Molecular Dynamics Simulation Reveals the Mechanism of Substrate Recognition by Lignin-Degrading Enzymes

TL;DR

This study uses molecular simulations to uncover how lignin-degrading enzymes recognize and break down complex lignin structures.

Contribution

The paper reveals a dual mechanism involving polar and aromatic residues in ligninolytic enzyme-substrate interactions.

Findings

Polar residues form hydrogen bonds to align substrates for bond cleavage.

Aromatic residues stabilize lignin conformations through hydrophobic interactions.

Virtual mutations showed aromatic residues are critical for binding, while polar residues control cleavage selectivity.

Abstract

Lignin, the most abundant aromatic biopolymer, represents a key renewable feedstock for sustainable biorefineries, yet its structural complexity poses a formidable challenge for enzymatic degradation. While ligninolytic enzymes such as laccases (LACs), lignin peroxidases (LiPs), and manganese peroxidases (MnPs) exhibit remarkable catalytic versatility, the molecular mechanisms underlying their ability to balance substrate specificity and structural flexibility remain unresolved. Here, we employed all-atom molecular dynamics (MD) simulations and virtual mutagenesis to dissect the dynamic interactions between these enzymes and lignin model compound (β-O-4-linked H-type dimers). Our simulations revealed a dual recognition mechanism in which polar residues (such as Asp, Glu, Arg and His) formed hydrogen bonds with hydroxyl and keto groups near catalytic cleavage sites, ensuring precise alignment for bond scission, while aromatic residues stabilized diverse lignin conformations via hydrophobic interactions with conserved aromatic rings. Conformational dynamics of active-site residues enabled adaptive adjustments to substrate heterogeneity, reconciling enzymatic specificity with structural promiscuity. Virtual mutation experiments further demonstrated that aromatic residues were indispensable for binding stability, whereas polar residues dictated cleavage-site selectivity. These findings provide atomic-scale insights into the catalytic mechanism of ligninolytic enzymes, with implications in the rational design of superior biocatalyst for lignin biorefineries.

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