The influence of the reactor wall attached biofilm on the nutrient removal performance was investigated in an open photobioreactor during long-term operation. Total nitrogen and phosphorus removal efficiencies were statistically similar between reactor with (reactor A) and without (reactor B) biofilm at the Hydraulic Retention Time (HRT) of 18, 13.5 and 9 days. When the HRT reduced to 8 days, total nitrogen and phosphorus removal efficiencies in the reactor A were 42.95 ± 5.11% and 97.97 ± 1.12%, respectively, while significant lower removal efficiencies (38.06 ± 5.80% for total nitrogen and 83.14 ± 8.16% for phosphorus) were obtained in the reactor B. The VSS concentrations throughout the test were statistically similar for the two reactors, with a mean value of 0.63 ± 0.25 g/l for reactor A and 0.69 ± 0.20 g/l for reactor B. This study indicated that the reactor wall attached biofilm supported high phosphorus and nitrogen removal, which may provide insight into the practical implementation of microalgae-based wastewater treatment.