Purification, characterisation and crystallisation of photosystem II from Thermosynechococcus elongatus cultivated in a new type of photobioreactor
| Autor: | Jan Kern, Bernhard Loll, Athina Zouni, C. Lüneberg, K.-D. Irrgang, Jacek Biesiadka, D. DiFiore |
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| Rok vydání: | 2005 |
| Předmět: |
Pheophytin
Photosystem II Biophysics Photobioreactor macromolecular substances Biology Cyanobacteria Thylakoids Biochemistry Cofactor composition chemistry.chemical_compound Pigment X-Ray Diffraction Biomass Chromatography High Pressure Liquid Thermosynechococcus elongatus Oxygen evolution activity Spectrophotometry Atomic Oxygen evolution Photosystem II Protein Complex Fast protein liquid chromatography Cell Biology Crystallisation Oxygen Crystallography Monomer chemistry Thylakoid visual_art visual_art.visual_art_medium Cell cultivation Electrophoresis Polyacrylamide Gel Crystallization Chromatography Liquid |
| Zdroj: | Biochimica et Biophysica Acta (BBA) - Bioenergetics. 1706(1-2):147-157 |
| ISSN: | 0005-2728 |
| DOI: | 10.1016/j.bbabio.2004.10.007 |
| Popis: | The thermophilic cyanobacterium Thermosynechococcus elongatus was cultivated under controlled growth conditions using a new type of photobioreactor, allowing us to optimise growth conditions and the biomass yield. A fast large-scale purification method for monomeric and dimeric photosystem II (PSII) solubilized from thylakoid membranes of this cyanobacterium was developed using fast protein liquid chromatography (FPLC). The obtained PSII core complexes (PSIIcc) were analysed for their pigment stoichiometry, photochemical and oxygen evolution activities, as well as lipid and detergent composition. Thirty-six chlorophyll a (Chla), 2 pheophytin a (Pheoa), 9+/- 1 beta-carotene (Car), 2.9+/-0.8 plastoquinone 9 (PQ9) and 3.8+/-0.5 Mn were found per active centre. For the monomeric and dimeric PSIIcc, 18 and 20 lipid as well as 145 and 220 detergent molecules were found in the detergent shell, respectively. The monomeric and dimeric complexes showed high oxygen evolution activity with 1/4 O(2) released per 37-38 Chla and flash in the best cases. Crystals were obtained from dimeric PSIIcc by a micro-batch method. They diffract synchrotron X-rays to a maximum resolution of 2.9-A, resulting in complete data sets of 3.2 A resolution. |
| Databáze: | OpenAIRE |
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