Enhancing Soluble Expression and Post-Translational Modification of Mussel Foot Protein in E. coli

Area of Science:

• Biotechnology
• Protein Engineering
• Materials Science

Background:

• Recombinant mussel foot proteins (Mfps) exhibit low solubility and stability, hindering large-scale production.
• Developing efficient systems for producing functional Mfps, particularly dopanized variants, is crucial for their application.

Purpose of the Study:

• To engineer a peptide tag that enhances the solubility of Mfps in Escherichia coli.
• To establish a scalable and cost-effective platform for producing functional, dopanized Mfps.

Main Methods:

• Engineered a 39-amino acid peptide tag, T7B9M3, to improve Mfps solubility in E. coli.
• Coexpressed tyrosinase with Mfps to enable dopanization.
• Assessed protein solubility, tyrosine hydroxylation efficiency, and substrate interactions.

Main Results:

• The T7B9M3 tag significantly increased Mfps solubility, achieving a titer of 365.96 mg/L with 98.75% solubility.
• The absence of net charged residues in the tag was key to solubility enhancement.
• Dopanized Mfps exhibited enhanced interactions with various substrates (glass, polypropylene, polyethylene).

Conclusions:

• The T7B9M3 tag provides a scalable solution for producing soluble Mfps.
• Coexpression of tyrosinase enables efficient dopanization, yielding functional Mfps with improved adhesion.
• This strategy offers a promising platform for low-cost, batch production of Mfps.