Phylogenetic relationships of three amino-acid-utilizing anaerobes, Selenomonas acidaminovorans, 'Selenomonas acidaminophila' and Eubacterium acidaminophilum, as inferred from partial 16S rDNA nucleotide sequences and proposal of Thermanaerovibrio acidaminovorans gen. nov., comb. nov. and Anaeromusa acidaminophila gen. nov., comb. nov.
(1/40)
16S rRNA gene sequences of three previously described amino-acid-fermenting anaerobes, Selenomonas acidaminovorans, 'Selenomonas acidaminophila' and Eubacterium acidaminophilum, were determined. All three were found to cluster within the Clostridium and related genera of the subphylum of the Gram-positive bacteria. The thermophile, S. acidaminovorans, formed an individual line of descent and was equidistantly placed between Dethiosulfovibrio peptidovorans and Anaerobaculum thermoterrenum (similarity of 85%), both of which also form single lines of descent. 'S. acidaminophila' was related to Clostridium quercicolum, a member of cluster IX, with a similarity of 90%, whereas E. acidaminophilum was closely related to Clostridium litorale (similarity of 96%) as a member of cluster XI. Based on the phylogenetic data presented in this report and the phenotypic descriptions of these bacteria published previously, it is recommended that S. acidaminovorans be transferred to a new genus, Thermanaerovibrio gen. nov., as Thermanaerovibrio acidaminovorans comb. nov. and 'Selenomonas acidaminophila' be transferred to a new genus, Anaeromusa gen. nov., as Anaeromusa acidaminophila comb. nov. Though the transfer of E. acidaminophilum to a new taxon is justified, this is not recommended until the taxonomic status of all the members of cluster XI has been reviewed. (+info)
Novel characteristics of Selenomonas ruminantium lysine decarboxylase capable of decarboxylating both L-lysine and L-ornithine.
(2/40)
Lysine decarboxylase (LDC; EC 4.1.1.18) of Selenomonas ruminantium is a constitutive enzyme and is involved in the synthesis of cadaverine, which is an essential constituent of the peptidoglycan for normal cell growth. We purified the S. ruminantium LDC by an improved method including hydrophobic chromatography and studied the fine characteristics of the enzyme. Kinetic study of LDC showed that S. ruminantium LDC decarboxylated both L-lysine and L-ornithine with similar Km and the decarboxylase activities towards both substrates were competitively and irreversibly inhibited by DL-alpha-difluoromethylornithine, which is a specific inhibitor of ornithine decarboxylase (EC 4.1.1.17). We also showed a drastic descent of LDC activity owing to the degradation of LDC at entry into the stationary phase of cell growth. (+info)
Molecular analysis of the microbial diversity present in the colonic wall, colonic lumen, and cecal lumen of a pig.
(3/40)
Random clones of 16S ribosomal DNA gene sequences were isolated after PCR amplification with eubacterial primers from total genomic DNA recovered from samples of the colonic lumen, colonic wall, and cecal lumen from a pig. Sequences were also obtained for cultures isolated anaerobically from the same colonic-wall sample. Phylogenetic analysis showed that many sequences were related to those of Lactobacillus or Streptococcus spp. or fell into clusters IX, XIVa, and XI of gram-positive bacteria. In addition, 59% of randomly cloned sequences showed less than 95% similarity to database entries or sequences from cultivated organisms. Cultivation bias is also suggested by the fact that the majority of isolates (54%) recovered from the colon wall by culturing were related to Lactobacillus and Streptococcus, whereas this group accounted for only one-third of the sequence variation for the same sample from random cloning. The remaining cultured isolates were mainly Selenomonas related. A higher proportion of Lactobacillus reuteri-related sequences than of Lactobacillus acidophilus- and Lactobacillus amylovorus-related sequences were present in the colonic-wall sample. Since the majority of bacterial ribosomal sequences recovered from the colon wall are less than 95% related to known organisms, the roles of many of the predominant wall-associated bacteria remain to be defined. (+info)
Identification of the amino acid residues conferring substrate specificity upon Selenomonas ruminantium lysine decarboxylase.
(4/40)
Lysine decarboxylase (LDC, EC 4.1.1.18) from Selenomonas ruminantium has decarboxylating activities towards both L-lysine and L-ornithine with similar K(m) and Vmax. Here, we identified four amino acid residues that confer substrate specificity upon S. ruminantium LDC and that are located in its catalytic domain. We have succeeded in converting S. ruminantium LDC to an enzyme with a preference in decarboxylating activity for L-ornithine when the four-residue of LDC were replaced by the corresponding residues of mouse ornithine decarboxylase (EC 4.1.1.17). (+info)
16S-23S rDNA spacer of Pectinatus, Selenomonas and Zymophilus reveal new phylogenetic relationships between these genera.
(5/40)
The 16S-23S rDNA spacer regions of two Pectinatus species, two Zymophilus species and one Selenomonas species were cloned after PCR amplification. The results of PCR amplification showed that these species had two types of spacer regions which differ in molecular size (long and short). Only the long spacer regions in these bacteria contained one or two tRNA genes (alanine and/or isoleucine). The spacer regions in these bacteria had a relatively high level of homology. Homology was particularly high for bacteria belonging to the same genus. Interestingly, the order of the two tRNA genes present in the long spacer regions of Pectinatus and Selenomonas was the reverse of that which had been previously reported for other bacteria. The results of spacer homology analysis and the order of the tRNA genes suggest that the taxonomic classification of anaerobic bacteria isolated from the brewing process should be re-examined. (+info)
Reclassification of Clostridium quercicolum as Dendrosporobacter quercicolus gen. nov., comb. nov.
(6/40)
Morphological features, genomic DNA base composition and 16S rDNA sequence similarities, as well as a distinct phospholipid pattern, whole-cell fatty acid distribution and the occurrence of the lipoquinone 'lipid F', indicate that Clostridium quercicolum belongs to the Sporomusa-Pectinatus-Selenomonas phyletic group and possesses only a remote relationship to members of the genus Clostridium sensu stricto. On the basis of these results, the new genus and combination Dendrosporobacter quercicolus gen. nov., comb. nov. are proposed. (+info)
Gene cloning and molecular characterization of lysine decarboxylase from Selenomonas ruminantium delineate its evolutionary relationship to ornithine decarboxylases from eukaryotes.
(7/40)
Lysine decarboxylase (LDC; EC 4.1.1.18) from Selenomonas ruminantium comprises two identical monomeric subunits of 43 kDa and has decarboxylating activities toward both L-lysine and L-ornithine with similar K(m) and V(max) values (Y. Takatsuka, M. Onoda, T. Sugiyama, K. Muramoto, T. Tomita, and Y. Kamio, Biosci. Biotechnol. Biochem. 62:1063-1069, 1999). Here, the LDC-encoding gene (ldc) of this bacterium was cloned and characterized. DNA sequencing analysis revealed that the amino acid sequence of S. ruminantium LDC is 35% identical to those of eukaryotic ornithine decarboxylases (ODCs; EC 4.1.1.17), including the mouse, Saccharomyces cerevisiae, Neurospora crassa, Trypanosoma brucei, and Caenorhabditis elegans enzymes. In addition, 26 amino acid residues, K69, D88, E94, D134, R154, K169, H197, D233, G235, G236, G237, F238, E274, G276, R277, Y278, K294, Y323, Y331, D332, C360, D361, D364, G387, Y389, and F397 (mouse ODC numbering), all of which are implicated in the formation of the pyridoxal phosphate-binding domain and the substrate-binding domain and in dimer stabilization with the eukaryotic ODCs, were also conserved in S. ruminantium LDC. Computer analysis of the putative secondary structure of S. ruminantium LDC showed that it is approximately 70% identical to that of mouse ODC. We identified five amino acid residues, A44, G45, V46, P54, and S322, within the LDC catalytic domain that confer decarboxylase activities toward both L-lysine and L-ornithine with a substrate specificity ratio of 0.83 (defined as the k(cat)/K(m) ratio obtained with L-ornithine relative to that obtained with L-lysine). We have succeeded in converting S. ruminantium LDC to form with a substrate specificity ratio of 58 (70 times that of wild-type LDC) by constructing a mutant protein, A44V/G45T/V46P/P54D/S322A. In this study, we also showed that G350 is a crucial residue for stabilization of the dimer in S. ruminantium LDC. (+info)
Competition among three predominant ruminal cellulolytic bacteria in the absence or presence of non-cellulolytic bacteria.
(8/40)
Competition among three species of ruminal cellulolytic bacteria - Fibrobacter succinogenes S85, Ruminococcus flavefaciens FD-1 and Ruminococcus albus 7 - was studied in the presence or absence of the non-cellulolytic ruminal bacteria Selenomonas ruminantium or Streptococcus bovis. Co-cultures were grown under either batch or continuous conditions and populations were estimated using species-specific oligonucleotide probes to 16S rRNA. The three cellulolytic species co-existed in cellobiose batch co-culture, but inclusion of either Sel. ruminantium or Str. bovis yielded nearly a monoculture of the non-cellulolytic competitor. In cellobiose chemostats, R. albus completely dominated the triculture, but R. flavefaciens became predominant over F. succinogenes and R. albus when Sel. ruminantium was co-inoculated into the chemostats. Similar effects on competition were observed in the presence of Str. bovis at a lower (0.021 h(-1)), but not at a higher (0.045 h(-1)) dilution rate. In cellulose batch co-cultures, R. albus was more abundant than both F. succinogenes and R. flavefaciens, regardless of the presence of the non-cellulolytic species. Co-existence among the three cellulolytic species was observed in almost all cellulose chemostats, but Sel. ruminantium altered the relative proportions of the cellulolytic species. R. albus and R. flavefaciens were found to produce inhibitors that suppressed growth of R. flavefaciens and F. succinogenes, respectively. These data indicate that interactions among cellulolytic bacteria, while complex, can be modified further by non-cellulolytic species. (+info)