A lipase-producing Yarrowia lipolytica strain YL1 was mutagenized by atmospheric room temperature plasma. Tributyrin plate method, p-NPP method and acid-base titration method were used to screen out the target strain C4 with high lipase production, and then its genetic stability was studied. The results showed that the optimal mutagenesis time of Y. lipolytica strain YL₁ was 60 s, and the lethality rate of the strain reached 97.45%. The lipase activity of the mutant strain C4 was 13.4 U/mL, which was 82.6% higher than the original strain, and it was genetically stable after multiple generations of culture. Compared with the original strain, the mutant strain lipase could increase the synthesis-conversion rate of vitamin A palmitate by 36.9%.