Recombinant major ampullate spidroin-particles as biotemplates for manganese carbonate mineralization

Abstract Mineral deposition in biological systems is often templated by organic matrices including proteins directing the nucleation and growth of bioceramics by interacting with early stage species of the mineralization process or coordinating specific facets of the forming crystal. Structurally, charged surface patches are a characteristic motif of biomineralization-associated proteins, which are able to accumulate and bind ions from the surrounding media and, therefore, initiate, promote or inhibit mineralization. Controlled protein engineering enables the manipulation and control of bioinspired in vitro precipitation systems, and thus not only opens prospects for the design of environmentally benign synthetic strategies towards hierarchically structured functional materials, but also enhances the understanding of fundamental interaction mechanisms in biomineralization processes. Here, two recombinant variants of the spider silk protein ADF4 were engineered with oppositely charged peptide tags. Both were processed into micrometer-sized particles and investigated for their influence on manganese carbonate mineralization. Micro- and nano-structured manganese carbonate represents an attractive material for diverse applications including catalysis and wastewater treatment. While both types of spider silk particles were incorporated into the mineral structure, the positively tagged proteins appeared to interact more strongly with the formed manganese carbonate crystals than their negatively charged counterparts. Combination of the spider silk particles and poly(acrylic acid) (PAA), a water-soluble structure-directing agent associated with the stabilization of amorphous precursor phases in carbonates, resulted in the formation of film-like non-equilibrium structures of MnCO3 entrapping the spider silk particles. With the aim to gain mechanistic insights and to elucidate the interaction between the different components involved in the mineralization process, we studied the interplay between PAA, positively or negatively tagged spider silk particles, and Mn(II) ions by time-resolved dynamic light scattering. The here used set-up affords the possibility to identify control strategies for the template-mediated mineralization of manganese carbonate.