Antioxidant activities of two metallothionein-like proteins from sweet potato (Ipomoea batatas [L.] Lam. `Tainong 57') storage roots and their synthesized peptides.
キーワード
概要
BACKGROUND
Metallothionein (MT) characterized by their low molecular weight and high cysteine content.
RESULTS
Two recombinant proteins of MT-I and MT-II overproduced in E. coli (M15) was purified by Ni2+-chelated affinity chromatography. The molecular mass of MT-I and MT-II are ca. 6,600 and 8,000 Da as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Total antioxidant status, DPPH radical scavenging activity, reducing power method, Fe2+-chelating ability, ferric thiocyanate (FTC) method, and protecting calf thymus DNA against hydroxyl radical-induced damage were studied. The MT-I and MT-II proteins with a concentration of 100 μg/mL exhibited the highest activity (expressed respectively as 61.72 ± 0.13 and 74.28 ± 1.15 μM Trolox equivalent antioxidative capacity, TEAC) in total antioxidant status test. Like total antioxidant status, DPPH radical scavenging activity, reducing power, Fe2+-chelating ability, FTC activity, and protecting calf thymus DNA against hydroxyl radical-induced damage all showed that MT-1 and MT-II proteins have antioxidant activities. In this study, we also found that antioxidant activities of MT-I and MT-II increased from 17% and 16% (0 h) to about 26% and 28% (24 h) after 24 h hydrolysis by trypsin. Smaller peptides increased the antioxidant activities. Four and three peptides, respectively, from MT-I and MT-II protein sequences for testing antioxidative activity were synthesized according to tryptic hydrolysis simulation. The obtained MSSGCK, CGSDCK, LTLEGSSEK, ATEGGHACK, CGNGCGGCK, and CDPCNCK showed IC50 values of 309.87, 1423.37, 3925.54, 561.32, 300.76, and 610.12 μM, respectively, when scavenging activity of DPPH radicals (%) was measured.
CONCLUSIONS
These findings mean that a cysteine residue is most important in antiradical activities. It was suggested that MT-I and MT-II might contribute their antioxidant activities against hydroxyl and peroxyl radicals.
