Suppression of N-glycan processing enzymes by deoxynojirimycin in tomato (Solanum lycopersicum) fruit

The present study investigated the potential of a small molecule inhibitor, 1-deoxynojirimycin (DNJ), to extend the shelf life of tomatoes. The optimum concentration of DNJ and the proper ripening stage for treatment were standardized using response surface methodology, following a central composite design. The concentration of DNJ used for the analysis was 0.15 mM, and 0.30 mM and the ripening stages of the tomato fruit analysed were immature green, mature green, breaker, ripen and over-ripen. Analysis of the influence of the DNJ treatment of the fruit using quadratic multiple regression models considering the factors colour, texture, and free sugars revealed significant responses. A DNJ concentration of 0.30 mM and fruit-ripening stage of mature green was found to be optimal for the treatment. DNJ-treatment maintained fruit firmness throughout ripening with a significant reduction in reducing sugar formation. Enzyme activity of the N-glycan processing enzymes involved in cell wall softening, alpha-mannosidase and beta-d-N-acetylhexosaminidase revealed a significant reduction in their activity by 2 and 3.5-fold, respectively. Down-regulation of expression of important ripening-related and softening process-associated genes, aminocyclopropane carboxylic synthase-4, aminocyclopropane carboxylic oxidase, polygalacturonase and pectin methylesterases at 4, 5, 6 and 5-fold, respectively, was also observed. The present results showed that the treatment of mature green tomato fruit with DNJ at a concentration of 0.30 mM can delay the ripening of the tomato fruit by inhibiting cell wall and N-glycan processing enzymes.