This study was conducted to evaluate the effect of cottonseed meal supplemented with lysine and enzyme on laying hens performance and egg quality. Eighty Hy-Line W-36 white Leghorns were allotted for 12 weeks in a 2 × 2 factorial experiment in a completely randomized design, including four treatments and five replications with four birds in each. Treatments included: basal diet + 1% lysine + 0% enzyme; basal diet + 1% lysine + 0.025% enzyme; basal diet + 2% lysine + 0% enzyme; and basal diet + 2% lysine + 0.025% enzyme. Protein content in the magnum tissue was not significantly affected by different levels of lysine and enzyme, although magnum Protein:RNA ratio increased with 2% of lysine as compared with 1%. Moreover, jejunum DNA's concentration was not significantly affected by lysine. Similarly, jejunum RNA:DNA ratio increased with 2% of lysine. Performance specificity significantly improved with 2% lysine and 0.025% enzyme. Diets supplemented with 2% lysine and 0.025% enzyme can improve performance, increase magnum protein synthesis and jejunum cell efficiency.