張晉源副教授研究團隊發表研究成果於J Am Chem Soc.
連結網址：https://pubmed.ncbi.nlm.nih.gov/41779877/
Abstract
The biosynthesis of α-cyclopiazonic acid (α-CPA) is notable for generating a complex pentacyclic scaffold using a minimal three-enzyme pathway. The final step, catalyzed by CpaO, converts linear β-CPA into α-CPA through an enigmatic oxidative cyclization. Here, we report the structural and mechanistic characterization of CpaO. X-ray crystallography reveals a three-domain architecture belonging to the flavin-dependent amine oxidase (FAO) superfamily, while CpaO represents a previously undescribed subfamily distinguished by an essential, covalently linked FAD (8α-N1-histidyl) and divergent substrate-binding domains. High-resolution structures of the CpaO/β-CPA complex, validated by mutagenesis, identify key active-site residues (His165, Trp317, Asp412, Tyr283) that anchor the substrate. Combined structural, mutational, and molecular dynamics analyses further suggest distinct yet cooperative roles for Tyr283 and Ser167 in modulating substrate access and subsequent binding. Derived from these data, we propose a stereospecific mechanism initiated by FAD-mediated hydride abstraction, which triggers a bicyclization cascade to form the final C and D rings. This study resolves a long-standing biosynthetic mystery and expands the catalytic repertoire of flavoenzymes, offering a template for the chemoenzymatic synthesis of complex indole alkaloids.
