The effects of pretreatment methods of wheat straw on sophorolipids (SLs) fermentation by Starmerella bombicola were investigated, and the fermentation parameters were optimized. Dilute acid pretreatment (DAP), sodium hydroxide pretreatment (SHP) or sulfur trioxide microthermal explosion pretreatment (STMEP) were chosen to pretreat wheat straw to enhance the following enzymatic saccharification. Then, the saccharification liquids were used for SLs fermentation, respectively. Glucose supplement and activated carbon detoxification were used to improve SLs production. The results showed that SHP was the most favorable method for enzymatic saccharification of wheat straw, and the glucose content reached 61.30 g/L, followed by STMEP and DAP, with glucose content of 48.33 g/L and 40.00 g/L, respectively. The total content of inhibitors in STMEP saccharification liquid was the lowest, followed by SHP and DAP. S. bombicola could directly use the above saccharification solution to produce SLs, but the fermentation characteristics were different. Compared to the control chemical synthesis medium, SHP and STMEP saccharification medium were more conducive to acidic sophorolipid (ASL) accumulation, and the yield increased by 74.27% and 92.33% to 100.45 g/L and 110.86 g/L. Glucose addition and activated carbon detoxification could further improve SLs production. For SHP saccharification medium, glucose addition combined with activated carbon detoxification further increased ASL production to 124.49 g/L; For STMEP saccharification medium, lactonic sophorolipid (LSL) production was further increased to 32.02 g/L, which was similar to the LSL fermentation level of chemical synthesis medium. Therefore, wheat straw had potential for SLs production, and different pretreatment methods and fermentation methods can be used to obtain different types of SLs. The present study is benificial to reduce the production cost of SLs and expand its application field.