Abstract. Rice straw is agricultural waste containing high potency to be treated to biogas. However, the usage of rice straw is still limited due to high lignin content that will cause low biodegradability. The aim of this research was to study the effect of pretreatment using NaOH and acetic acid to biogas production from rice straw. NaOH was varied from 2%w, 4%w, and 6%w; and acetic acid was varied from 0,075 M, 0,15 M dan 0,75 M. The rice straw was cut into 1 cm size and submerged for 30 minutes in NaOH and acetic acid solution. The rice straw then filtered and neutralized before sending to anaerobic digestion process using rumen fluid bacteria. Biogas produced was measured using water displacement method. The result showed that the optimum concentration of NaOH solution was 4%w that resulted in biogas volume of 21,1 ml/gTS. Meanwhile, the optimum concentration of acetic acid pretreatment was 0,075 M that produced biogas volume of 14,5 ml/gTS. These results suggest that pretreatment using NaOH solution is more effective for decreasing the lignin content from rice straw.