To develop bioelectronic devices and/or the structural analyses of proteins, two-dimensional crystallization of proteins is required. Heme proteins are particularly interesting due to electron transfer characteristics associated with the porphyrin-based prosthetic group. Cytochrome b(562) has been chosen as a model protein for the fabrication of two-dimensional crystals. Three approaches in making two-dimensional crystals of cytochrome b(562) are presented here : (1) the formation of monomolecular layer of cytochrome b(562) reconstituted with alkylated heme, (2) the control of surface charge density of cytochrome b(562) and (3) the enhancement of thermal stability both by means of protein engineering.