Laccase-catalyzed oxidative cross-linking of tyrosine and potato patatin- and lysozyme-derived peptides: Molecular and kinetic study.

Abstract

Laccase can catalyze the oxidative cross-linking of peptides, which is useful in the production of proteinaceous materials with enhanced functional properties. However, the kinetics and the pathway of this reaction remain unclear. In the present study, laccase-catalyzed oxidative cross-linking reaction was investigated through a combination of computational analysis, kinetic studies and end-product profiling using selected substrate models, including peptide AG-10 (AKKIVSDGNG) (without tyrosine) derived from lysozyme and tyrosine-containing peptide ST-10 (SYMTDYYLST) from potato protein (patatin), and tyrosine. Both laccases from Trametes versicolor (LacTv) and Coriolus hirsutus (LacCh) were used as biocatalysts. Laccase exhibited higher binding affinity and catalytic efficiency (kcat/Km) towards ST-10 and AG-10 than tyrosine. Among the laccases, LacCh showed higher kcat towards the substrate models than LacTv. Through the molecular docking, this result was attributed to the presence of the ASN206 at the cavity of LacCh. The end product profiles reveal the formation of homo-oligomers (> 5 units) of ST-10 in the reaction catalyzed by LacTv, while polymerization was favored by LacCh. These cross-linked products were identified to have a mix of oligo-tyrosine linkages. In contrast, the cross-linking of AG-10 required the presence of ferulic acid as mediator, which resulted in the formation of hetero-oligomers and polymers of AG-10. The knowledge obtained in the present study provide insight into an effective reaction for peptide cross-linking.