The present study investigated the application of photocatalytic process using TiO2 nanotubes (NTs) photocatalyst as an effective pre-treatment method for palm kernel expeller (PKE) in increasing its biodegradability to enhance the microalgal hydrogen production via dark fermentation. A series of TiO2 NTs photocatalysts were synthesized via employing electrochemical anodization of Ti foil at various voltages each in different types of electrolytes (choline chloride-ethylene glycol (ChCl-EG), choline chloride-urea (ChCl-U), choline chloride-glycerol (ChCl-Gly)). Increasing trend of average NTs diameters at 26.14 ± 1.30, 73.01 ± 1.94 and 122.00 ± 13.32 nm was observed for 25, 30 and 35 V in synthesizing TiO2 NTs using suitable ChCl-EG electrolyte, respectively. The high 35 V had resulted in noticeable pore ruptures within this TiO2 NTs photocatalyst. The PKE pre-treated by TiO2 NTs photocatalyst anodized at optimum 30 V in ChCl-EG electrolyte had demonstrated the highest releases of biochemical oxygen demand (BOD5) and soluble chemical oxygen demand (SCOD) concentrations at 594 and 607 mg/L, respectively. Moreover, the microalgal efficiency of BOD5 conversion and hydrogen yield were also consistently maintained from third cycle of TiO2 NTs photocatalyst's reutilizations, signifying the stability of TiO2 NTs photocatalyst. © 2023 Hydrogen Energy Publications LLC