caa a22 4500 445890134 CHVBK 20180317145613.0 cr unu---uuuuu 170323e20110901xx s 000 0 eng 10.1007/s11120-011-9679-4 doi (NATIONALLICENCE)springer-10.1007/s11120-011-9679-4 A novel and mild isolation procedure of chlorosomes from the green sulfur bacterium Chlorobaculum tepidum [Elektronische Daten] [Seiji Tokita, Keizo Shimada, Kazuyuki Watabe, Katsumi Matsuura, Mamoru Mimuro] In this article, we developed a new and mild procedure for the isolation of chlorosomes from a green sulfur bacterium Chlorobaculum tepidum. In this procedure, Fenna-Matthews-Olson (FMO) protein was released by long cold treatment (6°C) of cells under the presence of a chaotrope (2M NaSCN) and 0.6M sucrose. Chlorosomes were released by an osmotic shock of the cold-treated cells after the formation of spheroplasts without mechanical disruption. Chlorosomes were finally purified by a sucrose step-wise density gradient centrifugation. We obtained two samples with different density (20 and 23% sucrose band, respectively) and compared them by SDS-PAGE, absorption spectroscopy at 80K, fluorescence and CD spectroscopy at room temperature. Cells whose absorption maximum was longer than 750nm yielded higher amount of the 20% sucrose fraction than those having an absorption maximum shorter than 750nm. Springer Science+Business Media B.V., 2011 Isolation method nationallicence Chlorosomes nationallicence FMO protein nationallicence Low-temperature absorption nationallicence Chlorobaculum tepidum nationallicence BChl : Bacteriochlorophyll nationallicence FMO protein : Fenna-Matthews-Olson protein nationallicence EDTA : Ethylenediamine-N,N,N′,N′-tetraacetic acid nationallicence MOPS : 3-Morpholinopropanesulfonic acid nationallicence Tokita Seiji Department of Biology, Tokyo Metropolitan University, 192-0397, Tokyo, Japan aut Shimada Keizo Department of Biology, Tokyo Metropolitan University, 192-0397, Tokyo, Japan aut Watabe Kazuyuki Graduate School of Human and Environmental Studies, Kyoto University, 606-8501, Kyoto, Japan aut Matsuura Katsumi Department of Biology, Tokyo Metropolitan University, 192-0397, Tokyo, Japan aut Mimuro Mamoru Graduate School of Human and Environmental Studies, Kyoto University, 606-8501, Kyoto, Japan aut Photosynthesis Research Springer Netherlands 108/2-3(2011-09-01), 183-190 0166-8595 108:2-3<183 2011 108 11120 https://doi.org/10.1007/s11120-011-9679-4 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11120-011-9679-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Tokita Seiji Department of Biology, Tokyo Metropolitan University, 192-0397, Tokyo, Japan aut NATIONALLICENCE 700 1- Shimada Keizo Department of Biology, Tokyo Metropolitan University, 192-0397, Tokyo, Japan aut NATIONALLICENCE 700 1- Watabe Kazuyuki Graduate School of Human and Environmental Studies, Kyoto University, 606-8501, Kyoto, Japan aut NATIONALLICENCE 700 1- Matsuura Katsumi Department of Biology, Tokyo Metropolitan University, 192-0397, Tokyo, Japan aut NATIONALLICENCE 700 1- Mimuro Mamoru Graduate School of Human and Environmental Studies, Kyoto University, 606-8501, Kyoto, Japan aut NATIONALLICENCE 773 0- Photosynthesis Research Springer Netherlands 108/2-3(2011-09-01), 183-190 0166-8595 108:2-3<183 2011 108 11120 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer