Light-based control of metabolic flux through assembly of synthetic organelles
Light-based control of metabolic flux through assembly of synthetic organelles
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DOI：
10.1038/s41589-019-0284-8
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发表时间：
2019-04
影响因子：
14.8
通讯作者：
Evan M. Zhao;Nathan Suek;Maxwell Z. Wilson;E. Dine;N. Pannucci;Zemer Gitai;J. Avalos;Jared E. Toettcher-Jared-E
中科院分区：
文献类型：
--
作者：
Evan M. Zhao;Nathan Suek;Maxwell Z. Wilson;E. Dine;N. Pannucci;Zemer Gitai;J. Avalos;Jared E. Toettcher-Jared-E
为了最大限度地获得所需的产品，代谢工程师通常会将酶表达到高且恒定的水平。然而，永久的通路激活可能会产生不良后果，包括与基本通路的竞争和有毒中间体的积累。面对类似的挑战，自然代谢系统在一定条件下将酶划分为细胞器或翻译后诱导活性。在这里，我们报道光遗传控制可以用于扩展区隔化和动态控制酵母的工程代谢。我们描述了一套光遗传学工具来触发代谢活性酶簇的组装和拆卸。以脱氧紫罗兰素生物合成途径为模型系统，我们发现光开关聚类可以通过降低中间代谢物的浓度和减少竞争途径的通量，使产物形成增强6倍，产物特异性增强18倍。酶的诱导区室化进入合成细胞器可以用来控制工程代谢途径，限制中间体和有利于所需产物的形成。
To maximize a desired product, metabolic engineers typically express enzymes to high, constant levels. Yet, permanent pathway activation can have undesirable consequences including competition with essential pathways and accumulation of toxic intermediates. Faced with similar challenges, natural metabolic systems compartmentalize enzymes into organelles or post-translationally induce activity under certain conditions. Here we report that optogenetic control can be used to extend compartmentalization and dynamic control to engineered metabolisms in yeast. We describe a suite of optogenetic tools to trigger assembly and disassembly of metabolically active enzyme clusters. Using the deoxyviolacein biosynthesis pathway as a model system, we find that light-switchable clustering can enhance product formation six-fold and product specificity 18-fold by decreasing the concentration of intermediate metabolites and reducing flux through competing pathways. Inducible compartmentalization of enzymes into synthetic organelles can thus be used to control engineered metabolic pathways, limit intermediates and favor the formation of desired products.