Cereal Chem. 75(5):595-601In control dough, endogenous wheat lipase was inactive, because the triacylglycerol (TAG), 1,2-diacylglycerol (DAG 1,2 ), and 1,3-diacylglycerol (DAG 1,3 ) fractions of nonpolar lipids were not affected by mixing. Conversely, the free fatty acid (FFA) and monoacylglycerol (MAG) fractions decreased, mainly due to the oxidation of polyunsaturated fatty acids (PUFA) catalyzed by wheat lipoxygenase. Addition of exogenous lipase to flour (15 lipase units [LU] per gram of dry matter) resulted in substantial modification of nonpolar lipids during dough mixing. Due to the 1,3 specificity of the lipase used in this experiment, the TAG and DAG 1,3 fractions decreased, whereas the MAG and FFA fractions increased. The DAG 1,2 fraction increased at the beginning of mixing and decreased after 40 min of mixing. Moreover, part of the PUFA released by lipase activity was oxidized by wheat lipoxygenase, resulting in major losses of PUFA. Conversely, the net content of the saturated and monounsaturated fatty acids (SMUFA) remained constant, because the free SMUFA content increased primarily at the expense of the esterified forms. For a constant mixing time of 20 min, increasing the amount of lipase added to dough (from 2.5 to 25 LU/g of dry matter) resulted in a linear decrease in the TAG fraction and a linear increase in the SMUFA content in the FFA fraction. At the same time, the PUFA content of the FFA fraction increased only for additions of lipase to flour of >5 LU/g of dry matter, due to partial oxidation by wheat lipoxygenase.