An Intelligent Electrochemical Multi‐Enzyme Molecular Machine for Chiral Chemical

In vitro multi‐enzyme synthesis pathways harness the core elements of cellular synthesis while simplifying the complexities of cellular processes, facilitating the production of high‐value chemicals. However, these in vitro synthesis processes often operate like a “black box,” with limited monitoring of each reaction step, leading to a low substrate conversion efficiency. In this study, we present an intelligent multi‐enzyme molecular machine(iMEMM) as a model system for achieving the deracemization of D, L‐phosphinothricin (D, L‐PPT). The entire system leverages electrochemical power and enzyme‐catalyzed (cascade) reactions to establish substrate channel and enhance efficiency. By modularizing each reaction step and using electrochemical real‐time monitoring of the reaction process, a single‐step electrobiotransformation efficiency of up to 98% and a chiral target L‐PPT synthesis efficiency exceeding 99% have been achieved. This iMEMM eliminates the need for intermediate separation, enabling a "substrate in, product out" process. Our research paves the way for future green, intelligent, and automated biological manufacturing.