Marinococcus tarijensis sp. nov., a moderately halophilic bacterium isolated from a salt mine.

A Gram-stain-positive, coccoid-shaped, halophilic bacterium, strain SR-1(T), was isolated from a salt crystal obtained from a mine located in Tarija, Bolivia. The strain was investigated using a polyphasic approach. The optimum conditions for growth of strain SR-1(T) were reached at 5% (w/v) NaCl, pH 7.6 and 37-40 °C. The peptidoglycan contained meso-diaminopimelic acid as the diagnostic diamino acid. The isoprenoid quinone was MK-7. The major cellular fatty acids of strain SR-1(T) were anteiso-C(15:0), anteiso-C(17:0) and iso-C(16:0). The DNA G+C content of strain SR-1(T) was 48.6 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed a close relationship between strain SR-1(T) and Marinococcus halophilus JCM 2479(T) (99.7% 16S rRNA gene sequence similarity), Marinococcus halotolerans KCTC 19045(T) (99.4%) and Marinococcus luteus KCTC 13214(T) (99.8%). However, strain SR-1(T) also showed low levels of DNA-DNA relatedness with these reference strains (47, 61 and 58%, respectively). On the basis of phenotypic differences and DNA-DNA hybridization results, strain SR-1(T) is considered to represent a novel species of the genus Marinococcus, for which the name Marinococcus tarijensis sp. nov. is proposed. The type strain is SR-1(T) ( =LMG 26930(T) =CECT 8130(T)).

Genomic studies on nitrogen metabolism in Halomonas boliviensis: metabolic pathway, biochemistry and evolution.

Halomonas boliviensis LC1(T)=DSM 15516(T) is a halophilic bacterium that copiously produces osmolytes and polyesters. The growth of H. boliviensis is restricted when glutamate or glutamine is not included in its culture medium. The concentration of glutamate in the medium can regulate the production of either osmolytes or polyesters. However, genomic studies on the nitrogen assimilation have not been performed on H. boliviensis and other members of the family Halomonadaceae. Glutamate metabolism in H. boliviensis was discerned based on genome sequence analysis. The genome sequences of other Halomonadaceae members revealed similar enzymes to those found in H. boliviensis. H. boliviensis and H. elongata DSM 2581(T) acquired distinct glutamate dehydrogenase genes through horizontal gene transfer from a different bacterium. Two alleles of glutamine synthetase could be found in H. boliviensis, one of which was obtained from a thermophilic archaeon via horizontal gene transfer. Two subunits of glutamate synthase were also present in H. boliviensis. The small ß-subunit had a molecular weight of 52 kDa and was phylogenetically closely affiliated to proteins of other halomonads and Gammaproteobacteria. The large (161 kDa) α-subunit of the halomonads gathered in a separate phylogenetic group, hence glutamate synthase α-subunits of halomonads may be included a novel group of enzymes. Furthermore, putative enzymes obtained from the genome of H. boliviensis should permit complete glutamate metabolism. A similar metabolism should be followed by other halomonads. However, some phenotypic differences between halomonads, such as the ability to assimilate ammonia, resulted as a consequence of horizontal gene transfer. Each enzyme that forms part of the glutamate metabolism in prokaryotes evolved following a different pattern. Yet, most enzymes of halomonads diverged in phylogenetic clusters composed of Proteobacteria, as might be expected.

Evolutionary patterns of carbohydrate transport and metabolism in Halomonas boliviensis as derived from its genome sequence: influences on polyester production.

BACKGROUND: Halomonas boliviensis is a halophilic bacterium that is included in the γ-Proteobacteria sub-group, and is able to assimilate different types of carbohydrates. H. boliviensis is also able to produce poly(3-hydroxybutyrate) (PHB) in high yields using glucose as the carbon precursor. Accumulation of PHB by microorganisms is induced by excess of intracellular NADH.The genome sequences and organization in microorganisms should be the result of evolution and adaptation influenced by mutation, gene duplication, horizontal gen transfer (HGT) and recombination. Furthermore, the nearly neutral theory of evolution sustains that genetic modification of DNA could be neutral or selected, albeit most mutations should be at the border between neutrality and selection, i.e. slightly deleterious base substitutions in DNA are followed by a slightly advantageous substitutions. RESULTS: This article reports the genome sequence of H. boliviensis. The chromosome size of H. boliviensis was 4 119 979 bp, and contained 3 863 genes. A total of 160 genes of H. boliviensis were related to carbohydrate transport and metabolism, and were organized as: 70 genes for metabolism of carbohydrates; 47 genes for ABC transport systems and 43 genes for TRAP-type C4-dicarboxylate transport systems. Protein sequences of H. boliviensis related to carbohydrate transport and metabolism were selected from clusters of orthologous proteins (COGs). Similar proteins derived from the genome sequences of other 41 archaea and 59 bacteria were used as reference. We found that most of the 160 genes in H. boliviensis, c.a. 44%, were obtained from other bacteria by horizontal gene transfer, while 13% of the genes were acquired from haloarchaea and thermophilic archaea, only 34% of the genes evolved among Proteobacteria and the remaining genes encoded proteins that did not cluster with any of the proteins obtained from the reference strains. Furthermore, the diversity of the enzymes derived from these genes led to polymorphism in glycolysis and gluconeogenesis. We found further that an optimum ratio of glucose and sucrose in the culture medium of H. boliviensis favored cell growth and PHB production. CONCLUSIONS: Results obtained in this article depict that most genetic modifications and enzyme polymorphism in the genome of H. boliviensis were mainly influenced by HGT rather than nearly neutral mutations. Molecular adaptation and evolution experienced by H. boliviensis were also a response to environmental conditions such as the type and amount of carbohydrates in its ecological niche. Consequently, the genome evolution of H. boliviensis showed to be strongly influenced by the type of microorganisms, genetic interaction among microbial species and its environment. Such trend should also be experienced by other prokaryotes. A system for PHB production by H. boliviensis that takes into account the evolutionary adaptation of this bacterium to the assimilation of combinations of carbohydrates suggests the feasibility of a bioprocess economically viable and environmentally friendly.