The low RAP content, hot mixing conditions, and the addition of a high ratio of new bitumen and aggregates result in low economic and environmental benefits for current regeneration technologies. A bio-rejuvenated additive (BRA) that can fully (100%) regenerate the RAP without heating is proposed in this paper. To reveal the mechanisms of BRA-rejuvenated RAP, the effects of BRA on the chemical structure and molecular weight of the RAP were investigated using Fourier-transform infrared spectroscopy and gel permeation chromatography. The mechanical performance and water damage resistance of BRA-rejuvenated RAP were studied. Low contents of new bitumen or epoxy resin were suggested to increase the mechanical performance of 100% RAP. The results show that the 1.5% BRA-rejuvenated RAP had the best mechanical performance. The blending of BRA with recycled RAP is a completely physical process, without any chemical reactions. The molecular weight of BRA is lower than that of bitumen; it can substantially increase the content of light components in aged bitumen, and play the role of adjusting and restoring the balance of the components of aged bitumen. The mechanical performance of BRA-rejuvenated RAP is enhanced significantly by adding low dosages of new bitumen or epoxy resin.