Synsepalum dulcificum, also called the miracle fruit, which has the sweet-inducing activity can be used as additives in food, medicine and cosmetic industries. Some selected chemical properties of miracle fruit including percentage by weight, total anthocyanin, phenolic and antioxidant content of different parts of miracle fruit as well as physicochemical analysis of seed oil, nutritional elements of fruit juice were determined in this study. The results showed that miracle fruit contains a large amount of vitamin C (40.1 mg/100 g fresh fruit weight (FW)), phenolic content (625.57 mg GAE/100 g FW), high antioxidant capacity (457.3 μmol Trolox/100 g FW) and low total sugar content (5.6 g/100 g FW), suggesting that the fruit is healthy for human consumption. According to its fatty acid composition and Triacylglycerol (TAG) profile, miracle fruit seed oil is rich in oleic and palmitic acid.
Miracle fruit, Synsepalum dulcificum, contains a glycoprotein known as miraculin. After consuming this glycoprotein, sour foods taste sweet and the effect lasts for up to 4 h. With increasing demand for natural and "low-calorie" sweeteners, the use of miraculin as an additive is increasing enormously in the food, medicine and cosmetic industries. In this study, we used reverse micelles formed from a sodium di (2-ethylhexyl) sulfosuccinate (AOT)/isooctane system to purify miraculin from S. dulcificum. We studied factors affecting purification performance, such as surfactant (AOT) concentration and the pH of the crude during forward extraction. During backward extraction, we examined the effects of NaCl concentration, the pH of the aqueous phase and addition of isopropanol. We found that 0.1 mol/L AOT/isooctane solution mixed with crude extract at pH 8 during the forward extraction stage and 0.5 mol/L NaCl solution at pH 11 during the backward stripping stage were optimal purification conditions, from which 22% miraculin was recovered with a purity of 94.8%.
In this study, the methods for extraction and purification of miraculin from Synsepalum dulcificum were investigated. For extraction, the effect of different extraction buffers (phosphate buffer saline, Tris-HCl and NaCl) on the extraction efficiency of total protein was evaluated. Immobilized metal ion affinity chromatography (IMAC) with nickel-NTA was used for the purification of the extracted protein, where the influence of binding buffer pH, crude extract pH and imidazole concentration in elution buffer upon the purification performance was explored. The total amount of protein extracted from miracle fruit was found to be 4 times higher using 0.5M NaCl as compared to Tris-HCl and phosphate buffer saline. On the other hand, the use of Tris-HCl as binding buffer gave higher purification performance than sodium phosphate and citrate-phosphate buffers in IMAC system. The optimum purification condition of miraculin using IMAC was achieved with crude extract at pH 7, Tris-HCl binding buffer at pH 7 and the use of 300 mM imidazole as elution buffer, which gave the overall yield of 80.3% and purity of 97.5%. IMAC with nickel-NTA was successfully used as a single step process for the purification of miraculin from crude extract of S. dulcificum.
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