Dengue fever is a disease transmitted by Ae. aegypti. Controlling mosquitoes can use temephos. Temephos larvicides act primarily on the enzyme acetylcholinesterase (AChE) by inhibiting this enzyme. Temephos has drawbacks due to its high risk of resistance, so larvicides from natural ingredients such as pandan leaf extract (P. amaryllifolius) can be an alternative. The mechanism of action of P. amaryllifolius larvicide is still unclear. The purpose of thid study is to ascertain how LC₈₅ pandan leaf methanol extract affects AChE levels in Ae. aegypti. This is a true-experiment design with only a post-test control group research design. Tests were carried out by treating mosquito larvae with P. amaryllifolius LC₈₅ extract, aquades, and temephos for 24 hours and measuring AChE levels with ELISA Reader. The results showed that the AChE enzyme levels of P. amaryllifolius LC₈₅ extract had an average AChE enzyme level of 147.19 + 70.87 units/l. The AChE enzyme levels of larvae exposed to P. amaryllifolius LC₈₅, aquades, and temephos had a significant difference (p <0.05). P. amaryllifolius has potential as a larvicidal, with a mechanism of action as a neurotoxin.