POPULATION EXPLOSION, INDUSTRIALIZATION, DEPLETION OF FOSSIL FUELS AND THE RESULTED ENVIRONMENTAL POLLUTION CREATED A GREAT INTEREST TO FIND A RENEWABLE FUEL AS AN ALTERNATIVE TO CONVENTIONAL FUELS. BIOETHANOL FROM MICROALGAE, THE THIRD GENERATION OF BIOEHTANOL, IS ONE OF THE MOST PROMISING SOURCES OF ENERGY FOR FUTURE DEMANDS. MICROALGAE IS A CHEAP FEEDSTOCK WHICH HAS RAPID GROWTH RATE AND THE CAPABILITY OF GROWTH IN SEWAGE AS CULTURE MEDIA. THE IMPORTANT FEATURE OF MICROALGAE AS BIOETHANOL FEEDSTOCK IS THE MORE EASIER AND CHEAPER PRETREATMENT FOR SUGAR RELEASE COMPARED TO LIGNOCELLULOSIC FEEDSTOCKS. IN THIS STUDY, SEVERAL PRETREATMENT METHODS INCLUDING PHYSICAL, CHEMICAL AND PHYSICOCHEMICAL FOR MICROALGAE WERE INVESTIGATED. ULTRASONIC AND HEATING AS PHYSICAL, ACID AND LIME AS CHEMICAL AND COMBINED ULTRASONIC AND ACID AS PHYSICOCHEMICAL METHODS ARE EVALUATED IN THIS RESEARCH. AMONG THESE METHODS, PHYSICAL AND LIME PRETREATMENT WERE NOT EFFECTIVE. ACID PRETREATMENT CARRIED OUT BY SULFURIC ACID AND THE MAXIMUM YIELD OF PRETREATMENT WAS 97.3% WHICH IS CONSIDERABLY HIGH. ALSO, THE COMBINATION OF ACID AND ULTRASONIC METHOD IS ASSESSED FOR MICROALGAE PRETREATMENT. IN THIS METHOD, SUBSTRATE CONCENTRATION, ACID CONCENTRATION AND TIME OF HEATING PARAMETERS WERE OPTIMIZED.