In this study, we evaluated the methylene blue (MB) adsorption potential of non-treated and UV-C pretreated bacterial biomass from aqueous solution. The UV-C pretreatment denature the biomass and has increased overall functional groups when compared to non-treated biomass. The biosorbent was exposed to various pH, biomass dose, and contact time. The results showed that the dried and UV-C pretreated biomass effectively removed MB within 30 min. Dried and UV-C pretreated biomass of Bacillus pseudomycoides JH 2-2 showed a adsorption of 858.2 and 1072.4 mg/g at optimum conditions (pH: 9.0, contact time: 30 min, biomass dose: 1 g/L). Similarly, dried and UV-C pretreated biomass of Citrobacter freundii JH 11-2 showed an adsorption 868.3 and 954 mg/g at optimum conditions (pH: 9.0, contact time: 10 min, biomass dose: 1.5 g/L). The changes in the functional groups of UV-C pretreated biomass could be responsible for enhanced adsorption of MB. The results obtained have shown that non-treated and UV-C pretreated biomass has a high adsorption capacity for MB dye and can be used as a low-cost biosorbent in wastewater treatments.