Abstract

Adenosine triphosphate (ATP), the universal energy currency of life, has a central role in numerous biochemical reactions with potential for the synthesis of numerous high-value products. ATP can be regenerated by three types of mechanisms: substrate level phosphorylation, oxidative phosphorylation, and photophosphorylation. Current ATP regeneration methods are mainly based on substrate level phosphorylation catalyzed by one enzyme, several cascade enzymes, or in vitro synthetic enzymatic pathways. Among them, polyphosphate kinases and acetate kinase, along with their respective phosphate donors, are the most popular approaches for in vitro ATP regeneration. For in vitro artificial pathways, either ATP-free or ATP-balancing strategies can be implemented via smart pathway design by choosing ATP-independent enzymes. Also, we discuss some remaining challenges and suggest perspectives, especially for industrial biomanufacturing. Development of ATP regeneration systems featuring low cost, high volumetric productivity, long lifetime, flexible compatibility, and great robustness could be one of the bottom-up strategies for cascade biocatalysis and in vitro synthetic biology.

摘要

三磷酸腺苷 (ATP) 是生命的通用能量货币，在众多具有合成众多高价值产品潜力的生化反应中发挥着核心作用。ATP 可以通过三种机制再生：底物水平磷酸化、氧化磷酸化和光磷酸化。目前的 ATP 再生方法主要基于由一种酶、几种级联酶或体外合成酶途径催化的底物水平磷酸化。其中，多磷酸激酶和醋酸激酶以及它们各自的磷酸盐供体是体外ATP 再生最流行的方法。对于体外人工途径，可以实施无 ATP 或 ATP 平衡策略通过选择不依赖 ATP 的酶，通过智能途径设计。此外，我们还讨论了一些剩余的挑战并提出了一些观点，特别是对于工业生物制造。开发具有低成本、高容量生产率、长寿命、灵活兼容性和强大稳健性的 ATP 再生系统可能是级联生物催化和体外合成生物学的自下而上策略之一。