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通过对莲子中超氧化物歧化酶(Super Oxide Dimutase,SOD)进行分离提纯,证实其是Fe-SOD,并观察Fe-SOD的耐热性,为SOD的提取和应用开辟了新的思路【。方法】以热变性与有机溶剂沉淀联合法分离提纯莲子Fe-SOD;利用Fe-SOD活性受H2O2的抑制而不受氰化物的抑制的特性来鉴定;设置不同的温度观察提取额莲子SOD的耐热性。【结果】莲子提取酶液活性为90.522U/ml,SOD酶液对邻苯三酚自氧化的抑制率为96.3%。莲子SOD活性受H2O2的抑制而不受氰化物的抑制。在75℃时保温30min后,酶活力仍然保留60%以上,在100℃保温30min时酶活力保留35%。【结论】以热变性与有机溶剂沉淀联合法分离提纯莲子SOD并鉴定其为耐热性Fe-SOD。
Through the isolation and purification of superoxide dismutase (SOD) in lotus seeds, it was confirmed that it is Fe-SOD, and the heat resistance of Fe-SOD was observed, which opened a new way for the extraction and application of SOD. Methods The lotus seed Fe-SOD was isolated and purified by a combination of thermal denaturation and organic solvent precipitation. The activity of Fe-SOD was identified by the inhibition of H2O2 but not by the inhibition of cyanide. Different temperatures were used to extract the SOD of lotus seed Thermal. 【Result】 The results showed that the enzyme activity of lotus seed extract was 90.522 U / ml. The inhibition rate of SOD enzyme solution to autoxidation of pyrogallol was 96.3%. The lotus SOD activity is inhibited by H2O2 but not by cyanide. After incubated at 75 ℃ for 30min, the enzyme activity remained above 60%, and the enzyme activity retained 35% when incubated at 100 ℃ for 30min. 【Conclusion】 The lotus seed SOD was isolated and purified by thermal denaturation combined with organic solvent precipitation and identified as heat-tolerant Fe-SOD.