Collaborative Research: Understanding and exploiting the structure-function link between fatty acid biosynthesis and degradation enzymes for functionalized small molecule synthesis

商业上重要的产品通常是由小分子合成的。细菌和其他微生物擅长制造这种分子。许多是由脂肪和脂肪酸合成的。了解细胞如何制造和修饰脂肪将有助于确定如何利用微生物大量生产可再生燃料和化学物质。该项目还将帮助我们的社会了解通往可持续未来道路上的挑战和机遇。这包括为来自以少数民族人口为主的学区的本科生和高中生提供咨询和培训。这些努力将共同在具有国家战略重要性的领域提供学术和工业培训。通过表征酶/底物相互作用和来自各种生物体的酶的独特特征，将深入了解脂肪酸生物合成（FAB）和β -氧化（BOX）途径的进化分歧。采用非脱羧缩合和&amp；#946；-还原反应，将产生具有新功能的分子。II型FAB和BOX酶将互换使用，以充分利用每组酶的优势特性。将采用一种综合的方法，其中计算蛋白质建模和设计将通过FAB和BOX酶的分子建模来指导实验，这些酶具有广泛的生物合成可及化合物。该项目的具体目标是：1)II型FAB和BOX酶的计算蛋白质建模和设计；2) II型FAB和BOX酶的体外鉴定；3) II型FAB与BOX酶组合的体内构建与表征。生成中链（C5-C8）二羧酸和甲基支链羧酸，特别是二羧酸己二酸（C6）和亚酰基酸（C8）和甲基支链不饱和癸酸（C5）。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Commercially important products are often built up from small molecules. Bacteria and other microbes are proficient at producing such molecules. Many are synthesized from fats and fatty acids. Understanding how the cells make and modify fats will help determine how to make renewable fuels and chemicals in large amounts using microbes. The project will also help educate our society in the challenges and opportunities on the road to a sustainable future. This includes advising and training of undergraduate and high school students from school districts serving predominantly minority populations. Together, these efforts will provide academic and industrial training in areas of strategic national importance.Insight into the evolutionary divergence of fatty acid biosynthesis (FAB) and beta-oxidation (BOX) pathways will be sought by characterizing enzyme/substrate interactions and the unique characteristics of enzymes from various organisms. Using non-decarboxylative condensation and β-reduction reactions, molecules with novel functionalities will be generated. Type II FAB and BOX enzymes will be used interchangeably to capitalize on the advantageous characteristics of each set of enzymes. An integrated approach in which computational protein modeling and design will guide experimentation through molecular modeling of the FAB and BOX enzymes with a wide array of biosynthetically accessible compounds will be undertaken. The specific objectives of the project are: 1) computational protein modeling and design of type II FAB and BOX enzymes; 2) in vitro characterization of type II FAB and BOX enzymes; and 3) in vivo construction and characterization of combinations of type II FAB and BOX enzymes. Medium chain (C5-C8) dicarboxylic and methyl-branched carboxylic acids, specifically dicarboxylates adipic (C6) and suberic (C8) acids and methyl-branched unsaturated tiglic acid (C5) will be produced.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.