The inhibitory effect of onion extract on cassava leaf polyphenol oxidase was investigated. The polyphenol oxidase from cassava leaves was strongly inhibited by various anti-browning agents such as L-ascorbic acid and L-cysteine. The percentage of inhibition increased with the increased of anti-browning agents concentrations. The addition of heated onion extract exhibited a stronger inhibitory effect on cassava leaf polyphenol oxidase than the fresh onion extract. The highest percentage of inhibition was exhibited with heated onion extract in the presence of glucose and glycine, which was 87.18%. The onion extract inhibited the cassava leaf polyphenol oxidase non-competitively.