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

具有重要商业价值的产品通常是由小分子组成的。细菌和其他微生物擅长生产这种分子。许多是由脂肪和脂肪酸合成的。了解细胞如何制造和改变脂肪将有助于确定如何使用微生物大量制造可再生燃料和化学品。该项目还将帮助教育我们的社会在通往可持续未来的道路上面临的挑战和机遇。这包括为主要为少数民族人口服务的学区的本科生和高中生提供咨询和培训。这些努力将共同为国家战略重点领域提供学术和工业培训。通过表征酶/底物相互作用和各种生物体酶的独特特征，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。使用非脱羧缩合和#946;-还原反应，将产生具有新功能的分子。 II型FAB和BOX酶将可互换使用以利用每组酶的有利特征。将采用一种综合方法，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获得的化合物。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酶的体外表征;以及3）II型FAB和BOX酶的组合的体内构建和表征。中链（C5-C8）二羧酸和甲基支链羧酸，特别是二羧酸己二酸（C6）和辛二酸（C8）和甲基支链不饱和惕各酸（C5）将被生产。该奖项反映了NSF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.