In order to characterize lipid metabolism of yeasts during sufu manufacture, 23 isolates from sufu pehtze and acidic tofu whey were identified. Ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was used to analyse the lipid fingerprints of 7 strains of yeast, after the determination of lipase activities. It is suggested that lipid fingerprint mainly includes 2 kinds of sphingosine, 16 kinds of amide, 22 kinds of ester and 3 kinds of fatty acid. Kluyveromyces marxianus SP-1 and Candida (Pichia) norvegensis SP-5 show highest relative amount of 2-Amino-1,3,4-octadecanetrio and N, N-Dimethylsphingosine, which indicates potential for sphingolipids synthesis. Candida ethanolica ATW-1, Pichia kudriavzeviiSP-4 and Pichia kudriavzevii Y with higher cell-bound lipase activities have more abundant esters. PCA analysis further illustrates the lipid composition of these three strains are more similar to each other. The results indicate that the abundant lipids in yeasts may be closely related to cell-binding lipase.