Controlled release of oxygen from myoglobin was achieved by modulating autoxidation of oxymyoglobin using ascorbic acid as a reductant by temperature variation. Long-term storage, prompt release and re-storage of oxygen were also available with this system. Furthermore, 20 nm thick nanofilms composed of oxymyoglobin and type I collagen containing ascorbic acid could successfully show autoxidation of oxymyoglobin in response to environmental temperature. The ultrathin nanofilms will be useful as oxygen-controlled releasable scaffolds for tissue engineering application.