Flux Control at the Malonyl-CoA Node through Hierarchical Dynamic Pathway Regulation in Saccharomyces cerevisiae
Flux Control at the Malonyl-CoA Node through Hierarchical Dynamic Pathway Regulation in Saccharomyces cerevisiae
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DOI：
10.1021/acssynbio.5b00161
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发表时间：
2016-03-01
影响因子：
4.7
通讯作者：
Siewers, Verena
中科院分区：
文献类型：
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作者：
David, Florian;Nielsen, Jens;Siewers, Verena
在微生物宿主中建立异源途径，以高滴度和产量生产工业相关化学品，需要有效调整中心碳代谢，以确保流量导向目标产物。这可以通过代谢网络中关键分支点的调节来实现，在这里我们提出了一种动态调节途径通量和酶表达水平的新方法。该方法基于围绕关键途径中间体丙二酰辅酶A的分层动态控制系统。控制系统的上层确保下调脂肪酸生物合成中丙二酰辅酶A的内源性使用，从而导致该途径中间体的积累。控制系统的下层基于使用新型丙二酰辅酶A生物传感器来激活使用该代谢物生产3-羟基丙酸（3-HP）的异源途径的表达。丙二酰辅酶A传感器是基于枯草芽孢杆菌的FapR转录因子开发的，它展示了细菌代谢物传感器在酵母中的首批应用之一。双途径控制的引入使 3-HP 的产量增加了 10 倍，也可用于生产其他丙二酰辅酶 A 衍生产品。
The establishment of a heterologous pathway in a microbial host for the production of industrially relevant chemicals at high titers and yields requires efficient adjustment of the central carbon metabolism to ensure that flux is directed toward the product of interest. This can be achieved through regulation at key branch points in the metabolic networks, and here we present a novel approach for dynamic modulation of pathway flux and enzyme expression levels. The approach is based on a hierarchical dynamic control system around the key pathway intermediate malonyl-CoA. The upper level of the control system ensures downregulation of endogenous use of malonyl-CoA for fatty acid biosynthesis, which results in accumulation of this pathway intermediate. The lower level of the control system is based on use of a novel biosensor for malonyl-CoA to activate expression of a heterologous pathway using this metabolite for production of 3-hydroxypropionic acid (3-HP). The malonyl-CoA sensor was developed based on the FapR transcription factor of Bacillus subtilis, and it demonstrates one of the first applications of a bacterial metabolite sensor in yeast. Introduction of the dual pathway control increased the production of 3-HP by 10-fold and can also be applied for production of other malonyl-CoA-derived products.