Details

Autor(en) / Beteiligte

• Zwies, Constanze
• Vargas Rodríguez, Ángela María
• Naumann, Marcel
• Seifert, Franziska
• Pietzsch, Markus

Titel

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

Ist Teil von

• Protein expression and purification, 2024-07, Vol.219, p.106483-106483, Article 106483

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

2024

Quelle

MEDLINE

Beschreibungen/Notizen

• 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. [Display omitted] •Production of recombinant tag-free Mefp-3 using SUMO-fusion technology.•One-pot-strategy for fusion-tag removal and peptide-bound tyrosine hydroxylation.•In vitro application of microbial tyrosinase for efficient DOPA-modification.•DOPA-containing mMefp-3 displays excellent adsorption and tensile-shear strength.