Synthetic Cofactor-Linked Metabolic Circuits for Selective Energy Transfer
Synthetic Cofactor-Linked Metabolic Circuits for Selective Energy Transfer
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DOI：
10.1021/acscatal.6b03579
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发表时间：
2017-03-01
期刊：
影响因子：
12.9
通讯作者：
Zhao, Zongbao K.
中科院分区：
文献类型：
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作者：
Wang, Lei;Ji, Debin;Zhao, Zongbao K.
细胞能量传递过程可以类比为能量供应模块（ESM）和能量利用模块（EUM）之间的能量载体（EC）连接的代谢回路的运行。由于天然EC如还原型烟酰胺腺苷二核苷酸（NAD）连接多个能量传递模块和代谢回路，并且天然EC的形成通常与内源性物质的转化相耦合，因此选择性地传递能量是具有挑战性的。在这里，我们设计了合成的辅因子连接电路，用于路径选择性能量转移。我们将亚磷酸脱氢酶工程化为ESM以使用合成辅因子烟酰胺胞嘧啶二核苷酸（NCD）。我们构建不同的电路在体外通过结合不同的ESM，EUM，和EC，我们证明了能量转移过程是可控的，通过调整电路的每个组件的功能。更具体地说，我们表明，这是可能的，以驱动NCD连接的子系统，而留下NAD连接的反应几乎不受影响。当配备有这样的电路时，大肠杆菌细胞使用亚磷酸盐作为电子源来产生还原的NCD，其驱动丙酮酸的还原羧化以改善葡萄糖的苹果酸生产。总之，这项研究提供了一个机会，建立一个正交的能量转移系统的工程细胞工厂，并可用于设置一个额外的层的控制机制的生活。
A cellular energy-transfer process can be analogized as the running of an energy carrier (EC)-linked metabolic circuit between an energy supplying module (ESM) and an energy-utilizing module (EUM). Because natural EC such as the reduced nicotinamide adenosine dinucleotide (NAD) links multiple energy-transfer modules and metabolic circuits, and the formation of natural EC is routinely coupled with the transformation of endogenous substances, it is challenging to transfer energy selectively. Here we devise synthetic cofactor -linked circuits for pathway-selective energy transfer. We engineer phosphite dehydrogenase as ESM to use the synthetic cofactor nicotinamide cytosine dinucleotide (NCD). We construct diverse circuits in vitro by combining different ESM, EUM, and EC, and we demonstrate that an energy-transfer process is controllable by tuning the feature of each component of the circuit. More specifically, we show that it is possible to drive the NCD-linked subsystem while leaving the NAD-linked reaction virtually unaffected. When armed with such circuits, Escherichia coli cells used phosphite as the electron source to generate reduced NCD that drove reductive carboxylation of pyruvate for improved malate production from glucose. Together, this study provides an opportunity to establish an orthogonal energy-transfer system for engineering cell factories and may be used to set an additional layer of a control mechanism for life.