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.