Developing a Biomanufacturing Platform for the Site-Selective Functionalization and Structural Diversification of Cytochalasan-Based Carbon Skeletons

真菌可以利用一些高度协调的酶合成具有复杂结构的小分子。这些分子很难合成，它们可以帮助作物生产或对人类健康有益。该项目将确定真菌酶，有效地修改一个以上的底物在一个可预测的方式。此外，酶将被改造以扩大底物的范围。最终的结果将是一个以较低成本合成生物活性分子的生物制造平台。在这个项目中，学生将接受STEM核心技术的培训，重点是商业应用。细胞chalasans是植物毒性、细胞毒性和肌动蛋白结合的天然产物。由真菌产生，已经描述了超过400种变体。结构的多样性部分是由引入和修饰官能团的酶的灵活性来解释的。这些酶在结构上以位点选择性的方式重新排列核心碳骨架，通常在一个以上的底物上。基因组挖掘将被用于鉴定细胞chalasan剪裁酶。过量生产的菌株将用化学方法加以表征。转录因子过表达将被研究。靶向基因敲除将确认酶的功能和作用范围。这些酶将被改造以扩大它们的底物范围。合成生物学和代谢工程工具将被用来产生一个大规模生产和功能化的基于细胞chalasan的碳骨架的生物制造平台。该项目将为如何有效地设计酶和代谢途径以实现具有商业潜力的生物活性分子的高效和快速结构多样化提供基础知识。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。

Fungi can synthesize small molecules with complex structures using a number of highly coordinated enzymes. These molecules are difficult to make synthetically, and they can aid in crop production or have beneficial human health effects. This project will identify fungal enzymes that efficiently modify more than one substrate in a predictable way. Also, enzymes will be engineered to expand the range of substrates. A biomanufacturing platform to synthesize bioactive molecules at lower costs will be the end result. In this project, students will be trained in core STEM technologies, with a focus on commercial applications.Cytochalasans are phytotoxic, cytotoxic and actin-binding natural products. Produced by fungi, over 400 variants have been described. The structural diversity is partly explained by the flexibility of the enzymes that introduce and modify functional groups. These enzymes structurally rearrange the core carbon skeleton in a site-selective manner, often on more than one substrate. Genome mining will be used to identify cytochalasan tailoring enzymes. Overproducing strains will be characterized chemically. Transcription factor over-expression will be investigated. Targeted gene knock-out will confirm the function and scope of the enzymes. The enzymes will be engineered to expand their substrate range. Synthetic biology and metabolic engineering tools will be used to generate a biomanufacturing platform for large scale production and functionalization of cytochalasan-based carbon skeletons. This project will generate fundamental knowledge on how to effectively engineer enzymes and metabolic pathways for efficient and rapid structural diversification of bioactive molecules with commercial potential.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.