Alternative strategies for the recombinant synthesis, DOPA modification and analysis of mussel foot proteins – A case study for Mefp-3 from Mytilus edulis

• DOI: 10.1016/j.pep.2024.106483
• Publication Date: 2024-04-10T15:31:54Z
• Authors: Constanze Zwies; Ángela María Varg...; Marcel Naumann; Franziska Seifert; Markus Pietzsch
• Keywords: Hydroxylation; Blue mussel

Mussel foot proteins (Mfps) possess unique binding properties to various surfaces due the presence of L-3,4-dihydroxyphenylalanine (DOPA). Mytilus edulis protein-3 (Mefp-3) is one several in byssal adhesive plaque. Its localization at plaque-substrate interface approved that Mefp-3 plays a key role adhesion. Therefore, protein suitable for development innovative bio-based binders. However, recombinant Mfp-3s are mainly purified from inclusion bodies under denaturing conditions. Here, we describe robust and reproducible protocol obtaining soluble tag-free using SUMO-fusion technology. Additionally, microbial tyrosinase Verrucomicrobium spinosum was used vitro hydroxylation peptide-bound tyrosines first time. The highly hydroxylated Mefp-3, confirmed by MALDI-TOF-MS, exhibited excellent comparable commercial glue. These results demonstrate concerted simplified high yield production process Mfp3-based with on demand DOPA modification.