The anaerobic process is increasingly becoming a subject for many as it reduces greenhouse gas emissions and recovers carbon dioxide energy as methane. Even though these benefits are attainable, proper control and design of the process variables has to be done in order to optimize the system productivity and improve stability. The aim of this research was to optimize methane and biogas yields on the anaerobic co-digestion of sewage sludge using bio-chemical substrates as co-substrates. The first objective was to find the bio-chemical substrate that will generate the highest biogas and methane yields. The anaerobic digestion of these substrates was operated using 6 L digesters at 37.5℃. The substrate which generated the highest biogas and methane yield in the first batch experiment was then used for the second batch test. The objective was to optimize the anaerobic conditions (substrate to inoculum ratio, co-substrate concentration and temperature) in-order to optimize the biogas and methane yields. The second batch test was achieved using the conventional One-Factor-At-A-Time (OFAT) and the Design of Experiment (DOE) methods. Final analysis showed that the bio-chemical substrates could be substrates of interest to biogas generators. Amongst the substrates tested in the first batch experiment glycerol (Oleo-Chemical Product waste) generated the highest methane and biogas yields of 0.71 and 0.93 L. (g volatile solids added)-1, respectively. It was believed that glycerol contains significant amount of other organic substances such as lipids that have higher energy content than the other bio-chemical substrates, thus generating larger biogas and methane yields. Moreover, digestion of sewage sludge alone produced biogas yields of 0.19 L /g VS and 0.33 L/g COD, and methane yields of 0.16 L/g VS and 0.28 L/g COD. Generally, co-digestion yields were higher than digestion yields of sewage alone. Using the OFAT method the results of the second batch test on glycerol demonstrated highest amounts of volatile solids (VS) reduction, chemical oxygen demand (COD) reduction, biogas yield and methane yield of 99.7%, 100%, 0.94 L (g VS added)-1 and 0.75 L (g VS added)-1 at a temperature, substrate to inoculum ratio and glycerol volume of 50℃, 1 (on VS basis) and 10 mL, respectively. Above 22 mL and substrate to inoculum ratio of 1, the process was inhibited. The DOE results suggested that the highest methane and biogas yields were 0.75 and 0.94 L (g VS added)-1, respectively. These results were similar to the OFAT results, thus the DOE software may be used to define the biogas and methane yields equations for glycerol. In conclusion, anaerobic co-digestion of bio-chemical substrates as co-substrates on sewage sludge was successfully applied to optimize methane and biogas yields.