Taxonomy, phylogeny, and ecology of the heliobacteria

Heliobacteria are a recently discovered group of anoxygenic phototrophic bacteria, first described in 1983. Heliobacteria contain bacteriochlorophyll g, a pigment unique to species of this group, and synthesize the simplest photosynthetic complexes of all known phototrophs. Also, unlike all other phototrophs, heliobacteria lack a mechanism for autotrophy and produce endospores. Four genera of heliobacteria containing a total of 10 species are known. Species of the genera Heliobacterium, Heliobacillus, and Heliophilum grow best at neutral pH, whereas species of Heliorestis are alkaliphilic. Heliobacterium, Heliobacillus, and Heliophilum species form one phylogenetic clade of heliobacteria, while Heliorestis species form a second within the phylum Firmicutes of the domain Bacteria. Heliobacteria have a unique ecology, being primarily terrestrial rather than aquatic phototrophs, and may have evolved a mutualistic relationship with plants, in particular, rice plants. The genome sequence of the thermophile Heliobacterium modesticaldum supports the hypothesis that heliobacteria are “minimalist phototrophs” and that they may have played a key role in the evolution of phototrophic bacteria.     

Modulation of fluorescence in Heliobacterium modesticaldum cells

In what appears to be a common theme for all phototrophs, heliobacteria exhibit complex modulations of fluorescence yield when illuminated with actinic light and probed on a time scale of μs to minutes. The fluorescence yield from cells of Heliobacterium modesticaldum remained nearly constant for the first 10–100 ms of illumination and then rose to a maximum level with one or two inflections over the course of many seconds. Fluorescence then declined to a steady-state value within about one minute. In this analysis, the origins of the fluorescence induction in whole cells of heliobacteria are investigated by treating cells with a combination of electron accepters, donors, and inhibitors of the photosynthetic electron transport, as well as varying the temperature. We conclude that fluorescence modulation in H. modesticaldum results from acceptor-side limitation in the reaction center (RC), possibly due to charge recombination between P₈₀₀ ⁺ and A₀ ^?.     

The genome of Heliobacterium modesticaldum, a phototrophic representative of the Firmicutes containing the simplest photosynthetic apparatus

Despite the fact that heliobacteria are the only phototrophic representatives of the bacterial phylum Firmicutes, genomic analyses of these organisms have yet to be reported. Here we describe the complete sequence and analysis of the genome of Heliobacterium modesticaldum, a thermophilic species belonging to this unique group of phototrophs. The genome is a single 3.1-Mb circular chromosome containing 3,138 open reading frames. As suspected from physiological studies of heliobacteria that have failed to show photoautotrophic growth, genes encoding enzymes for known autotrophic pathways in other phototrophic organisms, including ribulose bisphosphate carboxylase (Calvin cycle), citrate lyase (reverse citric acid cycle), and malyl coenzyme A lyase (3-hydroxypropionate pathway), are not present in the H. modesticaldum genome. Thus, heliobacteria appear to be the only known anaerobic anoxygenic phototrophs that are not capable of autotrophy. Although for some cellular activities, such as nitrogen fixation, there is a full complement of genes in H. modesticaldum, other processes, including carbon metabolism and endosporulation, are more genetically streamlined than they are in most other low-G+C gram-positive bacteria. Moreover, several genes encoding photosynthetic functions in phototrophic purple bacteria are not present in the heliobacteria. In contrast to the nutritional flexibility of many anoxygenic phototrophs, the complete genome sequence of H. modesticaldum reveals an organism with a notable degree of metabolic specialization and genomic reduction.     

Carbon Flow of Heliobacteria Is Related More to Clostridia than to the Green Sulfur Bacteria

The recently discovered heliobacteria are the only Gram-positive photosynthetic bacteria that have been cultured. One of the unique features of heliobacteria is that they have properties of both the photosynthetic green sulfur bacteria (containing the type I reaction center) and Clostridia (forming heat-resistant endospores). Most of the previous studies of heliobacteria, which are strict anaerobes and have the simplest known photosynthetic apparatus, have focused on energy and electron transfer processes. It has been assumed that like green sulfur bacteria, the major carbon flow in heliobacteria is through the (incomplete) reductive (reverse) tricarboxylic acid cycle, whereas the lack of CO₂-enhanced growth has not been understood. Here, we report studies to fill the knowledge gap of heliobacterial carbon metabolism. We confirm that the CO₂-anaplerotic pathway is active during phototrophic growth and that isoleucine is mainly synthesized from the citramalate pathway. Furthermore, to our surprise, our results suggest that the oxidative (forward) TCA cycle is operative and more active than the previously reported reductive (reverse) tricarboxylic acid cycle. Both isotopomer analysis and activity assays suggest that citrate is produced by a putative (Re)-citrate synthase and then enters the oxidative (forward) TCA cycle. Moreover, in contrast to (Si)-citrate synthase, (Re)-citrate synthase produces a different isomer of 2-fluorocitrate that is not expected to inhibit the activity of aconitase.     

Characterization of new phototrophic heliobacteria and their habitats

Specific enrichment culture methods were used to isolate new phototrophic heliobacteria (anoxygenic phototrophic bacteria containing bacteriochlorophyll g) from various natural samples. A survey of terrestrial and aquatic habitats yielded heliobacteria only from soils, in particular rice soils, and from certain hot springs. Thirteen nonthermophilic and 7 thermophilic (capable of growth above 50) strains of heliobacteria were isolated from such habitats and characterized as to their basic cellular and nutritional properties. Phylogenetic studies of four strains showed them to be related to known species of heliobacteria. It is concluded that, unlike phototrophic purple and green bacteria, heliobacteria are primarily (if not exclusively) terrestrial, except for hot spring species. This suggests that the ecology of heliobacteria is fundamentally different from that of other anoxyphototrophs.     

Draft Genome Sequence of Rhodovulum sp. Strain PH10, a Phototrophic Alphaproteobacterium Isolated from a Soil Sample of Mangrove of Namkhana,India

We report the 4.8-Mb draft genome of Rhodovulum sp. strain PH10, a phototrophic bacterium belonging to class Alphaproteobacteria, isolated from a soil sample collected from the mangrove forest of Namkhana in India. This genome is the first from the genus Rhodovulum and will lead to a better understanding of the genes/pathways involved in activities like phototrophic growth and nitrogen fixation in this group of bacteria.     

Heliobacterium modesticaldum,sp. nov., a thermophilic heliobacterium of hot springs and volcanic soils

Enrichment cultures for heliobacteria at 50°C yielded several strains of a thermophilic heliobacterium species from Yellowstone hot spring microbial mats and volcanic soils from Iceland. The novel organisms grew optimally above 50°C, contained bacteriochlorophyll g, and lacked intracytoplasmic membranes. All isolates were strict anaerobes and grew best as photoheterotrophs, although chemotrophic dark growth on pyruvate was also possible. These thermophilic heliobacteria were diazotrophic and fixed N₂ up to their growth temperature limit of 56°C. Phylogenetic studies showed the new isolates to be specific relatives of Heliobacterium gestii and, as has been found in H. gestii, they produce heat-resistant endospores. The unique assemblage of properties found in these thermophilic heliobacteria implicate them as a new species of this group, and we describe them herein as a new species of the genus Heliobacterium, Heliobacterium modesticaldum.     

Nitrogen fixation and nitrogen metabolism in heliobacteria

Three species of anoxygenic phototrophic heliobacteria, Heliobacterium chlorum, Heliobacterium gestii, and Heliobacillus mobilis, were studied for comparative nitrogen-fixing abilities and regulation of nitrogenase. Significant nitrogenase activity (acetylene reduction) was detected in all species grown photoheterotrophically on N₂, although cells of H. mobilis consistently had higher nitrogenase activity than did cells of either H. chlorum or H. gestii. Nitrogen-fixing cultures of all three species of heliobacteria were subject to switch-off of nitrogenase activity by ammonia; glutamine also served to switch-off nitrogenase activity but only in cells of H. mobilis and H. gestii. Placing photosynthetically grown heliobacterial cultures in darkness also served to switch-off nitrogenase activity. Dark-mediated switch-off was complete in lactate-grown heliobacteria but in pyruvate-grown cells substantial rates of nitrogenase activity continued in darkness. In all heliobacteria examined ammonia was assimilated primarily through the glutamine synthetase/glutamate synthase (GS/GOGAT) pathway although significant levels of alanine dehydrogenase were present in extracts of cells of H. gestii, but not in the other species. The results suggest that heliobacteria, like phototrophic purple bacteria, are active N₂-fixing bacteria and that despite their gram-positive phylogenetic roots, heliobacteria retain the capacity to control nitrogenase activity by a switch-off type of mechanism. Because of their ability to fix N₂ both photosynthetically and in darkness, it is possible that heliobacteria are significant contributors of fixed nitrogen in their paddy soil habitat.     

The major carotenoid in all known species of heliobacteria is the C30 carotenoid 4,4′-diaponeurosporene, not neurosporene

The carotenoids of five species of heliobacteria (Heliobacillus mobilis, Heliophilum fasciatum, Heliobacterium chlorum, Heliobacterium modesticaldum, and Heliobacterium gestii) were examined by spectroscopic methods, and the C₃₀ carotene 4,4′-diaponeurosporene was found to be the dominant pigment; heliobacteria were previously thought to contain the C₄₀ carotenoid neurosporene. In addition, trace amounts of the C₃₀ diapocarotenes diapolycopene, diapo-ζ-carotene, diapophytofluene, and diapophytoene were also found. Up to now, diapocarotenes have been found in only three species of chemoorganotrophic bacteria, but not in phototropic organisms. Furthermore, the esterifying alcohol of bacteriochlorophyll g from all known species of heliobacteria was determined to be farnesol (C₁₅) instead of the usual phytol (C₂₀). Heliobacteria may be unable to produce geranylgeranyol (C₂₀). Received: 10 March 1997 / Accepted: 3 June 1997     

Comparative genomics of green sulfur bacteria

Eleven completely sequenced Chlorobi genomes were compared in oligonucleotide usage, gene contents, and synteny. The green sulfur bacteria (GSB) are equipped with a core genome that sustains their anoxygenic phototrophic lifestyle by photosynthesis, sulfur oxidation, and CO₂ fixation. Whole-genome gene family and single gene sequence comparisons yielded similar phylogenetic trees of the sequenced chromosomes indicating a concerted vertical evolution of large gene sets. Chromosomal synteny of genes is not preserved in the phylum Chlorobi. The accessory genome is characterized by anomalous oligonucleotide usage and endows the strains with individual features for transport, secretion, cell wall, extracellular constituents, and a few elements of the biosynthetic apparatus. Giant genes are a peculiar feature of the genera Chlorobium and Prosthecochloris. The predicted proteins have a huge molecular weight of 10⁶, and are probably instrumental for the bacteria to generate their own intimate (micro)environment.     

Global Landscape of Native Protein Complexes in Synechocystis sp. PCC 6803

Synechocystis sp. PCC 6803(hereafter: Synechocystis) is a model organism for studying photosynthesis, energy metabolism, and environmental stress. Although known as the first fully sequenced phototrophic organism, Synechocystis still has almost half of its proteome without functional annotations. In this study, by using co-fractionation coupled with liquid chromatographytandem mass spectrometry(LC-MS/MS), we define 291 multi-protein complexes, encompassing24,092 protein±protein interactions(PPIs...     

Molecular evidence that the capacity for endosporulation is universal among phototrophic heliobacteria

Although enrichment cultures for anoxygenic phototrophic heliobacteria commonly contain sporulating cells, once strains of heliobacteria are obtained in pure culture, they all but cease to sporulate. In fact, some species of heliobacteria have never been observed to sporulate. Thus, despite their phylogenetic connection to endospore-forming bacteria, the question of sporulation capacity in heliobacteria remains open. We have investigated this problem using PCR and Southern hybridization as tools and show here that all recognized species of heliobacteria tested, as well as several unclassified strains, contain homologs to the ssp genes of Clostridium and Bacillus species, genes that encode key sporulation-specific proteins. It can therefore be concluded that as a group, heliobacteria are likely all to be endospore-forming bacteria in agreement with their phylogenetic placement within the 'low GC' Gram-positive bacteria.     

Photosynthetic electron transport and anaerobic metabolism in purple non-sulfur phototrophic bacteria

Purple non-sulfur phototrophic bacteria, exemplifed byRhodobacter capsulatus andRhodobacter sphaeroides, exhibit a remarkable versatility in their anaerobic metabolism. In these bacteria the photosynthetic apparatus, enzymes involved in CO₂ fixation and pathways of anaerobic respiration are all induced upon a reduction in oxygen tension. Recently, there have been significant advances in the understanding of molecular properties of the photosynthetic apparatus and the control of the expression of genes involved in photosynthesis and CO₂ fixation. In addition, anaerobic respiratory pathways have been characterised and their interaction with photosynthetic electron transport has been described. This review will survey these advances and will discuss the ways in which photosynthetic electron transport and oxidation-reduction processes are integrated during photoautotrophic and photoheterotrophic growth.     

Heliophilum fasciatum gen. nov. sp. nov. and Heliobacterium gestii sp. nov.: endospore-forming heliobacteria from rice field soils

Two new taxa of phototrophic heliobacteria are described: Heliobacterium gestii sp. nov. and Heliophilum fasciatum gen. nov. sp. nov. Both organisms were isolated from dry paddy soils. Cells of H. gestii were motile spirilla; cells of H. fasciatum formed cell bundles that were motile as units. Both organisms produced endospores; H. gestii endospores contained dipicolinic acid and elevated levels of calcium. As with other heliobacteria, bacteriochlorophyll g was produced in both organisms and no intracytoplasmic photosynthetic membranes were observed. Growth of H. gestii and H. fasciatum occurred under both photoheterotrophic and chemotrophic conditions; nitrogen fixation also occurred in both organisms. H. gestii and H. fasciatum showed a phylogenetic relationship to the "low GC" line of gram-positive Bacteria, but H. fasciatum was distinct from H. gestii and all other heliobacteria. The ability of H. gestii and H. fasciatum to form endospores might be a significant ecological advantage for survival in their rice soil habitat. Received: 16 October 1995 / Accepted: 10 January 1996     

C-type cytochromes in the photosynthetic electron transfer pathways in green sulfur bacteria and heliobacteria

Green sulfur bacteria and heliobacteria are strictly anaerobic phototrophs that have homodimeric type 1 reaction center complexes. Within these complexes, highly reducing substances are produced through an initial charge separation followed by electron transfer reactions driven by light energy absorption. In order to attain efficient energy conversion, it is important for the photooxidized reaction center to be rapidly rereduced. Green sulfur bacteria utilize reduced inorganic sulfur compounds (sulfide, thiosulfate, and/or sulfur) as electron sources for their anoxygenic photosynthetic growth. Membrane-bound and soluble cytochromes c play essential roles in the supply of electrons from sulfur oxidation pathways to the P840 reaction center. In the case of gram-positive heliobacteria, the photooxidized P800 reaction center is rereduced by cytochrome c-553 (PetJ) whose N-terminal cysteine residue is modified with fatty acid chains anchored to the cytoplasmic membrane.     

BchY-Based Degenerate Primers Target All Types of Anoxygenic Photosynthetic Bacteria in a Single PCR

To detect anoxygenic bacteria containing either type 1 or type 2 photosynthetic reaction centers in a single PCR, we designed a degenerate primer set based on the bchY gene. The new primers were validated in silico using the GenBank nucleotide database as well as by PCR on pure strains and environmental DNA.Anoxygenic photosynthetic bacteria are diverse and important members of microbial communities (11, 13, 17, 20). There are five bacterial phyla containing anoxygenic phototrophs: Proteobacteria (purple bacteria), Chlorobi (green sulfur bacteria), Chloroflexi (green nonsulfur bacteria), Acidobacteria (“Candidatus Chloracidobacterium thermophilum” [7]), and Firmicutes (heliobacteria). While Heliobacterium modesticaldum, Chlorobi, and “Ca. Chloracidobacterium thermophilum” have a type 1 reaction center (RC1) similar to photosystem I in Cyanobacteria and higher plants, Chloroflexi and Proteobacteria possess a type 2 reaction center (RC2) similar to photosystem II of oxygenic phototrophs (7, 16).Primers based on pufM, the gene encoding the M subunit of RC2, have been widely used to detect phototrophic purple bacteria (1, 4, 12, 19). However, phototrophic bacteria that do not possess RC2 are not retrieved when pufM is used as the target. Achenbach and coworkers (1) developed primers targeting rRNA genes of Chlorobi, Chloroflexi, and heliobacteria, while Alexander and coworkers (2) have developed primers to specifically detect green sulfur bacteria (Chlorobi) by using 16S rRNA and fmoA as gene targets and applied these primers in environmental studies (3). No currently available primer set can simultaneously target phototrophs containing either RC1 or RC2.Since it is well established that both RC1- and RC2-containing anoxygenic phototrophs synthesize bacteriochlorophylls (BChls), we searched for a universal anoxygenic photosynthesis gene marker among all enzymes involved in BChl biosynthetic pathways. All known pathways for chlorophyll and BChl biosynthesis branch from the heme biosynthesis pathway at protoporphyrin IX and continue to chlorophyllide a (Chlide a) through the same intermediates (9). Chlide a is the branching point that separates chlorophyll and BChl biosynthetic pathways. Moreover, pathways for the synthesis of different BChls are also split at this stage: chlorophyllide oxidoreductase converts Chlide a to 3-vinyl-bacteriophyllide a, which is the precursor for BChls a, b, and g, while a yet unknown enzyme reduces Chlide a to 3-vinyl-bacteriophyllide d, a precursor for antenna BChls c, d, and e in Chlorobium spp. (9). Since 3-vinyl-bacteriophyllide a is the last common intermediate in the synthesis of BChl a and BChl g, and the latter is the only BChl in heliobacteria (14, 15), chlorophyllide oxidoreductase is the only enzyme that is (i) present in anoxygenic phototrophic bacteria and not in oxygenic phototrophs and (ii) common to all known anoxygenic phototrophic bacterial species (with the exception of “Ca. Chloracidobacterium thermophilum,” where the pathway for BChl synthesis is not yet known). Analyzing multiple alignments of the subunits of chlorophyllide oxidoreductase, we found that only the Y subunit (encoded by the BchY gene) had two conserved regions distinguishing this protein from its closest homologs; therefore, the bchY gene was chosen as a universal marker for anoxygenic photosynthesis.Due to likely codon variations coding identical amino acid sequences in different genomes (19), degenerate BchY primers were designed by reverse translation of two conserved regions of the BchY alignment (Fig. ​(Fig.1):1): bchY_fwd (5′-CCNCARACNATGTGYCCNGCNTTYGG-3′ [26 bases; 2,048 variants; corresponding amino acid sequence, PQTMCPAFG]) and bchY_rev (5′-GGRTCNRCNGGRAANATYTCNCC-3′ [23 bases; 4,096 variants; corresponding amino acid sequence, GE{I/M}FP{A/ V}DP]). Each primer had no more than two bases deviating from known bchY sequences in the GenBank nr database (except for H. modesticaldum) as well as to environmental BchY variants in the GenBank env_nr database. None of these deviations were located in the 3′ ends of the primers (see Tables S2 and S3 in the supplemental material). These primers, therefore, were predicted to amplify a wide diversity of bchY genes under nonstringent PCR conditions (50 to 52°C annealing temperature). The lengths of the expected PCR products were either 480 bp (for green sulfur, green nonsulfur bacteria, and heliobacteria) or 510 bp (for purple bacteria).Open in a separate windowFIG. 1.Multiple-amino-acid alignment of BchY proteins. Sequence abbreviations: R.den, Roseobacter denitrificans (gi|110677524); R.gel, Rubrivivax gelatinosus (gi|29893484); R.cap, Rhodobacter capsulatus (gi|114868); C.lit, Congregibacter litoralis KT 71 (gi|88706663); H.hal, Halorhodospira halophila (gi|121998388); C.aur, Chloroflexus aurantiacus (gi|163849328); C.tep, Chlorobium tepidum (gi|66576270); and H.mod, Heliobacterium modesticaldum (gi|167629410).In order to check primer specificity in silico, a screening procedure was developed. Putative primer sites (tags) for both the bchY_fwd and the bchY_rev primers were gathered from the GenBank nucleotide collection (nt) by BLAST with relaxed search conditions; the tags having mismatches at the 3′ end or more than five overall mismatches from their primer were filtered out, and the remaining tags were mapped to their sequences mimicking PCR primer annealing. Fragments ranging from 300 to 700 bp (virtual “PCR products”) were retrieved from GenBank and annotated (see Table S4 in the supplemental material). All bchY genes present in the GenBank nt database were virtually “amplified,” pointing to the robustness of the primers and our in silico PCR analysis. On the other hand, all nonspecific “amplicons” have major deviations from the primer sequences and would likely not be amplified by a real PCR. The same screening procedure was performed against the GenBank environmental nucleotide collection (env_nt) (see Table S5 in the supplemental material), and as in the case with the nt database, only bchY fragments were virtually “amplified.”The BchY primer set was validated using five key control organisms, including the RC2-containing the purple sulfur bacterium Allochromatium vinosum and the purple nonsulfur bacterium Rhodobacter capsulatus as well as the RC1-containing green sulfur bacterium Chlorobium limicola, green nonsulfur bacterium Chloroflexus aurantiacus, and the heliobacterium H. modesticaldum. Amplifications yielded the predicted products of 510 bp from the purple bacteria and 480 bp from the green sulfur and nonsulfur bacteria and H. modesticaldum. Negative-control Escherichia coli and Synechocystis sp. strain PCC 6803 did not yield amplification products when the bchY primers were used.The designed BchY primer set successfully amplified bchY genes from DNA obtained from both marine (East Mediterranean Sea) and freshwater (Lake Kinneret) environments (see Table S6 in the supplemental material for best BLASTX hits for selected sequenced fragments). These habitats were chosen for testing due to the previously reported wide diversity of their anoxygenic phototrophs (8, 10, 18, 19). A phylogenetic tree of bchY gene fragments amplified from both freshwater and marine DNA samples is shown in Fig. ​Fig.22.Open in a separate windowFIG. 2.BchY phylogenetic tree based on a maximum likelihood tree to which short sequences were added by ARB parsimony. The branches that appeared on the original maximum likelihood tree are shown with thicker lines. Bootstrap values greater than 50% are indicated next to the branches. Sequences obtained in this study are shown in bold. For reasons of clarity, not all BchY sequences retrieved are shown in the tree. For cases in which a BchY fragment was found in more than three clones, the numbers of clones are given in parentheses. Clones m21_2 and m21_3 are identical to the bchY gene of Hoeflea phototrophica strain DFL-43 (6); the m20_2 clone was identical to the bchY gene of Dinoroseobacter shibae (5).Our study underlines the utility of the bchY gene as a molecular marker for revealing genetic heterogeneity in phototrophic microbial populations. Using both wide-scale bioinformatic analysis and PCR on control strains and naturally occurring microbial community DNA, we have confirmed the specificity and coverage of the proposed degenerate BchY primers.     

Diversity of diazotrophs in the sediments of saline and soda lakes analyzed with the use of the nifH gene as a molecular marker

Phylogenetic analysis of the nifH genes, encoding the Fe protein of the nitrogenase enzymatic complex, was carried out for pure cultures of anoxygenic phototrophic bacteria of diverse origin, as well as for heterotrophic alkaliphilic sulfate reducers isolated from saline and soda lakes. Topology of the nitrogenase tree correlated with that of the 16S rRNA gene tree to a considerable degree, which made it possible to use the nifH gene as a molecular marker for investigation of diazotrophic bacterial communities in sediments of hyper saline and soda lakes. Although diazotrophs were revealed in all environmental samples, their phylogenetic diversity was relatively low. Sulfate-reducing deltaproteobacteria and photo- and chemotrophic gammaproteobacteria were predominant in integrated samples. Analysis of the upper sediment layers revealed predominance of phototrophic diazotrophs of various phyla, including purple sulfur and nonsulfur proteobacteria, green nonsulfur bacteria, heliobacteria, and cyanobacteria. Some phylotypes could not be identified, probably indicating the presence of bacterial groups which have not yet been studied by conventional microbiological techniques.     

Sequence of the Core Antenna Domain from the Anoxygenic Phototroph <Emphasis Type="Italic">Heliophilum fasciatum</Emphasis>: Implications for Diversity of Reaction Center Type I

Broad variation among anoxygenic reaction centers makes it essential to consider a wide variety when considering the origins of photosynthesis. The photosynthetic core antenna domain in the gene pshA from Heliophilum fasciatum was sequenced doubling the number of core sequences available from heliobacteria. The sequence shares a pattern of hydrophobicity and histidine residues with the core antenna domain of pshA from Heliobacillus mobilis. Sequence identity between the two pshA sequences was 68%, indicating heliobacterial reaction centers show similar diversity to photosystem I throughout cyanobacteria and plastids. Thus, the diversity of anoxygenic phototrophic reaction centers may be greater than previously thought.     

Energy metabolism of <Emphasis Type="Italic">Heliobacterium modesticaldum</Emphasis> during phototrophic and chemotrophic growth

Background  

Heliobacterium modesticaldum is a gram-positive nitrogen-fixing phototrophic bacterium that can grow either photoheterotrophically or chemotrophically but not photoautotrophically. Surprisingly, this organism is lacking only one gene for the complete reverse tricarboxylic acid (rTCA) cycle required for autotrophic carbon fixation. Along with the genomic information reported recently, we use multiple experimental approaches in this report to address questions regarding energy metabolic pathways in darkness, CO₂ fixation, sugar assimilation and acetate metabolism.