Separation of the Glycosylated Carotenoid Myxoxanthophyll from Synechocystis Salina by HPCCC and Evaluation of Its Antioxidant, Tyrosinase Inhibitory and Immune-Stimulating Properties

Abstract

Global demand for natural pigments has increased in the past few years. Myxoxanthophyll, a glycosylated monocyclic carotenoid, is a pigment that occurs naturally in cyanobacteria but no scalable isolation process has been developed to obtain it from its natural source to date. In this study, myxoxanthophyll was isolated from unicellular cyanobacterium Synechocystis salina (S. salina) using high-performance countercurrent chromatography (HPCCC), where the lower phase of the biphasic solvent system composed of n-heptane–ethanol–water (2:4:4, v/v/v) was used as a mobile phase, whereas its upper phase was the stationary phase. For the HPCCC isolation, a multi-injection method was developed, and four consecutive sample injections (70 mg each) were performed, obtaining, in total, 20 mg of myxoxanthophyll, which was finally purified with high-performance liquid chromatography (HPLC). Overall, a final myxoxanthophyll yield of 15 mg (98% purity) was obtained. The target pigment showed a weak antioxidant and tyrosinase inhibitory effect, and exhibited immune-stimulating properties by activating human granulocytes. The results presented here form a basis for the large-scale production of myxoxanthophyll, and show the potential benefits of this pigment for human health.

Production of Fucoxanthin from Phaeodactylum tricornutum Using High Performance Countercurrent Chromatography Retaining Its FOXO3 Nuclear Translocation-Inducing Effect

“…This last compound is more likely to be 13- or 13′- cis -fucoxanthin, in line with its hypsochromic shift peculiarity and the intensity of the cis peak (D B /D II : 47.0%), as earlier published [ 59 ]. The cis isomers of carotenoids have been shown to be commonly generated from (All- trans )-carotenoids by light and temperature effects and constitute no risk to human health [ 45 , 47 ].…”

“…The formed upper and lower phases were separated and used for the calculation of the K value using HPLC-DAD. The K value was calculated as the ratio between the fucoxanthin peak area in the upper phase to that of the lower phase [ 47 , 48 ]. The measurement of the settling time was performed as previously reported [ 47 ].…”

“…The K value was calculated as the ratio between the fucoxanthin peak area in the upper phase to that of the lower phase [ 47 , 48 ]. The measurement of the settling time was performed as previously reported [ 47 ]. The density differences between the two immiscible phases were calculated by weighting 1 mL of each phase using a microbalance [ 43 ].…”

“…Given that this technology lacks solid support, it has many advantages over the solid chromatographic techniques including large sample loading capacity, low risk of irreversible adsorption and sample denaturation, high sample recovery, low consumption of solvents and great operational versatility, since the roles of mobile and stationary phases can be exchanged during the chromatographic operation [ 40 , 41 , 42 ]. The capacity of countercurrent chromatography for obtaining valuable compounds from microalgae biomass has been widely demonstrated [ 42 , 43 , 44 , 45 , 46 , 47 , 48 ] and its application at industrial level is already a reality [ 49 ]. High-speed countercurrent chromatography (HSCCC) has been used for the isolation of fucoxanthin from edible brown macroalgae species such as Laminaria japonica , Undaria pinnatifida and Sargassum fusiforme [ 50 ] applying a single sample injection method.…”

Influence of Cultivation Conditions on the Sioxanthin Content and Antioxidative Protection Effect of a Crude Extract from the Vegetative Mycelium of Salinispora tropica

“…The glycosylated carotenoid myxoxanthophyll was also found in the cyanobacterium Synechocystis sp. [ 20 ], and exhibited immune-stimulating properties by activating human granulocytes [ 21 ].…”