The role of phospholipid modification initiated by phospholipase D (PLD) in enzymatic browning has been revoked through this study. Various alcohols and aldehydes were tried to read out their PLD controlling behaviour. Based on in-vitro results, reagents like hexanal and inositol were used to regulate PLD activity of litchi fruit stored at ambient temperature and their effects on fruit quality and physiological characteristics were also investigated. The results showed that combinatorial chemical treatment was successful in maintaining freshness of fruit through delayed physiological loss in weight and hence maintaining firmness. Combinatorial treated fruit had lower browning index than control by day 7. This novel treatment also maintained comparable levels of total phenolics and lowered the level of malondialdehyde. Evaluation of antioxidative enzymatic profile also confirmed the alleviation of oxidative stress of litchi fruit at ambient temperature. Thus, this strategy of enzyme regulation could play a vital role in overall quality maintenance of litchi fruit.