Aerobic anoxygenic phototrophs (AAPs) represent a relatively recently discovered unique BChl a-containing bacterial community. These organisms own a fully functional bacterial photosynthetic system. Their photosynthesis is operative only in the presence of oxygen. However, they are not able to grow autotrophically, they require a source of organic carbon for growth and metabolism. I have characterized the photosynthetic apparatus of Erythrobacter sp. NAP1, Roseobacter sp. COL2P and the strains Dor-2m, Dor-vul and Zun-kholbo. BChl a performs the function of the main light-harvesting pigment. Spheroidenone is the major carotenoid of the strains COL2P, Dor-2m, Dor-vul and Zun-kholbo. The NAP1 strain contains "non-photosynthetic" erythroxanthin sulfate, which is bound neither to the reaction center nor to the light-harvesting complex. Bacteriorubixanthinal, zeaxanthin and erythroxanthin are the main photosynthetic carotenoids of NAP1, coupled to the light- harvesting system. This system is composed of inner LH1 complex and reaction center, with no peripheral LH2 complexes. The photosynthetic unit size, in comparison with a typical anaerobic phototroph Rhodobacter sphaeroides, is limited due to the absence of LH2 antenna. The purified LH1-RC complex isolated from NAP1 is relatively thermostable,...

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Harvesting only 4 % of light striking the Earth could possibly fulfill present energy demands of a mankind. Chlorosome of green sulfur bacteria is re- garded as suitable light-harvesting system for photosynthesis imitation. This work presents comparison of absorption and hole burning spectra of artificially prepared aggregates similar to chlorosomes with different compositions in order to verify the proposed role of quinones in excitation quenching and its redox de- pendence. Absorption spectra at room and helium temperature showed a resem- blance between artificial aggregates and chlorosomes. Hole burning experiments verified the role of quinones in excitation quenching under aerobic conditions. Even more pronounced excitation quenching was observed under anaerobic con- ditions. Significant improvements of the original experimental set-up provided better experimental data which raised many further question that are worth trying to answer in the future.

Title: Theory of aggregates of photosynthetic molecules: spectroscopy, structure, transport phenomena Author: Ján Lalinský Institute: Institute of Physics of Charles University Supervisor of the master thesis: RNDr. Tomáš Mančal, PhD., Institute of Physics of Charles University Abstract. A theory of absorption of light in an isotropic solution of molecules was formulated on the basis of electromagnetic theory of par- ticles with attention to relativistic nature of their interaction and its im- pact on the spectrum of circular dichroism. Calculations of the absorption spectra of simple systems were performed to demonstrate the properties of the mathematical model. Also calculations of the absorption spectra of the models of the bacteriochlorophyll dimer which were proposed as possible basic unit in the lamellar model of the interior of a chlorosome were performed. The experimental spectra of solution of non-aggregated bacteriochlorophyll were used to fit the parameters of the mathematical model of the molecule and for these parameters the spectra of the pro- posed models of a dimer were calculated. It has been found that the new non-electrostatic terms in the description of the mutual interaction of the parts of the molecule are negligible for calculations of ordinary absorption spectrum, but they have...

The work presented in this thesis was aimed to investigate the structure-function relationship of several photosynthetic Light-Harvesting Complexes (LHCs) including Chlorophyll Protein 29 (CP29) and Light-Harvesting Complex II (LHCII) of green plants, and Fenna-Matthews-Olson (FMO) complex of green sulfur bacterium Chlorobium tepidum. Based on the exciton calculations, a model was proposed to the electronic excited states (EES) of both CP29 and LHCII complexes by incorporating a considerable part of the current information offered by structure determination, mutagenesis analysis and spectroscopy in the modeling. The essential parameters for characterizing the excited states, Qy dipole orientations and site energies were assigned by suggesting a model that can explain both the key features of the linear (polarized) absorption spectra and the time scales of the energy transfer processes in CP29 and LHCII. The idea of offering structural information through setting connection between the spectroscopy and the spectral simulations were supported by the presented results on CP29 and LHCII. New spectroscopic measurements (absorption, linear dichroism (LD) and circular dichroism), carried out at 4 K on the FMO complex were presented, and also the LD spectrum was corrected for the degree of orientation of the sample, in order to provide comparison of not only the shape but also the size of the simulated and experimental spectra. The EES structure of the FMO complex was studied by simulating the measured optical spectra with more realistic model than the previously applied models. Simulations have been carried out with a computer program based on exciton model, which includes inhomogeneous, homogeneous and lifetime broadenings explicitly.

The current research project investigated the anoxygenic phototrophic and metal(loid) transforming bacteria of hypersaline and deep ocean hydrothermal environments. The East German Creek brine springs, an unusual flowing hypersaline system, was enumerated using classical techniques. Subterranean sulfide supported purple sulfur and nonsulfur bacteria, but at the highly oxygenated surface, aerobic anoxygenic phototrophs (AAP) were numerically dominant (up to 16-36% of cultivable bacteria). Strains (EG8, EG13, EG17, EG19) with unusual phylogenetic affiliation and novel photosynthetic and metal(loid) reducing traits were described taxonomically. Chromocurvus halotolerans gen. nov., sp. nov. was proposed as a second example of a gammaproteobacterial AAP. It exhibited bent rod-shaped cells, unusual among AAP. Facultatively anaerobic Charonomicrobium ambiphototrophicum gen. nov., sp. nov. was capable of both aerobic and anaerobic anoxygenic photosynthesis, and incapable of photoautotrophy, distinguishing it from both AAP and purple nonsulfur bacteria. Roseovarius vanadiphilum sp. nov. surprisingly produced 4.5 times more biomass and 2 times more bacteriochlorophyll (BChl) at extremely high NaVO3 concentration (7.5 g/l) than in metal-free medium. A second novel metabolic mode, anaerobic respiration on the toxic metalloid tellurate, was described for a relative of non-phototrophic Shewanella frigidimarina (ER-Te-48), from deep ocean hydrothermal vent Paralvinella worms at Explorer Ridge in the Pacific Ocean. Other strains respired on SeO32- (ER-Se-17L), VO3- (ER-V-6), and VO43- (AV-V-25). These organisms provided the first examples of anaerobic respiration on Te, Se and V at hydrothermal vents. High level resistance of AAP to metal(loid)s prompted investigation of the influence of TeO32- on photosynthetic pigment production in species including Erythromicrobium ramosum (from a terrestrial hydrothermal system) and Erythrobacter litoralis (from a hypersaline supralittoral system). Tellurite enhanced photosynthetic pigment production up to 3.4 times, consistent with an antioxidant carotenoid-based defense mechanism. However, in E. litoralis BChl precursors such as Mg protoporphyrin or its monomethyl ester also accumulated, indicating biosynthetic pathway interruption. In hydrothermal and hypersaline ecosystems, largely devoid of eukaryotic phototrophs but often enriched in metal(loid)s, AAP and metal(loid) reducers are key modulators of nutrient and toxin availability. The presented results on their ecology, physiology and biochemistry have important implications for theoretical understanding of extreme environments and hold potential for biotechnological applications.

The current research project investigated the anoxygenic phototrophic and metal(loid) transforming bacteria of hypersaline and deep ocean hydrothermal environments. The East German Creek brine springs, an unusual flowing hypersaline system, was enumerated using classical techniques. Subterranean sulfide supported purple sulfur and nonsulfur bacteria, but at the highly oxygenated surface, aerobic anoxygenic phototrophs (AAP) were numerically dominant (up to 16-36% of cultivable bacteria). Strains (EG8, EG13, EG17, EG19) with unusual phylogenetic affiliation and novel photosynthetic and metal(loid) reducing traits were described taxonomically. Chromocurvus halotolerans gen. nov., sp. nov. was proposed as a second example of a gammaproteobacterial AAP. It exhibited bent rod-shaped cells, unusual among AAP. Facultatively anaerobic Charonomicrobium ambiphototrophicum gen. nov., sp. nov. was capable of both aerobic and anaerobic anoxygenic photosynthesis, and incapable of photoautotrophy, distinguishing it from both AAP and purple nonsulfur bacteria. Roseovarius vanadiphilum sp. nov. surprisingly produced 4.5 times more biomass and 2 times more bacteriochlorophyll (BChl) at extremely high NaVO3 concentration (7.5 g/l) than in metal-free medium. A second novel metabolic mode, anaerobic respiration on the toxic metalloid tellurate, was described for a relative of non-phototrophic Shewanella frigidimarina (ER-Te-48), from deep ocean hydrothermal vent Paralvinella worms at Explorer Ridge in the Pacific Ocean. Other strains respired on SeO32- (ER-Se-17L), VO3- (ER-V-6), and VO43- (AV-V-25). These organisms provided the first examples of anaerobic respiration on Te, Se and V at hydrothermal vents. High level resistance of AAP to metal(loid)s prompted investigation of the influence of TeO32- on photosynthetic pigment production in species including Erythromicrobium ramosum (from a terrestrial hydrothermal system) and Erythrobacter litoralis (from a hypersaline supralittoral system). Tellurite enhanced photosynthetic pigment production up to 3.4 times, consistent with an antioxidant carotenoid-based defense mechanism. However, in E. litoralis BChl precursors such as Mg protoporphyrin or its monomethyl ester also accumulated, indicating biosynthetic pathway interruption. In hydrothermal and hypersaline ecosystems, largely devoid of eukaryotic phototrophs but often enriched in metal(loid)s, AAP and metal(loid) reducers are key modulators of nutrient and toxin availability. The presented results on their ecology, physiology and biochemistry have important implications for theoretical understanding of extreme environments and hold potential for biotechnological applications.