Mussel adhesive proteins, including the 20-plus variants of foot protein type 3 (fp-3), have been suggested as potential environmentally friendly adhesives for use in aqueous conditions and in medicine. Here we report the novel production of a recombinant Mytilus galloprovincialis foot protein type 3 variant A (Mgfp-3A) fused with a hexahistidine affinity ligand in Escherichia coli and its approximately 99% purification with affinity chromatography. Recombinant Mgfp-3A showed a superior purification yield and better apparent solubility in 5% acetic acid (prerequisites for large-scale production and practical use) compared to those of the previously reported recombinant M. galloprovincialis foot protein type 5 (Mgfp-5). The adsorption abilities and adhesion forces of purified recombinant Mgfp-3A were compared with those of Cell-Tak (a commercial mussel extract adhesive) and recombinant Mgfp-5 using quartz crystal microbalance analysis and modified atomic force microscopy, respectively. These assays showed that the adhesive ability of recombinant Mgfp-3A was comparable to that of Cell-Tak but lower than that of recombinant Mgfp-5. Collectively, these results indicate that recombinant Mgfp-3A may be useful as a commercial bioadhesive or an adhesive ingredient in medical or underwater environments.
in Wiley InterScience (www.interscience.wiley.com).Mussel adhesive proteins (MAPs) have a potential as environmentally friendly adhesives for use under aqueous conditions. MAPs maybe of particular value in medical applications. We previously reported the functional expression of recombinant foot protein type 5 (fp-5) and foot protein type 3A (fp-3A), both of which have significant adhesion abilities, in Escherichia coli. However, these proteins were produced at low levels because of post-induction cell growth inhibition, and the proteins showed poor post-purification solubility. Here, we design and produce a new type of recombinant MAP, fp-353, that is a fusion protein with fp-3A at each terminus of fp-5. Because fp-353 formed inclusion bodies, host cell growth inhibition did not occur. In addition, the solubility of MAP fp-353 after purification was significantly enhanced, permitting the preparation of a viscous concentrated glue solution for large-scale adhesion strength measurements. Together with large-scale cowhide adhesion measurements and cell-adhesion analyses, we successfully demonstrated that fusion mussel protein fp-353 has potential as a practical alternative bioadhesive.
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