Recombinant production of chimeric protein Mfp-3-GvpA in yeast Pichia pastori

Document Type : Research Article

Authors

• nazanin bolghari ¹
• Hadi Hbibollahi ¹
• masoumeh anvari ²
• hossin shahsavarani ³

¹ Department of Biology, Rasht branch, Islamic Azad University, Rasht,Iran.

² Department of Biology, Rasht Branch, Islamic Azad University, Rasht, Iran

³ Department of Cell and Molecular Sciences, Faculty of Life Sciences and Biotechnology, Shahid Beheshti Univ., Tehran, Iran.

Abstract

Mussels secrete protein-based polymers mainly mussel foot proteins (Mfps), enabling them to anchor to various surfaces in a saline, intertidal zone. Using Mfp proteins as novel water-resistant adhesive materials have been suggested for biomedical purposes due to their unique features including large abundance of catechol aligned with amphiphilic and ionic properties. Despite the more promising characteristics of foot proteins, they have not been widely exploited for biomedical or even industrial applications, since farming the mussels are not commercially viable due to ecological issues and their highly territorial nature. Present study aimed to examine engineering of recombinant synthesis of Mfp3 protein fused to another marine based curli protein, gas vesicle protein A(GvpA) in nonpathogenic yeast, Pichia pastoris, expression system. To this end, sequences of the Mfp3 and GvpA were extracted from the NCBI database and were inserted into the pPICZ A vector in order to obtain an efficient expression level of chimeric protein. Obtained vectors were transformed into E. coli TOP10F' for multiplication and then linearized plasmid transformed into Pichia pastoris for protein expression. The best expression level obtained after 96 hours incubation with methanol induction. In conclusion, active recombinant adhesive Mfp3-GvpA fused protein were successfully expressed in P. pastoris suggesting potential expression system for future bio-adhesive or any fused protein production.

Keywords

• Mytilus edulis
• Mfp-3
• GvpA
• Pichia pastoris
• Protein expiration

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