Glycine Synthesis From Methanol by a Cofactor‐Neutral In Vitro Multienzyme Cascade

The global demand for amino acids continues to rise, yet traditional synthesis methods face environmental and safety challenges. To contribute to a sustainable future vision, we herein report a novel cofactor‐neutral in vitro multienzyme cascade that directly converts methanol, a renewable C1 feedstock, into glycine under mild conditions. A two‐step strategy achieves 82.2% methanol‐to‐glycolaldehyde and 86.0% glycolaldehyde‐to‐glycine conversion, respectively. At 500 mM methanol, it yields 18.8 mM glycine after 6 h, which represents a 26.8‐fold improvement over the one‐pot approach. These results demonstrate the potential of modular in vitro multienzyme catalysis for efficient C1‐to‐amino acid transformation and establish a foundation for the synthesis of many other complex nitrogen‐containing molecules.