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，细菌就会产生聚合物。细菌碳储存聚合物被称为PHA。PHA是可生物降解的，并显示出有用的材料特性。从细菌中恢复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。这将极大地简化回收过程，并将增加从天然原料中提取聚合物的产量。来自一种新型嗜热菌的酶将执行无细胞合成。该系统的生产率将得到表征。附近部落学院的本科生将被招募参与这项研究。作为这个项目的一部分，他们将得到一系列专业开发在线研讨会的支持。这个项目将建立一个概念验证，以重新连接作为胞外产品的PHA的合成。要做到这一点，预计会有几个中间步骤。首先，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，将农业原料一步强化生物转化为PHA前体。下一步是使用模型菌株中的耐热酶将这些酸进行无细胞酶聚合生成PHA。最后，使用绿色离子液体提纯合成的生物聚合物。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。

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.