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

当细胞和生物系统暴露在致命性压力(如热应激)下时，它们会对诱导的各种潜在功能做出反应，如特定蛋白质的快速合成，包括一组热休克蛋白(HSPs)，受损蛋白质的重折叠，蛋白质跨膜转移和释放，以惰性不溶形式形成蛋白质聚集体等。细胞、蛋白质和膜的这些应激反应或刺激反应功能已经被用双水相分配法等在不同水平上进行了定量分析，发现是由部分变性的蛋白质与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所介导。这些功能已经被成功地用于生物生产和生物分离过程，主要是将它们与应激诱导的自组装结构相结合，如刺激响应聚合物、双水相系统(ATPs)和脂质体。当培养的细胞或蛋白质混合物溶液暴露在特定的热应激条件下，脂质体、刺激响应聚合物和/或ATP存在时，目标蛋白质可以被选择性地捕获、重折叠、移位和恢复，这取决于施加的应激水平和蛋白质。

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.