In this study, Auxenochlorella protothecoides (AP-CK) was selected due to its reported high growth potential in sterilized black and odorous water (SBOW). In order to improve the resource utilization level of microalgae for wastewater treatment, AP-CK was mutated using 12C6+ heavy-ion beam irradiation, and a high lipid-containing mutant (AP-34#) was isolated and further evaluated to treat original black and odorous water (OBOW). Compared with the wild type, the maximum removal rates of COD, NH4+-N and TP of the mutant increased by 8.12 ± 0.33%, 10.43 ± 0.54% and 11.97 ± 0.16%, respectively, while maximum dissolved oxygen content increased from 0 to 4.36 ± 0.25 mg/L. Besides, the mutant lipid yield increased by 115.87 ± 3.22% over the wild type in OBOW. The fatty acid profile of AP-34# grown in SBOW and OBOW showed higher proportion of saturated fatty acids (C16:0 and C18:0) and valuable polyunsaturated fatty acids (mainly C20:5n3 and C22:6n3) which are more suitable for biodiesel production and value-added products, respectively. This work provides a new perspective on improving the characteristics of microalgae and an innovative approach for resource-based microalgae wastewater treatment through bioremediation of black and odorous water.