Strains Of Green Sulfur Bacteria Research Articles

Osmoadaptative accumulation of N ɛ-acetyl-β-lysine in green sulfur bacteria and Bacillus cereus CECT 148T

The compatible solute N(ɛ)-acetyl-β-lysine (NeABL), thus far considered unique to methanogenic Archaea, has been found to accumulate in several strains of green sulfur bacteria (GSB) and Bacillus cereus CECT 148(T) under salt stress. A similar mixture of compatible solutes including trehalose, α-glutamate, β-glutamate and NeABL has been detected in salt-tolerant GSB strains of different phylogenetic branches. The ability of B. cereus to synthesize this compound was predicted from available genomic data, and nuclear magnetic resonance analyses of cultures grown in salt-containing media indicated that NeABL was present in the solute pools of osmotically challenged cells. The present results describe for the first time in the bacterial domain the use of this compound for osmoadaptation.

Mutational analysis of three bchH paralogs in (bacterio-)chlorophyll biosynthesis in Chlorobaculum tepidum

The first committed step in the biosynthesis of (bacterio-)chlorophyll is the insertion of Mg2+ into protoporphyrin IX by Mg-chelatase. In all known (B)Chl-synthesizing organisms, Mg-chelatase is encoded by three genes that are homologous to bchH, bchD, and bchI of Rhodobacter spp. The genomes of all sequenced strains of green sulfur bacteria (Chlorobi) encode multiple bchH paralogs, and in the genome of Chlorobaculum tepidum, there are three bchH paralogs, denoted CT1295 (bchT), CT1955 (bchS), and CT1957 (bchH). Cba. tepidum mutants lacking one or two of these paralogs were constructed and characterized. All of the mutants lacking only one of these BchH homologs, as well as bchS bchT and bchH bchT double mutants, which can only produce BchH or BchS, respectively, were viable. However, attempts to construct a bchH bchS double mutant, in which only BchT was functional, were consistently unsuccessful. This result suggested that BchT alone is unable to support the minimal (B)Chl synthesis requirements of cells required for viability. The pigment compositions of the various mutant strains varied significantly. The BChl c content of the bchS mutant was only approximately 10% of that of the wild type, and this mutant excreted large amounts of protoporphyrin IX into the growth medium. The observed differences in BChl c production of the mutant strains were consistent with the hypothesis that the three BchH homologs function in end product regulation and/or substrate channeling of intermediates in the BChl c biosynthetic pathway.

Phylogenetic position of three strains of green sulfur bacteria

) are aphysiologically homogeneous group forming a sepa-rate phylogenetic branch isolated from other bacteria(phylum Chlorobi). Until recently, their taxonomy wasbased on phenotypic characteristics (morphologicaland biochemical properties) [1, 2]. Comparative analy-sis of the sequences of the 16S rRNA genes and the fmogenes, encoding FMO protein, which occurs exclu-sively in Chlorobiaceae representatives, led to substan-tial reorganization of the taxonomy of this group ofbacteria [3]. As a result, strains previously assigned toone species may have been transferred to several differ-ent species and even genera.The culture collection of the Department of Micro-biology, Moscow State University, includes severalstrains of green sulfur bacteria previously assigned toChl. limicola forma thiosulfatophilum (strains C, X,and L, or KM MGU 319, 320, and 321, respectively).These have been used in studies of various aspects ofmetabolism of green sulfur bacteria [4–8]; therefore,determination of their phylogenetic positions bychemotaxonomic and molecular biological methods istopical.Using the method described in [9], we determinedthe fatty acid compositions of strains C, X, and L(table); such data are rather informative but scarce forgreen sulfur bacteria. Strains L, X, and C were found tocontain a considerable amount of palmitoleic (16:1 ω7),palmitic (16:0) and myristic (14:0) acids, whereas the18:1 acids (oleic and cis-vaccenic) were virtuallyabsent. Such a fatty acid pattern is characteristic ofChlorobiaceae representatives [2, 3, 10].In order to perform comparative analysis of 16SrRNA genes, their fragments were amplified usingstandard bacterial primers as described in [11]. As aresult, sequences of more than 1300 nucleotides weredetermined for strains C, X, and L (and deposited inGenBank: Chb. parvum strain L, EF560699; Chb. lim-naeum strain X, EF560700; and Chb. limnaeumstrain C, EF560701).According to the preliminary phylogenetic screen-ing performed with BLAST (http://www.ncbi.nlm.nih.gov/blast), all the strains studied belonged to thesubdivision of green sulfur bacteria. For more detailedphylogenetic analysis, the 16S rRNA gene sequences ofthe strains studied and reference bacterial species werealigned using the CLUSTLW v1.75 program. Phyloge-netic trees were constructed using different algorithmsimplemented in the TREECONW software package[12]. In the constructed phylogenetic tree (Fig. 1), allthree strains studied fell into phylogenetic cluster 4,

Biogeochemical processes in the saline meromictic Lake Kaiike, Japan: implications from molecular isotopic evidences of photosynthetic pigments

Stable carbon and nitrogen isotopic compositions were determined for individual photosynthetic pigments isolated and purified from the saline meromictic Lake Kaiike, Japan, to investigate species-independent biogeochemical processes of photoautotrophs in the natural environment. In the anoxic monimolimnion and benthic microbial mats, the carbon isotopic compositions of BChls e and isorenieratene related to brown-coloured strains of green sulfur bacteria are substantially ( approximately 10 per thousand) depleted in (13)C relative to those found in the chemocline. In conjunction with 16S rDNA evidence reported previously, it strongly suggests that Pelodyctyon luteolum inhabited and photosynthesized in the anoxic monimolimnion and benthic microbial mats by using (13)C-depleted regenerated CO(2). By contrast, both Chl a and BChl a in the monimolimnion and microbial mats have similar isotopic compositions as they do in the chemocline, implying that the source organisms live only in the chemocline. In the chemocline, the nitrogen isotopic compositions of BChl e homologues ranges from -7.7 to-6.5 per thousand, whereas that of BChl a is -2.1 per thousand. These isotopic compositions suggest that green sulfur bacteria Chlorobium phaeovibrioides would conduct nitrogen fixation in the chemocline, whereas purple sulfur bacteria Halochromatium sp. and cyanobacteria Synechococcus sp. may assimilate nitrite.

Vertical distributions of stable isotopic compositions and bacteriochlorophyll homologues in suspended particulate matter in saline meromictic Lake Abashiri

We determined chloropigment composition as well as stable carbon and nitrogen isotopic compositions of dissolved and particulate species in saline meromictic Lake Abashiri. We observed a sharp peak of bacteriochlorophyll e in a narrow redox boundary zone and the upper monimolimnion, indicating a dense population of brown-colored strains of green sulfur bacteria around the chemocline. Nitrogen isotopic records of particulate nitrogen and dissolved ammonium suggested that the green sulfur bacteria in the redox boundary zone assimilated either ammonium or dinitrogen through the nitrogen fixation pathway. In the anoxic monimolimnion, several lines of evidence suggest that a major portion of particulate organic matter originated from the overlying mixolimnion and redox boundary zone.

Novel bacteriochlorophyll e structures and species-specific variability of pigment composition in green sulfur bacteria.

The relative composition of bacteriochlorophyll (BChl) homologs in five different strains of brown-colored green sulfur bacteria was investigated by HPLC-MS/MS and NMR analyses. In addition, the effect of incubation light intensities on homolog distribution was studied in one of the strains (strain Dagow III). A total of 23 different BChl e structures were detected and comprise four homologous porphyrin ring systems and eight different esterifying alcohols. Several BChl e structures are novel. These include a C-8 ethyl, C-12 methyl [E, M] BChl e(F) homolog which was identified by (1)H-NMR analyses of the isolated, main farnesyl homologs (BChl e(F)). In addition, five previously unknown homolog series with dodecanol, pentadecenol, tetradecanol, hexadecenol and phytol as the esterifying alcohols were detected. The composition of BChl e homologs from the five strains of green sulfur bacteria differed with respect to the relative abundance of the homologs (BChl e(F) : 25.6-67.0% of total BChl e content in stationary cultures). In strain Dagow III, the abundance of BChl e(F) homologs decreased upon entry into the stationary phase. In all free-living strains, the abundance of BChl e(F) was increased when the relative carotenoid content was low. The present results provide a detailed picture of pigment composition in chlorosomes and thus will help to elucidate their structure and function. Furthermore, the newly discovered BChl e molecules are valuable biomarkers for the study of the occurrence and metabolism of green sulfur bacteria in past and present ecosystems.

Phylogeny of the PscB reaction center protein from green sulfur bacteria.

The reaction center (RC) of green sulfur bacteria has iron-sulfur clusters as terminal acceptors and is related to the Type I RC found in Heliobacter sp. and in Photosystem I (PS I) of green plants and cyanobacteria. Degenerate primers were used to retrieve the genes coding for one of the RC proteins, PscB, from 11 strains of green sulfur bacteria. PCR using the same primers gave no product with a second group of strains and the protein from these strains did not crossreact with antibodies raised against purified PscB from the first group, suggesting the presence of a high degree of variability. The sequences shared a high degree of similarity in the region coding for the binding motif for the 4Fe-4S centers. However, the N-terminal portion of the deduced protein sequences was highly variable and contained a highly positively charged, low-complexity region with repeated tetrapeptides with two alanines flanked by proline or lysine. The PscB sequences obtained fell into two major groups, and the results suggested a lack of correlation between the pigmentation of the chlorosome antenna system and the reaction center protein. There is also a lack of correlation between the grouping of the pscB sequences and the phylogeny deduced from 16S rRNA.

Phylogeny and molecular fingerprinting of green sulfur bacteria.

The 16S rDNA sequences of nine strains of green sulfur bacteria (Chlorobiaceae) were determined and compared to the four known sequences of Chlorobiaceae and to sequences representative for all eubacterial phyla. The sequences of the Chlorobiaceae strains were consistent with the secondary structure model proposed earlier for Chlorobium vibrioforme strain 6030. Similarity values > 90.1% and Knuc values < 0.11 indicate a close phylogenetic relatedness among the green sulfur bacteria. As a group, these bacteria represent an isolated branch within the eubacterial radiation. In Chlorobiaceae, a similar morphology does not always reflect a close phylogenetic relatedness. While ternary fission is a morphological trait of phylogenetic significance, gas vesicle formation occurs also in distantly related species. Pigment composition is not an indicator of phylogenetic relatedness since very closely related species contain different bacteriochlorophylls and carotenoids. Two different molecular fingerprinting techniques for the rapid differentiation of Chlorobiaceae species were investigated. The 16S rDNA fragments of several species could not be separated by denaturing gradient gel electrophoresis. In contrast, all strains investigated during the present work gave distinct banding patterns when dispersed repetitive DNA sequences were used as targets in PCR. The latter technique is, therefore, well suited for the rapid screening of isolated pure cultures of green sulfur bacteria.