Sucrose metabolism was followed in developing fruit of domesticated cherry tomato (Lycopersicon esculentum var. cerasiforme Alef.). The high amounts of reducing sugars were consistently linked to high soluble acid invertase (EC 22.214.171.124), whereas sucrose synthase (EC 126.96.36.199) followed the same pattern of sucrose levels and reached a peak of activity during early stage of maturation and then decreased to near nil. In comparison, sucrose phosphate synthase (EC 188.8.131.52) activity remain relatively constant throughout development. Thus, sucrose synthase and acid invertase, rather than sucrose phosphate synthase, are the critical enzymes regulating sucrose accumulation in tomatoes. Cultivated cherry tomato sucrose synthase (UDP-glucose: D-fructose 2-glucosyltransferase) was purified to homogeneity by ammonium sulfate precipitation, anion exchange chromatography on DEAE-Toyopreal 650, and gel filtration on Sephadex G-200. Further purification to homogeneity resulted from a single band from SDS-PAGE. The enzyme was identified as a homotetramer with a total molecular mass of 370 kDa and subunits of 92 kDa. The enzyme showed maximum activity for the cleavage and synthesis of sucrose was at pH 7.0 and 8.0, respectively, and the optimum temperature was 40°C in both directions for HEPES-KOH buffer. The enzymatic reaction followed typical Michaelis–Menten kinetics, with the following parameters: Km (fructose),7.4; Km (UDP-glucose), 0.2612; Km (sucrose), 33.24; Km (UDP), 0.0946. The enzyme was very sensitive to inhibition by heavy metals.
Seasonal fluctuations of carbohydrate levels and compositions and the activities of related enzymes of three cultivated tomato (Lycopersicon esculentum Mill. cv. Lady First, Momotaro, and Minicarol) cultivars were examined at 45-days interval with seven different sowing in the relatively warm climate of Japan. Fruits picked on early winter to spring seasons had higher sugar concentrations compared to hot season. Fructose and glucose in nearly equal amounts were the predominant sugar in all the seasons. Sucrose was present in trace quantities, but cherry cultivar Minicarol accumulated higher levels than the other two large-fruited types.
Acid invertase (EC 184.108.40.206) was highest at red stage during December to April, while fruit matured during May to August had lowest activity. The activity levels of soluble invertase were predominant compared to cell wall-bound fraction. The sucrose synthase (EC 220.127.116.11) showed highest activity in rapidly growing fruits followed by a very low activity with fruit maturation. Sucrose synthase showed the higher activity during November to February, and almost low activity during all the experimental periods. The sucrose phosphate synthase (EC 18.104.22.168) also showed higher activity during October to February, but the activity levels did not change drastically throughout the fruit development. The results substantiate the conclusion that, in all the planting seasons, acid invertase is a principal enzyme in the process of tomato fruit ripening and during early stage of tomato fruit development, sucrose synthase is the dominant enzyme, which, in turn, plays a part in regulating the translocation of sucrose into the fruit.