Primary recovery of miraculin from miracle fruit, Synsepalum dulcificum by AOT reverse micellar system

Abstract

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%.

Effect of Key Parameters on Jacalin Extraction from Aqueous Phase into Anionic Reverse Micelles

“…Further increase of AOT concentration beyond this value resulted in a decrease of protein transfer and finally leveling off at 40 to 80 mM. The observed result is in line with another study which also describes an increment in the forward extraction efficiency with increasing surfactant concentration followed by a decrease in the extraction efficiency during forward extraction of miraculin from the crude extract of Synsepalum dulcificum using AOT reverse micelles [30].…”

“…Jacalin is a 66 kDa medium size protein, therefore, a higher pH-pI value is needed for an effective transfer to AOT reverse micelles. Similar studies dealt with AOT reverse micelles have also reported higher recovery of biomolecules at low pH, indicating that pH value below isoelectric is preferred for forward extraction [29][30].…”

Miracle fruit: An alternative sugar substitute in sour beverages

“…Moreover, it showed high antioxidant activity (Du, Shen, Zhang, Prinyawiwatkul, & Xu, 2014). Studies have investigated developing miraculin into an alternative sweetener (Bartoshuk, Gentile, Moskowitz, & Meiselman, 1974;Wong & Kern, 2011;Igarashi et al 2013), its molecular mechanisms (Bartoshuk, 1987;Dzendolet, 1969;Paladino, Costantini, Colonna, & Facchiano, 2008Yamamoto et al 2006), and the engineering of other plants to produce miraculin (Sun, Cui, Ma, & Ezura, 2006;Sun, Kataoka, Yano, & Ezura, 2007;He at al. 2015).…”

Pharmaceutical and Therapeutic Potential of Miraculin and Miracle Berry

“…They macerate leaves and fruits of miracle plant with ethanol and then evaporate at room temperature. 17 He, et al, 30 modified the extraction method for miraculin from miracle berry as described by Theerasilp and Kurihara., 22 In this method, lyophilized pulp powder was suspended with water and homogenized. The homogenate was centrifuged and the sediment was homogenized in 0.5 M sodium chloride solution.…”

“…22,30 Miraculin has been purified by using various methods like solvent extraction method from polar to non-polar and from non-polar to polar in succession, 9 ion-exchange chromatography technique, 10 immobilizedmetal affinity chromatography (IMAC), 31 and reverse micelle system. 30 The solvent extraction method is tedious and the purity of miraculin is relatively low. High purity miraculin was obtained by ion-exchange chromatography but this is time-consuming, inefficient and costly 10 .…”