This study aims to obtain the best lignin content, ligninase enzyme activity and in vitro digestibility value of fermented sugarcane shoots using Pleurotus ostreatus and Aspergillus oryzae at different fermentation times. The research process is divided into 2 stages. Stage 1 treatment samples include A1B1 = Sugarcane shoots fermented with Pleurotus ostreatus for 14 days; A1B2 = Sugarcane shoots fermented with Pleurotus ostreatus for 21 days; A1B3 = Sugarcane shoots fermented with Pleurotus ostreatus for 28 days; A2B1 = Sugarcane shoots fermented with Aspergillus oryzae for 14 days; A2B2 = Sugarcane shoots fermented with Aspergillus oryzae for 21 days; A2B3 = Sugarcane shoots fermented with Aspergillus oryzae for 28 days. The design used is a completely randomized design with a factorial pattern. Stage 2 treatment samples cover A = Sugarcane shoots fermented with Pleorotus ostreatus for 21 days; B = Sugarcane shoots fermented with Pleorotus ostreatus for 28 days; C = Sugarcane shoots fermented with Aspergillus oryzae for 21 days; D = Fermented sugarcane shoots with Aspergillus oryzae for 28 days. The design used was a randomized block design. The results showed that there was no interaction between the type of mold and the fermentation time on the lignin content (P>0.05), but there was an interaction with the enzyme activity of Laccase, LiP and MnP (P<0.05). Also, there were significant differences in the digestibility of protein, cellulose, hemicellulose, VFA and NH3 (P<005), however the digestibility of DM, OM, ADF, NDF and rumen fluid pH had no significant difference (P>0.05). It was concluded that sugarcane shoots fermented with Pleurotus ostreatus mold for 28 days got the best results with the value as follow, lignin content (11.55%), CP digestibility (57.90%), cellulose digestibility (50.25%), Hemicellulose digestibility (62.65%), Laccase enzyme activity (2.68 U/mL), LiP enzyme activity (19.44 U/mL), VFA (111.67 mM) and NH3 (10.48 mg/100 mL).