Tomato leaf miner, Tuta absoluta (Meyrick) is a key and dangerous pest of Solanaceaus plant in Iran. Larva of the pest cause damage on different parts of host plants including leaves, stems and fruits. Excessive use of chemical insecticides quickly leads to pest resistance. Using protein-based compoinds such as the cholesterol oxidase enzyme to control pests is a good alternative to chemical insecticides. In this research, the effect of cholesterol oxidase enzyme on preference of tomato leaf miner was investigated.The three varieties of Newton, Izmir and Ergon were treated with cholesterol oxidase enzyme at 0.08, 0.6 and 0.2 mg/L during three phases of 30, 45 and 60 days of phenology. The amount of eggs laid out after 72 hours was evaluated. The results showed that by increasing the enzyme concentration ovulation decreased. Also the highest egg number was found on the Ergon variety and 0.08 mg/L and lowest egg number was found on the Newton varity on the concentration of 0.6 mg/L and 30-day phenology. This experiment was carried out in five replications and a light period: 16: 8 and 60 ± 5% relative humidity and 25 ± 5 ° C in greenhouse conditions, the results suggest that the cholestrol oxidase enzyme has the ability to control and repatriation tomato leaf miner. Also, the results so far point to the health of this compound for humans and consumer animals, because the building of this enzyme, like other proteins, is degraded and inactive under high temperature conditions, and its recommended to protect the invironment and health of user.