Temperature triggered switching of enzymatic activity via polymer bound, thermoresponsive inhibitor motifs

温度通过聚合物结合的热响应抑制剂基序触发酶活性的切换

• 批准号: 385965031
• 负责人: Dr. Stefan Reinicke
• 金额: --
• 依托单位: Fraunhofer-Institut für Angewandte Polymerforschung (IAP)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2017
• 资助国家: 德国
• 起止时间: 2016-12-31 至 2019-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/385965031?language=en
• 关键词: Temperature; triggered; switching; enzymatic; activity

The main goal is to establish a new concept for temperature controllable enzymatic activity. This concept, comparable to an "on/off"-switch principle, relies on a reversible but almost complete deactivation of an enzyme by the crossing of a particular threshold temperature. For that, we want to make use of the inhibition effect of certain low molecular weight organic substances towards selected enzymes. These inhibiting structural motifs are supposed to be derivatized in a way, that they can be covalently attached to a thiolactone containing, thermoresponsive polymer. By reaching the cloud point of the respective polymer, these motifs can be reversibly removed from the solution volume, making it possible to switch enzymatic activity on and off using a temperatur trigger. The proposed concept can be used in temperature indicator systems but is also potentially useful for a temperature controlled regulation of enzyme cascades in biocatalysis (selective, reversible deactivation of single enzymes within the reaction cascade). At the same time, it widens the application spectrum of functional, responsive polymers and thereby emphasizes the importance of novel strategies for post polymerization modification. Within the proposed project, at least three enzyme/inhibitor combinations, that have been selected by a prior screening of respective databases, are supposed to be utilized in order to demonstrate the broad applicability of this new concept. Tasks to be accomplished include the provision of the selected enzymes, the synthesis of the derivatized inhibitors, the verification of the inhibtion effect after derivatization and binding to the thermoresponsive polymer and investigations on and optimization of the temperature switch effect by structural variation of the utilized polymers. Finally, at least one of the previously optimized systems is supposed to be utilized to generate of photometric indicator that displays the exceedance of a certain, defined threshold temperature as well as the duration of this exceedance.

主要目的是建立一个新的概念，温度可控的酶活性。这一概念类似于"开/关"开关原理，它依赖于通过越过特定的阈值温度而使酶可逆但几乎完全失活。为此，我们希望利用某些低分子量有机物质对选定酶的抑制作用。这些抑制结构基序被认为是衍生化的方式，它们可以共价连接到含硫内酯的温敏聚合物。通过达到相应聚合物的浊点，这些基序可以从溶液体积中可逆地去除，使得可以使用温度触发器打开和关闭酶活性。所提出的概念可用于温度指示系统，但也可能用于生物催化中的酶级联的温度控制调节（反应级联内的单个酶的选择性可逆失活）。同时，它拓宽了功能性、响应性聚合物的应用范围，从而强调了聚合后改性新策略的重要性。在拟议的项目中，至少有三种酶/抑制剂的组合，已经选择了各自的数据库的事先筛选，应该被用来证明这一新概念的广泛适用性。要完成的任务包括提供选定的酶，衍生化的抑制剂的合成，衍生化和结合到温敏聚合物后的抑制作用的验证，以及通过所用聚合物的结构变化对温度开关作用的研究和优化。最后，假定利用先前优化的系统中的至少一个来生成光度指示器，该光度指示器显示某个限定的阈值温度的持续时间以及该持续时间的持续时间。