Research on Preparationand Application of Stress-Responsive Nano-Colloid

应力响应纳米胶体的制备及应用研究

• 批准号: 08555183
• 负责人: KUBOI Ryoichi
• 金额: $ 4.93万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08555183/
• 关键词: Stress Response; Bio-Separation; Reverse Micelle; Percolation; Refolding; Protein; Hydrophobic Interaction; Liposome; 分離・精製; 分子集合体; 膜透過; 機能材料; 会合コロイド; シャペロニン; 疎水性

The stress responsive functions, for example synthesis of specific proteins including a set of heat shock proteins, refolding of damaged proteins, translocation arid release of proteins across membranes, and formation of protein aggregates in an inert insoluble form etc., of biological nano-colloid such as cells, membranes, and proteins have been quantitatively analyzed by using the percolation method, aqueous two-phase partitioning method, and immobilized liposome chromatographyin the various levels focusing on the transient and weak molecular interactions between proteins and lipid membranes or reverse micelles under various stress conditions. It was found that the formation of a new transient structure between partly denatured proteins and reverse micelles or liposomes played the key role in the above stress resposive functions. It was found to be driven by the fluidity of nano-colloidal interface combined with the transient hydrophobic interaction between them and could be mediated and controlled by kinds of stressors, their combinations, and stress levels. These quantitative results have been successfully utilized for the design and development of novel and efficient protein refolding and bio-separation processes.

生物纳米胶体的应激响应功能，例如包括一组热休克蛋白在内的特定蛋白质的合成、受损蛋白质的重折叠、蛋白质跨膜易位和释放、以及惰性不溶形式的蛋白质聚集体的形成等采用渗滤法、水两相分配法、固定化脂质体层析等方法对细胞、细胞膜、蛋白质等物质进行不同水平的定量分析，重点关注蛋白质与脂膜之间瞬时、微弱的分子相互作用或反向相互作用。不同应力条件下的胶束。研究发现，部分变性蛋白质与反胶束或脂质体之间形成的新的瞬时结构在上述应激反应功能中起关键作用。研究发现，它是由纳米胶体界面的流动性以及它们之间的瞬时疏水相互作用驱动的，并且可以通过各种压力源、它们的组合和压力水平来介导和控制。这些定量结果已成功用于设计和开发新型高效的蛋白质重折叠和生物分离过程。

项目成果

Hiroshi Umakoshi, Ryoichi Kuboi, and Isao Komasawa: "Control of Partitioning Behaviors of Bacterial Cells and Characterization of Their Surface Properties in Aqueous Two-Phase Systems" J.Ferment.Bioeng.84 (6). 572-578 (1997)

Hiroshi Umakoshi、Ryoichi Kuboi 和 Isao Komasawa：“水性两相系统中细菌细胞分配行为的控制及其表面性质的表征”J.Ferment.Bioeng.84 (6)。

Dong-Pyo,Hong etal.: "Extraction of Proteins and Polymers Using Reversed Micelles and Percolation Process." Korean J.Chem.Eng.14(5). 334-340 (1997)

Dong-Pyo, Hong 等人：“使用反胶束和渗滤过程提取蛋白质和聚合物。”

Hiroshi Umakoshi, Toshinori Shimanouchi, and Ryoichi Kuboi: "Selective Separation Process of Proteins Based on the Heat Stress-Induced Translocation across Phospholipid Membranes." J.Chromatogr.B. 711. 111-116 (1998)

Hiroshi Umakoshi、Toshinori Shimanouchi 和 Ryoichi Kuboi：“基于热应激诱导的磷脂膜易位的蛋白质选择性分离过程”。

Ryoichi Kuboi: "Refolding of Carbonic Anhydrase Assisted by 1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine Liposomes," Biotechnol.Prog.13. 828-836 (1997)

Ryoichi Kuboi：“1-棕榈酰-2-油酰-sn-甘油-3-磷酸胆碱脂质体辅助碳酸酐酶的重折叠”，Biotechnol.Prog.13。

Hiroshi Umakoshi etal.: "Selective Separation Process of Proteins Based on the Heat Stress-Induced Translocation across Phospholipid Membranes." J.Chromatogr.B.711. 111-116 (1998)

Hiroshi Umakoshi 等人：“基于热应激诱导的磷脂膜易位的蛋白质选择性分离过程。”