The objective of the present study is to develop a rapid and convenient method to determine antioxidative activity. It was determined by the inhibition capacity on the hemoglobin-catalyzed peroxidation of linoleic acid. The appropriate conditions for reaction of 4 mM linoleic acid were 0.002% hemoglobin at 37 degrees C for 10 min. Adding methanol to the reaction mixture at <20% showed no significant effect on the peroxidation of linoleic acid. Products formed from hemoglobin-catalyzed peroxidation of linoleic acid were 9- and 13-hydroperoxyoctadecadienoic acid at a ratio of approximately 50:50. Eight synthetic antioxidants were assayed for their antioxidative activity; all of them showed linear response to the logarithm of their concentration. Antioxidative activity from different plant samples was also examined. Tea, ginger, chrysanthemum, and roselle showed higher antioxidative activity. Either hydrophobic or hydrophilic antioxidants were able to be assayed with this method within 15 min.
The objective of the present study was to purify and characterize the lipoxygenase (LOX) from banana leaf (Giant Cavendishii, AAA), an unutilized bioresource. LOX was extracted, isolated, and purified 327-fold using 25-50% saturation of ammonium sulfate fractionation, hydroxyapatite column separation, and gel filtration on Superdex 200. The molecular mass of the purified LOX was 85 kDa, K(m) was 0.15 mM, and V(max) was 2.4 microM/min.mg using linoleic acid as substrate. Triton X-100 was required in the extraction medium; otherwise, no LOX activity was detected. LOX activity increased with the concentration of Triton X-100 with an optimum at 0.1%. The optimal pH of the purified LOX from banana leaf was 6.2, and optimal temperature was 40 degrees C. The LOX showed the highest reactivity toward 18:2 followed by 18:3 and 20:4. A very low reaction rate was observed toward 20:5 and 22:6. On the basis of retention time in normal phase HPLC, the products of 18:2 or 18:3 catalyzed by purified LOX were hydroperoxyoctadecadienoic acid or hydroperoxyoctadecatrienoic acid. It seems that 9-LOX is the predominant enzyme in banana leaf. Banada leaf dried at 110 degrees C for 2 h developed algal aroma. Banana leaf extract stored at 10 degrees C for 12 h formed an oolong tea-like flavor. Banana leaf extract reacted with 18:2 or soybean oil pretreated with bacterial lipase produced green and melon-like aroma, whereas the same reaction with 18:3 produced a sweet, fruity, cucumber-like flavor note.
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