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目的:研究锌胁迫诱导小球藻类金属硫蛋白的抗氧化活性。方法:小球藻经60μmol/L锌诱导胁迫培养8d,离心收集藻细胞;超声波破碎细胞,离心,上清液经凝胶层析柱(葡聚糖G-75)及脱盐柱(葡聚糖G-25)分离、纯化,得到锌结合金属蛋白;经光谱特征分析,确认为类金属硫蛋白(Zn-MT-like)。对Zn-MT-like的抗氧化活性进行比较研究,包括基于Fenton反应体系的羟自由基(·OH),1,1-二苯基苦基苯肼(DPPH)产生的自由基和邻苯三酚自氧化产生的超氧阴离子(·O2-)自由基的清除能力。结果:小球藻诱导产生的Zn-MT-like对这3种自由基均有较强的清除作用,其清除羟自由基、DPPH自由基和超氧阴离子自由基的半数抑制率浓度(IC50)分别为13.12、2.23和48.59mg/L;其抗羟自由基、DPPH自由基和超氧阴离子自由基的能力高于谷胱苷肽和兔肝锌诱导的金属硫蛋白(Zn-MTs)。结论:小球藻锌结合类金属硫蛋白具有显著的抗氧化活性能力。
Objective: To study the anti-oxidative activity of ZnS in chlorella induced by zinc stress. Methods: The chlorella was cultured under the stress of 60μmol / L zinc for 8 days. The cells were collected by centrifugation. The cells were sonicated and centrifuged. The supernatant was separated by gel column (dextran G-75) and desalting column G-25) to obtain the zinc-binding metalloprotein. After spectral characterization, it was confirmed as Zn-MT-like. The antioxidant activities of Zn-MT-like were compared, including hydroxyl radical (· OH) based on Fenton reaction system, free radicals produced by 1,1-diphenylpicrylphenylhydrazine (DPPH) Scavenging ability of superoxide anion (· O2-) radical produced by phenol autoxidation. Results: Zn-MT-like induced by Chlorella vulgaris had a strong scavenging effect on these three free radicals. The half-inhibitory concentration (IC50) of hydroxyl radical scavenging, DPPH free radical and superoxide anion radical scavenging activity of Zn- Which were 13.12, 2.23 and 48.59 mg / L, respectively. The ability of anti-hydroxyl radical, DPPH radical and superoxide anion radical was higher than that of glutathione and rabbit hepatic zinc-induced metallothionein (Zn-MTs). Conclusion: Chlorella zinc-binding metallothionein has significant antioxidant activity.