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坛紫菜中R-藻红蛋白的色谱纯化及性质研究

Chromatographic Purification and Characteristic of R-phycoerythrin from Porphyra Haitanensis

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【作者】 陈小强龚兴国史锋潘远江

【Author】 CHEN Xiao-qiang, GONG Xing-guo, SHI Feng(Department of Biotechnology,Zhejiang Univesity, Hangzhou 310027) and PAN Yuan-jiang (Department of Chemistry,Zhejiang Univesity, Hangzhou 310027)

【机构】 浙江大学生命科学学院生物技术系浙江大学理学院化学系 杭州310027杭州310027杭州310027

【摘要】 从坛紫菜(Porphyrahaitanensis)藻胆蛋白粗提液中分离纯化R 藻红蛋白(R phycoerythrin,R PE),经过两次不同柱长规格的自制羟基磷灰石柱层析和一次DEAE SepharoseFastFlow离子交换层析,获得了较高纯度的R PE(A565nm A280nm>5.3);测定了R PE的紫外可见吸收光谱和荧光光谱,R PE的吸收峰在565和498nm,在540nm有一吸收肩,荧光发射峰在573nm;用SDS PAGE测定组成R PE各亚基的相对分子质量,α β亚基约为18~20kDa,γ亚基约为35kDa;对R PE在不同的pH和温度环境协同影响下的荧光稳定性进行了研究,摸索了R PE的长期有效保存的方法并探讨了(NH4)2SO4的保护机理。

【Abstract】 R-Phycoerythrin was isolated and purified from red alga, Porphyra haitanensis. After successive hydroxylapatite (HA) column chromatography and DEAE-Sepharose Fast Flow ion-exchange column chromatography separations,the purity(A565 nm/A280 nm) was more than 5.3. Absorption spectra of purified R-PE showed characteristic peaks at 565 nm and 498 nm, and a shoulder at 540 nm, and fluorescence emission spectra displayed a peak at 573 nm. The relative molecular weights of subunits consisting in R-PE were detected by SDS-PAGE, α/β subunit is about 18~20 kDa,and γ about 35 kDa; R-PE stability was investigated using a large range of pH cooperated with various temperatures, and the effect of (NH42SO4 to long-time storage for R-PE was also studied.

【关键词】 R藻红蛋白色谱纯化光谱
【Key words】 R-phycoerythrinChromatographyPurificationSpectrum
  • 【文献出处】 分析试验室 ,Analytical Laboratory , 编辑部邮箱 ,2004年11期
  • 【分类号】O657.8
  • 【被引频次】11
  • 【下载频次】298
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