Papaya is widely grown in tropical and subtropical climates due to its high yield potential and high returns. The vital hormone melatonin, regulating various biological processes in plants, is eco-friendly and less harmful to humans than other chemicals. This study aims to enhance the quality and antioxidant enzyme activities and lessen postharvest senescence in papaya cv. CO 8 fruits during both ambient (32 ± 2 °C and 55 ± 5% RH) and cold storage (10 ± 2 °C and 90–95% RH) as exogenous melatonin (EMT) is applied in varying concentrations. An optimum melatonin dose of 1.5 mM was applied as a pre-harvest spray 15 days before harvest and a postharvest dip proved effective in prolonging shelf life (under ambient it prolonged to day 9 and under cold storage up to 28 days) and delaying ripening and softening. Exogenous melatonin application enhanced antioxidant activity, reduced weight loss, maintained firmness, delayed ripening enzymes, and lowered ethylene and CO2 levels. For instance, control fruits had weight losses between 7.42% and 10.09%, while fruits treated with 1.5 mM melatonin showed 5.74% and 9.06% weight loss under ambient and cold storage, respectively. In conclusion, applying EMT (1.5 mM) could be an economically viable and environmentally benign way to lessen senescence after harvest and preserve the qualities of the papaya fruit during ambient and cold storage.