Vulcanimicrobium alpinus gen. nov. sp. nov., the first cultivated representative of the candidate phylum "Eremiobacterota", is a metabolically versatile aerobic anoxygenic phototroph.

The previously uncultured phylum "Candidatus Eremiobacterota" is globally distributed and often abundant in oligotrophic environments. Although it includes lineages with the genetic potential for photosynthesis, one of the most important metabolic pathways on Earth, the absence of pure cultures has limited further insights into its ecological and physiological traits. We report the first successful isolation of a "Ca. Eremiobacterota" strain from a fumarolic ice cave on Mt. Erebus volcano (Antarctica). Polyphasic analysis revealed that this organism is an aerobic anoxygenic photoheterotrophic bacterium with a unique lifestyle, including bacteriochlorophyll a production, CO2 fixation, a high CO2 requirement, and phototactic motility using type IV-pili, all of which are highly adapted to polar and fumarolic environments. The cells are rods or filaments with a vesicular type intracytoplasmic membrane system. The genome encodes novel anoxygenic Type II photochemical reaction centers and bacteriochlorophyll synthesis proteins, forming a deeply branched monophyletic clade distinct from known phototrophs. The first cultured strain of the eighth phototrophic bacterial phylum which we name Vulcanimicrobium alpinus gen. nov., sp. nov. advances our understanding of ecology and evolution of photosynthesis.

Reticulibacter mediterranei gen. nov., sp. nov., within the new family Reticulibacteraceae fam. nov., and Ktedonospora formicarum gen. nov., sp. nov., Ktedonobacter robiniae sp. nov., Dictyobacter formicarum sp. nov. and Dictyobacter arantiisoli sp. nov., belonging to the class Ktedonobacteria.

The aerobic, Gram-positive, mesophilic Ktedonobacteria strains, Uno17T, SOSP1-1T, 1-9T, 1-30T and 150040T, formed mycelia of irregularly branched filaments, produced spores or sporangia, and numerous secondary metabolite biosynthetic gene clusters. The five strains grew at 15-40 °C (optimally at 30 °C) and pH 4.0-8.0 (optimally at pH 6.0-7.0), and had 7.21-12.67 Mb genomes with 49.7-53.7 mol% G+C content. They shared MK9(H2) as the major menaquinone and C16 : 1-2OH and iso-C17 : 0 as the major cellular fatty acids. Phylogenetic and phylogenomic analyses showed that Uno17T and SOSP1-9T were most closely related to members of the genus Dictyobacter, with 94.43-96.21 % 16S rRNA gene similarities and 72.16-81.56% genomic average nucleotide identity. The strain most closely related to SOSP1-1T and SOSP1-30T was Ktedonobacter racemifer SOSP1-21T, with 91.33 and 98.84 % 16S rRNA similarities, and 75.13 and 92.35% average nucleotide identities, respectively. Strain 150040T formed a distinct clade within the order Ktedonobacterales, showing <90.47 % 16S rRNA gene similarity to known species in this order. Based on these results, we propose: strain 150040T as Reticulibacter mediterranei gen. nov., sp. nov. (type strain 150 040T=CGMCC 1.17052T=BCRC 81202T) within the family Reticulibacteraceae fam. nov. in the order Ktedonobacterales; strain SOSP1-1T as Ktedonospora formicarum gen. nov., sp. nov. (type strain SOSP1-1T=CGMCC 1.17205T=BCRC 81203T) and strain SOSP1-30T as Ktedonobacter robiniae sp. nov. (type strain SOSP1-30T=CGMCC 1.17733T=BCRC 81205T) within the family Ktedonobacteraceae; strain Uno17T as Dictyobacter arantiisoli sp. nov. (type strain Uno17T=NBRC 113155T=BCRC 81116T); and strain SOSP1-9T as Dictyobacter formicarum sp. nov. (type strain SOSP1-9T=CGMCC 1.17206T=BCRC 81204T) within the family Dictyobacteraceae.

High cellulolytic potential of the Ktedonobacteria lineage revealed by genome-wide analysis of CAZymes.

Traditionally, filamentous fungi and actinomycetes are well-known cellulolytic microorganisms that have been utilized in the commercial production of cellulase enzyme cocktails for industrial-scale degradation of plant biomass. Noticeably, the Ktedonobacteria lineage (phylum Chloroflexi) with actinomycetes-like morphology was identified and exhibited diverse carbohydrate utilization or degradation abilities. In this study, we performed genome-wide profiling of carbohydrate-active enzymes (CAZymes) in the filamentous Ktedonobacteria lineage. Numerous CAZymes (153-290 CAZymes, representing 63-131 glycoside hydrolases (GHs) per genome), including complex mixtures of endo- and exo-cellulases, were predicted in 15 available Ktedonobacteria genomes. Of note, 4-28 CAZymes were predicted to be extracellular enzymes, whereas 3-29 CAZymes were appended with carbohydrate-binding modules (CBMs) that may promote their binding to insoluble carbohydrate substrates. This number far exceeded other Chloroflexi lineages and were comparable to the cellulolytic actinomycetes. Six multi-modular extracellular GHs were cloned from the thermophilic Thermosporothrix hazakensis SK20-1T strain and heterologously expressed. The putative endo-glucanases of ThazG5-1, ThazG9, and ThazG12 exhibited strong cellulolytic activity, whereas the putative exo-glucanases ThazG6 and ThazG48 formed weak but observable halos on carboxymethyl cellulose plates, indicating their potential biotechnological application. The purified recombinant ThazG12 had near-neutral pH (optimal 6.0), high thermostability (60°C), and broad specificity against soluble and insoluble polysaccharide substrates. It also represented described a novel thermostable bacterial ß-1,4-glucanase in the GH12 family. Together, this research revealed the underestimated cellulolytic potential of the Ktedonobacteria lineage and highlighted its potential biotechnological utility as a promising microbial resource for the discovery of industrially useful cellulases.

Dictyobacter vulcani sp. nov., belonging to the class Ktedonobacteria, isolated from soil of the Mt Zao volcano.

An aerobic, Gram-stain-positive, mesophilic Ktedonobacteria strain, W12T, was isolated from soil of the Mt Zao volcano in Miyagi, Japan. Cells were filamentous, non-motile, and grew at 20-37 °C (optimally at 30 °C), at pH 5.0-7.0 (optimally at pH 6.0) and with <2 % (w/v) NaCl on 10-fold diluted Reasoner's 2A (R2A) medium. Oval-shaped spores were formed on aerial mycelia. Strain W12T hydrolysed microcrystalline cellulose and xylan very weakly, and used d-glucose as its sole carbon source. The major menaquinone was MK-9, and the major cellular fatty acids were C16 : 1 2-OH, iso-C17 : 0, summed feature 9 (10-methyl C16 : 0 and/or iso-C17 : 1ω9c) and anteiso-C17 : 0. Cell-wall sugars were mannose and xylose, and cell-wall amino acids were d-glutamic acid, glycine, l-serine, d-alanine, l-alanine, ß-alanine and l-ornithine. Polar lipids were phosphatidylinositol, phosphatidylglycerol, diphosphatidylglycerol, an unidentified glycolipid and an unidentified phospholipid. Strain W12T has a genome of 7.42 Mb with 49.7 mol% G+C content. Nine copies of 16S rRNA genes with a maximum dissimilarity of 1.02 % and 13 biosynthetic gene clusters mainly coding for peptide products were predicted in the genome. Phylogenetic analysis based on both 16S rRNA gene and whole genome sequences indicated that strain W12T represents a novel species in the genus Dictyobacter. The most closely related Dictyobacter type strain was Dictyobacter alpinus Uno16T, with 16S rRNA gene sequence similarity and genomic average nucleotide identity of 98.37 % and 80.00 %, respectively. Herein, we propose the name Dictyobacter vulcani sp. nov. for the type strain W12T (=NBRC 113551T=BCRC 81169T) in the bacterial class Ktedonobacteria.

Formation of sporangiospores in Dictyobacter aurantiacus (class Ktedonobacteria in phylum Chloroflexi).

Currently, actinomycetes and myxobacteria are the only bacteria believed to form sporangia. Here, we describe a sporangium-forming process identified in Dictyobacter aurantiacus strain S27T belonging to the class Ktedonobacteria in the phylum Chloroflexi. Microscopic observations showed that strain S27T forms a substrate mycelium and subsequently produces globose or subglobose terminal sporangia arising from the vegetative mycelia through short stalk cells. This morphogenetic differentiation is similar to that seen in members of Actinoplanes belonging to the class Actinobacteria. However, unlike in Actinoplanes, motile spores could not be observed. This is the first report of the existence of a bacterium, other than actinomycetes and myxobacteira, with a complex morphogenetic differentiation that forms sporangia and is an important microbiological discovery.

Gandjariella thermophila gen. nov., sp. nov., a new member of the family Pseudonocardiaceae, isolated from forest soil in a geothermal area.

Six mycelium-forming actinomycete strains were isolated from forest soil near the Cisolok geysers in West Java, Indonesia. The 16S rRNA gene sequences of these strains showed high similarity to members of genera in the family Pseudonocardiaceae with values less than 96.0 %, and most closely related to the genus Thermotunica, T. guangxiensis AG2-7T(94.6-95.2 % similarity). The type strain, designated SL3-2-4T, was aerobic, thermophilic, Gram-stain-positive that formed branched, non-fragmented substrate mycelia and unbranched aerial mycelia with long-chain, oval-shaped spores on International Streptomyces Project (ISP) 3 medium. It produced light-orange substrate mycelia and light-orange diffusible pigments on ISP 3 medium with 2 % gellan gum, grown at 30-55 °C, with optimum growth at 45 °C. The pH range for growth was 4.0-8.0, with optimum growth at pH 7.0. Strain SL3-2-4T was able to hydrolyze casein, esculin, gelatin, guanine, hypoxanthine, starch, L-tyrosine, and xanthine, but not adenine, carboxymethyl-cellulose, cellulose, chitin, Tween 20, or xylan. The major fatty acid was iso-C16 : 0, and the major menaquinone was MK-8 (H4). The detected polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidyl-N-methylethanolamine, unidentified aminophospholipids, unidentified glycolipids, and unidentified phospholipids. The cell wall hydrolysate of SL3-2-4T contained meso-2,4-diaminopimelic acid. The whole cell sugars were arabinose and galactose. The DNA G+C content was 71.6 mol%. Phenotypic features and phylogenetic data differentiated SL3-2-4T from members of the family Pseudonocardiaceae. Therefore, the strain SL3-2-4T is proposed as a representative of a novel species in a novel genus, Gandjariella thermophila gen. nov., sp. nov. The type strain is SL3-2-4T (=UICC B-83T=NRRL B-67478T=InaCC A981T).

Genome Features and Secondary Metabolites Biosynthetic Potential of the Class Ktedonobacteria.

The prevalence of antibiotic resistance and the decrease in novel antibiotic discovery in recent years necessitates the identification of potentially novel microbial resources to produce natural products. Ktedonobacteria, a class of deeply branched bacterial lineage in the ancient phylum Chloroflexi, are ubiquitous in terrestrial environments and characterized by their large genome size and complex life cycle. These characteristics indicate Ktedonobacteria as a potential active producer of bioactive compounds. In this study, we observed the existence of a putative "megaplasmid," multiple copies of ribosomal RNA operons, and high ratio of hypothetical proteins with unknown functions in the class Ktedonobacteria. Furthermore, a total of 104 antiSMASH-predicted putative biosynthetic gene clusters (BGCs) for secondary metabolites with high novelty and diversity were identified in nine Ktedonobacteria genomes. Our investigation of domain composition and organization of the non-ribosomal peptide synthetase and polyketide synthase BGCs further supports the concept that class Ktedonobacteria may produce compounds structurally different from known natural products. Furthermore, screening of bioactive compounds from representative Ktedonobacteria strains resulted in the identification of broad antimicrobial activities against both Gram-positive and Gram-negative tested bacterial strains. Based on these findings, we propose the ancient, ubiquitous, and spore-forming Ktedonobacteria as a versatile and promising microbial resource for natural product discovery.

Tengunoibacter tsumagoiensis gen. nov., sp. nov., Dictyobacter kobayashii sp. nov., Dictyobacter alpinus sp. nov., and description of Dictyobacteraceae fam. nov. within the order Ktedonobacterales isolated from Tengu-no-mugimeshi, a soil-like granular mass of micro-organisms, and emended descriptions of the genera Ktedonobacter and Dictyobacter.

Three mesophilic, Gram-stain-positive, aerobic bacterial strains, designated Uno3T, Uno11T and Uno16T, were isolated from a soil-like granular micro-organism mass (termed Tengu-no-mugimeshi) collected from Tsumagoi, Gunma, Japan. They grow at 11-37 °C and pH 4.0-8.0, form branched mycelia, and have a G+C content between 49.4-50.3 mol%. The major menaquinone and fatty acid of Uno3T are MK-9 and iso-C16 : 0, respectively, whereas Uno11T and Uno16T share MK-9 (H2) and C16 : 1-2OH. The major cell-wall sugars are mannose (Uno3T and Uno11T) and glucose (Uno16T). Phylogenetic analysis based on 16S rRNA gene sequences indicated that these three strains belong to the order Ktedonobacterales and are most closely related to Dictyobacter aurantiacus S-27T (sequence similarity of 91.3, 96.4 and 95.5 %). Average nucleotide identity values were <79.9 % among Uno11T, Uno16T and D. aurantiacus S-27T, well below the 95-96 % species circumscription threshold. Based on phenotypic features and phylogenetic positions, we propose that Uno3T represents a novel genus and species, Tengunoibacter tsumagoiensis gen. nov., sp. nov. (type strain Uno3T=NBRC 113152T=LMG 30471T=BCRC 81113T) within the new family Dictyobacteraceae fam. nov. Strains Uno11T and Uno16T are also considered to represent novel species: Dictyobacterkobayashii sp. nov. (type strain Uno11T=NBRC 113153T=LMG 30472T=BCRC 81114T) and Dictyobacteralpinus sp. nov. (type strain Uno16T=NBRC 113154T=BCRC 81115T). We also propose an emended description of the genus Dictyobacter, classifying it within family Dictyobacteraceae, and provide emended descriptions of the genera Dictyobacter and Ktedonobacter.

Thermogemmatispora aurantia sp. nov. and Thermogemmatispora argillosa sp. nov., within the class Ktedonobacteria, and emended description of the genus Thermogemmatispora.

Two thermophilic, aerobic, Gram-stain-positive Ktedonobacteria strains, A1-2T and A3-2T, were isolated from geothermal soil in Japan. The strains formed orange-coloured colonies on 10-fold diluted Reasoner's 2A medium, followed by formation of branched aerial mycelium with multiple grape-like spores. Both strains hydrolysed casein, carboxymethyl cellulose, starch, chitin and xylan, but did not liquify gelatin. Strain A1-2T utilised sucrose and gellan gum and was inhibited by inositol, while strain A3-2T utilised only gellan gum and was not inhibited by inositol. The DNA G+C contents of strain A1-2T and A3-2T were 63.2 and 63.1 mol%, respectively. Chemotaxonomic data (major fatty acid, iso-C17 : 0; major menaquinone, MK-9(H2); cell-wall amino acids, ornithine, serine, glycine, glutamic acid, alanine and ß-alanine; polar lipids, phosphatidylinositol, phosphatidylglycerol, diphosphatidylglycerol, one unidentified lipid, one unidentified phosphoglycolipid and three unidentified glycolipids; major cell-wall sugars, mannose, arabinose and xylose) indicate that both strains belong to the genus Thermogemmatispora. 16S rRNA gene sequence analysis indicated that strain A1-2 T was most closely related to the type strains of Thermogemmatispora onikobensis (97.7 % sequence similarity), and that strain A3-2T was most closely related to the type strains of Thermogemmatispora carboxidivorans(97.2%), but DNA-DNA hybridization shows relatedness values of <67 % with previously described type strains. Moreover, 16S rRNA gene sequence similarity and DNA-DNA relatedness between strain A1-2T and strain A3-2T were 96.0 and 33.4%, respectively, suggesting that the two strains are genetically distinct. The two strains are proposed as Thermogemmatispora aurantia sp. nov. and Thermogemmatispora argillosa sp. nov.

Dictyobacter aurantiacus gen. nov., sp. nov., a member of the family Ktedonobacteraceae, isolated from soil, and emended description of the genus Thermosporothrix.

A mesophilic, Gram-stain-positive, spore-forming bacterium that formed branched mycelia was isolated from paddy soil in Gunung Salak (Mount Salak), West Java, Indonesia. This strain, designated S-27T, grew at temperatures between 20 and 37 °C; the optimum growth temperature was 25 to 30 °C, and no growth was observed at 15 or 45 °C. The pH range for growth was pH 3.5 to 8.6; the optimum pH was 6.0, and no growth was observed at pH 3.0 or 9.2. Strain S-27T was able to hydrolyse polysaccharides such as starch, cellulose and xylan. The G+C content of the DNA of strain S-27T was 55.7 mol%. The major fatty acids were iso-C17 : 0 and C16 : 1 2-OH, and the major menaquinone was MK-9 (H2). The cell wall of strain S-27T contained d-glutamic acid, glycine, l-alanine, d-alanine, l-ornithine and ß-alanine in a molar ratio of 1.0 : 1.6 : 1.4 : 0.6 : 0.9 : 1.1. The polar lipids consisted of phosphatidylglycerol, phosphatidylinositol and two glycolipids. The major cell-wall sugar was arabinose. Detailed phylogenetic analysis based on 16S rRNA gene sequences indicated that strain S-27T belongs to the order Ktedonobacterales and is most closely related to Ktedonobacter racemifer SOSP1-21T (89.6 % sequence identity). On the basis of its chemotaxonomic and phenotypic features and phylogenetic position, we concluded that strain S-27T represents a novel genus and species, for which we propose the name Dictyobacter aurantiacus gen. nov., sp. nov. The type strain of Dictyobacter aurantiacus is strain S-27T (=NBRC 109595T=InaCC B312T). Emendation of the description of the genus Thermosporothrix is also provided.

Noviherbaspirillum denitrificans sp. nov., a denitrifying bacterium isolated from rice paddy soil and Noviherbaspirillum autotrophicum sp. nov., a denitrifying, facultatively autotrophic bacterium isolated from rice paddy soil and proposal to reclassify Herbaspirillum massiliense as Noviherbaspirillum massiliense comb. nov.

Thirty-nine denitrifying bacterial strains closely related to one another, represented by strains TSA40T and TSA66T, were isolated from rice paddy soils. Strains TSA40T and TSA66T were Gram-stain-negative, slightly curved rod-shaped, and motile by means of polar flagella. They were able to reduce nitrate, nitrite and nitrous oxide, but unable to fix atmospheric N2. While strain TSA66T was able to grow autotrophically by H2-dependent denitrification, strain TSA40T could not. Phylogenetic analysis suggested that they belong to the family Oxalobacteraceae, the order Burkholderiales in the class Betaproteobacteria. Major components in the fatty acids (C16 : 0, C17 : 0 cyclo, C18 : 1ω7c and summed feature 3) and quinone (Q-8) also supported the affiliation of strains TSA40T and TSA66T to the family Oxalobacteraceae. Based on 16S rRNA gene sequence comparisons, strains TSA40T and TSA66T showed the greatest degree of similarity to Herbaspirillum massiliense JC206T, Noviherbaspirillum malthae CC-AFH3T, Noviherbaspirillum humi U15T, Herbaspirillum seropedicae Z67T and Paucimonas lemoignei LMG 2207T, and lower similarities to the members of other genera. Average nucleotide identity values between the genomes of strain TSA40T, TSA66T and H. massiliense JC206T were 75-77 %, which was lower than the threshold value for species discrimination (95-96 %). Based on the 16S rRNA gene sequence analysis in combination with physiological, chemotaxonomic and genomic properties, strains TSA40T (=JCM 17722T=ATCC TSD-69T) and TSA66T (=JCM 17723T=DSM 25787T) are the type strains of two novel species within the genus Noviherbaspirillum, for which the names Noviherbaspirillum denitrificans sp. nov. and Noviherbaspirillum autotrophicum sp. nov. are proposed, respectively. We also propose the reclassification of Herbaspirillum massiliense as Noviherbaspirillum massiliense comb. nov.

Diversity of Ktedonobacteria with Actinomycetes-Like Morphology in Terrestrial Environments.

Bacteria with an actinomycetes-like morphology have recently been discovered, and the class Ktedonobacteria was created for these bacteria in the phylum Chloroflexi. They may prove to be a valuable resource with the potential to produce unprecedented secondary metabolites. However, our understanding of their diversity, richness, habitat, and ecological significance is very limited. We herein developed a 16S rRNA gene-targeted, Ktedonobacteria-specific primer and analyzed ktedonobacterial amplicons. We investigated abundance, diversity, and community structure in forest and garden soils, sand, bark, geothermal sediment, and compost. Forest soils had the highest diversity among the samples tested (1181-2934 operational taxonomic units [OTUs]; Chao 1 estimate, 2503-5613; Shannon index, 4.21-6.42). A phylogenetic analysis of representative OTUs revealed at least eight groups within unclassified Ktedonobacterales, expanding the known diversity of this order. Ktedonobacterial communities markedly varied among our samples. The common mesic environments (soil, sand, and bark) were dominated by diverse phylotypes within the eight groups. In contrast, compost and geothermal sediment samples were dominated by known ktedonobacterial families (Thermosporotrichaceae and Thermogemmatisporaceae, respectively). The relative abundance of Ktedonobacteria in the communities, based on universal primers, was ≤0.8%, but was 12.9% in the geothermal sediment. These results suggest that unknown diverse Ktedonobacteria inhabit common environments including forests, gardens, and sand at low abundances, as well as extreme environments such as geothermal areas.

Draft Genome Sequence of Thermogemmatispora onikobensis NBRC 111776T, an Aerial Mycelium- and Spore-Forming Thermophilic Bacterium Belonging to the Class Ktedonobacteria.

Here, we report the draft genome sequence of Thermogemmatispora onikobensis NBRC 111776T, an aerial mycelium- and spore-forming thermophilic bacterium belonging to the class Ktedonobacteria The genome contains five biosynthetic gene clusters coding for secondary metabolites, such as terpene, thiopeptide, lantipeptide, nonribosomal peptide, and lassopeptide, suggesting the potential to produce secondary metabolites.

Paenibacillus cisolokensis sp. nov., isolated from litter of a geyser.

A Gram-stain-positive, endospore-forming, aerobic and thermophilic bacterium, designated strain LC2-13AT, was isolated from Cisolok geyser, West Java, Indonesia, at 50 °C. The isolate was rod-shaped and motile by means of peritrichous flagella. The major cellular fatty acids were iso-C16 : 0, C16 : 0 and anteiso-C15 : 0 and the major quinone was menaquinone 7. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. The genomic DNA G+C content was 56.6 mol% and the major diagnostic diamino acid in the cell-wall peptidoglycan was meso-diaminopimelic acid. Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain LC2-13AT is related most closely to Paenibacillus kobensis DSM 10249T (94.86 % similarity), Paenibacillus tarimensis SA-7-6T (94.77 %) and Paenibacillus barengoltzii SAFN-016T (94.77 %). On the basis of phenotypic, chemotaxonomic and phylogenetic evidence, strain LC2-13AT is affiliated to the genus Paenibacillus, but could be distinguished from recognized species of this genus. A novel species with the name Paenibacillus cisolokensis sp. nov. is thus proposed. The type strain is LC2-13AT (=UICC B-42T=NRRL B-65368T=DSM 101873T).

Thermosporothrix narukonensis sp. nov., belonging to the class Ktedonobacteria, isolated from fallen leaves on geothermal soil, and emended description of the genus Thermosporothrix.

A thermophilic, Gram-stain-positive, spore-forming bacterium that formed branched vegetative and aerial mycelia was isolated from fallen leaves on geothermal soil. This strain, designated F4T, grew at temperatures between 30 and 60 °C; optimum growth temperature was 50 °C, whereas no growth was observed below 28 °C or above 65 °C. The pH range for growth was 4.9-9.5; the pH for optimum growth was 7.0, but no growth was observed at pH below 4.4 or above 10.0. Strain F4T was able to hydrolyse polysaccharides such as cellulose, xylan, chitin and starch. The G+C content in the DNA of strain F4T was 52.5 mol%. The major fatty acid was iso-C17 : 0 and the major menaquinone was MK-9 (H2). The cell wall of strain F4T contained glutamic acid, serine, glycine, alanine and ornithine in a molar ratio of 1.0:1.5:1.4:1.8:0.7. The polar lipids of this strain consisted of phosphatidylglycerol, diphosphatidylglycerol, phosphatidylinositol, one unknown phospholipid, three unknown glycolipids and two unknown lipids. The cell-wall sugar was mannose. Detailed phylogenetic analysis based on 16S rRNA gene sequences indicated that strain F4T belongs to the genus Thermosporothrix, and that it was related most closely to Thermosporothrix hazakensis SK20-1T (98.7 % similarity). DNA-DNA hybridization showed relatedness values of less than 15 % with the type strain of Thermosporothrix hazakensis. On the basis of phenotypic features and phylogenetic position, strain F4T is considered to represent a novel species, Thermosporothrix narukonensis sp. nov. The type strain is F4T(=NBRC 111777T=BCCM/LMG 29329T).

Identification and biosynthesis of new acyloins from the thermophilic bacterium Thermosporothrix hazakensis SK20-1(T).

Two new acyloin compounds were isolated from the thermophilic bacterium Thermosporothrix hazakensis SK20-1(T) . Genome sequencing of the bacterium and biochemical studies identified the thiamine diphosphate (TPP)-dependent enzyme Thzk0150, which is involved in the formation of acyloin. Through extensive analysis of the Thzk0150-catalyzed reaction products, we propose a putative reaction mechanism involving two substrates: 4-methyl-2-oxovalerate as an acyl donor and phenyl pyruvate as an acyl acceptor.

Sphingobacterium thermophilum sp. nov., of the phylum Bacteroidetes, isolated from compost.

A Gram-negative bacterium, designated CKTN2(T), was isolated from compost. Cells of strain CKTN2(T) were strictly aerobic rods. The isolate grew at 20-50 °C (optimum 40-45 °C), but not below 15 °C or above 52 °C, and at pH 5.9-8.8 (optimum pH 7.0), but not below pH 5.4 or above pH 9.3. The DNA G+C content was 40.3 mol%. The predominant menaquinone was MK-7. The major fatty acids were iso-C15 : 0 (45.2 %), iso-C17 : 0 3-OH (11.1 %) and C18 : 0 (14.5 %). Analysis of the 16S rRNA gene sequence of strain CKTN2(T) revealed that it is a member of the genus Sphingobacterium and is most closely related to Sphingobacterium alimentarium DSM 22362(T) (93.2 % 16S rRNA gene sequence similarity). Strain CKTN2(T) could be distinguished from its closest phylogenetic relatives by different phenotypic characteristics. According to the phenotypic and genotypic characteristics, strain CKTN2(T) represents a novel species of the genus Sphingobacterium, for which the name Sphingobacterium thermophilum sp. nov. is proposed. The type strain is CKTN2(T) ( = JCM 17858(T)  = KCTC 23708(T)).

Thermovum composti gen. nov., sp. nov., an alphaproteobacterium from compost.

A Gram-stain-positive thermophilic bacterium, designated strain Nis3(T), was isolated from compost. The strain grew at 23-57 °C (optimum, 50 °C); no growth was observed below 15 or above 60 °C. The pH range for growth was 5.9-8.8 (optimum, 7.0); no growth was observed below pH 5.4 or above pH 9.3. The DNA G+C content of strain Nis3(T) was 63.4 mol%. The dominant quinone type was ubiquinone Q-10. The major fatty acids were C(18:1)ω7c, C(19:0)ω8c cyclo and C(18:0). The polar lipids comprised phosphatidylcholine, phosphatidylglycerol, phosphatidylethanolamine, hydroxyphosphatidylethanolamine, phosphatidylinositol, phosphatidylmonomethylethanolamine, an unknown glycolipid and a ninhydrin-positive phospholipid. 16S rRNA gene sequence analysis assigned this bacterium to the family Phyllobacteriaceae in the Alphaproteobacteria but it shared less than 95.2% sequence similarity with other members of the family. The chemotaxonomic and phenotypic characteristics of strain Nis3(T) differed in some respects from those of members of the family Phyllobacteriaceae. Therefore, strain Nis3(T) is considered to represent a novel species of a new genus in the family Phyllobacteriaceae, for which the name Thermovum composti gen. nov., sp. nov. is proposed. The type strain is Nis3(T) ( = JCM 17863(T) = KCTC 23707(T)).