Development of new technology for removing viruses from water environment

水环境病毒去除新技术开发

• 批准号: 16360260
• 负责人: OMURA Tatsuo
• 金额: $ 9.66万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16360260/
• 关键词: Virus-Binding Proteins; Pathogenic viruses; Water environment; Immobilization; Polylysine-tag; ポリリジンタグ; PCR; ELISA; 配向固定化; グルタルアルデヒド; ポリオウイルス; クローニング

Virus-binding proteins (VBPs) are bacterial proteins to which human viruses preferentially adsorb in water environment. VBPs were discovered from a bacterial culture derived from activated sludge with affinity chromatography assay in which a viral capsid peptide was used as a ligand. The purpose of this study was to develop a novel technology using VBPs for virus removal from environmental water. Achievements described below were obtained during the 3-year study duration.At the first year of the study duration, the scale-up of VBP production and purification was conducted in order to obtain large amount of VBP. A handmade column for VBP purification was constructed, which made it possible to obtain about 10 times larger volume of VBP than conventional procedure.At the second year, gene of hexalysine-tagged VBP (LysTagVBP), which has 6-consecutive lysine residues in its C terminus, was fabricated with PCR, and used for production of LysTagVBP with E. coli. Then, polylysine region in LysTagVBP was coupled with the surface of glass particles using silane coupling reagent and glutaraldehyde. The creation of LysTagVBP directly linked to the effective immobilization of VBP on carrier surface.At the third year of the study duration, the virus-removing column in which LysTagVBP was immobilized was constructed, and virus removal efficiency with the column was evaluated. As a result, about 40% of viruses were reduced during the first 1-hour operation, and 70% of viruses were removed after the 3-hour operation. The development of virus removal technology using VBPs is a totally new challenge, and the achievements with regards to the utilization of VBPs as viral adsorbent would be useful in the farther application of VBPs to virus removal from water.

病毒结合蛋白（VBP）是人类病毒在水环境中优先吸附的细菌蛋白。 VBP 是通过亲和层析法从活性污泥衍生的细菌培养物中发现的，其中病毒衣壳肽用作配体。本研究的目的是开发一种利用 VBP 从环境水中去除病毒的新技术。在为期3年的研究期间，取得了以下成果。 在研究期间的第一年，进行了VBP生产和纯化的放大，以获得大量的VBP。构建了用于VBP纯化的手工柱，这使得获得比常规方法大约10倍体积的VBP成为可能。第二年，六赖氨酸标记的VBP基因（LysTagVBP），其具有6个连续的赖氨酸残基C 末端，通过 PCR 制备，并用于大肠杆菌生产 LysTagVBP。然后，使用硅烷偶联剂和戊二醛将LysTagVBP中的聚赖氨酸区域与玻璃颗粒表面偶联。 LysTagVBP的创建直接关系到VBP在载体表面的有效固定化。在研究进行的第三年，构建了固定有LysTagVBP的病毒去除柱，并评估了该柱的病毒去除效率。结果，在最初1小时的操作中，约40%的病毒被减少，在3小时的操作后，70%的病毒被清除。利用VBPs开发病毒去除技术是一个全新的挑战，利用VBPs作为病毒吸附剂的成果将有助于VBPs在水中病毒去除方面的进一步应用。

项目成果

ファージディスプレイ法を用いた凝集阻害誘因タンパク質をコードする藻類遺伝子の分離

使用噬菌体展示方法分离编码聚集抑制诱导蛋白的藻类基因

• 发表时间: 2006
• 期刊: 環境工学研究論文集 43
• 作者: K.Mizuta;H.Izumi;R.Ichiki;T.Kaneko;市来 龍大;R.Ichiki;T.Kaneko;市來 龍大;市來 龍大;市來 龍大;R.Ichiki;R.Ichiki;市來 龍大;市來 龍大;金子 俊郎;市來 龍大;市來 龍大;R.Ichiki;Grant S.Hansman;Daisuke Sano;Daisuke Sano;Daisuke Sano;Daisuke Sano;佐野大輔
• 通讯作者: 佐野大輔

Detection of infectious pathogenic viruses in water and wastewater samples from urbanized areas

城市化地区水和废水样品中传染性致病病毒的检测

• 发表时间: 2004
• 期刊: Water Science and Technology 50(1)
• 作者: K.Mizuta;H.Izumi;R.Ichiki;T.Kaneko;市来 龍大;R.Ichiki;T.Kaneko;市來 龍大;市來 龍大;市來 龍大;R.Ichiki;R.Ichiki;市來 龍大;市來 龍大;金子 俊郎;市來 龍大;市來 龍大;R.Ichiki;Grant S.Hansman;Daisuke Sano;Daisuke Sano;Daisuke Sano;Daisuke Sano;佐野大輔;Daisuke Sano;You Ueki;Tomoko Takaara;You Ueki;Daisuke Sano;佐野大輔;Daisuke Sano;Daisuke Sano;Daisuke Sano;佐野大輔;Daisuke Sano
• 通讯作者: Daisuke Sano

Human Sapovirus in Clams, Japan

日本蛤蜊中的人类沙波病毒

• 发表时间: 2007
• 期刊: Emerging Infectious Diseases 13・4(in press)
• 作者: Norio Onikura;Naohiko Takeshia;Seiichi Matsui;Seiro Kimura;Grant S.Hansman;Grant S.Hansman
• 通讯作者: Grant S.Hansman

Virus-binding proteins recovered from bacterial culture derived from activated sludge by affinity chromatography assay using a viral capsid peptide

• DOI: 10.1128/aem.70.6.3434-3442.2004
• 发表时间: 2004-06-01
• 期刊: APPLIED AND ENVIRONMENTAL MICROBIOLOGY
• 影响因子: 4.4
• 作者: Sano, D;Matsuo, T;Omura, T
• 通讯作者: Omura, T

アデノウイルス吸着タンパク質(Adenovirus-Binding Protein : ADVBP)の活性汚泥細菌からの分離

从活性污泥细菌中分离腺病毒结合蛋白 (ADVBP)

• 发表时间: 2004
• 期刊: 環境工学研究論文集 41
• 作者: K.Mizuta;H.Izumi;R.Ichiki;T.Kaneko;市来 龍大;R.Ichiki;T.Kaneko;市來 龍大;市來 龍大;市來 龍大;R.Ichiki;R.Ichiki;市來 龍大;市來 龍大;金子 俊郎;市來 龍大;市來 龍大;R.Ichiki;Grant S.Hansman;Daisuke Sano;Daisuke Sano;Daisuke Sano;Daisuke Sano;佐野大輔;Daisuke Sano;You Ueki;Tomoko Takaara;You Ueki;Daisuke Sano;佐野大輔;Daisuke Sano;Daisuke Sano;Daisuke Sano;佐野大輔
• 通讯作者: 佐野大輔