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Development of virus removal technology using virus-binding proteins

Research Project

Project/Area Number 13555145
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Civil and environmental engineering
Research InstitutionTohoku University

Principal Investigator

OMURA Tatsuo  Tohoku University, Graduate School of Engineering, Professor, 大学院・工学研究科, 教授 (30111248)

Co-Investigator(Kenkyū-buntansha) WATANABE Toru  Tohoku University, Graduate School of Engineering, Research Associate, 大学院・工学研究科, 助手 (10302192)
KURIHARA Kazue  Tohoku University, Institute of Multidisciplinary Research for Advanced Materials, Professor, 多元物質科学研究所, 教授 (50252250)
Project Period (FY) 2001 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥13,500,000 (Direct Cost: ¥13,500,000)
Fiscal Year 2003: ¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 2002: ¥4,700,000 (Direct Cost: ¥4,700,000)
Fiscal Year 2001: ¥6,500,000 (Direct Cost: ¥6,500,000)
KeywordsVirus-Binding Protein / Pathogenic viruses / Treated Wastewater / Affinity chromatography / Cloning / Oriented immobilization / Poly Lysine-tag / Aminosilane / ウイルス吸着タンパク質(Virus-Binding Protein : VBP) / グルタルアルデヒド / ELISA / VBP遺伝子 / ゲノムDNAライブラリ / コロニーハイブリダイゼーション / VBPクローニング / ポリオウイルスI型 / 抗原決定基 / イオン交換クロマトグラフィ / ELISA法 / 活性汚泥細菌 / 尿素
Research Abstract

The contamination of water environments by pathogenic viruses has raised concerns about outbreaks of viral Infectious diseases in our society. Because conventional water and wastewater treatment systems are not effective enough to inactivate or remove pathogenic viruses, a new technology for virus removal needs to be developed. In this project, the virus-binding proteins (VBPs) in a bacterial culture derived from activated sludge were successfully recovered. The recovery of VBPs was achieved by applying extracted crude proteins from a bacterial culture to an affinity column in which a custom-made peptide of capsid protein from poliovirus type 1 (PV1) was immobilized as a ligand. VBPs exhibited the ability to adsorb infectious particles of PV1 as determined by ELISA. The adsorption of the VBPs to the viral peptide in the affinity column occurred with a strong attractive force able to overcome the electrostatic repulsive force. Two-dimensional electrophoresis revealed that the isolated VBPs include a number of proteins, and their molecular weights were widely distributed but smaller than 100kDa. Homology searches for the N termini against all protein sequences in NCBI database showed that the isolated VBPs were newly discovered proteins. The VBP gene of activated sludge bacteria was isolated, and the cloning system of the VBP was established. The isolation of the VBP gene from DNA libraries for activated sludge bacteria was achieved with the colony hybridization technique. It was confirmed that E. coli BL21 transformed by pRSET carrying the isolated VBP gene could extensively produce
the VBP clones. ELISA revealed that the VBP clone exhibited the binding ability with intact particles of PV1. The VBP cloning system developed in this study would make it possible to produce a mass volume of the VBP, and to utilize them as a new material of the viral adsorbent.

Report

(4 results)
  • 2003 Annual Research Report   Final Research Report Summary
  • 2002 Annual Research Report
  • 2001 Annual Research Report
  • Research Products

    (30 results)

All Other

All Publications (30 results)

  • [Publications] Takahiro Matsuo, Daisuke Sano, Kensuke Fukushi, Tatsuo Omura: "Adsorption of Poliovirus 1 and Adenovirus 41 to Virus-Binding Proteins"Proceeding of Asian Waterqual 2003, IWA-Asia Pacific Regional Conference. (CD-ROM). (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Daisuke Sano, Kensuke Fukushi, Yasuko Yoshida, Tatsuo Omura: "Detection of enteric viruses in municipal sewage sludge by a combination of the enzymatic virus elution method and RT-PCR"Water Research. 37(14). 3490-3498 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Taro Antsuki, Daisuke Sano, Tatsuo Omura: "Functional metal-binding proteins by metal-stimulated bacteria for the development of an innovative metal removal technology"Water Science and Technology. 47(10). 109-115 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Syujiro Hayashi, Takashi Abe, Nobuyuki Higashi, Masazo Niwa, Kazue Kurihara: "Polyelectroyte Brush Layers Studied by Surface Forces Measurement : Dependence on pH and Salt Concentrations and Scaling"Langmuir. 18. 3932-3944 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Kazue Kurihara: "Nanostructuring of Liquids at Solid-Liquid Interfaces"Progr.Colloid Polym.Sci.. 121. 49-56 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Masashi Mizukami, Masashi Moteki, Kazue Kurihara: "Hydrogen-Bonded Macrocluster Formation of Ethanol on Silica Surfaces in Cyclohexane"J.Am.Chem.Soc.. 124. 12889-12897 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Takahiro Matsuo, Daisuke Sano, Kensuke Fukushi, Tatsuo Omura: "Adsorption of Poliovirus 1 and Adenovirus 41 to virus-Binding Proteins"Proceeding of Asian Waterqual 2003, IWA-Asia oacific Regional Conference. (CD-ROM). (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Daisuke Sano, Kensuke Fukushi, Yasuko Yoshida, Tatsuo Omura: "Detection of enteric viruses in municipal sewage sludge by a combination of the enzymatioc virus elution method and RT-PCR"Water Research. 37(14). 3490-3498 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Taro Antsuki, Daisuke Sano, Tatsuo Omura: "Functional metal-binding proteins by metal-stimulated bacteria for the development of an innovative metal removal technology"Water Science and Technology. 47(10). 109-115 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Syujiro Hayashi, Takeshi Abe, Nobuyuki Higashi, Masazo Niwa, Kazue Kurihara: "Polyelectroyte Brush Layers Studied by Surface Forces Measurement : Dependence on pH and Salt Concentrations and Scaling"Lanfmuir. 18. 3932-3944 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Kazue Kurihara: "Nanostructuring of Liquids at Solid-Liquid Interfaces"Progr. Colloid Polym.Sci.. 121. 49-56 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Masashi Mizukami, Masashi Moteki, Kazue Kurihara: "Hydrogen-Bonded Macrocluster Formation of Ethanol on Silica Surfaces in Cyclohexane"J.Am.Chem.Soc.. 124. 12889-12897 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] T.Suzuki, Y-W.Zhang, T.Koyama, D.Y.Sasaki, K.Kurihara: "Direct Observation of Specific Interaction between Enzyme-Substrate Complexes Using Colloidal Probe Atomic Force Microscopy"Chemistry Letters. (in press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Daisuke Sano, Takahiro Matsuo, Tatsuo Omura: "Virus-binding proteins recovered from bacterial culture derived from activated sludge by the affinity chromatography using a viral capsid peptide"Applied and Environmental Microbiology. (in press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Daisuke Sano, Kensuke Fukushi, Yasuko Yoshida, Tatsuo Omura: "Detection of enteric viruses in municipal sewage sludge by a combination of the enzymatic virus elution method and RT-PCR"Water Research. 37・14. 3490-3498 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Taro Antsuki, Daisuke Sano, Tatsuo Omura: "Functional metal-binding proteins by metal-stimulated bacteria for the development of an innovative metal removal technology"Water Science and Technology. 47・10. 109-115 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] K.Kurihara, Y.Nakagawa, M.Mizukami: "Hydrogen-bonded Macrocluster Formation of Ethylene Glycol on Silica Surfaces in Ethylene Glycol-Cyclohexane Binary Liquids"Chemistry Letters. 32・1. 84-85 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] M.Mizukami, K.Kurihara: "Hydrogen-Bonded Macrocluster Formation of 1-Propanol and 2-Pronanol on Silica Surfaces"Aust.J.Chem. 56. 1071-1080 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 松尾崇宏, 佐野大輔, 大村達夫: "アフィニティクロマトグラフィによる活性汚泥からのウイルス吸着タンパク質(Virus-Binding Protein : VBP)の分離と特性評価"環境工学研究論文集. 39. 345-353 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 松尾崇宏, 佐野大輔, 大村達夫: "活性汚泥からのウイルス吸着タンパク質(Virus-Binding Protein : VBP)の分離およびその吸着能評価"第57回土木学会年次学術講演会講演集. 57. 335-336 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Taro Antsuki, Daisuke Sano, Tatsuo Omura: "Functional Metal-Binding Proteins by Metal-stimulated Bacteria for the Development of an Innovative Metal Removal Technology"Water Science and Technology. (in press). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Syujiro Hayashi, Takashi Abe, Nobuyuki Higashi, Masazo Niwa, Kazue Kurihara: "Polyelectroyte Brush Layers Studied by Surface Forces Measurement : Dependence on pH and Salt Concentrations and Scaling"Langmuir. 18. 3932-3944 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Kazue Kurihara: "Nanostructuring of Liquids at Solid-Liquid Interfaces"Progr. Colloid Polym. Sci.. 121. 49-56 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Masashi Mizukami, Masashi Moteki, Kazue Kurihara: "Hydrogen-Bonded Macrocluster Formation of Ethanol on Silica Surfaces in Cyclohexane"J. Am. Chem. Soc.. 124. 12889-12897 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] T.Antsuki, D.Sano, T.Omura: "Functional Metal-Binding Proteins by Metal-stimulated Bacteria for the Development of an Innovative Metal Removal Technology"3rd World Water Congress of International Water Association. (発売予定). (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] 佐野大輔, 長山恒紀, 大村達夫: "アフィニティークロマトグラフィによる活性汚泥中タンパク質からのVirus Binding Protein(VBP)の分離"土木学会第56回年次学術講演会講演集. 第7巻. 374-375 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 安附太郎, 佐野大輔, 大村達夫: "重金属親和性を有する活性汚泥中タンパク質の検索"土木学会第56回年次学術講演会講演集. 第7巻. 456-457 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Y.Nakai, K.Kurihara: "Photoinduced Long-Range Attraction between Spiropiran Monolayers Studied by Surface Forces Measurement,"Studies in Surface Science and Catalysis. Vol.132. 869-872 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] K.Kurihara: "Nanostructuring of Liquids at the Solid-Liquid Interfaces"Progr. Colloid Polym. Sci. (in press). (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] 宮原隆、栗原和枝: "表面力測定による表面キャラクタリゼーション"日本接着学会誌. 第37巻. 36-41 (2001)

    • Related Report
      2001 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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