Methane production from marine microalgae Isochrysis galbana was assessed before and after mechanical and chemical pretreatments. Mechanical pretreatment resulted in a 61.7% increase in soluble Chemical Oxygen Demand. Different hydrolysis conditions were evaluated by varying temperature - T, sulfuric acid concentration - AC and biomass suspension concentration (measured as particulate COD - CODp) using an experimental design. The most significant interaction occurred between AC and T and the hydrolysis condition that showed the best result in the anaerobic digestion step was the condition at 40<SUP>o</SUP>C with addition of 0.2% (v/v) acid for 16h (9.27LCH<SUB>4</SUB>/kgVS). The low methane yields were attributed to inhibitory sodium concentrations for anaerobic digestion. Eliminating inhibitory sodium in the anaerobic digestion by biomass prewashing, there was a 71.5% increase in methane yield for biomass after acid hydrolysis, demonstrating the need for pretreatment and reduction in sodium concentration in the anaerobic digestion.