Horticultural Sciences Department, University of Florida, Gainesville, FL, USA.
Mathematics and Computer Science Division, Argonne National Laboratory, Argonne, IL, USA.
Metab Eng. 2017 Nov;44:150-159. doi: 10.1016/j.ymben.2017.10.006. Epub 2017 Oct 10.
The necessarily sharp focus of metabolic engineering and metabolic synthetic biology on pathways and their fluxes has tended to divert attention from the damaging enzymatic and chemical side-reactions that pathway metabolites can undergo. Although historically overlooked and underappreciated, such metabolite damage reactions are now known to occur throughout metabolism and to generate (formerly enigmatic) peaks detected in metabolomics datasets. It is also now known that metabolite damage is often countered by dedicated repair enzymes that undo or prevent it. Metabolite damage and repair are highly relevant to engineered pathway design: metabolite damage reactions can reduce flux rates and product yields, and repair enzymes can provide robust, host-independent solutions. Herein, after introducing the core principles of metabolite damage and repair, we use case histories to document how damage and repair processes affect efficient operation of engineered pathways - particularly those that are heterologous, non-natural, or cell-free. We then review how metabolite damage reactions can be predicted, how repair reactions can be prospected, and how metabolite damage and repair can be built into genome-scale metabolic models. Lastly, we propose a versatile 'plug and play' set of well-characterized metabolite repair enzymes to solve metabolite damage problems known or likely to occur in metabolic engineering and synthetic biology projects.
代谢工程和代谢合成生物学对途径及其通量的高度关注,往往使人们忽视了途径代谢物可能经历的破坏性酶促和化学副反应。尽管这些副反应在历史上被忽视和低估,但现在已知它们存在于整个代谢过程中,并在代谢组学数据集中产生(以前神秘的)峰。现在也知道,代谢物损伤通常会被专门的修复酶所抵消,这些酶可以消除或防止损伤。代谢物损伤和修复与工程途径设计密切相关:代谢物损伤反应会降低通量速率和产物产量,而修复酶可以提供稳健的、与宿主无关的解决方案。在介绍了代谢物损伤和修复的核心原则之后,我们使用案例研究来记录损伤和修复过程如何影响工程途径的有效运行——特别是那些异源的、非天然的或无细胞的途径。然后,我们回顾了如何预测代谢物损伤反应,如何预测修复反应,以及如何将代谢物损伤和修复纳入基因组尺度的代谢模型。最后,我们提出了一套通用的“即插即用”的经过良好表征的代谢物修复酶,以解决代谢工程和合成生物学项目中已知或可能出现的代谢物损伤问题。