ABSTRACT
Chlorobium tepidum is a Gram-negative bacterium of the green sulfur phylum (Chlorobia). Chlorobia are obligate anaerobic photolithoautotrophs that are widely distributed in aquatic environments where anoxic layers containing reduced sulfur compounds are exposed to light. The envelope of C. tepidum is a complex organelle composed of the outer membrane, the periplasm-peptidoglycan layer, and the cytoplasmic membrane. In addition to the outer and plasma membranes, C. tepidum contains chlorosomes attached to the cytoplasmic side of the plasma membrane. Each cellular compartment has a unique set of proteins, called sub-proteome. An important aim of proteome analysis is to study the level of the expressed genes and their response to environmental changes. Membrane protein studies are of primary importance to understand how nutrients are transported inside the cell, how toxic molecules are exported, and the mechanisms of photosynthesis and energy metabolism.
Subject(s)
Bacterial Proteins/analysis , Cell Membrane/metabolism , Chlorobium/metabolism , Electrophoresis, Polyacrylamide Gel/methods , Membrane Proteins/analysis , Proteome/analysis , Chlorobium/cytology , Chlorobium/drug effects , Detergents/pharmacology , Proteomics , Solubility/drug effectsABSTRACT
The effects of exogenous vitamin B12 on the green sulfur photosynthetic bacterium Chlorobium (Chl.) tepidum were examined. Wild-type cells and mutant cells lacking a gene CT0388 (denoted as VB0388) of Chl.tepidum were grown in liquid cultures containing different concentrations of vitamin B12. The VB0388 cells hardly grew in vitamin B12-limited media, indicating that the product of CT0388 actually played an important role in vitamin B12 biosynthesis in Chl. tepidum. Both wild-type and VB0388 cells in vitamin B12-limited media exhibited absorption bands and CD signals at the Qy region that were shifted to a shorter wavelength than those of cells grown in normal media. BChl c isomers that had S-stereochemistry at the 3(1)-position tended to increase in Chl. tepidum grown in vitamin B12-limited media.
Subject(s)
Bacterial Proteins/analysis , Bacterial Proteins/chemistry , Bacteriochlorophylls/analysis , Bacteriochlorophylls/chemistry , Chlorobium/drug effects , Chlorobium/metabolism , Light-Harvesting Protein Complexes/chemistry , Mutation/genetics , Vitamin B 12/pharmacology , Chlorobium/chemistry , Chlorobium/genetics , Chromatography, High Pressure Liquid , Circular Dichroism , Dose-Response Relationship, Drug , Molecular Conformation , Protein Isoforms/analysis , Protein Isoforms/chemistry , SpectrophotometryABSTRACT
Green sulfur photosynthetic bacteria Chlorobium (Chl.) vibrioforme (DSM 263 strain and NCIB 8327 substrain possessing BChl-c) and Chl. tepidum (ATCC 49652) were photoautotrophically grown in liquid cultures containing different concentrations of sodium sulfide (Na2S). BChl-c homologs possessing a methyl group at the 12-position tended to increase in cells of the two strains of Chl. vibrioforme cultured under high Na2S concentrations. In contrast, the Na2S concentration in liquid cultures did not affect the relative composition of BChl-c homologs in Chl. tepidum. 8-Propyl-12-methyl([P,M])-BChl-c homolog, which has been little observed in usual cultivations, could be isolated by reverse-phase high-performance liquid chromatography from the cells of Chl. vibrioforme grown under high Na2S contents. The [P,M]-BChl-c homolog has the R-configuration at the 3(1)-position, which was determined by 1H-NMR analyses.
Subject(s)
Bacterial Proteins/chemistry , Bacteriochlorophylls/chemistry , Chlorobium/drug effects , Chlorobium/metabolism , Sulfides/pharmacology , Bacterial Proteins/metabolism , Bacteriochlorophylls/metabolism , Chlorobium/growth & development , Chromatography, High Pressure Liquid , Culture Media/chemistry , Culture Media/pharmacology , Magnetic Resonance Spectroscopy , Molecular StructureABSTRACT
Two sub-strains of the anoxygenic photosynthetic green sulfur bacterium Chlorobium vibrioforme NCIB 8327 were derived from the same clone and could be discriminated only by their possession of either bacteriochlorophyll (BChl) c or d as the major pigment in the peripheral light-harvesting antenna system, chlorosome (Saga Y et al. (2003) Anal Sci 19: 1575-1579). In the presence of a proper amount of oxygen in the initial culture medium, the BChl d strain showed longer retardation on its growth initiation than the BChl c strain, indicating that the latter was advantageous for survival under aerobic light conditions which produced reactive oxygen species in vivo. The result would be ascribable to the difference of the midpoint potentials between two kinds of chlorosomes formed by self-aggregates of BChl c and d as measured by their fluorescence quenching.
