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

真菌可以使用许多高度协调的酶来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。该项目将确定以可预测的方式有效地修饰不止一种底物的真菌酶。此外，将对酶进行改造，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。细胞松弛素是具有植物毒性、细胞毒性和肌动蛋白结合的天然产物。由真菌产生的，已经描述了400多种变体。这种结构多样性的部分原因是引入和修饰官能团的酶的灵活性。这些酶以选择性的方式在结构上重排核心碳骨架，通常是在不止一种底物上。基因组挖掘将被用于鉴定细胞松弛素剪裁酶。将对高产菌株进行化学表征。将对转录因子的过度表达进行调查。靶向基因敲除将确认酶的功能和范围。这些酶将经过改造，以扩大其底物范围。合成生物学和代谢工程工具将被用来产生一个生物制造平台，用于大规模生产和功能化基于细胞松弛素的碳骨架。这个项目将产生关于如何有效地设计酶和代谢途径的基础知识，以高效和快速地实现具有商业潜力的生物活性分子的结构多样化。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势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.