Construction and Characterization of B850-Only LH2 Energy Transfer System in Purple Bacteria
LI Kai1, ZHAO Chun-gui1*, YUE Hui-ying1, YANG Su-ping1*, QU Yin-bo2, JIAO Nian-zhi3
1. Department of Bioengineering and Biotechnology, Huaqiao University, Xiamen 361021, China 2. State Key Laboratory of Microbial Technology, Shandong University, Ji’nan 250100, China 3. State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen 361005, China
Abstract：To seek microscopic molecular mechanism of energy transfer and complex reconstitution in the photosynthesis, the conditions for construction of B850-only peripheral light-harvesting complex (LH2) and their properties were investigated using absorption, fluorescence spectroscopy, molecular sieve chromatography, ultrafiltration and sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) from the purple bacteria. The results indicated that bacteriochlorophylls (BChl) of B800 incubated in 10 mmo·L-1 Tris-HCl (pH 8.0) buffer are selectively released from their binding sites of LH2 of Rhodobacter azotoformans (A-LH2) by 0.08% (W/V) SDS. B850-only A-LH2 was constructed after removing free BChl mixing with 10% methyl alcohol by ultrafiltration. B850 BChl was released after A-LH2 was incubated for 240 min in dark at room temperature (RT). While BChl of B800 incubated in pH 1.9 buffer were selectively released from their binding sites of LH2 of Rhodopseudomonas palustris (P-LH2). The authors acquired two components using molecular sieve chromatography. Free BChl of one component was not removed and self-assembled to P-LH2. The other removed free BChl and B850-only P-LH2 was constructed. B850 unchanged after P-LH2 was incubated. P-LH2 α and β subunits have different molecular weights, but those of A-LH2 are in the contrary. It is concluded that B850-only P-LH2 is more stable than A-LH2. The enigmatic split of the B800 absorption band was not observed in these LH2, but we acquired two kinds of B800-released LH2 from Rhodopseudomonas palustris. The authors’ results may provide a new light to separate homogeneous Apoprotein LH2.
李 凯1，赵春贵1*，岳慧英1，杨素萍1*，曲音波2，焦念志3 . 紫细菌B800缺失LH2能量传递模型的构建及性质 [J]. 光谱学与光谱分析, 2015, 35(04): 875-880.
LI Kai1, ZHAO Chun-gui1*, YUE Hui-ying1, YANG Su-ping1*, QU Yin-bo2, JIAO Nian-zhi3 . Construction and Characterization of B850-Only LH2 Energy Transfer System in Purple Bacteria . SPECTROSCOPY AND SPECTRAL ANALYSIS, 2015, 35(04): 875-880.
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