2000 Fiscal Year Annual Research Report

生体分子アーキテクチャーの構築およびその応用

Research Project

Project/Area Number	10145102
Research Institution	TOKYO UNIVERSITY OF AGRICULTURE AND TECHNOLOGY
Principal Investigator	松永是東京農工大学, 工学部, 教授 (10134834)
Co-Investigator(Kenkyū-buntansha)	小畠英理東京工業大学, 生命理工学部, 助教授 (00225484) 後藤雅宏九州大学, 工学部, 助教授 (10211921) 小宮山真東京大学, 大学院・工学系研究科, 教授 (50133096)
Keywords	分子アーキテクチャー / 磁性細菌粒子 / モレキュラーインプリント法 / 人工レセプター / 界面活性剤-酵素複合 / バイオターゲティング / 脂質修飾抗体結合タンパク質 / イムノリポソーム
Research Abstract	1)磁性細菌粒子膜局在性タンパク質の解析からMms16タンパク質が単離され、塩基配列が決定された。MagA、Mms16をアンカー分子として用い、プロテインA、エストロゲンレセプターを粒子表面にディスプレイし、構築された分子アーキテクチャーを用いたエストロゲン様化学物質、糖尿病マーカー物質の同時検出を行うことが可能となった。また、磁性細菌粒子膜タンパク質の除去を行うことにより、in vitro系におけるMagA-ルシフェラーゼ融合タンパク質の挿入効率が向上することが示された。 2)モレキュラーインプリント法で得られた架橋シクロデキストリン高分子のHPLC担体への応用について検討を行った。インプリント・シクロデキストリン高分子をシリカゲル上に固定化しHPLC担体を作製したところ、ステロイド類、ペプチド・抗生物質の分離が可能であった。実用に耐えうるカラムであることを明らかにし、HPLC担体として有用であることを示した。 3)ガン細胞へのターゲッティング性を高めるため、糖質界面活性剤を含むハイブリッド型リポソームを作製し、ガン細胞表面に存在するコンカナバリンAとの相互作用について評価を行った。グルコース含有ハイブリッドリポソームはコンカナバリンAと特異的に結合し、その様子は可逆的であることが明らかとなった。 4)脂質修飾抗体結合タンパク質の設計にあたり目的とした、脂質膜への分子の導入および、膜表面での分子機能の発現をイムノリポソームの機能として確認した。脂質修飾タンパク質としての機能であるタンパク質表面への分子の集積は、抗体分子との複合体の状態を維持したまま膜表面に集積されていることが明らかとなった。

Research Products

(29 results)

All Other

All Publications (29 results)

[Publications] T.Matunaga: "Cloning and characterization of a genempsA,encoding a protein associated with intracellular magnetic particles from Magnetospirillum sp. strain AMB-1."Biochem.Biophys.Res.Commun.. 268. 932-937 (2000)
[Publications] Y.Okamura: "Two-dimensional analysis of proteins specific to the bacterial particle membrane from Magnetospirillum sp.AMB 1."Appl.Biochem.Biotechnol.. 84-86. 441-446 (2000)
[Publications] H.Takeyama: "Discrimination between atlantic and pacific subspecies of northern bluefin tuna (Thunnus thynnus) by magnetic-capture hybridization using bacterial magnetic particles."Mar.Biotechnol.. 2. 309-313 (2000)
[Publications] T.Tanaka: "Fully automated chemiluminescence immunoassay of insulin using antibody-protein A-bacterial magnetic particle complexes."Anal.Chem.. 72. 3518-3522 (2000)
[Publications] T.Matsunaga: "Production of luciferase-magnetic particle complex by recombinant Magnetospirillum sp.AMB-1."Biotech.Bioeng.. 70. 704-709 (2000)
[Publications] T.Matsunaga: "Molecular mechanism of magnet formation in bacteria."J Biosci Bioeng.. 90. 1-13 (2000)
[Publications] A.T.Wahyudi: "Isolation of Magnetospirillum magneticum AMB-1 mutants defective in magnetic particles synthesis by transposon mutagenesis."Appl.Biochem.Biotechnol.. (in press).
[Publications] C.D.Yang: "Synthesis of bacterial magnetic particles during cell cycle of Magnetospirillum magneticum AMB-1."Appl.Biochem.Biotechnol.. (in press).
[Publications] T.Matsunaga: "Fluorescent detection of cyanobacterial DNA using bacterial magnetic particles on a MAG microarray."Biotechnol.Bioeng.. (in press).
[Publications] R.Sato: "Development of a high performance and rapid immunoassay for food allergen using antibody conjugated bacterial magnetic particles and fully-automated system."Appl.Biochem.Biotechnol.. (in press).
[Publications] C.D.Yang: "Optimization of iron feeding and stability of plasmid for production of recombinant bacterial magnetic paricles by Magnetospirillum magneticum AMB-1 in fed-batch culture."J.Biosci.Bioeng.. (in press).
[Publications] C.D.Yang: "Effects of growth medium composition, iron sources and atmospheric oxygen concentrations on production of luciferase-bacterial magnetic particle complex by a recombinant Magnetospirillum magneticum AMB-1."Enzyme Microb.Technol.. (in press).
[Publications] T.Matsunaga: "16SrRNA-targeted identification of cyanobacterial genera using oligonucleotide-probes immobilized on bacterial magnetic particles."J.Appl.Phycol.. (in press).
[Publications] H.Asanuma: "Tailor-made receptors by molecular imprinting."Adv.Mater.. 12. 1019-1030 (2000)
[Publications] H.Asanuma: "Molecular imprinting of cyclodextrin in water for the recognition of nano-meter-scaled guests."Ana.Chim.Acta.. (in press).
[Publications] N.Kamiya: "Surfactant-HRP complex catalytically active in organic media."Biotechnol.Prog.. 16. 52-58 (2000)
[Publications] S.Okazaki: "Surfactant-protease complex as a novel biocatalyst for peptide synthesis in hydrophilic organic solvents"Enzyme Microb.Technol.. 26. 159-164 (2000)
[Publications] S.Okazaki: "Characterization and catalytic property of surfactant-laccase complex in organic media"Biotechnol.Prog.. 16. 583-588 (2000)
[Publications] Y.Sugimura: "A study on the surface hydrophobicity of lipases."Biochem.Engin.J.. 5. 123-128 (2000)
[Publications] S.Okazaki: "Surfactant-lactoperoxidase complex catalytically active in organic media."Biochem.Eng.J.. 6. 103-108 (2000)
[Publications] M.Goto: "Reversed micelles as novel protein refolding media."ACS Symposium Series 740 : Surfactant-Based Separation. 22. 374-383 (2000)
[Publications] S.Kanno: "Assembling of engineered IgG-binding protein on gold surface for high oriented antibody immobilization."J.Biotechnol.. 76. 207-214 (2000)
[Publications] E.Kobatake: "Aizawa : Immunoassay systems based on immunoliposomes consisting of genetically engineered single-chain antibody."Snsors and Actuators.. B65. 42-45 (2000)
[Publications] T.Sugihara: "Genetically synthesized antibody-binding protein self-assembled on hydrophobic matrix."Bioconjugate Chem.. 11. 789-794 (2000)
[Publications] 松永是: "磁性細菌におけるマグネタイト微粒子の形成機構月刊海洋Vol.32,No.6"海洋出版. 7 (2000)
[Publications] 松永是: "「DNAチップ」化学と教育48巻6号"社団法人日本化学会. 2 (2000)
[Publications] 松永是: "「バイオ産業・事業の創造的開発強化研究会シリーズ(6)食品・環境への応用」技術と経済,402"(社)科学技術と経済の会. 14 (2000)
[Publications] 松永是: "「生物磁石を利用した癌治療!」実験医学(9月号)"羊土社. 3 (2000)
[Publications] 松永是: "「細胞表面タンパク質検出のための生体分子認識ナノプローブの開発」-マイクロマシン技術に関する研究助成-第6回研究成果報告書"財団法人マイクロマシンセンター. 5 (2000)

2000 Fiscal Year Annual Research Report

生体分子アーキテクチャーの構築およびその応用

Principal Investigator

松永 是 東京農工大学, 工学部, 教授 (10134834)

Research Products

[Publications] T.Matunaga: "Cloning and characterization of a genempsA,encoding a protein associated with intracellular magnetic particles from Magnetospirillum sp. strain AMB-1."Biochem.Biophys.Res.Commun.. 268. 932-937 (2000)

[Publications] Y.Okamura: "Two-dimensional analysis of proteins specific to the bacterial particle membrane from Magnetospirillum sp.AMB 1."Appl.Biochem.Biotechnol.. 84-86. 441-446 (2000)

[Publications] H.Takeyama: "Discrimination between atlantic and pacific subspecies of northern bluefin tuna (Thunnus thynnus) by magnetic-capture hybridization using bacterial magnetic particles."Mar.Biotechnol.. 2. 309-313 (2000)

[Publications] T.Tanaka: "Fully automated chemiluminescence immunoassay of insulin using antibody-protein A-bacterial magnetic particle complexes."Anal.Chem.. 72. 3518-3522 (2000)

[Publications] T.Matsunaga: "Production of luciferase-magnetic particle complex by recombinant Magnetospirillum sp.AMB-1."Biotech.Bioeng.. 70. 704-709 (2000)

[Publications] T.Matsunaga: "Molecular mechanism of magnet formation in bacteria."J Biosci Bioeng.. 90. 1-13 (2000)

[Publications] A.T.Wahyudi: "Isolation of Magnetospirillum magneticum AMB-1 mutants defective in magnetic particles synthesis by transposon mutagenesis."Appl.Biochem.Biotechnol.. (in press).

[Publications] C.D.Yang: "Synthesis of bacterial magnetic particles during cell cycle of Magnetospirillum magneticum AMB-1."Appl.Biochem.Biotechnol.. (in press).

[Publications] T.Matsunaga: "Fluorescent detection of cyanobacterial DNA using bacterial magnetic particles on a MAG microarray."Biotechnol.Bioeng.. (in press).

[Publications] R.Sato: "Development of a high performance and rapid immunoassay for food allergen using antibody conjugated bacterial magnetic particles and fully-automated system."Appl.Biochem.Biotechnol.. (in press).

[Publications] C.D.Yang: "Optimization of iron feeding and stability of plasmid for production of recombinant bacterial magnetic paricles by Magnetospirillum magneticum AMB-1 in fed-batch culture."J.Biosci.Bioeng.. (in press).

[Publications] C.D.Yang: "Effects of growth medium composition, iron sources and atmospheric oxygen concentrations on production of luciferase-bacterial magnetic particle complex by a recombinant Magnetospirillum magneticum AMB-1."Enzyme Microb.Technol.. (in press).

[Publications] T.Matsunaga: "16SrRNA-targeted identification of cyanobacterial genera using oligonucleotide-probes immobilized on bacterial magnetic particles."J.Appl.Phycol.. (in press).

[Publications] H.Asanuma: "Tailor-made receptors by molecular imprinting."Adv.Mater.. 12. 1019-1030 (2000)

[Publications] H.Asanuma: "Molecular imprinting of cyclodextrin in water for the recognition of nano-meter-scaled guests."Ana.Chim.Acta.. (in press).

[Publications] N.Kamiya: "Surfactant-HRP complex catalytically active in organic media."Biotechnol.Prog.. 16. 52-58 (2000)

[Publications] S.Okazaki: "Surfactant-protease complex as a novel biocatalyst for peptide synthesis in hydrophilic organic solvents"Enzyme Microb.Technol.. 26. 159-164 (2000)

[Publications] S.Okazaki: "Characterization and catalytic property of surfactant-laccase complex in organic media"Biotechnol.Prog.. 16. 583-588 (2000)

[Publications] Y.Sugimura: "A study on the surface hydrophobicity of lipases."Biochem.Engin.J.. 5. 123-128 (2000)

[Publications] S.Okazaki: "Surfactant-lactoperoxidase complex catalytically active in organic media."Biochem.Eng.J.. 6. 103-108 (2000)

[Publications] M.Goto: "Reversed micelles as novel protein refolding media."ACS Symposium Series 740 : Surfactant-Based Separation. 22. 374-383 (2000)

[Publications] S.Kanno: "Assembling of engineered IgG-binding protein on gold surface for high oriented antibody immobilization."J.Biotechnol.. 76. 207-214 (2000)

[Publications] E.Kobatake: "Aizawa : Immunoassay systems based on immunoliposomes consisting of genetically engineered single-chain antibody."Snsors and Actuators.. B65. 42-45 (2000)

[Publications] T.Sugihara: "Genetically synthesized antibody-binding protein self-assembled on hydrophobic matrix."Bioconjugate Chem.. 11. 789-794 (2000)

[Publications] 松永是: "磁性細菌におけるマグネタイト微粒子の形成機構 月刊海洋Vol.32,No.6"海洋出版. 7 (2000)

[Publications] 松永是: "「DNAチップ」化学と教育48巻6号"社団法人日本化学会. 2 (2000)

[Publications] 松永是: "「バイオ産業・事業の創造的開発強化研究会シリーズ(6)食品・環境への応用」技術と経済,402"(社)科学技術と経済の会. 14 (2000)

[Publications] 松永是: "「生物磁石を利用した癌治療!」実験医学(9月号)"羊土社. 3 (2000)

[Publications] 松永是: "「細胞表面タンパク質検出のための生体分子認識ナノプローブの開発」-マイクロマシン技術に関する研究助成-第6回研究成果報告書"財団法人マイクロマシンセンター. 5 (2000)

松永是東京農工大学, 工学部, 教授 (10134834)

[Publications] 松永是: "磁性細菌におけるマグネタイト微粒子の形成機構月刊海洋Vol.32,No.6"海洋出版. 7 (2000)