In practice, primary and secondary sludge are fed into anaerobic digestion. However, the microbial cell exists in secondary sludge are an unfavorable substrate for biodegradation. Thermal pretreatment is proved to increase the bioavailability of organic and improve the biodegradation subsequently. During low thermal pretreatment, both intracellular (within the microbial cell) and extracellular (within the polymeric network) materials were extracted. This process increases the bioavailability meaning that organic compounds are accessible to the microorganisms for their degradation. This research aims to investigate the effect of thermal pretreatment on domestic mixed sludge disintegration. Domestic mixed sludge was thermally treated at 70°C for various holding times. The pre-thermally treated domestic mixed sludge was measured for protein and carbohydrates following the Lowry Method, and Phenol-Sulphuric Acid Method respectively. DR 6000 UV-Vis spectrophotometer, DRB200 Reactor (digester) and COD vial (TNT plus 822) were used for COD determination, based on Reactor Digestion Method approved by USEPA. The results showed that the organic matter in domestic mixed sludge is efficiently solubilised during thermal treatment organic matter. The higher soluble yield for each monitored parameter determined in this study indicated that low thermal pretreatment improve bioavailability.