FMSG: Bio: RAMP-Bio-CAFe: Routing Advanced Manufacturing of BioPolymer(s): Capturing Agri-feedstocks in Cell free Factories

当有多余的碳时，细菌会产生聚合物。细菌碳储存聚合物被称为pha。pha是可生物降解的，具有有用的材料特性。从细菌中回收pha是一个主要障碍。该项目的主要目标是开发一种无细胞合成PHA的方法。这将大大简化回收过程，并将提高从天然原料中获得聚合物的收率。来自一种新型嗜热细菌的酶将进行无细胞合成。该系统的生产力将被描述。附近部落学院的本科生将被招募来参与这项研究。作为该项目的一部分，他们将得到一系列专业发展在线研讨会的支持。该项目将建立一个概念验证，以重新连接pha合成作为细胞外产品。要做到这一点，预计需要几个中间步骤。首先是通过设计一种非模式微生物菌株，从未加工的玉米秸秆中合成C6-C12羟基酸，从而将农业材料转化为PHA前体的单步巩固生物转化。下一步是使用来自模型菌株的耐热酶将这些酸无细胞酶聚合成PHA。最后，利用绿色离子液体对合成的生物聚合物进行纯化。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。

Bacteria produce polymers when there is excess carbon. Bacterial carbon storage polymers are referred to as PHAs. PHAs are biodegradable and exhibit useful material properties. Recovering PHAs from bacteria is a major hurdle. The primary objective of the project is to develop a cell-free method for PHA synthesis. This would greatly simplify the recovery process and would increase the yield of the polymer from natural feedstocks. Enzymes from a novel thermophilic bacteria will perform the cell-free synthesis. The productivity of that system will be characterized. Undergraduates from nearby tribal colleges will be recruited to participate in the research. They will be supported by a series of professional development online workshops developed as part of this project.This project will establish a proof-of-concept to rewire the synthesis of PHAs as an extracellular product. To accomplish this, several intermediate steps are anticipated. First is the single-step consolidated bioconversion of agri-materials into PHA precursors by engineering a non-model microbial strain to synthesize C6-C12 hydroxy acids from unprocessed corn stover. Next is cell-free enzymatic polymerization of these acids into PHA using thermostable enzymes from the model strain. Finally, purification of the synthesized biopolymers using green ionic liquids.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.