Spatial enzyme assembly for microbial pharmaceutical synthesis

Microbial synthesis provides a sustainable and efficient platform for pharmaceutical production, primarily through heterologous enzyme expression in engineered cell factories. However, this approach often suffers from intermediate diffusion, metabolic flux competition, and cytotoxic accumulation. Inspired by naturally evolved pathways where enzymes are spatially organized to enhance catalytic specificity and efficiency, artificial spatial assembly tools have been developed to overcome these limitations. This review comprehensively summarizes recent advances in such systems - including rational peptide linkers, peptide-protein interaction domains, nucleic acid scaffolds, and self-assembling elements - applied to diverse pharmaceuticals like antitumor drugs, antimalarials, and nutraceuticals. We further discuss key challenges and future directions to advance efficient and sustainable biosynthesis via spatial engineering.