Introduction: Inhibition of the cholinesterase’s function leads to paralysis and death. This mechanism is served as a common mode of action of insecticide. The three tropical seaweeds, namely Bryopsis pennata, Padina australis and Sargassum binderi were reported for its potential mosquito larvicidal effect. In the present study, these seaweeds were evaluated for their potential as a cholinesterase inhibitor in the mechanism of larvicidal action. Methods: Ace- tylcholinsterase (AChE) inhibition assay was carried out based on the colorimetric method using a microplate reader. Phytochemical content of the seaweed extracts was screened by using liquid chromatography-mass spectroscopy (LC-MS). Results: Green seaweed B. pennata showed the strongest inhibition effect towards in vitro AChE by using
tissue homogenates of Aedes aegypti (IC50 value = 0.84 mg mL ) and Aedes albopictus as the enzyme source (IC
-1
value = 0.92 mg mL-1). The pattern of Lineweaver-Burk plots revealed that B. pennata was a mixed type inhibitor of
AChE, as the readings of Km, Vmax, Ki and Ki’, indicates that it had a strong inhibition ability with high binding affin- ity towards both free enzyme and enzyme-substrate complex. Conclusion: These findings suggest the compound(s) in
B. pennata extract serves as a promising source that could be developed into a mosquito larvicidal agent with AChE inhibition effect.