To meet the increasing demands of sophorolipids as biosurfactants and bioactive compounds, it is necessary to obtain higher and more specific sophorolipid-producing strains. One sophorolipid-producing strain, Wickerhamiella domercqiae var. sophorolipid CGMCC 1576 (Y-2A), was mutated by low-energy nitrogen ion beam implantation. Eighteen mutants produced 20 % more sophorolipids than the wild strain, and one mutant, N3-18, produced the highest yield of sophorolipids, 104 g/l, in a shaking flask, which increased by 84.71 % than the wild strain, and further elevated to 135 g/l in a 5-l bioreactor. High performance liquid chromatography analysis showed that the composition of every sophorolipid mixture from different strains was similar, while the contents of most components from mutants were higher than that from the wild strain. Two mutants, N1-32 and N3-18, produced more acidic sophorolipid components; three lactonic sophorolipid molecules with good anticancer activities were greatly enhanced in several mutants, especially monoacetylated lactonic sophorolipid with a C18 monounsaturated fatty acid, which were enhanced by 153 and 211 % in strains N1-32 and N3-18. Low-energy nitrogen ion beam implantation was efficient for obtaining a variety of high and specific sophorolipid-producing mutants to be applied in food, cosmetic, environmental, and pharmaceutical sectors.