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The charge separation and excited states of antenna bacteriochlorophyll in membrane fragments of the recently discovered photosynthetic bacterium Helobacterium chlorum were studied by absorbance-difference spectroscopy. Formation ofsinglet excited states of bacteriochlorophyll g with a lifetime of 200 ps or less was observed as the disappearance of the ground state absorption bands. From the absorbancedifference spectra, it was concluded that the primary photochemical reaction consists of the transfer of an electron from the primary donor P-798 to a possibly bacteriochlorophyll c-like pigment absorbing at 670 nm. Electron transfer to the secondary acceptor occurred with a time constant of about 500 ps. The midpoint potential of this acceptor (between -450 and -560 mV) and the absence of sigifcant absorbance changes in the near-infrared upon its reduction suggest that this acceptor is an iron-sulfur center. It is concluded that the primary photochemistry of H. chlorum is similar to that of green sulfur bacteria.The recently discovered photosynthetic bacterium Heliobacterium chlorum (1) has an unusual pigment composition and structure. Instead of bacteriochlorophyll (BChl) a or b, it contains BChl g (2) and a pigment absorbing at 670 nm, and it does not appear to possess either chlorosomes, like green bacteria, or invaginations of the cytoplasmic membrane, like purple bacteria. Nothing is known about its mechanism of photosynthetic electron transport, except that illumination results in (I) the bleaching of a pigment absorbing at 798 nm that may serve as primary electron donor and (ih) the oxidation of a c-type cytochrome (3).Recently we have shown that picosecond absorptiondifference spectroscopy of isolated membranes of the green sulfur bacterium Prosthecochloris aestuarii provides information not only on the properties of the excited states generated in the antenna but also on the components involved in the primary charge separation in this bacterium (4). In the present communication we report the results of a picosecond study of membranes of H. chlorum. Evidence will be presented that the primary photochemistry in this bacterium is similar to that in green sulfur bacteria.MATERIALS AND METHODS H. chlorum was grown anaerobically in medium 112 of the American Type Culture Collection (1) containing 2.5 mM ascorbate. Cells were harvested by centrifugation and resuspended in a buffer of pH 8.0 containing 10 mM Tris, 10 mM ascorbate, and 2 mM dithiothreitol. Membrane fragments were prepared by sonication followed by centrifugation for 15 min at 27,000 x g to remove large cell fragments. The preparation was stored in the dark at 50C before use.For measurements of absorption changes under continuous illumination, the samples were used without further additions. The absorbance changes were measured as in ref.5. Suitable interference and absorbance filters were used to select the actinic illumination and to protect the photomultiplier from stray actinic light. The picosecond absorbancedifference measurements wer...

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Flash-induced absorbance changes in the picosecond and nanosecond time range have been measured in the 'Dl-D2-cyt b-559 complex' isolated from photosystem II membranes. The results indicate the efficient formation of the primary radical pair P680' pheophytin-, which had a lifetime of about 36 ns, and the presence of unconnected chlorophyll in this preparation. It is concluded that the complex contains the active photosystem II reaction center, and that this reaction center contains at most 4 chlorophyll a molecules.