Display omitted •Mono-digestion system of Pennisetum hybrid failed in long-term operation.•The stable organic loading rate (OLR) increased 1-fold by adding bioaugmentation.•Maximum volumetric biogas yield of 1.95 m3/(m3·d) obtained at OLR of 4.0 g VS/m3·d.•Dramatically increase in protein- and amino acid-utilizing bacteria observed in R2.•The syntrophism of Syntrophomonas and methanogens has positive effect on stability. In this study, bioaugmentation with methanogenic propionate-utilizing enrichment was investigated as a method to improve the mono-digestion performance of Pennisetum hybrid in a semi-continuous mode. The effect of bioaugmentation on the microbial community was analyzed as well. The results demonstrate that the steady-state organic loading rate (OLR) of the bioaugmented reactor increased to 4.0 g VS/(L·d) with a volumetric biogas production of 1.95 ± 0.17 m3/(m3·d). In contrast, the non-bioaugmented reactor failed at an OLR of 2.0 g VS/(L·d) accompanied with the accumulation of volatile fatty acids (VFAs). The results of whole genome pyrosequencing analysis suggest that the decrease in relative abundance of syntrophic butyrate and propionate oxidizers, such as Syntrophomonas, Syntrophobacter, and Syntrophorhabdus, reduced the conversion efficiency of butyrate and propionate which leads to the accumulation of butyrate and propionate, influencing the performance of the mono-digestion reactor. Conversely, in the bioaugmented reactor, the higher density of protein- and amino acid-utilizing bacteria, such as Proteiniphilum, Thermovirga, and Lutaonella, as well as the syntrophic association of Syntrophomonas spp. coupled with the methanogens Methanosarcina and Methanocella has a positive effect on system stability and performance.