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去污剂对裙带菜色素-蛋白质复合物电泳分离效果及其性质的影响
李爱芬, 陈敏, 周百成
中国科学院海洋研究所 青岛266071
摘要:
于1998年3月在青岛市太平角采集裙带菜孢子体,通过破碎、离心制备类囊体膜。用去污剂SDS、DOC、DMG和DIG增溶裙带菜类囊体膜,经PAGE分离色素-蛋白质复合物,并测定其光谱特性。结果表明,电泳分离效果与所用的去污剂的种类有关。去污剂不同,分离到的同一类型复合物的吸收光谱有差异,捕光色素-蛋白质复合物的数目、种类及吸收光谱明显不同。电泳中产生的游离色素的多少也随去污剂种类而异。在4种去污剂中,以DMG和DIG增溶的效果最好,经SDS-PAGE分出6种色素-蛋白质复合物,游离色素较少。
关键词:  裙带菜  去污剂  凝胶电泳  色素-蛋白质复合物  吸收光谱  荧光光谱
DOI:
分类号:
基金项目:国家“九五”攻关资助项目,96-C01-05-01号
EFFECT OF DETERGENTS ON ELECTROPHORETIC SEPARATION AND PROPERTIES OF PIGMENT-PROTEIN COMPLEXES FROM A BROWN ALGA UNDARIA PINNATIFIDA
LI Ai-Fen, CHEN Min, ZHOU Bai-Cheng
Institute of Oceanology,The Chinese Academy of Sciences,Qingdao 266071
Abstract:
The thylakiod membranes isolated from a brown alga Undaria pinnatifida were solubilized by detergents sodium dodecyl sulfate (SDS), deoxycholate (DOC), decanoyl-N-methylglucamide (DMG) and digitonin (DIG) at different concentrations (SDS:Chl=10:1, DOC:Chl=60:1, DMG:Chl=20:1, DIG:Chl =50:1). The pigment-protein complexes were separated by the SDS-PAGE, and their spectrum characteristics were determined. The results of PAGE separation varied with detergents. DOC solubilization yielded 10 pigment-containing bands and resulted in a large number of free pigments. SDS solubilization yielded only 5 pigment-containing bands and lots of free pigment were produced. Comparing with SDS and DOC, DMG and DIG solubilization resulted in 7 pigment-containing bands and only a little free pigment. When thylakoid membranes were solubilized by different kinds of detergents and separated by SDS-PAGE, the same pigment-protein complexes had different absorption and fluorescence emission spectra. The absorption maxima of chlorophyll-a in PSI complexes were different in blue and red lights, and its short-wavelength fluorescence emission maxima at 77K also varied with detergent. PSI I complexes treated with DMG or DIG had absorption of fucoxinthin. The light-harvesting pigment-protein complexes (LHC) were also different in number, type and absorption spectrum. Two Chl a/c-protein complexes were resolved by SDS and four by DOC, but no fucoxinthin-Chl a/c-protein complexes (FCPC) were separated by SDS or DOC. DMG or DIG solubilization can yield three types of LHC, of which were two FCPC and one Chl a/c-protein complexes. According to the PAGE results, the best separation was obtained by using DMG or DIG, in which 6 pigment-protein complexes were resolved and a little free pigment were produced.
Key words:  Undaria pinnatifida, Detergent, SDS-PAGE, Pigment-protein complexes, Absorption spectra, Fluorescence emissions spectra
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