<p>Food waste (FW) is rich in biodegradable organic matter, making it a promising source of renewable bioenergy through anaerobic digestion (AD). Typically, anaerobic reactors are inoculated with granular sludge from other reactors because of its excellent sedimentation and specific methanogenic activity (SMA). However, in developing countries, flocculent sludge with its lower sedimentation and SMA is more commonly available. This study evaluated two strategies to condition flocculent inoculum: first, the pre-addition of a rapidly degradable carbon source (sucrose) at 35&#xa0;°C, and second, acclimatization at 35&#xa0;°C and 55&#xa0;°C without substrate addition. The research assessed their effects on key metabolic activities hydrolytic (SHA), acidogenic (SAA), and methanogenic (SMA) as well as methane production via biochemical methane potential (BMP) assays and process kinetics. Sucrose addition resulted in the highest SAA (5.7 gCOD_Glucose gVS⁻¹ d⁻¹) and process disturbances in a buffering system with a bicarbonate alkalinity/total alkalinity (BA/TA) ratio &gt; 0.7. Temperature acclimatization at 35&#xa0;°C emerged as the superior strategy, enhancing both SHA (0.14 gCOD_Glucose gVS⁻¹ d⁻¹) and SMA (0.052 gCOD_CH₄ gVS⁻¹ d⁻¹) while improving buffer capacity (BA/TA ratio between 0.5 and 0.7). This improved microbial balance increased the hydrolysis rate (<i>k</i>_<i>h</i>: 0.09 d⁻¹) and the consumption of volatile fatty acids (<i>k</i>_<i>VFA</i>: 0.97 d⁻¹), leading to a 31% boost in methane production (113 mLCH₄ gVS⁻¹) compared to the control. Overall, AD of FW proved most efficient at 35&#xa0;°C. In future studies, molecular biology analyses should be incorporated. In addition, economic and environmental aspects of the impact of the strategies during their sustainable large-scale implementation should be included.</p>