Anaerobic digestion (AD) is an attractive bioprocess for waste treatment and energy recovery through methane-rich biogas production. Under temperate to cold climate, the implementation of AD for low-organic load wastewater treatment has been limited to date, due to the energetic and economic cost of maintaining optimal mesophilic temperature. Hence, we aim at (i) exploring the biotechnological potential of a microbial inoculum from Antarctic soils and sediments to run AD at low temperatures; and (ii) evaluating the effect of temperature over a psychrophilic-mesophilic range on both methane production rates and microbial community composition. Methane production stimulated by acetate amendment was detected from 5 to 37 °C, with a maximum at 25 °C, corresponding to the highest relative abundance of methanogenic archaea (c. 21.4% of the total community). From 5 to 25 °C, the predominant methanogen was Methanosaeta, while it shifted to Methanocorpusculum at 30 °C. Compared with an industrial mesophilic sludge, the relative methane production rate at 5 °C (compared to the maximum) was 40% greater in the Antarctic inoculum. Microbial communities from permanently cold Antarctic sediments efficiently produce methane at low temperatures revealing a biotechnological potential for the treatment of low-organic load residues in cold regions.