Termitomycesfungi associated with fungus-growing termites are the edible mushrooms and can produce useful chemicals, enzymes, and volatile organic compounds (VOCs) that have both fuel and biological potentials. To this purpose, we examined theTermitomycesmycelial growth performance on various substrates, clarified lignocellulose-degrading enzyme activity, and also identified the VOCs produced byTermitomyces. Our results indicated that the optimal nutrition and condition requirements for mycelial growth ared-sorbitol,d-(+)-glucose, andd-(-)-fructose as carbon sources; peptone as well as yeast extract and ammonium tartrate as nitrogen sources; and Mn2+, Na+, and Mg(2+)as metal ions with pH range from 7.0 to 8.0. Besides, the orthogonal matrix method results revealed that the ideal composition for mycelial growth is 20 g/Ld-(-)-fructose, 5 g/L yeast extract, 0.5 g/L Mg2+, and pH = 7. We also screened various substrates composition for the activity of lignocellulose-degrading enzymes, i.e., lignin peroxidase, manganese peroxidase, beta-glucosidase, a-l-arabinofuranosidase, and laccase. Furthermore, we identified 37 VOCs using GC-MS, and the most striking aspect was the presence of a big series of alcohols and acids, collectively constituted about 49% of the total VOCs. Ergosta-5, 8, 22-trien-3-ol, (3.beta.,22E) was the most plenteous compound constituted 30.369%. This study hopes to establish a better understanding for researchers regardingTermitomyces heimiicultivation on a large scale for the production of lignocellulosic enzymes and some fungal medicine.