In this study, we focused on natural water-soluble antioxidants from Tetraselmis suecica (T. suecica) and Chlorella ellipsoidea (C. ellipsoidea). They were prepared by enzymatic digestion using five carbohydrases (Viscozyme, Celluclast, AMG, Termamyl and Ultraflo) and five proteases (Protamex, Alcal...
In this study, we focused on natural water-soluble antioxidants from Tetraselmis suecica (T. suecica) and Chlorella ellipsoidea (C. ellipsoidea). They were prepared by enzymatic digestion using five carbohydrases (Viscozyme, Celluclast, AMG, Termamyl and Ultraflo) and five proteases (Protamex, Alcalase, Flavourzyme, Neutrase, and Kojizyme), and the potential antioxidant activity of each was assessed. Most enzymatic digests from T. suecica had a higher radical scavenging activity than those from C. ellipsoidea. Among the enzymatic digests, Kojizyme digest from T. suecica exhibited the highest effect on DPPH radical scavenging. Viscozyme (30.2%) and Neutrase (34.6%) digests from T. suecica exhibited higher hydroxyl radical scavenging activity. Kojizyme digest from T. suecica (81.5%) had strong alkyl radical scavenging activity. Neutrase (61.9%) and Kojizyme (61.5%) digest from T. suecica possessed the highest effects on hydrogen peroxide scavenging. Among the tested samples, Neutrase (TN) and Kojizyme (TK) digests from T. suecica showed the highest antioxidant activity (DPPH, alkyl radical, hydrogen peroxide). Therefore, TN and TK digests were selected for use in the further experiments. Those digests showed enhanced cell viability against $H_2O_2$-induced oxidative damage, and relatively good hydrogen peroxide scavenging activity in an African green monkey kidney (Vero) cell line. These results suggested that an enzymatic digestion will be an effective way for the production of a potential water-soluble antioxidant from a microalgae, T. suecica.
In this study, we focused on natural water-soluble antioxidants from Tetraselmis suecica (T. suecica) and Chlorella ellipsoidea (C. ellipsoidea). They were prepared by enzymatic digestion using five carbohydrases (Viscozyme, Celluclast, AMG, Termamyl and Ultraflo) and five proteases (Protamex, Alcalase, Flavourzyme, Neutrase, and Kojizyme), and the potential antioxidant activity of each was assessed. Most enzymatic digests from T. suecica had a higher radical scavenging activity than those from C. ellipsoidea. Among the enzymatic digests, Kojizyme digest from T. suecica exhibited the highest effect on DPPH radical scavenging. Viscozyme (30.2%) and Neutrase (34.6%) digests from T. suecica exhibited higher hydroxyl radical scavenging activity. Kojizyme digest from T. suecica (81.5%) had strong alkyl radical scavenging activity. Neutrase (61.9%) and Kojizyme (61.5%) digest from T. suecica possessed the highest effects on hydrogen peroxide scavenging. Among the tested samples, Neutrase (TN) and Kojizyme (TK) digests from T. suecica showed the highest antioxidant activity (DPPH, alkyl radical, hydrogen peroxide). Therefore, TN and TK digests were selected for use in the further experiments. Those digests showed enhanced cell viability against $H_2O_2$-induced oxidative damage, and relatively good hydrogen peroxide scavenging activity in an African green monkey kidney (Vero) cell line. These results suggested that an enzymatic digestion will be an effective way for the production of a potential water-soluble antioxidant from a microalgae, T. suecica.
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문제 정의
The objective of this study was to investigate the antioxidant properties of enzymatic digests using five carbohydrases (Viscozyme, Celluclast, AMG, Termamyl and Ultraflo) and five proteases (Protamex, Kojizyme, Neutrase, Flavourzyme and Alcalase) from Tetraselmis suecica and Chlorella ellipsoidea in order to understand the usefulness of these algae in food and pharmaceutical industry.
제안 방법
Hydrogen peroxide scavenging assay by DCF-DA
As Neutrase (TN) and Kojizyme (TK) digests from T. suecica possessed the highest radical scavenging activity, this digests was further evaluated for its H2O2 scavenging activity by DCF-DA. The effect of the TN and TK on H2O2 scavenging activity by DCFH-DA is exhibited in Fig.
03% at the concentration of 200 μg/mL, and the scavenging activities increased when increasing the sample concentration. As the TN and TK generated in this study evidenced such good H2O2 radical scavenging activity, this samples was evaluated further with regard to its protective effects against H2O2-induced cellular damage. The protective effects of TN and TK on cell viability in H2O2-induced Vero cells were measured via an MTT assay.
The exposure of cultured cells to H2O2 results in an imbalance in energy metabolism and the deleterious effects of hydroxyl and peroxyl radicals on membrane lipids and proteins. Therefore, in this study, we investigated scavenging activity of the TN and TK after the administration of H2O2 treatment in Vero cell lines. The DCF-DA method was used to detect the intracellular hydrogen peroxide level (36).
이론/모형
Proximate chemical composition of freeze-dried samples was determined according to the AOAC methods (17). Crude lipid content was determined by Soxhlet method and crude protein content was determined by Kjeldhal method. Ash content was determined by calcinations in furnace at 550℃, the moisture content was determined by keeping in a dry oven at 105℃ for 24 hr and Crude carbohydrate content was determined by phenol-sulfuric acid reaction (absorbance at 480 nm, using glucose as the calibration standard)
DPPH radical scavenging activity was measured using the method described by Nanjo et al. (19). Methanol solution of 60 μL of each sample (or methanol itself as control) was added to 60 μL of DPPH (60 μmol/L) in methanol.
Hydrogen peroxide scavenging activity was determined according to the method of Muller (22). A 100 μL of 0.
Proximate chemical composition of freeze-dried samples was determined according to the AOAC methods (17). Crude lipid content was determined by Soxhlet method and crude protein content was determined by Kjeldhal method.
Therefore, in this study, we investigated scavenging activity of the TN and TK after the administration of H2O2 treatment in Vero cell lines. The DCF-DA method was used to detect the intracellular hydrogen peroxide level (36). DCF-DA diffuses into cells, where it is hydrolyzed by intracellular esterase to polar 2′,7′-dichlorodihydrofluorescein.
suecica on scavenging intracellular hydrogen peroxide. The intracellular hydrogen peroxide generated was detected by DCF-DA method. T-N; Neutrase digest from T.
Total polyphenolic compounds in the digests were determined with Folin-Ciocalteu reagent according to the method of Chandler and Dodds (18) using gallic acid as a standard phenolic compound. Sample (1 mL) was mixed with 1 mL of 95% ethanol, 5 mL of distilled water and 0.
성능/효과
ellipsoidea. All the enzymatic digests of T. suecica except AMG digest exhibited more than 40% H2O2 scavenging activity, and proteases-treated digests exhibited greater effects than carbohydrases-treated digests, suggesting that proteases have more capability to liberate H2O2 scavenging compounds. Specifically, the Neutrase and Kojizyme digests evidenced more profound activities (～61.
ellipsoidea is shown in Table 3. As shown in the results, remarkable scavenging effects of all the enzymatic digests were observed in alkyl radical scavenging assay, compared to the other scavenging assays. Several digests yielded approximately 75% scavenging activities.
Enzymatic digests exhibited appreciable antioxidative activities. In conclusion, Tetraselmis suecica and Chlorella ellipsoidea exhibited antioxidant potential against ROS and it would be a potential candidate as a natural water-soluble antioxidative source, which can be applied in food or food related industries.
DPPH is stable free radical and accepts an electron or hydrogen radical to become a stable diamagnetic molecule, and has often been used as a substrate to evaluate the antioxidative activity of natural compounds (30). In this study, DPPH free radical scavenging activity of proteases digests from T. suecica and C. ellipsoidea was relatively higher than the digests by treated with carbohydrases. Especially, Kojizyme digests of T.
(32) reported a high correlation between DPPH radical scavenging activities and total polyphenolics. In this study, Neutrase digests of T. suecica showed a lower DPPH free radical scavenging activity than Kojizyme digest, although that contained as much phenolic compounds as the Kojizyme digests. It is thought that other bioactive components, such as proteins, polysaccharides and different kinds of pigments.
Despite a variety of biological functions of microalgae, effective application of microalgae is still limited because of their inefficient cultivation and high production cost. Therefore, in this study, we selected two marine microalgae, Tetraselmis suecica and Chlorella ellipsoidea that can be massively cultivated and low production cost. Also, it has been reported that Tetraselmis suecica and Chlorella ellipsoidea exhibits antimicrobial activity and functional biomaterial (14,15).
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