This study aimed to isolate a novel archaeon capable of simultaneously producing methane and bioflocculants. The isolated methanogen was identified as Methanosarcina spelaei RK-23 by molecular characterization. Strain RK-23 grew with different carbon sources, but acetate was found to be the most beneficial carbon source for the production of methane and bioflocculants. Moreover, RK-23 produced methane and bioflocculants within an initial pH range of 5.0-8.0, with an optimal pH for methane production of 6.5 and flocculating activity of 7.0. In addition, strain RK-23 exhibited tolerance to high-salinity conditions and produced methane and bioflocculants within 0-1.2 M NaCl, with an optimum NaCl concentration of 0.3 M. The methane production and flocculating activity of strain RK-23 under optimal culture conditions were 17.4 mmol methane/mol acetate and 95.6%, respectively. MBF-23 mostly consisted of 83.5% carbohydrates and 14.3% proteins. Furthermore, the bridging of adsorption is believed to be the main bioflocculation mechanism in this study. (C) 2020 Hydrogen Energy Publications LLC. Published by Elsevier Ltd. All rights reserved.