Proposal of new screening method of adhesive probiotic lactic acid bacteria by enzymic activity

• Project/Area Number: 22780241
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Zootechnical science/Grassland science
• Research Institution: Miyagi University
• Principal Investigator: KINOSHITA Hideki 宮城大学, 食産業学部, 助教 (50533288)
• Co-Investigator(Renkei-kenkyūsha): SAITO Tadao (00118358) ; ISHIDA Mitsuharu (40151386)
• Project Period (FY): 2010 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2012: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2011: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2010: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 乳酸菌 / 腸管付着性 / 付着因子 / アドヘシン / 酵素活性 / プロバイオティクス / ムチン / 熱ショックタンパク質 / GroEL / 酵素 / スクリーニング / グリセルアルデヒド-3-リン酸脱水素酵素

Research Abstract

Our aim was to construct a new screening method for highly adhesive lactobacilli based on an enzymatic activity without using the adhesion test. Thirty lactobacilli isolates from human intestinal tissues were measured for cell surface GAPDH activity. GAPDH activities were detected in all samples from 18 h cultures. This suggests GAPDH is universally expressed on the bacterial cell surfaces from many lactobacilli. Thenew screening method using GAPDH enzymatic activity without an adhesion test may be possible due to the positive correlation of GAPDH activity with adhesion of lactobacilli.Moreover, the heat shock protein, GroEL, was also detected in many lactobacilli. This suggests GAPDH and GroEL are a very important protein not only in the cytoplasm but also on the cell surface and may be a universal adhesin for lactobacilli.

Research Products

• [Journal Article] Proposal of screening method for intestinal mucus adhesive lactobacilli using the enzymatic activity of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (2013)
  • Author(s): H. Kinoshita, S. Imoto, Y. Suda, M. Ishida, M. Watanabe, Y. Kawai, H. Kitazawa, K. Miura, A. Horii, T. Saito
  • Journal Title: Animal Science Journal Volume: 84巻 Pages: 150-158
  • Peer Reviewed
• [Journal Article] Proposal of screening method for intestinal mucus adhesive lactobacilli using the enzymatic activity of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (2013)
  • Author(s): Kinoshita H, Imoto S, Suda Y, Ishida M, Watanabe M, Kawai Y, Kitazawa H, Miura K, Horii A, Saito T.
  • Journal Title: Anim Sci J. Volume: 84 Issue: 2 Pages: 150-158
  • DOI: 10.1111/j.1740-0929.2012.01054.x
  • Peer Reviewed
• [Journal Article] An Adhesin-Like Protein, Lam29, from Lactobacillus mucosae ME-340 Binds to Histone H3 and Blood Group Antigensin Human Colonic Mucus、Bioscience (2012)
  • Author(s): M. Watanabe, H.Kinoshita, IN. Huang, K. Eguchi, T. Tsurumi, Y. Kawai, H. Kitazawa, K. Kimura, N. Taketomo, D. Kikuchi, T. Sase, K. Miura, H. Ogawa, C. Shibata, A. Horii, T. Saito
  • Journal Title: Biotechnology, and Biochemistry Volume: 76巻 Pages: 1655-1660
  • URL: http://dx.doi.org/10.1271/bbb.120150
  • Peer Reviewed
• [Journal Article] 有用乳酸菌の単離と腸管付着メカニズムの解明に関する研究 (2012)
  • Author(s): 木下英樹
  • Journal Title: ミルクサイエンス Volume: 61巻 Pages: 217-228
  • NAID: 130003398722
  • URL: http://ci.nii.ac.jp/naid/40019556814
• [Journal Article] An Adhesin-Like Protein, Lam29, from <i>Lactobacillus mucosae</i> ME-340 Binds to Histone H3 and Blood Group Antigens in Human Colonic Mucus (2012)
  • Author(s): Watanabe M, Kinoshita H, Huang IN, Eguchi K, Tsurumi T, Kawai Y, Kitazawa H, Kimura K, Taketomo N, Kikuchi D, Sase T, Miura K, Ogawa H, Shibata C, Horii A, Saito T.
  • Journal Title: Bioscience, Biotechnology, and Biochemistry Volume: 76 Issue: 9 Pages: 1655-1660
  • DOI: 10.1271/bbb.120150
  • NAID: 130004137955
  • ISSN: 0916-8451, 1347-6947
  • Peer Reviewed
• [Journal Article] MRS寒天培地と変法LBS寒天培地における乳酸菌の選択的単離能の比較 (2011)
  • Author(s): 木下英樹,井本瞬,須田義人,石田光晴
  • Volume: 60巻
  • Pages: 171-176
  • NAID: 130003398707
  • URL: http://ci.nii.ac.jp/naid/40019173279
  • Peer Reviewed
• [Journal Article] MRS寒天培地と変法LBS寒天培地における乳酸菌の選択的単離能の比較 (2011)
  • Author(s): 木下英樹, 井本瞬, 須田義人, 石田光晴
  • Journal Title: ミルクサイエンス誌 Volume: 60 Pages: 171-176
  • NAID: 130003398707
  • Peer Reviewed
• [Journal Article] Identification of a new adhesin-like protein from Lactobacillus mucosae ME-340 with specific affinity to the human blood group A and B antigens (2010)
  • Author(s): M. Watanabe, H. Kinoshita, M. Nitta, R. Yukishita, Y. Kawai, K. Kimura, N. Taketomo, Y. Yamazaki, Y. Tateno, K. Miura, A. Horii, H. Kitazawa and T. Saito
  • Journal Title: Journal of applied Microbiology Volume: 109巻 Issue: 3 Pages: 927-935
  • DOI: 10.1111/j.1365-2672.2010.04719.x
  • Peer Reviewed
• [Journal Article] プロバイオティック乳酸菌の酵素活性を有する腸管付着因子とその腸管付着メカニズム (2010)
  • Author(s): 木下英樹
  • Journal Title: 宮城大学食産業学部紀要 Volume: 4巻 Pages: 45-50
  • NAID: 120006015011
  • URL: http://ci.nii.ac.jp/naid/40017144924
  • Peer Reviewed
• [Journal Article] Identification of a new adhesin-like protein from Lactobacillus mucosae ME-340 with specific affinity to the human blood group A and B antigens (2010)
  • Author(s): M.Watanabe, H.Kinoshita, et al
  • Journal Title: Journal of Applied Microbiology Volume: 109 Pages: 927-935
  • Peer Reviewed
• [Presentation] Lactobacillus mucosae由来アドへシン様タンパク質(Lam29)の新規ヒト大腸粘液由来結合因子の探索および同定 (2012)
  • Author(s): 渡辺真通, 木下英樹,川井泰,北澤春樹,三浦康,柴田近,堀井明,木村勝紀,竹友直生,齋藤忠夫
  • Organizer: 日本畜産学会第115回大会
  • Place of Presentation: 名古屋(名古屋大学東山キャンパス)
  • Year and Date: 2012-03-28
• [Presentation] Lactobacillus mucosae由来アドヘシン様タンパク質(Lam29)の新規ヒト大腸粘液由来結合因子の探索および同定 (2012)
  • Author(s): 渡辺真通、木下英樹, ら
  • Organizer: 日本畜産学会第115回大会
  • Place of Presentation: 名古屋大学
  • Year and Date: 2012-03-28
• [Presentation] Discovery of a new adhesin-like protein from Lactobacillus mucosae ME-340 with binding activity to human blood group A and B antigens (2010)
  • Author(s): M. Watanabe, H. Kinoshita, M. Nitta, R. Yukishita, M. Haruta, K. Eguchi, Y. Kawai, K. Kimura, N. Taketomo, K. Miura, A. Horii, H. Kitazawa, T. Saito
  • Organizer: The 14th AAAP Animal Science Congress (AAAP2010)
  • Place of Presentation: National Pingtung University of Science and Technology、台湾
  • Year and Date: 2010-08-27
• [Presentation] Search for new Bifidobacterium adhesins with specific binding activity to human colonic mucin (2010)
  • Author(s): R. Yukishita, M. Watanabe, M. Nitta, H. Kinoshita, Y. Kawai, K. Miura, A. Horii, J. Z. Xiao, T. Yaeshima, K. Iwatsuki, H. Kitazawa, T. Saito
  • Organizer: The 14th AAAP Animal Science Congress (AAAP2010)
  • Place of Presentation: National Pingtung University of Science and Technology、台湾
  • Year and Date: 2010-08-27
• [Presentation] Discovery of a new adhesin-like protein from Lactobacillus mucosae ME-340 with binding activity to human blood group A and B antigens (2010)
  • Author(s): Watanabe, Kinoshita, et al
  • Organizer: The 14th AAAP Animal Science Congress
  • Place of Presentation: 台湾(屏東科学技術大学)
  • Year and Date: 2010-08-27
• [Presentation] Search for new Bifidobacterium adhesins with specific binding activity to human colonic mucin (2010)
  • Author(s): Yukishita, Kinoshita, et al
  • Organizer: The 14th AAAP Animal Science Congress
  • Place of Presentation: 台湾(屏東科学技術大学)
  • Year and Date: 2010-08-27
• [Presentation] A new method to screen for lactobacilli adhesive to human colonic mucin (2010)
  • Author(s): H. Kinoshita, S. Imoto, Y. Suda, M. Ishida, M. Watanabe, M. Nitta, R. Yukishita, Y. Kawai, H. Kitazawa, K. Miura, A. Horii, T. Saito
  • Organizer: The 14th AAAP Animal Science Congress (AAAP2010)
  • Place of Presentation: National Pingtung University of Science and Technology、台湾
  • Year and Date: 2010-08-24
• [Presentation] A new method to screen for lactobacilli adhesive to human colonic mucin (2010)
  • Author(s): Kinoshita, et al
  • Organizer: The 14th AAAP Animal Science Congress
  • Place of Presentation: 台湾(屏東科学技術大学)
  • Year and Date: 2010-08-24
• [Presentation] GAPDH 酵素活性を指標とした腸管付着性乳酸菌の新規スクリーニング法の検討 (2010)
  • Author(s): 木下英樹 ,井本瞬, 須田義人,石田光晴,三浦康,堀井明,渡辺真通,新田将光,雪下玲奈, 川井泰,北澤春樹,齋藤忠夫
  • Organizer: 日本乳酸菌学会 2010 年度大会
  • Place of Presentation: 仙台(フォレスト仙台)
  • Year and Date: 2010-07-27
• [Presentation] GAPDH酵素活性を指標とした腸管付着性乳酸菌の新規スクリーニング法の検討 (2010)
  • Author(s): 木下, ら
  • Organizer: 日本乳酸菌学会2010年度大会
  • Place of Presentation: フォレスト仙台
  • Year and Date: 2010-07-27
• [Presentation] ヒト ABO 式血液型抗原を介したプロバイオティック乳酸菌の腸管付着性機構に関する研究 (2010)
  • Author(s): 渡辺真通,新田将光,雪下玲奈,春田美由樹,江口慧,川井泰,北澤春樹,齋藤忠夫,木下英樹,三浦康,堀井明,木村勝紀,竹友直生
  • Organizer: 日本乳酸菌学会 2010 年度大会
  • Place of Presentation: 仙台(フォレスト仙台)
  • Year and Date: 2010-07-26
• [Presentation] ビフィズス菌の新規ヒト腸管付着性因子(アドヘシン)の探索と付着機構の解明 (2010)
  • Author(s): 雪下玲奈,渡辺真通,新田将光,木下英樹,川井泰,北澤春樹,三浦康,堀井明,清水(肖)金忠,八重島智子,岩附慧二,齋藤忠夫
  • Organizer: 日本乳酸菌学会 2010 年度大会
  • Place of Presentation: 仙台(フォレスト仙台)
  • Year and Date: 2010-07-26
• [Presentation] ヒトABO式血液型抗原を介したプロバイオティック乳酸菌の腸管付着性機構に関する研究 (2010)
  • Author(s): 渡辺、木下, ら
  • Organizer: 日本乳酸菌学会2010年度大会
  • Place of Presentation: フォレスト仙台
  • Year and Date: 2010-07-26
• [Presentation] ビフィズス菌の新規ヒト腸管付着性因子(アドヘシン)の探索と付着機構の解明 (2010)
  • Author(s): 雪下、木下, ら
  • Organizer: 日本乳酸菌学会2010年度大会
  • Place of Presentation: フォレスト仙台
  • Year and Date: 2010-07-26
• [Book] 乳酸菌とビフィズス菌のサイエンス (2010)
  • Author(s): 木下英樹、齋藤忠夫
  • Publisher: 京都大学学術出版会
• [Book] 乳酸菌とビフィズス菌のサイエンス (2010)
  • Author(s): 木下, ら
  • Total Pages: 668
  • Publisher: 京都大学学術出版会
• [Remarks]
  • URL: http://www.myu-syoku.jp/university/faculty/Introduction/index/introductionID/51