘味タンパク質 / ソーマチン / X線結晶構造解析 / 室温構造解析 / 苦味抑制 / thaumatin / sweet taste / bitter taste / X-ray crystallography / XFEL / atomic resolution / room temperature / radiation damage / taste / sweetness / bitterness / x-ray crystallization
Outline of Final Research Achievements

The structure of recombinant thaumatin II was determined at the resolution of 0.99Å. The flexible conformations in two critical residues were important for interaction with sweet taste receptors. We introduced a grease-matrix carrier for protein microcrystals and obtained the structure of thaumatin under ambient conditions by Serial femtosecond X-ray crystallography.
We successfully obtained a protein with enhanced sweetness by removing negative charges on the interacting side of thaumatin. The complex model between sweet receptor and thaumatin confirmed that each of the positively charged residues critical for sweetness is close to a receptor residue of opposite charge to yield optimal electrostatic interaction. We prepared the HEK cells stably expressing bitter receptors and ascertained the effect of the masking of bitterness by thaumatin.

Report

(4 results)
  • 2015 Annual Research Report   Final Research Report ( PDF )
  • 2014 Research-status Report
  • 2013 Research-status Report
  • Research Products

    (31 results)

All 2016 2015 2014 2013 Other

All Int'l Joint Research (1 results) Journal Article (9 results) (of which Int'l Joint Research: 3 results,  Peer Reviewed: 8 results,  Open Access: 4 results,  Acknowledgement Compliant: 1 results) Presentation (18 results) (of which Invited: 2 results) Book (1 results) Remarks (2 results)

  • [Int'l Joint Research] フェデリコ2世ナポリ大学(イタリア)

    • Related Report
      2015 Annual Research Report
  • [Journal Article] A Hypersweet Protein: Removal of The Specific Negative Charge at Asp21 Enhances Thaumatin Sweetness.2016

    • Author(s)
      Masuda, T., Ohta, K., Ojiro, N., Murata, K., Mikami, B., Tani, F., Temussi, P.A., and Kitabatake, N.
    • Journal Title

      Sci. Rep.

      Volume: 6 Issue: 1

    • DOI

      10.1038/srep20255

    • NAID

      120005702814

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] Precipitation of sword bean proteins by heating and addition of magnesium chloride in a crude extract.2016

    • Author(s)
      Nishizawa, K., Masuda, T., Takenaka, Y., Masui, H., Tani, F., and Arii, Y.
    • Journal Title

      Biosci. Biotechnol. Biochem.

      Volume: 29 Issue: 8 Pages: 1-9

    • DOI

      10.1080/09168451.2016.1164587

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Redox-coupled proton transfer mechanism in nitrite reductase revealed by femtosecond crystallography.2016

    • Author(s)
      Fukuda Y, Tse KM, Nakane T, Nakatsu T, Suzuki M, Sugahara M, Inoue S, Masuda T, Yumoto F, Matsugaki N, Nango E, Tono K, Joti Y, Kameshima T, Song C, Hatsui T, Yabashi M, Nureki O, Murphy ME, Inoue T, Iwata S, Mizohata E.
    • Journal Title

      PNAS

      Volume: 113 Issue: 11 Pages: 2828-2933

    • DOI

      10.1073/pnas.1517770113

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Native sulfur/chlorine SAD phasing for serial femtosecond crystallography.2015

    • Author(s)
      Nakane T, Song C, Suzuki M, Nango E, Kobayashi J, Masuda T, Inoue S, Mizohata E, Nakatsu T, Tanaka T, Tanaka R, Shimamura T, Tono K, Joti Y, Kameshima T, Hatsui T, Yabashi M, Nureki O, Iwata S, Sugahara M.
    • Journal Title

      Acta Crystallogr D Biol Crystallogr.

      Volume: 71 Issue: 12 Pages: 2519-25

    • DOI

      10.1107/s139900471501857x

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Grease matrix as a versatile carrier of proteins for serial crystallography.2015

    • Author(s)
      Sugahara M, Shimamura T, et al.
    • Journal Title

      Nat Methods.

      Volume: 12 Issue: 1 Pages: 61-63

    • DOI

      10.1038/nmeth.3172

    • NAID

      120005682565

    • Related Report
      2014 Research-status Report
    • Peer Reviewed
  • [Journal Article] Atomic structure of recombinant thaumatin II reveals flexible conformations in two residues critical for sweetness and three consecutive glycine residues.2014

    • Author(s)
      Masuda, T., Mikami, B., Tani, F.
    • Journal Title

      Biochimie.

      Volume: 106 Pages: 33-38

    • DOI

      10.1016/j.biochi.2014.07.016

    • NAID

      120005682566

    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] 甘味タンパク質の構造機能相関、ソーマチンから見えてきたこと2014

    • Author(s)
      桝田哲哉
    • Journal Title

      化学と生物

      Volume: 52 Pages: 23-32

    • NAID

      130004808931

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [Journal Article] Five amino acid residues in cysteine-rich domain of human T1R3 were involved in the response for sweet-tasting protein, thaumatin.2013

    • Author(s)
      Masuda T., Taguchi, W., Sano, A., Ohta, K., Kitabatake, N., and Tani, F.
    • Journal Title

      Biochimie

      Volume: 95 Issue: 7 Pages: 1502-1505

    • DOI

      10.1016/j.biochi.2013.01.010

    • NAID

      120005293582

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [Journal Article] 甘味タンパク質ソーマチンと甘味受容体との応答特性2013

    • Author(s)
      桝田哲哉、谷史人
    • Journal Title

      生体の科学、増大特集 細胞表面受容体

      Volume: 64 Pages: 432-433

    • Related Report
      2013 Research-status Report
  • [Presentation] 甘味タンパク質ソーマチンの甘味強化2016

    • Author(s)
      桝田 哲哉、太田 圭介、小代 奈央子、村田 一輝、三上 文三、谷 史人、Temussi Piero、 北畠 直文
    • Organizer
      日本農芸化学会
    • Place of Presentation
      札幌コンベンションセンター
    • Year and Date
      2016-03-28
    • Related Report
      2015 Annual Research Report
  • [Presentation] シリアル結晶学における重原子誘導体を用いた位相決定2015

    • Author(s)
      山下 恵太郎、潘 東青、長谷川 和也、菅原 道泰、村井 智洋、小段 篤史、溝端 栄一、鈴木 守、桝田 哲哉、平田 邦生、加藤 博章、吾郷 日出夫、熊坂 崇、山本 雅貴、中津 亨
    • Organizer
      日本結晶学会
    • Place of Presentation
      大阪府立大学
    • Year and Date
      2015-10-17
    • Related Report
      2015 Annual Research Report
  • [Presentation] 甘味タンパク質の甘味発現機構2015

    • Author(s)
      桝田 哲哉、北畠 直文、三上 文三、谷 史人
    • Organizer
      食品酵素化学研究会
    • Place of Presentation
      京都大学
    • Year and Date
      2015-09-12
    • Related Report
      2015 Annual Research Report
  • [Presentation] 甘味タンパク質ソーマチン Asp21 変異体の構造活性相関2015

    • Author(s)
      村田 一輝、桝田 哲哉、三上 文三、谷 史人
    • Organizer
      日本食品科学工学会
    • Place of Presentation
      京都大学
    • Year and Date
      2015-08-27
    • Related Report
      2015 Annual Research Report
  • [Presentation] 高分解能 X 線結晶構造解析による甘味タンパク質ソーマチンと苦味物質の相互作用解析2015

    • Author(s)
      大久保 喬平、桝田 哲哉、三上 文三、谷 史人
    • Organizer
      日本食品科学工学会
    • Place of Presentation
      京都大学
    • Year and Date
      2015-08-27
    • Related Report
      2015 Annual Research Report
  • [Presentation] 甘味タンパク質の甘味発現機構2015

    • Author(s)
      桝田 哲哉
    • Organizer
      日本食品科学工学会
    • Place of Presentation
      京都大学
    • Year and Date
      2015-08-27
    • Related Report
      2015 Annual Research Report
    • Invited
  • [Presentation] 甘味タンパク質ソーマチンの連続フェムト秒結晶構造解析2015

    • Author(s)
      桝田 哲哉、南後 恵理子、菅原 道泰、鈴木 守、登野 健介、城地 保昌、亀島 敬、Park Jaehyun、Song Changyong、初井 宇記、矢橋 牧名、大久保 喬平、三上 文三、北畠 直文、谷 史人、岩田 想
    • Organizer
      日本農芸化学会
    • Place of Presentation
      岡山大学
    • Year and Date
      2015-03-28
    • Related Report
      2014 Research-status Report
  • [Presentation] SACLAにおけるシリアルフェムト秒X線結晶構造解析2014

    • Author(s)
      田中 智之、南後 恵理子、菅原 道泰、鈴木 守、桝田 哲哉、山下 鮎美、田中 里枝、登野 健介、城地 保昌、亀島 敬、Jaehyun Park、Changyong Song、初井 宇記、矢橋 牧名、潘 東青、島村 達郎、溝端 栄一、北郷 悠、岩田 想
    • Organizer
      日本結晶学会
    • Place of Presentation
      東京大学
    • Year and Date
      2014-11-01 – 2014-11-03
    • Related Report
      2014 Research-status Report
  • [Presentation] 卵白リゾチームの甘味特性2014

    • Author(s)
      桝田 哲哉、北畠 直文、谷 史人
    • Organizer
      食品酵素化学研究会
    • Place of Presentation
      大阪府立大学I-site難波
    • Year and Date
      2014-08-30
    • Related Report
      2014 Research-status Report
    • Invited
  • [Presentation] 甘味タンパク質ソーマチン Asp21 変異体の構造活性相関2014

    • Author(s)
      村田 一輝、桝田 哲哉、三上 文三、谷 史人
    • Organizer
      日本食品科学工学会
    • Place of Presentation
      中村学園大学
    • Year and Date
      2014-08-28 – 2014-08-30
    • Related Report
      2014 Research-status Report
  • [Presentation] 非可食部に由来する不溶性多糖類ナノファイバーの物性解析2014

    • Author(s)
      二村 健太、阿部 賢太郎、桝田 哲哉、松宮 健太郎、南部 優子、松村 康生、矢野 浩之、谷 史人
    • Organizer
      日本食品科学工学会
    • Place of Presentation
      中村学園大学
    • Year and Date
      2014-08-28 – 2014-08-30
    • Related Report
      2014 Research-status Report
  • [Presentation] XFELを用いた創薬ターゲット蛋白質の迅速構造解析法の開発2014

    • Author(s)
      南後 恵理子、田中 智之、鈴木 守、桝田 哲哉、山下 鮎美、田中 里枝、登野 健介、城地 保昌、亀島 敬、Jaehyun Park、Changyong Song、初井 宇記、矢橋 牧名、潘 東青、島村 達郎、溝端 栄一、北郷 悠、石谷 隆一郎、湯本 史明、岩田 想、
    • Organizer
      日本蛋白質科学会年会
    • Place of Presentation
      ワークピア横浜
    • Year and Date
      2014-06-27
    • Related Report
      2014 Research-status Report
  • [Presentation] Structure-sweetness relationships in the sweet-tasting protein, thaumatin2014

    • Author(s)
      Masuda, T., Sano, A., Murata, K., Ohta, K., and Tani, F.
    • Organizer
      Food Structure Functionality Symposium
    • Place of Presentation
      Amsterdam, The Netherlands.
    • Year and Date
      2014-04-01
    • Related Report
      2014 Research-status Report
  • [Presentation] Structure-sweetness relationships in the sweet-tasting protein, thaumatin2014

    • Author(s)
      Masuda, T., Sano, A., Murata, K., Ohta, K., and Tani, F
    • Organizer
      Food Structure and Functionality Forum Symposium
    • Place of Presentation
      Amsterdam
    • Related Report
      2013 Research-status Report
  • [Presentation] 甘味タンパク質ソーマチンの甘味発現機構の解明

    • Author(s)
      佐野 文音、村田 一輝、桝田 哲哉、三上 文三、谷 史人
    • Organizer
      日本農芸化学会、関西支部大会
    • Place of Presentation
      京都大学
    • Related Report
      2013 Research-status Report
  • [Presentation] 甘味タンパク質ソーマチンと甘味受容体との応答特性

    • Author(s)
      桝田 哲哉、田口 若奈、佐野 文音、太田 圭介、 北畠 直文、 谷 史人
    • Organizer
      日本栄養・食糧学会、近畿支部大会
    • Place of Presentation
      滋賀県立大学
    • Related Report
      2013 Research-status Report
  • [Presentation] 甘味タンパク質ソーマチンの高分解能構造解析

    • Author(s)
      桝田 哲哉、村田 一輝、佐野 文音、三上 文三、北畠 直文、谷 史人
    • Organizer
      甘味タンパク質ソーマチンの高分解能構造解析、日本農芸化学会
    • Place of Presentation
      県立広島大学
    • Related Report
      2013 Research-status Report
  • [Presentation] 甘味タンパク質ソーマチンの苦味抑制作用

    • Author(s)
      桝田 哲哉、塚原 礼恵、太田 圭介、北畠 直文、谷 史人
    • Organizer
      日本食品科学工学会
    • Place of Presentation
      実践女子大学
    • Related Report
      2013 Research-status Report
  • [Book] 化学2016

    • Author(s)
      桝田哲哉
    • Total Pages
      2
    • Publisher
      化学同人
    • Related Report
      2015 Annual Research Report
  • [Remarks] 甘味タンパク質の高甘味度化に成功 -低カロリータンパク質性甘味料の更なる有効利用に期待-

    • URL

      http://www.kyoto-u.ac.jp/ja/research/research_results/2015/160203_1.html

    • Related Report
      2015 Annual Research Report
  • [Remarks] 甘味タンパク質の高甘味度化に成功 -低カロリータンパク質性甘味料の更なる有効利用に期待-

    • URL

      http://www.spring8.or.jp/ja/news_publications/press_release/2016/160204_2/

    • Related Report
      2015 Annual Research Report

URL: 

Published: 2014-07-25   Modified: 2019-07-29  

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