Development of Novel Bio-Production and Separation Processes Based on Self-Organiizing Structure Formation

基于自组织结构形成的新型生物生产和分离工艺的开发

• 批准号: 09650828
• 负责人: KUBOI Ryoichi
• 金额: $ 2.18万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09650828/
• 关键词: Self-assembly; Aqueous Two-Phase; Stress; Hydrophobicity; Stimuli; Translocation; Separation; Protein; 流動性; 水性ニ相; シャペロン; 酵素; 構造; 細胞膜

When cells and biological systems are exposed to lethal stresses, for example, heat stress, theyrespond to inducevarious latent functions, such as rapid synthesis of specific proteins including a set of heat shock proteins(HSPs), refolding of damaged proteins, translocation and release of proteins across membranes, formation of protein aggregates in an inert insoluble form etc.. These stress responsive or stimuli-responsive functions of cells, proteins, and membraneshave been quantitatively analyzed by the aqueous two-phase partitioning method etc. in the various levels and found to be triggered by the formation of the new transient structure between partly denaturedproteins and HSPs or cell membranes/liposomes, driven by the common principle of self-organIzation, and mediated by the transient hydrophobic Interaction among them. These functions have been successfully utilized for bio-production and bio-separation processes mainly integrating them with the stress induced self-assembled structure, such as stimuli-responsive polymers, aqueous two-phase systems (ATPS), and liposomes. When the cultivated cells or the protein mixture solution was exposed to the specific heat stress condition in the presence of liposomes, stimuli-responsive polymers and/or ATPS, the target protein could be trapped, refolded, translocated, and recovered selectively depending on the applied stress level and proteins.

当细胞和生物系统暴露于致命应激（例如热应激）时，它们会响应诱导各种潜在功能，例如特定蛋白质（包括一组热休克蛋白（HSP））的快速合成、受损蛋白质的重折叠、易位和释放蛋白质跨膜，形成惰性不溶形式的蛋白质聚集体等。细胞、蛋白质和膜的这些应激响应或刺激响应功能已通过水两相分配方法进行定量分析等，发现是由部分变性蛋白质和热休克蛋白或细胞膜/脂质体之间形成新的瞬时结构触发的，由自组织的共同原理驱动，并由它们之间的瞬时疏水相互作用介导。这些功能已成功用于生物生产和生物分离过程，主要将它们与应力诱导的自组装结构相结合，例如刺激响应聚合物、水性两相系统（ATPS）和脂质体。当培养的细胞或蛋白质混合物溶液在脂质体、刺激响应聚合物和/或ATPS存在下暴露于特定的热应激条件时，目标蛋白质可以根据应用选择性地被捕获、重折叠、易位和回收。压力水平和蛋白质。

项目成果

Hiroshi Umakoshi et al.: "Model system for heat-induced translocation of cytophasmic β-galactocidase across phospholipid bilayer membrane" Biotechnol.Prog.14(2). 218-225 (1998)

Hiroshi Umakoshi 等人：“细胞质 β-半乳糖苷酶跨磷脂双层膜热诱导易位的模型系统”Biotechnol.Prog.14(2) (1998)。

Hiroshi Umakoshi, et al.: "Control of Partitioning of Bacterial Cells and Characterization of Their Surface Properties in Agueous Two-Phase Systems" J.Ferment.Bioeng.84(No.6). 572-578 (1997)

Hiroshi Umakoshi 等人：“水两相系统中细菌细胞分配的控制及其表面特性的表征”J.Ferment.Bioeng.84（No.6）。

Makoto Yoshimoto et al.: "Immbilized hiposome Chromatography for Studies of Protein-Membrane Interactions and Refolding of Penatured Bovine Carbonic Anlydrase" J.Chromatogr.B.712. 59-71 (1998)

Makoto Yoshimoto 等人：“固定化海脂质体色谱法用于研究蛋白-膜相互作用和修饰牛碳酸酐酶的重折叠”J.Chromatogr.B.712。

Makoto Yoshimoto et al.: "Immobilized liposome chromatography for studies of protein-membrane interactions and refolding of denatured bovine carbonic anhydrase" J.Chromatogr.B. 712. 59-71 (1998)

Makoto Yoshimoto 等人：“用于研究蛋白质-膜相互作用和变性牛碳酸酐酶重折叠的固定化脂质体色谱”J.Chromatogr.B.

Hiroshi Umakoshi and Ryoichi Kuboi: "Enzyme Release from Heat-Stressed Cell Membranes as a Function of Hydrophobicity Evaluated by Using Aqueous Two-Phase Systems." J.Chromatogr.B. 711. 217-222 (1998)

Hiroshi Umakoshi 和 Ryoichi Kuboi：“使用水性两相系统评估热应激细胞膜的酶释放与疏水性的关系。”