In this study, the modified hydrophobin, engineered for biomimetic mineralization, has been employed as a structure-directing agent for mineralization of calcium carbonate. For the first time amphiphilic calcium carbonate particles have been obtained, using engineered proteins. The mineral microparticles have been characterized by optical microscopy, scanning electron microscopy (SEM) and X-ray diffraction (XRD). While mineralization in the presence of non-modified hydrophobin results in polymorph mineral structures, uniform microspheres with an average particle diameter of one micron are obtained by employing hydrophobin which has been modified with an additional ceramophilic protein sequence. Owing to the tri-functionality of the modified hydrophobin (hydrophilic, hydrophobic and ceramophilic), the obtained mineral microparticles exhibit amphiphilic properties. Potential applications are in the areas of functional fillers and pigments, like biomedical and composite materials. Pickering emulsions have been prepared as a demonstration of the emulsion-stabilizing properties of the obtained amphiphilic mineral microspheres. The structure-directing effects of the studied engineered hydrophobins are compared with those of synthetic polymers (i.e. polycarboxylates) used as crystallization and scaling inhibitors in industrial applications.
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