Structure Determination of Natural Product Biosynthesis Enzymes
Tao, Yizhi (Jane); Phillips, Jr, George N
Doctor of Philosophy
Natural products (NPs) frequently display elaborate molecular frameworks challenging biologists to ascertain the genetic origin and catalytic logic affording intricate NP biosynthesis. Beyond the basic research inspired by multiplicity of NPs discovered thus far, the biological activities accompanying the unprecedented chemical architectures has generated incredible momentum for the pharmaceutical industries to exploit the unusual modes of action and design strategies for NP synthesis. Such efficacious NP derived or inspired therapeutics are best exemplified by paclitaxel (taxol), lovastatin and penicillin for the treatment of cancer, high-cholesterol-lowering and bacterial infections respectively (1–3). Total synthesis remains the primary strategy for drug and derivative production, however characterization of NP biosynthetic enzymes bears immense promise for their utility in NP production. Tremendous advancement in genetic sequencing technology has enabled scientists to decipher biosynthetic gene clusters (BGCs) facilitating interrogation of the synthetic and regulatory protein machinery. Structural and functional pursuit of NP associated proteins sheds light on the molecular mechanisms governing intricate regiospecific and stereospecific modifications laying the foundation for future bioengineering and synthetic biology initiatives. This thesis work describes the CalS10, DynU16 and AtmM structures determined by X-ray crystallography and kinetic parameters expanding our enzymatic toolkit for NP biosynthesis.
natural product biosynthesis; protein crystallography; enediyne; natural product