Ribulose Monophosphate Shunt Provides Nearly All Biomass and Energy Required for Growth of E. coli,ACS Synthetic Biology

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Ribulose Monophosphate Shunt Provides Nearly All Biomass and Energy Required for Growth of E. coli
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2018-05-14 00:00:00 , DOI: 10.1021/acssynbio.8b00093
Hai He ₁ , Christian Edlich-Muth ₁ , Steffen N. Lindner ₁ , Arren Bar-Even ₁

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The ribulose monophosphate (RuMP) cycle is a highly efficient route for the assimilation of reduced one-carbon compounds. Despite considerable research, the RuMP cycle has not been fully implemented in model biotechnological organisms such as Escherichia coli, mainly since the heterologous establishment of the pathway requires addressing multiple challenges: sufficient formaldehyde production, efficient formaldehyde assimilation, and sufficient regeneration of the formaldehyde acceptor, ribulose 5-phosphate. Here, by efficiently producing formaldehyde from sarcosine oxidation and ribulose 5-phosphate from exogenous xylose, we set aside two of these concerns, allowing us to focus on the particular challenge of establishing efficient formaldehyde assimilation via the RuMP shunt, the linear variant of the RuMP cycle. We have generated deletion strains whose growth depends, to different extents, on the activity of the RuMP shunt, thus incrementally increasing the selection pressure for the activity of the synthetic pathway. Our final strain depends on the activity of the RuMP shunt for providing the cell with almost all biomass and energy needs, presenting an absolute coupling between growth and activity of key RuMP cycle components. This study shows the value of a stepwise problem solving approach when establishing a difficult but promising pathway, and is a strong basis for future engineering, selection, and evolution of model organisms for growth via the RuMP cycle.

中文翻译：

核糖一磷酸分流器提供了大肠杆菌生长所需的几乎所有生物量和能量

核糖一磷酸（RuMP）循环是同化还原一碳化合物的高效途径。尽管进行了大量研究，但RuMP循环尚未在模型生物技术生物（如大肠杆菌）中完全实施，这主要是因为该途径的异源建立需要解决多个挑战：甲醛生产充分，甲醛同化有效以及甲醛受体充分再生， 5-磷酸核糖。在这里，通过从肌氨酸氧化高效生产甲醛和从外源木糖高效生产5-磷酸核糖，我们抛开了其中两个问题，使我们能够专注于通过建立有效的甲醛同化的特殊挑战。RuMP分流器，RuMP循环的线性变体。我们已经产生了缺失菌株，其生长在不同程度上取决于RuMP分流器的活性，从而逐渐增加了合成途径活性的选择压力。我们的最终菌株取决于RuMP分流器的活性，从而为细胞提供几乎所有的生物量和能量需求，从而在关键RuMP循环组分的生长和活性之间呈现出绝对的耦合。这项研究显示了逐步建立问题的方法在建立困难但有希望的途径时的价值，并且为通过RuMP循环进行生长的模型生物的未来工程，选择和进化奠定了坚实的基础。

更新日期：2018-05-14

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