Fast protein liquid chromatography system for protein purification

所有生物都有专门的分子，称为酶，来执行特定的生物功能。许多酶已经进化到使用自由基(高活性的孤电子)来执行生物转化。自由基的优势在于它们的高活性，因此能够进行化学上困难且需要大量能量的反应。然而，自由基不分青红皂白地反应，这可能会破坏酶。酶是动态系统；因此，内部运动的时间必须精确地与化学事件同步，以防止自由基自行熄灭。研究实验室的目标是研究酶如何通过大规模的结构重排来协调自由基化学。为了做到这一点，实验室将研究一种含有维生素B12衍生物的酶，它用来通过打破一种不寻常的碳-钴键来启动自由基化学。然后，自由基被传播到另一种由维生素衍生的分子(维生素B6)，在遥远的位置依偎在酶中。快速反应光谱技术以及类似于B12的新化合物的化学合成将使深入研究酶的动力学和动态行为成为可能。通过这项研究，将得到基于自由基的催化模型，该模型可应用于其他酶系统。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目的的其他化学转化。然而，要研究一种单独的酶，首先必须将其与细菌细胞中的其他数千种蛋白质和酶分离。KTA FPLC等高压液相色谱系统就是为实现这一目的而设计的。该系统是完全自动化的，能够以高分辨率、快速和可重复性地分离蛋白质。该系统在工业和学术界得到了广泛的应用，而在KTA系统上获得专业知识的HQP将为未来的研究和开发事业做好更好的准备。UBC Okanagan目前没有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.