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富碲钝顶螺旋藻中含碲藻胆蛋白的纯化及其体外抗氧化活性

Purification and in vitro antioxidant activities of tellurium-containing phycobiliproteins from tellurium-enriched Spirulina platensis.

作者信息

Yang Fang, Wong Ka-Hing, Yang Yufeng, Li Xiaoling, Jiang Jie, Zheng Wenjie, Wu Hualian, Chen Tianfeng

机构信息

Department of Chemistry, Jinan University, Guangzhou, People's Republic of China.

Department of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, Hong Kong, People's Republic of China.

出版信息

Drug Des Devel Ther. 2014 Oct 9;8:1789-800. doi: 10.2147/DDDT.S62530. eCollection 2014.

Abstract

Tellurium-containing phycocyanin (Te-PC) and allophycocyanin (Te-APC), two organic tellurium (Te) species, were purified from tellurium-enriched Spirulina platensis by a fast protein liquid chromatographic method. It was found that the incorporation of Te into the peptides enhanced the antioxidant activities of both phycobiliproteins. With fractionation by ammonium sulfate precipitation and hydroxylapatite chromatography, Te-PC and Te-APC could be effectively separated with high purity, and Te concentrations were 611.1 and 625.3 μg g(-1) protein in Te-PC and Te-APC, respectively. The subunits in the proteins were identified by using MALDI-TOF-TOF mass spectrometry. Te incorporation enhanced the antioxidant activities of both phycobiliproteins, as examined by 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid assay. Moreover, Te-PC and Te-APC showed dose-dependent protection on erythrocytes against the water-soluble free radical initiator 2,2'-azo(2-asmidinopropane)dihydrochloride-induced hemolysis. In the hepatoprotective model, apoptotic cell death and nuclear condensation induced by tert-butyl hydroperoxide in HepG2 cells was significantly attenuated by Te-PC and Te-APC. Taken together, these results suggest that Te-PC and Te-APC are promising Te-containing proteins with application potential for treatment of diseases related to oxidative stress.

摘要

通过快速蛋白质液相色谱法从富碲钝顶螺旋藻中纯化出含碲藻蓝蛋白(Te-PC)和别藻蓝蛋白(Te-APC)这两种有机碲(Te)物种。研究发现,碲掺入肽中增强了两种藻胆蛋白的抗氧化活性。通过硫酸铵沉淀和羟基磷灰石色谱分级分离,可有效分离出高纯度的Te-PC和Te-APC,Te-PC和Te-APC中碲的浓度分别为611.1和625.3 μg g(-1)蛋白质。使用基质辅助激光解吸电离飞行时间串联质谱法鉴定蛋白质中的亚基。如通过2,2'-联氮-双-(3-乙基苯并噻唑啉-6-磺酸)测定法所检测,碲掺入增强了两种藻胆蛋白的抗氧化活性。此外,Te-PC和Te-APC对红细胞免受水溶性自由基引发剂2,2'-偶氮(2-脒基丙烷)二盐酸盐诱导的溶血具有剂量依赖性保护作用。在肝保护模型中,Te-PC和Te-APC显著减轻了叔丁基过氧化氢在HepG2细胞中诱导的凋亡细胞死亡和核浓缩。综上所述,这些结果表明Te-PC和Te-APC是有前景的含碲蛋白,在治疗与氧化应激相关疾病方面具有应用潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/46ef/4199980/6079d5aa598e/dddt-8-1789Fig2.jpg

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