Cordyceps coffee, containing Cordycepin and -glucan, was developed to improve quality and functionality of coffee. We evaluated its biological activities and quality characteristics. We treated green coffee beans with mixed extracts from three medicinal mushrooms, Cordyceps mushroom (Cordyceps militaris), Phellinus mushroom (Phellinus linteus), and Chaga mushroom (Inonotus obliquus). Both control [27.00 mg gallic acid equivalent (GAE)/g] and Cordyceps coffee (37.34 mg GAE/g) had higher contents of polyphenol than the Cordyceps mushroom raw materials (23.17 mg GAE/g). 1,1-Diphenyl-2-picrylhydrazyl scavenging activity was higher for both the control and Cordyceps coffee (80.56% and 82.21 %, respectively) compared with Cordyceps mushrooms (62.89%). -Glucan content was determined by the enzyme method; the raw Cordyceps, Phellinus, and Chaga mushrooms contained 3.79%, 7.06%, and 8.57% -glucan, respectively. However, -glucan and total glucan contents were much lower in Cordyceps coffee (2.03% and 3.11%, respectively) than in the raw mushrooms. The amount of Cordycepin, as determined by high performance liquid chromatograph, was 2,274.70 mg/kg for the Cordyceps coffee and 11,533.22 mg/kg for the Cordyceps mushrooms. Through flavor pattern analysis using the electronic nose system, we showed Cordyceps coffee kept the original coffee aroma (similar as the control), and this was not obstructed by the off-flavor of the Cordyceps mushrooms. In conclusion, this study suggests that Cordyceps coffee may be a novel functional coffee containing the functional components Cordycepin and -glucan.