Currently, algal fuel research has commenced to shift toward genetically engineered mutants able to express and excrete desired products directly into the culture. In this study, a mutant strain of Chlamydomonas reinhardtii, engineered for bisabolene (alternative biodiesel) excretion, was cultivated at different illumination and temperatures to investigate their effects on cell growth and bisabolene production. Moreover, a kinetic model was constructed to identify the desirable conditions for biofuel synthesis. Three original contributions were concluded. First, this work confirmed that bisabolene was partially synthesized independently of biomass growth, indicating its feasibility for continuous production. Second, it was found that while bisabolene synthesis was independent of light intensity, it was strongly affected by temperature, resulting in conflicting desirable conditions for cell growth and product synthesis. Finally, through model prediction, optimal operating conditions were identified for mutant growth and bisabolene synthesis. This study therefore paves the way toward chemostat production and process scale-up.