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。
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 (NH_4)_2SO_4 to long-time storage for R-PE was also studied.
出处
《分析试验室》
CAS
CSCD
北大核心
2004年第11期5-9,共5页
Chinese Journal of Analysis Laboratory
关键词
R-藻红蛋白
色谱
纯化
光谱
R-phycoerythrin
Chromatography
Purification
Spectrum
