Purification, characterisation and crystallisation of photosystem II from Thermosynechococcus elongatus cultivated in a new type of photobioreactor.
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.
Kern
J
J
Max-Volmer-Laboratory for Biophysical Chemistry and Biochemistry, Technical University Berlin, 10623 Berlin, Strasse des 17. Juni 135, Germany.
Loll
B
B
Lüneberg
C
C
DiFiore
D
D
Biesiadka
J
J
Irrgang
K-D
KD
Zouni
A
A
D018533:Biomass; D002851:Chromatography, High Pressure Liquid; D002853:Chromatography, Liquid / Q000379:methods; D003460:Crystallization; D000458:Cyanobacteria / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D010100:Oxygen / Q000378:metabolism; D045332:Photosystem II Protein Complex / Q000302:isolation & purification*; D013054:Spectrophotometry, Atomic; D020524:Thylakoids / Q000737:chemistry*; D014961:X-Ray Diffraction
D045332:Photosystem II Protein Complex; D010100:Oxygen