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