[1S00e] 01. ASBA - Part 3

5:20 PM - 5:35 PM JST(8:20 AM - 8:35 AM UTC)

[1ASBA-15] Decoding Remote Homology: A Novel Heterometallic MCO Subverting Traditional Paradigms for Biocatalytic Skeletal Editing

○Yajie Wang¹ (1. Westlake University (China))

Keywords:

Biocatalysis,Protein Engineering

Remote homologs, the "dark matter" of the protein universe, evade detection by traditional alignment tools, leaving their biological significance and structure-function relationships largely unexplored. To address this, we developed a computational framework based on ESM-1b. By fine-tuning the model to capture deep evolutionary and structural features of target enzymes, we mapped candidate sequences against a high-performing parent enzyme using semantic similarity, significantly accelerating the discovery of superior and unconventional biocatalysts. Applying this strategy to blue multicopper oxidases (MCOs), we identified a highly divergent novel enzyme, Bfre, featuring a previously unreported Cu-Mn Heterodinuclear active center that fundamentally overturns the traditional trinuclear copper cluster of classical MCOs. Mechanistic studies reveal that this unique cofactor endows Bfre with a stronger oxidative driving force and higher electron transfer efficiency, achieving an approximately 215-fold activity leap over the reference enzyme EcLac. Furthermore, we leveraged Bfre to achieve the first enzyme-mediated skeletal editing, enabling direct, one-step transformation of phenolic and indole derivatives into functionalized tropones and quinoline analogues through exogenous single-carbon insertion. This unprecedented biocatalytic transformation yielded novel molecules with potent antibacterial efficacy against multidrug-resistant bacteria. Building on these findings, we are now pursuing a comprehensive investigation of remote homology sequences through integrated mining, evolutionary analysis, and structure-function characterization, with particular focus on enzymes that represent key bottlenecks in both scientific research and industrial applications.