A dynamic membrane (DM) is a biofilm that forms on a support material, acting as a filter to retain high-density biomass. This study aims to explain the effects of alginate immobilization on DM formation during dark-H-2 fermentation. Galactose is used as a model substrate. Heat-treated anaerobic sludge, with and without immobilization, is used for an inoculum for two identical lab-scale DM bioreactors (DMBR). The DMBRs are continuously operated for more than 40 days by changing the hydraulic retention time (HRT) from 12 to 3 h. Biomass retention and H-2 production performance are significantly improved at an HRT of 3 h with immobilization. The alginate-added bioreactor shows higher extracellular polymeric substance content both in the mixed liquor and the DM. At an HRT of 3 h with immobilization, the fraction of Sporolactobacillus spp. and Lactobacillus spp. increases, possibly contributing to DM formation. However, lactic-acid concentration does not increase, implying it can be further consumed by the dominant bacteria, Clostridium butyricum. (C) 2019 Hydrogen Energy Publications LLC. Published by Elsevier Ltd. All rights reserved.