Haematococcus pluvialis microalgae have emerged as a prevalent source of antioxidants in cosmetics and nutritional products. Additionally, numerous researchers have posited the potential of this microalgae to produce fatty acid methyl esters (FAME). Nevertheless, the optimization of the production of FAME from H. pluvialis oil has not been investigated. In this study, the transesterification reaction of H. pluvialis bio-oil was optimized using the response surface methodology, resulting in optimal experimental conditions for an oil to methanol ratio of 1:4.17, at a temperature of 80 °C, with a reaction time of 47 min. The resulting FAME was found to not comply with the biodiesel standard in terms of the content of polyunsaturated fatty acids (6.02%), as well as kinematic viscosity (7.02 mm2/s). Further study is required to reduce these parameters in order to ensure biodiesel quality and compliance with the standard. Nevertheless, its high flash point value of 150 °C and its high thermal stability within the temperature range of 211–290 °C suggest the potential for utilization as a biolubricant.