The utilization of semi-simultaneous saccharification and fermentation (SSSF) as the novel configuration has resulted in enhanced succinic acid (SA) production from lignocellulose biomass by Actinobacillus succinogenes. The effect of inoculum concentration, biomass type, substrate concentration, and fermentation configuration on SA production was examined in this study. The pre-hydrolysis process was applied to the pre-treated biomass for 6 h to facilitate the simultaneous saccharification and fermentation (SSF) process, which was then carried out for 48 h to achieve the SSSF configuration. According to the results, the production of SA from oil palm empty fruit bunch (OPEFB) through SSF and SSSF was 0.93 and 1.18 g/L and from sugarcane bagasse (SB) was 0.98 and 1.19 g/L, respectively. Results revealed, SSSF resulted in a 21–26% higher SA concentration compared to SSF. Furthermore, the concentration of the inoculum and substrate significantly affected the generation of SA from OPEFB but not for SB. According to this study, SSSF significantly enhanced SA production from lignocellulose biomass compared to SSF. © The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature 2025.