Pseudohaliea rubra CM41_15aT was deposited in the DSMZ by the Laboratoire Arago, Université Pierre et Marie Curie (Banyuls-sur-Mer, France) under the conditions of a Material Transfer Agreement. For routine cultivation all strains were grown in SYPHC medium at 28°C [15]. Replacing of pyruvate in SYPHC medium with 10 mM DL-malate
resulted in SYMHC medium. SYM medium was obtained, if the supplementary amino acids L-histidine and L-cysteine were omitted. The preparation of defined media for growth on single carbon sources and the generation of various gas atmospheres in batch cultures has been described elsewhere [15, 18]. A 40 W incandescent bulb was Caspase Inhibitor VI clinical trial used as light source for the Go6983 Determination of growth curves in the light. For the illumination of cultures with light of distinct wavelengths LED lamps were used emitting blue,
green and red visible light with peak wavelengths of 627, 518 and 466 nm, respectively. All used chemicals were obtained from Sigma-Aldrich (Taufkirchen, Germany) and complex nutrients from DIFCO BBL (Becton Dickinson; Heidelberg, Germany). Determination of growth, cellular pigmentation and cytochromes The absorbance values of growing cultures were determined in a Thermo Scientific BioMate 6 split beam UV/visible spectrophotometer selleckchem using 1 cm light path disposable cuvettes and water as blank. The A660nm reading was used to estimate the cell density. The cellular dry weight of grown cultures was determined by overnight freeze-drying of cell pellets harvested by centrifugation. Expression of the light-harvesting complex in L. syltensis was estimated by determining the A870nm to A660nm ratio, for cultures of C. litoralis and C. halotolerans a ratio of A880nm to A660nm was used and for P. rubra a ratio of A820nm to A660nm. Photosynthetic pigments were extracted from wet cell pellets PAK5 using a mixture of
acetone/methanol (7:2) as described previously [15]. The concentrations of bacteriopheophytin a, bacteriochlorophyll a and spirilloxanthin in the acetone/methanol extracts were determined from the absorbance values obtained at 747, 771 and 475 nm, respectively, using the spectral reconstruction method of van der Rest and Gingras [31]. The detection and identification of various cytochrome types was done as reported previously [15]. Semiquantitative detection of transcripts using PCR RNA was isolated from cultures of C. litoralis DSM 17192T that were grown to early stationary phase under various incubation conditions. A culture volume equivalent to a cell suspension of one ml with an A660nm of approx. 1.0 was diluted with two volumes of RNAprotect Bacteria Reagent (Qiagen; Hilden, Germany), then cells were harvested by centrifugation.
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