微生物辅因子工程研究进展

Research Progress of Microbial Cofactor Engineering

微生物细胞中的大部分酶促反应都需要各种辅因子的参与,辅因子平衡对维持细胞内的生化反应稳态非常重要,辅因子供应不足会导致细胞生长和化合物生产的紊乱。近年来,辅因子在生化反应过程中的关键作用备受关注,但由于其价格较昂贵、稳定性差,因此限制了辅因子工程的发展。合成生物学和代谢工程的发展为辅因子的可持续供应提供了可行的解决方案,多种加强辅因子供应的策略有效地推动了目标化合物的生物合成。其中,烟酰胺类辅因子NAD(P)^(+)、NAD(P)H是微生物代谢过程中最常见的氧化还原辅因子,它们在所有生物体内作为重要的电子受体或供体推动合成与分解代谢反应,对维持胞内氧化还原动态平衡起着决定性作用。从NAD(P)H的主要来源和NAD(P)^(+)/NAD(P)H的平衡对天然产物生物合成中的影响出发,重点从三个不同维度讨论辅因子工程策略,综述代谢途径调节、外源氧化还原酶的引入、蛋白质工程等多种辅因子再生策略的最新研究进展及应用,展望辅因子代谢工程在生物合成中的未来发展方向。

Various cofactors are required in the process of most enzyme-catalyzed reaction of cells.Cofactor balance is very important to maintain the homeostasis of biochemical reaction in cells.However,insufficient supply of cofactors will lead to the disorder of cell growth and compound production.In recent years,the key role of cofactors in the biochemical reaction process has attracted the attention of researchers,but most cofactors are expensive and have poor stability.These disadvantages have limited the potential application of cofactor engineering.The development of synthetic biology and metabolic engineering provides feasible solutions for the sustainable supply of cofactors,and multiple strategies to strengthen the supply of cofactors effectively promote the biosynthesis of target compounds.Among them,nicotinamide cofactors NAD(P)^(+)and NAD(P)H are the most common redox cofactors in the process of microbial metabolism.They are important electron receptors or donors in all organisms,promote synthesis and catabolism reactions,and play a decisive role in maintaining the dynamic balance of intracellular redox.Starting from the main sources of NAD(P)H and the influence of NAD(P)^(+)/NAD(P)H balance in the synthesis of natural products,cofactor engineering strategies were reviewed from three different dimensions.This paper introduces the latest research progress and application of multiple cofactor regeneration strategies through metabolic pathway regulation,introduction of exogenous oxidoreductase,and protein engineering.The future development prospects of cofactor metabolic engineering in biosynthesis is also discussed.