Unlocking Applications of Cell-Free Biotechnology through Enhanced Shelf Life and Productivity of E. coli Extracts
Unlocking Applications of Cell-Free Biotechnology through Enhanced Shelf Life and Productivity of E. coli Extracts
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DOI：
10.1021/acssynbio.9b00433
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发表时间：
2020-04-17
影响因子：
4.7
通讯作者：
Oza, Javin P.
中科院分区：
文献类型：
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作者：
Gregorio, Nicole E.;Kao, Wesley Y.;Oza, Javin P.
无细胞蛋白合成（CFPS）是一种平台生物技术，具有广泛的应用前景。然而，由于CFPS所需的水细胞提取物的货架稳定性差，现场应用仍然有限。冻干大肠杆菌提取物提高了保质期，但仍然不足以在室温下延长储存。为了解决这一限制，我们以组合的方式绘制了十种具有四种不同作用机制的低成本添加剂的化学空间，以确定能够稳定冻干细胞提取物的配方。我们报告了三个关键发现：(1)独特的添加剂配方，可保持细胞提取物在4℃和23℃下的充分生产力；(2)提高萃取效率近2倍的添加剂配方；(3)一种机器学习算法，为未经过实验测试的添加剂配方的稳定效果提供预测能力。这些发现提供了一个简单和低成本的进展，使CFPS在生物制造领域具有成本竞争力。
Cell-free protein synthesis (CFPS) is a platform biotechnology that enables a breadth of applications. However, field applications remain limited due to the poor shelf-stability of aqueous cell extracts required for CFPS. Lyophilization of E. coli extracts improves shelf life but remains insufficient for extended storage at room temperature. To address this limitation, we mapped the chemical space of ten low-cost additives with four distinct mechanisms of action in a combinatorial manner to identify formulations capable of stabilizing lyophilized cell extract. We report three key findings: (1) unique additive formulations that maintain full productivity of cell extracts stored at 4 degrees C and 23 degrees C; (2) additive formulations that enhance extract productivity by nearly 2-fold; (3) a machine learning algorithm that provides predictive capacity for the stabilizing effects of additive formulations that were not tested experimentally. These findings provide a simple and low-cost advance toward making CFPS field-ready and cost-competitive for biomanufacturing.