Fast protein liquid chromatography system for protein purification

所有生物都有专门的分子，称为酶，以执行特定的生物功能。许多酶已经进化到使用自由基（高活性孤电子）来进行生物转化。 自由基的优点在于它们的高反应性，因此能够进行化学上困难且需要大量能量的反应。然而，自由基反应不加选择，这可能会破坏酶。 酶是动态系统;因此，内部运动的时间必须与化学事件精确地同步，以防止自由基本身熄灭。 该研究实验室的目标是研究酶如何通过大的结构重排来协调自由基化学。 为此，该实验室将研究一种含有维生素B12衍生物的酶，该酶通过破坏一种不寻常的碳钴键来引发自由基化学反应。 然后，自由基传播到另一种维生素衍生分子（维生素B6），位于酶的远处。快速反应光谱技术沿着化学合成的新化合物类似B12将允许深入调查的动力学和动力学行为的酶。 从这项研究中，基于自由基的催化模型将被导出，可以应用于其他酶系统。这项研究还将使我们能够重新设计酶，以进行适用于药物合成或其他工业用途的其他化学转化。 然而，要研究单个酶，首先必须将其与细菌细胞中其他数千种蛋白质和酶分离。 一个高压液相色谱系统，如ÄKTA FPLC就是为此而设计的。 该系统是完全自动化的，能够以高分辨率，快速和可重复地分离蛋白质。 该系统在工业界和学术界广泛使用，HQP谁获得的专业知识，在ÄKTA系统将更好地为未来的研究和开发事业做好准备。 UBC Okanagan目前还没有一个<$KTA FPLC或类似的系统。

All living things have specialized molecules, called enzymes, to carry out specific biological functions. A number of enzymes have evolved to use radicals (highly reactive lone electrons) to perform biological transformations. The advantage of radicals lies in their highly reactive nature, and therefore ability to carry out reactions that are chemically difficult and require a lot of energy. However, radicals react indiscriminately, which can damage the enzyme. Enzymes are dynamic systems; hence, the timing of internal motions must be exquisitely synchronized with chemical events to prevent the radical extinguishing itself. The goal of the research laboratory is to study how enzymes orchestrate radical chemistry with large structural rearrangements. To do this, the laboratory will study an enzyme that contains a vitamin B12-derivative, which it uses to initiate radical chemistry through breakage of an unusual carbon-cobalt bond. The radicals are then propagated to another vitamin-derived molecule (vitamin B6), nestled in the enzyme at a distant location. Fast-reaction spectroscopic techniques along with chemical synthesis of novel compounds similar to B12 will allow for an in-depth investigation into the kinetic and dynamic behaviour of the enzyme. From this research, models for radical-based catalysis will be derived that can be applied to other enzyme systems. The research will also enable us to re-engineer enzymes to carry out other chemical transformations that are applicable to drug synthesis or other industrial purposes. However, to study an individual enzyme, it first must be separated from all of the other thousands of proteins and enzymes in the bacterial cell. A high-pressure liquid chromatography system, like the ÄKTA FPLC is designed to do just that. The system is fully automated and is able to separate proteins at high resolution, quickly and reproducibly. The system is widely used in industry and academia, and HQP who gain expertise on the ÄKTA system will be better prepared for future careers in research and development. An ÄKTA FPLC, or a comparable system, is currently not available at UBC Okanagan.