A ferredoxin adsorbed hetero self-assembled layer was fabricated on chemically modified Au substrate, 4-Aminothiophenol (4-ATP) was deposited onto Au substrate and then N-succinimidyl-3-[2-pyridyldithio] propionate (SPDP) was adsorbed on the 4-ATP layer, since SPDP was used as a bridging molecule for ferredoxin adsorption, Ferredoxin/SPDP/4-ATP structured hetero layer was constructed because of strong chemical binding of ferredoxin, SPDP, and 4-ATP, The surface of the ferredoxin-adsorbed SPDP/4-ATP layer was observed by scanning tunneling microscopy, The hetero film formation was verified by surface plasmon resonance measurement. The current flow and rectifying property based on the scanning tunneling spectroscopy I-V characteristics was achieved in the proposed hetero layer. Thus, the hetero layer structure of ferredoxin functioned as a molecular diode with rectifying property, The proposed molecular diode can be usefully applied for the development of molecular scale electronic devices.