Abstract
This is the first report to purify and characterize POX in shoots of buckwheat species. POX was partially purified from Tartary buckwheat shoots by 48.0 fold with a final yield of 9.07%. During ion-exchange and gel filtration chromatography,only one peak corresponding to POX activity was found. The molecular weight of POX was determined to be 37.5 kDa using gel filtration chromatography. The optimal pH of POX activity was 5.5 (guaiacol, quercetin) and 5.0 (ABTS). The Km of POX activity was 22.3 mM (guaiacol), 6.3 mM (ABTS) and 0.92 mM (quercetin). In contrast, the Km for quercetin in the presence of Fe3+ions was two orders of magnitude less (0.018 mM) than that in its absence. The stability of POX activity was increased in the presence of trivalentmetal ions, even after 186 h in solution. POX activity was retained by 83.6% and 56.1% in the presence of 1 mMFe3+and Al3+ions, respectively, whereasit was completely inactivated in their absence. To the best of our knowledge, this is thefirst study to detail the activation and stabilization of POX activity in relation to trivalent metal ions.
Highlights
Buckwheat is a rich source of functional polyphenols such as rutin and quercetin [1,2,3,4]
Tartary buckwheat (Fagopyrum tataricum) shoots contain a substantial amount of peroxidase (POX) activity, the characteristics and physiological roles of which have not been studied
POX in Tartary buckwheat shoot was purified by ammonium sulfate precipitation, cation-exchange chromatography and gel filtration (Table 1)
Summary
Buckwheat is a rich source of functional polyphenols such as rutin and quercetin [1,2,3,4]. Tartary buckwheat (Fagopyrum tataricum) shoots contain a substantial amount of peroxidase (POX) activity, the characteristics and physiological roles of which have not been studied. D. Mikami et al / Journal of Agricultural Chemistry and Environment 2 (2013) 59-64 tein have been reported to contain metal ions, but Ca2+ sometimes inhibited POX activities [23]. Mikami et al / Journal of Agricultural Chemistry and Environment 2 (2013) 59-64 tein have been reported to contain metal ions, but Ca2+ sometimes inhibited POX activities [23] These reports revealed the importance concerning the effects of metal ions on POX activity in studying the physiological roles of POX. We investigated the effects of metal ions on purified POX activities as well
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