Cellulomonas fulva sp. nov., isolated from oil-contaminated soil.

A yellow-coloured, Gram-stain-positive, motile, aerobic and rod-shaped bacteria, designated DKR-3T, was isolated from oil-contaminated experimental soil. Strain DKR-3T could grow at pH 5.0-10.5 (optimum, pH 7.0-8.5), at 10-40 °C (optimum, 25-32 °C) and tolerated 3.5 % of NaCl. Phylogenetic analyses based on its 16S rRNA gene sequence indicated that strain DKR-3T formed a lineage within the family Cellulomonadaceae and was clustered with members of the genus Cellulomonas. Strain DKR-3T had highest 16S rRNA gene sequence similarities to Cellulomonas gelida DSM 20111T (98.3 %), Cellulomonas persica JCM 18111T (98.2 %) and Cellulomonas uda DSM 20107T (97.8 %). The predominant respiratory quinone was tetrahydrogenated menaquinone with nine isoprene units [MK-9(H4)]. The principal cellular fatty acids were anteiso-C15 : 0, C16 : 0 and anteiso-C17 : 0. The major polar lipids were diphosphatidylglycerol and phosphatidylglycerol. The cell-wall diamino acid was l-ornithine whereas rhamnose and glucose were the cell-wall sugars. The DNA G+C content was 74.2mol %. The genome of strain DKR-3T was 3.74 Mb and contained three putative biosynthetic gene clusters. The average nucleotide identity and digital DNA-DNA hybridization relatedness values between strain DKR-3T and its phylogenetically related members were below the species threshold values. Based on a polyphasic study, strain DKR-3T represents a novel species belonging to the genus Cellulomonas, for which the name Cellulomonas fulva sp. nov. is proposed. The type strain is DKR-3T (=KACC 22071T=NBRC 114730T).

Cellulomonas citrea sp. nov., isolated from paddy soil.

Two Gram-stain-positive, facultatively anaerobic, motile, aerobic, rod-shaped and non-spore-forming actinobacteria, strains AO-9T and AO-18, were isolated from paddy soil collected from Daejeon, Republic of Korea. Colonies were smooth, lemon-yellow and circular and 0.5-0.8×2.0-2.4 µm in diameter after 3 days of incubation at 28 °C on tryptic soy agar. Phylogenetic analyses based on 16S rRNA gene sequences revealed that strains AO-9T and AO-18 belonged to the genus Cellulomonas, showing the highest sequence similarities to Cellulomonas marina FXJ8.089T (96.6 %), Cellulomonas endophytica SYSUP0004T (96.5 %), Cellulomonas gelida DSM 20111T (96.2 %), Cellulomonas uda DSM 20107T (96.1 %), Cellulomonas rhizosphaerae NEAU-TCZ24T (96.1 %), Cellulomonas composti TR7-06T (96.0 %), Cellulomonas persica JCM 18111T (96.0 %) and less than 96 % to other closely related species. The DNA-DNA hybridization values between strains AO-9T and AO-18 were 87 %. The average nucleotide identity and digital DNA-DNA hybridization values between strain AO-9T and type strains of related species of the genus Cellulomonas were 84.0-85.8 % and 20.3-20.9 %, respectively. The major cellular fatty acids are anteiso-C15:0 (49.9 %), C14:0 (12.9 %) and iso-C14:0 (12.1 %). The predominant isoprenoid quinone was MK-9 (H4). The polar lipid profile consists of diphosphatidylglycerol, phosphatidylglycerol and one unidentified lipid. The DNA G+C content was 72.9 mol%. Based on its distinctive phenotypic, phylogenetic and chemotaxonomic characteristics, the two strains are considered to represent novel species of the genus Cellulomonas, for which the name Cellulomonas citrea sp. nov. is proposed. The type strain is AO-9T (=KACC 19069T=NBRC 112523T).

Cellulomonas endophytica sp. nov., isolated from Gastrodia elata Blume.

A Gram-stain-positive, facultatively anaerobic and non-motile strain, designated SYSUP0004T, was isolated from the tubers of Gastrodia elata Blume collected from Yunnan Province, PR China. The 16S rRNA gene sequence result showed that the strain SYSUP0004T shared low similarity (97.7 %) with the type strain of Cellulomonas marina. SYSUP0004T grew at pH 6.0-9.0 (optimum, pH 8.0), temperature 4-30 °C (optimum, 28 °C) and could tolerate NaCl up to 4 % w/v (optimum in the absence of NaCl). The cell-wall peptidoglycan type was A4ß with an interpeptide bridge l-ornithine-d-glutamic acid. Cell-wall sugars were mannose, ribose, glucose, galactose and fucose. The menaquinone was MK-9(H4). The major fatty acids were anteiso-C15:0, anteiso-C15 : 1 A, C16 : 0 and anteiso-C17 : 0. The polar lipids of SYSUP0004T were diphosphatidylglycerol, unidentified phosphoglycolipid, phosphatidylinositol mannosides and unidentified glycolipid. The genomic DNA G+C content was 76.5 %. The average nucleotide identity values between SYSUP0004T and members of the genus Cellulomonas were below the cut-off level (95-96 %) recommended as the ANI criterion for interspecies identity. Thus, based on the above results strain SYSUP0004T represents a novel species of the genus Cellulomonas, for which the name Cellulomonas endophytica sp. nov. is proposed. The type strain, SYSUP0004T (=KCTC 49025T=CGMCC 1.16405T).

Cellulomonas shaoxiangyii sp. nov., isolated from faeces of Tibetan antelope (Pantholops hodgsonii) on the Qinghai-Tibet Plateau.

Two Gram-stain-positive, catalase-positive and oxidase-negative, aerobic, non-motile, cellobiose-utilizing, short-rod-shaped strains (Z28T and Z29) were isolated from faeces of Tibetan antelope (Pantholops hodgsonii) collected on the Qinghai-Tibet Plateau. Strain Z28T shared 98.1, 98.0, 97.8 and 97.4 % 16S rRNA gene similarity, 24.1, 22.8, 23.2 and 26.3 % digital DNA-DNA hybridization relatedness and 80.8, 80.0, 80.7 and 80.9 % average nucleotide identity values with Cellulomonas oligotrophica DSM 24482T, Cellulomonas flavigena DSM 20109T, Cellulomonas iranensis DSM 14785T and Cellulomonas terrae JCM 14899T, respectively. Results from further phylogenetic analyses based on the 16S rRNA gene and 148 core genes indicated that strains Z28T and Z29 were closest to C. oligotrophica DSM 24482T and C. flavigena DSM 20109T, but clearly separated from the currently recognized species of the genus Cellulomonas. The genomic DNA G+C content of strain Z28T was 75.3 mol%. The major cellular fatty acids were anteiso-C15 : 0, anteiso-C15 : 1 A, C16 : 0 and anteiso-C17 : 0. Ribose and mannose were detected as the whole-cell sugars. The major respiratory quinone was MK-9(H4) and ornithine was the diamino acid of the cell wall. The polar lipids present in strain Z28T were phosphatidylethanolamine, five phospholipids, two aminophospholipids, aminolipid and three unidentified lipids. Comparison of phenotypic and phylogenetic features between the two strains and the related organisms revealed that Z28T and Z29 represent a novel species of the genus Cellulomonas, for which the name Cellulomonas shaoxiangyii sp. nov. is proposed. The type strain is Z28T (=CGMCC 1.16477T=DSM 106200T).

Cellulomonas telluris sp. nov., an endoglucanase-producing actinobacterium isolated from Badain Jaran desert sand.

A Gram-stain-positive, aerobic bacterium, designated CPCC 204705T, was isolated from a desert soil sample, collected from the Badain Jaran desert. Growth of strain CPCC 204705T was observed at pH 6.0-8.0 and 15-37 °C, with optimal growth at 28 °C and pH 7.0. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain CPCC 204705T belonged to the genus Cellulomonas, showing the highest similarity (98.54 %) of 16S rRNA gene sequence to Cellulomonas oligotrophica JCM 17534T. The peptidoglycan type was A4ß, containing d-ornithine and d-glutamic acids as diagnostic amino acids. Rhamnose and galactose were detected in the whole-cell hydrolysate as diagnostic sugars. The major cellular fatty acids were anteiso-C15 : 0, anteiso-C15 : 1A, C14 : 0 and C16 : 0. The major menaquinone was MK-9 (H4) and the polar lipid system contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol mannoside, one unidentified lipid, one unidentified aminolipid and two unidentified aminophospholipids. The DNA-DNA hybridization value between strain CPCC 204705T and C. oligotrophica JCM 17534T was 7.1±0.4 %, and the value of average nucleotide identity between these two strains was 79.8 %. The DNA G+C content of strain CPCC 204705T was 75.4 mol%. Based on the results of physiological experiments, chemotaxonomic data, phylogenetic analysis and DNA-DNA hybridization value, strain CPCC 204705T should be classified as a novel Cellulomonas species. The name Cellulomonas telluris sp. nov. is proposed, with strain CPCC 204705T (=DSM 105430T=KCTC 39974T) as the type strain.

Cellulomonas algicola sp. nov., an actinobacterium isolated from a freshwater alga.

An actinomycete strain, TKZ-21T, was isolated from a freshwater alga (Chetophoraceae) collected from the Takizawa River, Yamanashi, Japan, and examined using a polyphasic taxonomic approach. Cells were Gram-stain positive, aerobic, non-sporulating, motile, and coccoid or short rod-shaped. The strain grew in the presence of 0-4 % (w/v) NaCl, between pH 6-9.4, and over a temperature range of 15-40 °C, with optimum growth at 30 °C. The peptidoglycan type of strain TKZ-21T was A4ß, containing l-ornithine as diagnostic diamino acid and d-glutamic acid as the interpeptide bridge. The predominant menaquinone was MK-9(H4). The polar lipids were diphosphatidylglycerol, phosphatidylglycerol, ninhydrin-positive glycolipid, and unidentified phospholipids. The major cellular fatty acids were anteiso-C15 : 0 and anteiso-C17 : 0, and the DNA G+C content was 75.6 mol%. On the basis of 16S rRNA gene sequence analysis, strain TKZ-21T was closely related to Cellulomonas fimi (98.5 % sequence similarity) and Cellulomonas biazotea (98.3 %). The genome orthoANI value between strain TKZ-21T and C. biazotea and C. fimi were 84.7 and 84.2 %, respectively. On the basis of fatty acid and MALDI-TOF MS profile analysis, phylogenetic analyses, genomic analysis, and phenotypic data, it is proposed that the isolate be classified as a representative of a novel species of the genus Cellulomonas, with the name Cellulomonas algicola sp. nov. The type strain is TKZ-21T (=NBRC 112905T=TBRC 8129T).

Cellulomonas aurantiaca sp. nov., isolated from a soil sample from a tangerine field.

A Gram-stain positive, facultatively aerobic, motile and rod-shaped bacterial strain, designated THG-SMD2.3T, was isolated from a soil sample collected in a tangerine field, Republic of Korea. According to the 16S rRNA gene sequence comparisons, the isolate was identified as a member of the genus Cellulomonas and to be closely related to Cellulomonas fimi ATCC 484T (98.5%), Cellulomonas biazotea DSM 20112T (98.3%), Cellulomonas chitinilytica X.bu-bT (98.0%), Cellulomonas xylanilytica XIL11T (97.2%), Cellulomonas humilata ATCC 25174T (97.1%) and Cellulomonas composti TR7-06T (97.0%). The 16S rRNA gene sequence similarities with other current species of the genus Cellulomonas were in the range 95.4-96.6%. Catalase and oxidase tests were found to be positive. The DNA G+C content was determined to be 73.0 mol%. DNA-DNA hybridization values between strain THG-SMD2.3T and C. fimi ATCC 484T, C. biazotea DSM 20112T, C. chitinilytica X.bu-bT, C. xylanilytica XIL11T, C. humilata ATCC 25174T and C. composti TR7-06T were 58.1 ± 1.6%, 56.7 ± 0.8%, 30.3 ± 1.6%, 22.8 ± 1.6%, 19.9 ± 1.6%, and 13.5 ± 3.0%, respectively. Strain THG-SMD2.3T was also found to be able to grow at 20-42 °C, at 0-3% NaCl and at pH 5.5-10. The major fatty acids were identified as anteiso-C15:0, iso-C15:0, anteiso-C17:0 and iso-C14:0. The predominant menaquinone was identified as tetrahydrogenated menaquinones with nine isoprene units [MK-9(H4)]. The polar lipids were found to be diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, two unidentified aminolipids and two unidentified phospholipids. Based on these phenotypic, genotypic and phylogenetic characterisations strain THG-SMD2.3T (= KACC 19341T = CGMCC 1.16303T) is concluded to represent a novel species of the genus Cellulomonas, for which the name Cellulomonas aurantiaca sp. nov. is proposed.

Cellulomonas rhizosphaerae sp. nov., a novel actinomycete isolated from soil.

A novel actinomycete, designated strain NEAU-TCZ24T, was isolated from soil and characterized using a polyphasic approach. The results of phylogenetic analysis based on the 16S rRNA gene sequence indicated that the organism should be assigned to the genus Cellulomonas and formed a stable clade with its closest relatives Cellulomonas terrae JCM 14899T (98.4 % 16S rRNA gene sequence similarity), Cellulomonas xylanilytica JCM 14281T (97.9 %) and Cellulomonas humilata JCM 11945T (97.7 %). The major menaquinones were identified as MK-9(H4) and MK-8(H4). The phospholipid profile was found to contain diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositolmannoside, a ninhydrin-positiveglycolipid, an unidentified phosphoglycolipid, an unidentified phospholipid and an unidentified lipid. The major fatty acids were identified as anteiso-C15 : 0, C18 : 1ω9c, C16 : 0 and anteiso-C17 : 0. Moreover, morphological and chemotaxonomic properties of NEAU-TCZ24T also confirmed the affiliation of the isolate to the genus Cellulomonas. However, multilocus sequence analysis based on five other house-keeping genes (gyrB, rpoB, recA, relA and atpD), DNA-DNA relatedness, physiological and biochemical data indicated that NEAU-TCZ24T could be distinguished from its closest relatives. Therefore, it is proposed that NEAU-TCZ24T represents a novel species of the genus Cellulomonas, for which the name Cellulomonas rhizosphaerae sp. nov. is proposed. The type strain is NEAU-TCZ24T (=CCTCC AA 2018042T=JCM 32383T).

Cellulomonas macrotermitis sp. nov., a chitinolytic and cellulolytic bacterium isolated from the hindgut of a fungus-growing termite.

To investigate the symbiotic roles of the gut microbiota in the fungus-growing termite Macrotermes barneyi, a novel strain with chitinolytic and cellulolytic activity, designated strain an-chi-1T, was isolated from the hindgut of M. barneyi. Strain an-chi-1T grows optimally at 28-30 °C, pH 8.0 in PYG medium. On the basis of 16S rRNA gene sequence analysis, this isolate belongs to the genus Cellulomonas with high sequence similarity to Cellulomonas iranensis (99.4%), followed by Cellulomonas flavigena (98.4%), Cellulomonas phragmiteti (97.4%), Cellulomonas oligotrophica (97.2%) and Cellulomonas terrae (97.0%). The DNA-DNA relatedness between an-chi-1T and the type strains of C. iranensis and C. flavigena DSM20109T are 35.4% and 23.7%, respectively. The major cellular fatty acids are anteiso-C15:0 and C14:0. The polar lipid profile consists of diphosphatidylglycerol, phosphatidylinositol mannosides, phosphatidylinositol dimannosides and one unidentified phospholipid. The cell-wall sugar is ribose. The peptidoglycan contains glutamic acid, aspartic acid and alanine. The DNA G+C content is 67.3 mol%. Based on its distinctive phenotypic, phylogenetic, and chemotaxonomic characteristics, an-chi-1T represents a novel species of the genus Cellulomonas, for which the name Cellulomonas macrotermitis sp. nov. is proposed. The type strain is an-chi-1T (= JCM 31923T = CICC 24195T).

Identification of clinically relevant Corynebacterium strains by Api Coryne, MALDI-TOF-mass spectrometry and molecular approaches.

We evaluated the Bruker Biotyper matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) for the identification of 97 Corynebacterium clinical in comparison to identification strains by Api Coryne and MALDI-TOF-MS using 16S rRNA gene and hypervariable region of rpoB genes sequencing as a reference method. C. striatum was the predominant species isolated followed by C. amycolatum. There was an agreement between Api Coryne strips and MALDI-TOF-MS identification in 88.65% of cases. MALDI-TOF-MS was unable to differentiate C. aurimucosum from C. minutissimum and C. minutissimum from C. singulare but reliably identify 92 of 97 (94.84%) strains. Two strains remained incompletely identified to the species level by MALDI-TOF-MS and molecular approaches. They belonged to Cellulomonas and Pseudoclavibacter genus. In conclusion, MALDI-TOF-MS is a rapid and reliable method for the identification of Corynebacterium species. However, some limits have been noted and have to be resolved by the application of molecular methods.

Cellulomonas pakistanensis sp. nov., a moderately halotolerant Actinobacteria.

A rod-shaped, motile, facultatively anaerobic and moderately halotolerant plant-growth-promoting actinobacterial strain, designated NCCP-11(T), was isolated from paddy grains. To delineate its taxonomic position, the strain was subjected to a polyphasic characterization. Cells of strain NCCP-11(T) grew at 10-37 °C (optimum 28-32 °C), at pH 6-9 (optimum pH 7) and in 0-12% (w/v) NaCl (optimum 1-2%) in broth medium. Based on 16S rRNA gene sequence analysis, strain NCCP-11(T) showed highest similarity to the type strains of Cellulomonas hominis (98.99%) and Cellulomonas denverensis (98.09 %) and less than 97 % with other closely related taxa. The chemotaxonomic data [major menaquinone: MK-9(H4); cell-wall peptidoglycan: type A4ß; major fatty acids: anteiso-C15 : 0, C16 : 0, C14 : 0 and anteiso-C17 : 0; major polar lipids: diphosphatidylglycerol, phosphatidylinositol, phosphatidylinositolmannosides and two unknown polar lipids] also supported the affiliation of strain NCCP-11(T) to the genus Cellulomonas. The level of DNA-DNA relatedness between strain NCCP-11(T) and the two type strains mentioned above was less than 42.7%. On the basis of DNA-DNA relatedness, physiological and biochemical characteristics and phylogenetic position, strain NCCP-11(T) can be differentiated from species of the genus Cellulomonas with validly published names and thus represents a novel species, for which the name Cellulomonas pakistanensis sp. nov. is proposed. The type strain is NCCP-11(T) ( = DSM 24792(T) = JCM 18755(T) = KCTC 19798(T)).

Cellulomonas marina sp. nov., isolated from deep-sea water.

A bacterial strain FXJ8.089(T) was isolated from deep-sea water collected from the southwest Indian Ocean (49° 39' E 37° 47' S) at a depth of 2800 m, and its taxonomic position was investigated by a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain FXJ8.089(T) belonged to the genus Cellulomonas and had the highest similarities with Cellulomonas oligotrophica (96.9 %) and Cellulomonas aerilata (96.6 %). It contained MK-9(H4) as the predominant menaquinone. The polar lipids were diphosphatidylglycerol and phosphatidylinositol mannosides. The cell-wall peptidoglycan type was A4ß with an interpeptide bridge L-Orn-D-Glu. The cell-wall sugars were glucose, mannose and ribose. The DNA G+C content was 70.3 mol%. The strain also showed a number of physiological and biochemical characteristics that were distinct from the closely related species. Based on phenotypic and genotypic data, strain FXJ8.089(T) (= CGMCC 4.6945(T) = DSM 24960(T)) represents a novel species of the genus Cellulomonas, for which the name Cellulomonas marina sp. nov. is proposed.

The genome sequences of Cellulomonas fimi and "Cellvibrio gilvus" reveal the cellulolytic strategies of two facultative anaerobes, transfer of "Cellvibrio gilvus" to the genus Cellulomonas, and proposal of Cellulomonas gilvus sp. nov.

Actinobacteria in the genus Cellulomonas are the only known and reported cellulolytic facultative anaerobes. To better understand the cellulolytic strategy employed by these bacteria, we sequenced the genome of the Cellulomonas fimi ATCC 484(T). For comparative purposes, we also sequenced the genome of the aerobic cellulolytic "Cellvibrio gilvus" ATCC 13127(T). An initial analysis of these genomes using phylogenetic and whole-genome comparison revealed that "Cellvibrio gilvus" belongs to the genus Cellulomonas. We thus propose to assign "Cellvibrio gilvus" to the genus Cellulomonas. A comparative genomics analysis between these two Cellulomonas genome sequences and the recently completed genome for Cellulomonas flavigena ATCC 482(T) showed that these cellulomonads do not encode cellulosomes but appear to degrade cellulose by secreting multi-domain glycoside hydrolases. Despite the minimal number of carbohydrate-active enzymes encoded by these genomes, as compared to other known cellulolytic organisms, these bacteria were found to be proficient at degrading and utilizing a diverse set of carbohydrates, including crystalline cellulose. Moreover, they also encode for proteins required for the fermentation of hexose and xylose sugars into products such as ethanol. Finally, we found relatively few significant differences between the predicted carbohydrate-active enzymes encoded by these Cellulomonas genomes, in contrast to previous studies reporting differences in physiological approaches for carbohydrate degradation. Our sequencing and analysis of these genomes sheds light onto the mechanism through which these facultative anaerobes degrade cellulose, suggesting that the sequenced cellulomonads use secreted, multidomain enzymes to degrade cellulose in a way that is distinct from known anaerobic cellulolytic strategies.

Cellulomonas soli sp. nov. and Cellulomonas oligotrophica sp. nov., isolated from soil.

Two novel bacterial strains, designated Kc1(T) and Kc5(T), were isolated from soil in Japan. Cells of the novel strains were Gram-reaction-positive, aerobic or facultatively anaerobic, motile rods. Phylogenetic analyses based on 16S rRNA gene sequences indicated that both strains belonged to the genus Cellulomonas. The 16S rRNA gene sequences of strains Kc1(T) and Kc5(T) showed closest similarity to that of Cellulomonas terrae DB5(T) (98.1 % and 98.4 % similarity, respectively), and the 16S rRNA gene similarity between the two novel strains was 97.8 %. In both strains, the major menaquinone was MK-9(H(4)), the predominant polar lipids were diphosphatidylglycerol and phosphatidylinositol mannosides, and the peptidoglycan contained ornithine and glutamic acid. Cell-wall sugars were identified as rhamnose, galactose and mannose in strain Kc1(T) and rhamnose and glucose in strain Kc5(T). The DNA G+C contents of strains Kc1(T) and Kc5(T) were 73.6 mol% and 75.8 mol%, respectively. Based on the chemotaxonomic and physiological data and the results of DNA-DNA hybridizations, the two strains represent two novel species within the genus Cellulomonas, for which the names Cellulomonas soli sp. nov. (type strain Kc1(T) =DSM 24484(T) =JCM 17535(T)) and Cellulomonas oligotrophica sp. nov. (type strain Kc5(T) =DSM 24482(T) =JCM 17534(T)) are proposed.

Cellulomonas carbonis sp. nov., isolated from coal mine soil.

A Gram-positive, aerobic, motile, rod-shaped bacterium, designated strain T26(T), was isolated from subsurface soil of Tianjin coal mine, China. Colonies were yellow-white, convex, circular, smooth and non-transparent on R2A agar. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain T26(T) was closely related to members of the genus Cellulomonas and a member of the genus Actinotalea with 96.8-94.7% and 96.7% gene sequence similarities, respectively. The peptidoglycan type of strain T26(T) was A4ß, containing l-ornithine-d-glutamic acid as the interpeptide bridge. The cell-wall sugars were rhamnose, galactose, xylose and inositol. The major fatty acids (>10%) were anteiso-C(15:0) (33.6%), anteiso-C(15:1) A (22.1%), C(16:0) (14.4%) and C(14:0) (12.1%). The predominant respiratory quinone was MK-9(H(4)) and the genomic DNA G+C content was 74.4 mol%. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol-mannosides and phosphatidylinositol. Comparison of phenotypic and phylogenetic characteristics between strain T26(T) and related organisms revealed that the new isolate represented a novel species of the genus Cellulomonas, for which the name Cellulomonas carbonis sp. nov. is proposed. The type strain is T26(T) ( = CGMCC 1.10786(T) = KCTC 19824(T) = CCTCC AB2010450(T)).

Cellulomonas phragmiteti sp. nov., a cellulolytic bacterium isolated from reed (Phragmites australis) periphyton in a shallow soda pond.

An alkalitolerant and moderately halophilic strain, designated KB23(T), characterized by optimal growth at pH 8.0-9.0 and in the presence of 5-7 % (w/v) NaCl, was isolated from a reed (Phragmites australis) periphyton sample originating from an extremely shallow, alkaline soda pond located in Hungary. Cells of strain KB23(T) were Gram-stain-positive, motile straight rods. Strain KB23(T) was facultatively anaerobic, catalase-positive, oxidase-negative and contained peptidoglycan type A4ß (L-Orn-D-Asp). MK-9(H4) was the predominant isoprenoid quinone and anteiso-C(15 : 0), C(16 : 0) and anteiso-C(15 : 1) were the major cellular fatty acids. The DNA G+C content of strain KB23(T) was 74.8 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that this strain belongs to the genus Cellulomonas and that it is related most closely to Cellulomonas flavigena DSM 20109(T) (97.35 % similarity), Cellulomonas terrae DB5(T) (96.81 %), Cellulomonas iranensis O(T) (96.75), Cellulomonas chitinilytica X.bu-b(T) (96.60 %), Cellulomonas persica I(T) (96.53 %), Cellulomonas composti TR7-06(T) (96.45 %), Cellulomonas biazotea DSM 20112(T) (96.34 %) and Cellulomonas fimi DSM 20113(T) (96.20 %). According to these results, together with DNA-DNA hybridization and physiological data, strain KB23(T) is considered to represent a novel species of the genus Cellulomonas, for which the name Cellulomonas phragmiteti sp. nov. is proposed. The type strain is KB23(T) ( = DSM 22512(T)  = NCAIM B002303(T)).

Cellulomonas aerilata sp. nov., isolated from an air sample.

A Gram-positive, aerobic, motile, coccoid or short rod-shaped bacterium, 5420S-23(T), was isolated from an air sample collected in the Republic of Korea. According to phylogenetic analysis based on 16S rRNA gene sequences, strain 5420S-23(T) revealed 97.5, 97.3, 97.3 and 97.2 % similarity, respectively, to Cellulomonas biazotea DSM 20112(T), Cellulomonas cellasea DSM 20118(T), Cellulomonas fimi DSM 20113(T) and Cellulomonas chitinilytica X.bu-b(T). The peptidoglycan type of strain 5420S-23(T) was A4beta, containing l-ornithine-d-glutamic acid. The cell-wall sugars were galactose, glucose and xylose. The major fatty acids were anteiso-C(15 : 0) (49.7 %) and C(16 : 0) (20.0 %). The major menaquinone was MK-9(H(4)) and major polar lipids were diphosphatidylglycerol and phosphatidylglycerol. The DNA G+C content was 74 mol%. The results of DNA-DNA hybridization with strains of closely related Cellulomonas species, in combination with chemotaxonomic and physiological data, demonstrated that isolate 5420S-23(T) represents a novel Cellulomonas species, for which the name Cellulomonas aerilata sp. nov. is proposed, with strain 5420S-23(T) (=KACC 20692(T) =DSM 18649(T)) as the type strain.

Cellulomonas chitinilytica sp. nov., a chitinolytic bacterium isolated from cattle-farm compost.

A bacterial strain, designated X.bu-b T, with chitin-, xylan-, cellulose- and starch-degrading activities, was isolated from compost at a cattle farm near Daejeon, Republic of Korea. The strain comprised Gram-positive, aerobic or facultatively anaerobic, non-motile, rod-shaped bacteria. On the basis of an analysis of 16S rRNA gene sequences, the phylogenetic position of X.bu-b T was within the genus Cellulomonas, and the strain exhibited relatively high sequence similarities with respect to Cellulomonas biazotea DSM 20112T (98.1 %), C. cellasea DSM 20118T (98.1 %), C. fimi DSM 20113T (98.0 %), C. terrae DB5T (97.9 %), C. humilata ATCC 25174T (97.7 %), C. xylanilytica XIL11 T (97.5 %), C. uda DSM 20107T (97.4 %), C. gelida DSM 20111 T (97.3 %), C. iranensis OT (97.3 %) and C. flavigena DSM 20109T (97.0 %). The phylogenetic distance from other Cellulomonas species with validly published names was greater than 3 % (i.e. less than 97.0 % sequence similarity). Chemotaxonomic data also supported the classification of strain X.bu-b T within the genus Cellulomonas: L-ornithine was the cell-wall diamino acid, anteiso-C15:0 and anteiso-C17:0 were the major fatty acids, rhamnose, galactose, xylose and ribose were the cell-wall sugars, MK-9(H4) was the predominant menaquinone and diphosphatidylglycerol and phosphatidylglycerol were present in the polar lipids. The G+C content of the genomic DNA was 73.6 mol%. DNA-DNA hybridization experiments showed that the values for DNA-DNA relatedness between strain X.bu-b T and the phylogenetically closest neighbours were below 23 %. On the basis of its phenotypic properties and phylogenetic distinctiveness, strain X.bu-b T represents a novel species of the genus Cellulomonas, for which the name Cellulomonas chitinilytica sp. nov. is proposed. The type strain is X.bu-b T (=KCTC 19133T =DSM 17922T).

Characterization and action patterns of two beta-1,4-glucanases purified from cellulomonas uda CS1-1.

Two beta-1,4-glucanases (DI and DIII fractions) were purified to homogeneity from the culture filtrate of a cellulolytic bacteria, Cellulomonas sp. CS1-1, which was classified as a novel species belonging to Cellulomonas uda based on chemotaxanomic and phylogenetic analyses. The molecular mass was estimated as 50,000 Da and 52,000 Da for DI and DIII, respectively. Moreover, DIII was identified as a glycoprotein with a pI of 3.8, and DI was identified as a non-glycoprotein with a pI of 5.3. When comparing the ratio of the CMC-saccharifying activity and CMC-liquefying activity, DI exhibited a steep slope, characteristic of an endoglucanase, whereas DIII exhibited a low slope, characteristic of an exoglucanase. The substrate specificity of the purified enzymes revealed that DI efficiently hydrolyzed CMC as well as xylan, whereas DIII exhibited a high activity on microcrystalline celluloses, such as Sigmacells. A comparison of the hydrolysis patterns for pNP-glucosides (DP 2-5) using an HPLC analysis demonstrated that the halosidic bond 3 from the nonreducing end was the preferential cleavage site for DI, whereas bond 2, from which the cellobiose unit is split off, was the preferential cleavage site for DIII. The partial N-terminal amino acid sequences for the purified enzymes were 1Ala-Gly-Ser-Thr-Leu-Gln-Ala-Ala-Ala-Ser-Glu-Ser-Gly-Arg-Tyr15- for DI and 1Ala-Asp-Ser-Asp-Phe-Asn-Leu-Tyr-Val-Ala-Glu-Asn-Ala-Met-Lys15- for DIII. The apparent sequences exhibited high sequence similarities with other bacterial beta-1,4-glucanases as well as beta-1,4-xylanases.

Cellulomonas composti sp. nov., a cellulolytic bacterium isolated from cattle farm compost.

A bacterial strain, TR7-06(T), which has cellulase and beta-glucosidase activities, was isolated from compost at a cattle farm near Daejeon, Republic of Korea. It was a Gram-positive, aerobic or facultatively anaerobic, non-motile, rod-shaped bacterium. Phylogenetic analysis based on 16S rRNA gene sequences showed that this strain belongs to the genus Cellulomonas, with highest sequence similarity to Cellulomonas uda DSM 20107(T) (98.5 %). Cell wall analysis revealed the presence of type A4beta, L-orn-D-Glu peptidoglycan. The cell-wall sugars detected were mannose and glucose. The predominant menaquinone was MK-9(H(4)); MK-8(H(4)) was detected in smaller quantities. The major fatty acids were anteiso-C(15 : 0), C(16 : 0), C(14 : 0) and C(18 : 0). The polar lipids detected were diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylinositol. The results of DNA-DNA hybridization and physiological and biochemical tests clearly demonstrated that TR7-06(T) represents a novel species. The combined genotypic and phenotypic data show that strain TR7-06(T) (=KCTC 19030(T)=NBRC 100758(T)) merits description as the type strain of a novel Cellulomonas species, Cellulomonas composti sp. nov.