Nocardioides panaciterrulae sp. nov., isolated from soil of a ginseng field, with ginsenoside converting activity

A Gram-positive, coccoid to rod-shaped, non-spore-forming bacterium, designated Gsoil 958^T, was isolated from soil of a ginseng field located in Pocheon province in South Korea. This bacterium was characterized in order to determine its taxonomic position by using a polyphasic approach. Strain Gsoil 958^T was observed to grow well at 25–30 °C and at pH 7.0 on R2A and nutrient agar without NaCl supplementation. Strain Gsoil 958^T was determined to have β-glucosidase activity and the ability to transform ginsenoside Rb₁ (one of the dominant active components of ginseng) to F₂ via gypenoside XVII and Rd. On the basis of 16S rRNA gene sequence similarity, strain Gsoil 958^T was shown to belong to the family Nocardioidaceae and related most closely to Nocardioides koreensis MSL-09^T (97.6 % 16S rRNA gene sequence similarity), Nocardioides aquiterrae GW-9^T (97.0 %), and Nocardioides sediminis MSL-01^T (97.0 %). The sequence similarities with other validly named species within the genus Nocardioides were less than 96.8 %. Strain Gsoil 958^T was characterized chemotaxonomically as having LL-2,6-diaminopimelic acid in the cell-wall peptidoglycan, MK-8(H₄) as the predominant menaquinone, and iso-C_16:0, iso-C_16:1 H, iso-C_14:0, iso-C_15:0 were identified as the major fatty acids. The G + C content of genomic DNA was determined to be 70.8 mol %. The chemotaxonomic properties and phenotypic characteristics supported the affiliation of strain Gsoil 958^T to the genus Nocardioides. The results of both physiological and biochemical tests allowed for differentiation of strain Gsoil 958^T from the recognized Nocardioides species. Therefore, strain Gsoil 958^T is considered to represent a novel species of the genus Nocardioides, for which the name Nocardioides panaciterrulae sp. nov. is proposed, with the type strain Gsoil 958^T (KACC 14271^T = KCTC 19471^T = DSM 21350^T).     

β-Glucosidase from Penicillium aculeatum hydrolyzes exo-, 3-O-, and 6-O-β-glucosides but not 20-O-β-glucoside and other glycosides of ginsenosides

A novel β-glucosidase from Penicillium aculeatum was purified as a single 110.5-kDa band on SDS–PAGE with a specific activity of 75.4 U?mg^?1 by salt precipitation and Hi-Trap Q HP and Resource Q ion exchange chromatographies. The purified enzyme was identified as a member of the glycoside hydrolase 3 family based on its amino acid sequence. The hydrolysis activity for p-nitrophenyl-β-d-glucopyranoside was optimal at pH 4.5 and 70 °C with a half-life of 55 h. The enzyme hydrolyzed exo-, 3-O-, and 6-O-β-glucosides but not 20-O-β-glucoside and other glycosides of ginsenosides. Because of the novel specificity, this enzyme had the transformation pathways for ginsenosides: Rb₁?→?Rd?→?F₂?→?compound K, Rb₂?→?compound O?→?compound Y, Rc?→?compound Mc₁?→?compound Mc, Rg₃?→?Rh₂?→?aglycone protopanaxadiol (APPD), Rg₁?→?F₁, and Rf?→?Rh₁?→?aglycone protopanaxatriol (APPT). Under the optimum conditions, the enzyme converted 0.5 mM Rb_2, Rc, Rd, Rg₃, Rg₁, and Rf to 0.49 mM compound Y, 0.49 mM compound Mc, 0.47 mM compound K, 0.23 mM APPD, 0.49 mM?F₁, and 0.44 mM APPT after 6 h, respectively.     

Flavobacterium kyungheensis sp. nov., isolated from soil of a ginseng field

A Gram-staining negative, strictly aerobic, motile by gliding, non-spore-forming, pale yellow pigmented and rod-shaped bacterium designated strain THG-107^T was isolated from soil of a ginseng field on Ganghwa Island in the Republic of Korea and its taxonomic position was investigated by using a polyphasic study. Growth of strain THG-107^T was found to occur at 4–37 °C (optimum, 20–30 °C), at pH 5.5–10 (optimum, pH 7.0) and in the presence of 0–1 % (w/v) NaCl (optimum, absence) on R2A agar. On the basis of 16S rRNA gene sequence similarity, strain THG-107^T was shown to belong to the family Flavobacteriaceae and was related to Flavobacterium denitrificans ED5^T (99.1 % similarity). The G+C content of the genomic DNA was determined to be 34.2 mol%. These results are consistent with characteristics of members of the genus Flavobacterium. The only isoprenoid quinone detected in strain THG-107^T was menaquinone-6 (MK-6) and the major polyamine was identified as homospermidine (82.9 %). The major polar lipid detected was phosphatidylethanolamine and the major cellular fatty acids were identified as iso-C_15:0 (26.3 %), iso-C_17:0 3OH (12.6 %) and summed feature 3 (comprising C_16:1 ω7c and/or C_16:1 ω6c; 11.6 %). Flexirubin-type pigments were found to be present. Strain THG-107^T has β-glucosidase activity to convert ginsenosides Rb₁ and Rd into Gyp17 and F₂. DNA-DNA hybridization with F. denitrificans ED5^T was 52 %. Strain THG-107^T could be distinguished from F. denitrificans ED5^T and the other species of the genus Flavobacterium by its phylogenetic and genetic distinctiveness and by several phenotypic properties. Therefore, strain THG-107^T is considered to represent a novel species in the genus Flavobacterium, for which the name Flavobacterium kyungheensis sp. nov. is proposed (type strain THG-107^T = KACC 16219^T = LMG 26575^T).     

Characterization of the ginsenoside-transforming recombinant β-glucosidase from Actinosynnema mirum and bioconversion of major ginsenosides into minor ginsenosides

This study focused on the cloning, expression, and characterization of ginsenoside-transforming recombinant β-glucosidase from Actinosynnema mirum KACC 20028^T in order to biotransform ginsenosides efficiently. The gene, termed as bglAm, encoding a β-glucosidase (BglAm) belonging to the glycoside hydrolase family 3 was cloned. bglAm consisted of 1,830 bp (609 amino acid residues) with a predicted molecular mass of 65,277 Da. This enzyme was overexpressed in Escherichia coli BL21(DE3) using a GST-fused pGEX 4T-1 vector system. The recombinant BglAm was purified with a GST·bind agarose resin and characterized. The optimum conditions of the recombinant BglAm were pH 7.0 and 37 °C. BglAm could hydrolyze the outer and inner glucose moieties at the C3 and C20 of the protopanaxadiol-type ginsenosides (i.e., Rb₁ and Rd, gypenoside XVII) to produce protopanaxadiol via gypenoside LXXV, F₂, and Rh₂(S) with various pathways. BglAm can effectively transform the ginsenoside Rb₁ to gypenoside XVII and Rd to F₂; the K _m values of Rb₁ and Rd were 0.69?±?0.06 and 0.45?±?0.02 mM, respectively, and the V _max values were 16.13?±?0.29 and 51.56?±?1.35 μmol min^?1 mg^?1 of protein, respectively. Furthermore, BglAm could convert the protopanaxatriol-type ginsenoside Re and Rg₁ into Rg₂(S) and Rh₁(S) hydrolyzing the attached glucose moiety at the C6 and C20 positions, respectively. These various ginsenoside-hydrolyzing pathways of BglAm may assist in producing the minor ginsenosides from abundant major ginsenosides.     

Bioconversion of ginsenosides Rb(1), Rb(2), Rc and Rd by novel β-glucosidase hydrolyzing outer 3-O glycoside from Sphingomonas sp. 2F2: cloning, expression, and enzyme characterization

A new β-glucosidase gene (bglSp) was cloned from the ginsenoside converting Sphingomonas sp. strain 2F2 isolated from the ginseng cultivating filed. The bglSp consisted of 1344 bp (447 amino acid residues) with a predicted molecular mass of 49,399 Da. A BLAST search using the bglSp sequence revealed significant homology to that of glycoside hydrolase superfamily 1. This enzyme was overexpressed in Escherichia coli BL21 (DE3) using a pET21-MBP (TEV) vector system. Overexpressed recombinant enzymes which could convert the ginsenosides Rb₁, Rb₂, Rc and Rd to the more pharmacological active rare ginsenosides gypenoside XVII, ginsenoside C-O, ginsenoside C-Mc₁ and ginsenoside F₂, respectively, were purified by two steps with Amylose-affinity and DEAE-Cellulose chromatography and characterized. The kinetic parameters for β-glucosidase showed the apparent K_m and V_max values of 2.9 ± 0.3 mM and 515.4 ± 38.3 μmol min⁻¹ mg of protein⁻¹ against p-nitrophenyl-β-d-glucopyranoside. The enzyme could hydrolyze the outer C3 glucose moieties of ginsenosides Rb₁, Rb₂, Rc and Rd into the rare ginsenosides Gyp XVII, C-O, C-Mc₁ and F₂ quickly at optimal conditions of pH 5.0 and 37 °C. A little ginsenoside F₂ production from ginsenosides Gyp XVII, C-O, and C-Mc₁ was observed for the lengthy enzyme reaction caused by the side ability of the enzyme.     

<Emphasis Type="Italic">Maribacter marinus</Emphasis> sp. nov. isolated from a deep-sea seamount

A Gram-stain negative, rod-shaped, aerobic strain, designated YC973^T, was isolated from a seamount near the Yap Trench in the tropical western Pacific. Phylogenetic analysis based on its 16S rRNA gene sequence showed that strain YC973^T is related to the genus Maribacter and has high 16S rRNA gene sequence similarity to Maribacter orientalis KMM 3947^T (97.6%). The predominant cellular fatty acids were iso-C_15:0, iso-C_15:1 G and an unidentified fatty acid of equivalent chain-length 13.565. The polar lipid profile contained phosphatidylethanolamine and five unidentified lipids. The genomic DNA G+C content of strain YC973^T was 36.1 mol%. On the basis of the evidence presented in this study, strain YC973^T represents a novel species of the genus Maribacter, for which we propose the name Maribacter marinus sp. nov. (type strain YC973^T = KACC 19025^T = CGMCC 1.16328^T).     

Pacificamonas flava gen. nov., sp. nov., a Novel Member of the Family Sphingomonadaceae Isolated from the Southeastern Pacific

Strain JLT2015^T was isolated from surface seawater of the Southeastern Pacific. The strain was Gram-negative, aerobic, motile by gliding, and rod shaped. The dominant fatty acids were C_18:1ω7c, C_16:0, and C_16:1ω7c. The major respiratory ubiquinone was Q-10, and the predominant polyamine pattern was spermidine. The components of the polar lipid profile were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, and sphingoglycolipid. The DNA G+C content was 64.2 %. Phylogenetic analysis based on 16S rRNA gene sequence revealed strain JLT2015^T belonged to belong to the family Sphingomonadaceae, exhibiting 94.7 % 16S rRNA gene sequence similarity with Novosphingobium pentaromativorans. On the basis of the taxonomic data presented, together with phylogenetic and genetic characteristics, strain JLT2015^T is considered to represent a novel genus, for which the name Pacificamonas flava gen. nov., sp. nov. is proposed. The type strain of Pacificamonas flava is JLT2015^T (=LMG27364^T = CGMCC1.12401^T).     

Bioconversion of ginsenoside Rc into Rd by a novel α-l-arabinofuranosidase, Abf22-3 from Leuconostoc sp. 22-3: cloning, expression, and enzyme characterization

A novel α-l-arabinofuranosidase (Abf22-3) that could biotransform ginsenoside Rc into Rd was obtained from the ginsenoside converting Leuconostoc sp. strain 22-3, isolated from the Korean fermented food kimchi. The gene, termed abf22-3, consisting of 1,527 bp and encoding a protein with a predicted molecular mass of 58,486 Da was cloned into the pMAL-c2x (TEV) vector. A BLAST search using the Abf22-3’s amino acid sequence revealed significant homology to that of family 51 glycoside hydrolases. The over-expressed recombinant Abf22-3 in Escherichia coli BL21 (DE3) catalyzed the hydrolysis of the arabinofuranoside moiety attached to the C-20 position of ginsenoside Rc under optimal conditions of pH 6.0 and 30 °C. This result indicated that Abf22-3 selectively converts ginsenoside Rc into Rd, but did not catalyze the hydrolysis of glucopyranosyl groups from Rc or other ginsenosides such as Rb₁ and Rb₂. Over-expressed recombinant enzymes were purified by two steps with amylose-affinity and DEAE-cellulose chromatography and then characterized. The kinetic parameters for α-l-arabinofuranosidase showed apparent K_m and V_max values of 0.95 ± 0.02 μM and 1.2 ± 0.1 μmol min^?1 mg of protein^?1 against p-nitrophenyl-α-l-arabinofuranoside, respectively. Using a purified MBP–Abf22-3 (10 μg/ml), 0.1 % of ginsenoside Rc was completely converted to ginsenoside Rd within 20 min.     

Glaciecola aquimarina sp. nov., a gammaproteobacterium isolated from coastal seawater

A Gram-negative, aerobic, non-motile and ovoid or rod-shaped bacterial strain, GGW-M5^T, was isolated from seawater on the southern coast in Korea, and its taxonomic position was investigated by using a polyphasic approach. Strain GGW-M5^T grew optimally at pH 7.0–8.0, at 30 °C and in the presence of 2 % (w/v) NaCl. A neighbour-joining phylogenetic tree based on 16S rRNA gene sequences revealed that strain GGW-M5^T belonged to the genus Glaciecola, joining the cluster comprising the type strains of G. agarilytica, G. arctica, G. chathamensis, G. mesophila, G. polaris and G. psychrophila, with which it exhibited 16S rRNA gene sequence similarity values of 95.9–96.7 %. Strain GGW-M5^T exhibited sequence similarity values of 93.2–94.8 % to the type strains of the other Glaciecola species. Strain GGW-M5^T contained Q-8 as the predominant ubiquinone and C_16:1 ω7c and/or iso-C_15:0 2-OH, C_16:0 and C_14:0 2-OH as the major fatty acids. Major polar lipids were phosphatidylglycerol, phosphatidylethanolamine and diphosphatidylglycerol. The DNA G+C content was 42.4 mol%. Differential phenotypic properties, together with the phylogenetic distinctiveness, demonstrated that strain GGW-M5^T could be distinguished from other Glaciecola species. On the basis of the data presented, strain GGW-M5^T is considered to represent a novel species of the genus Glaciecola, for which the name Glaciecola aquimarina sp. nov. is proposed. The type strain is GGW-M5^T (=KCTC 32108^T = CCUG 62918^T).     

Haloplanus litoreus sp. nov. and Haloplanus ruber sp. nov., from a marine solar saltern and an aquaculture farm,respectively

Two halophilic archaea, strains GX21^T and R35^T, were isolated from a marine solar saltern and an aquaculture farm in China, respectively. Cells of the two strains were observed to be pleomorphic, flat, to contain gas vesicles, stain Gram-negative and produce red-pigmented colonies. Strain GX21^T was found to be able to grow at 25–50 °C (optimum 37 °C), at 2.6–4.8 M NaCl (optimum 3.4 M NaCl), at 0.05–1.0 M MgCl₂ (optimum 0.1 M MgCl₂) and at pH 6.0–8.5 (optimum pH 6.5) while strain R35^T was found to be able to grow at 25–45 °C (optimum 37 °C), at 2.1–4.8 M NaCl (optimum 3.1 M NaCl), at 0–0.7 M MgCl₂ (optimum 0.03 M MgCl₂) and at pH 5.5–9.5 (optimum pH 6.5–7.0). The cells of both isolates were observed to lyse in distilled water. The minimum NaCl concentrations that prevented cell lysis were determined to be 15 % (w/v) for strain GX21^T and 12 % (w/v) for strain R35^T. The major polar lipids of the two strains were identified as phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate, one major glycolipid and a minor lipid chromatographically identical to sulfated mannosyl glucosyl diether and mannosyl glucosyl diether, respectively. 16S rRNA gene sequence analysis revealed that strains GX21^T and R35^T show 97.1 % sequence similarity to each other and are closely related to Haloplanus aerogenes TBN37^T (96.8 and 95.8 %), Haloplanus vescus RO5-8^T (96.7 and 96.1 %), Haloplanus salinus YGH66^T (96.4 and 95.8 %) and Haloplanus natans JCM 14081^T (96.3 and 95.4 %). The rpoB′ gene similarity between strains GX21^T and R35^T is 90.5 % and show 88.5–90.8 % similarity to the Haloplanus species with validly published names. The DNA G+C content of strain GX21^T and R35^T were determined to be 65.8 and 66.0 mol%, respectively. The DNA–DNA hybridization values between strain GX21^T and strain R35^T, and the two strains with the Haloplanus species with validly published names, showed less than 50 % DNA–DNA relatedness. It was concluded that strain GX21^T (=CGMCC 1.10456^T = JCM 17092^T) and strain R35^T (=CGMCC 1.10594 ^T = JCM 17271^T) represent two new species of Haloplanus, for which the names Haloplanus litoreus sp. nov. and Haloplanus ruber sp. nov. are proposed.     

Saccharopolyspora dendranthemae sp. nov. a halotolerant endophytic actinomycete isolated from a coastal salt marsh plant in Jiangsu, China

A halotolerant actinomycete strain, designated strain KLBMP 1305^T, was isolated from a salt marsh plant Dendranthema indicum (Linn.) Des Moul collected from the coastal region of Nantong, Jiangsu Province, in east China and was studied in detail for its taxonomic position. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain KLBMP 1305^T is a member of the genus Saccharopolyspora. The 16S rRNA gene sequence similarity indicated that strain KLBMP 1305^T was most closely related to ‘Saccharopolyspora pathumthaniensis’ S582^T (99.31 %), ‘Saccharopolyspora endophytica’ YIM 61095^T (99.17 %) and Saccharopolyspora tripterygii YIM 65359^T (99.15 %); similarity to other type strains of the genus Saccharopolyspora was <97.2 %. The organism had chemical and morphological features consistent with its classification in the genus Saccharopolyspora such as meso-diaminopimelic acid as the diagnostic diamino acid in the cell wall peptidoglycan and arabinose and galactose as the diagnostic sugars. The predominant menaquinone was MK-9(H₄). The polar lipids detected were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, an unknown glycolipid and an unknown lipid. The major fatty acids were iso-C_16:0, iso-C_15:0, anteiso-C_15:0, anteiso-C_17:0 and sum in feature 8 (18:1ω7c/18:1ω6c)_. The G+C content of the genomic DNA of the type strain was 68.7 mol%. DNA–DNA relatedness data, together with phenotypic differences, clearly distinguished the isolate from its closest relatives. On the basis of these phenotypic and genotypic data, the isolate represents a novel species, for which the name Saccharopolyspora dendranthemae sp. nov. is proposed. The type strain is KLBMP 1305^T (=KCTC 19889^T = NBRC 108675^T).     

Salinicola peritrichatus sp. nov., isolated from deep-sea sediment

A Gram stain-negative, aerobic and rod-shaped bacterium, strain DY22^T, was isolated from a deep-sea sediment collected from the east Pacific Ocean. The isolate was found to grow in the presence of 0–20.0 % (w/v) NaCl and at pH 4.5–8.5; optimum growth was observed with 0.5–2.0 % (w/v) NaCl and at pH 5.0–7.0. Chemotaxonomic analysis showed the presence of ubiquinone-9 as predominant respiratory quinone and C_16:0, C_19:0 ω8c cyclo and C_12:0 3-OH as major cellular fatty acids. The genomic DNA G+C content was determined to be 59.6 mol%. Comparative 16S rRNA gene sequence analysis revealed that the novel isolate belongs to the genus Salinicola. Strain DY22^T exhibited the closest phylogenetic affinity to the type strain of Salinicola salarius with 97.2 % sequence similarity and less than 97 % sequence similarity with respect to other Salinicola species with validly published names. The DNA–DNA reassociation values between strain DY22^T and S. salarius DSM 18044^T was 52 ± 4 %. On the basis of phenotypic, chemotaxonomic and genotypic data, strain DY22^T represents a novel species of the genus Salinicola, for which the name Salinicola peritrichatus sp. nov. (type strain DY22^T = CGMCC 1.12381^T = JCM 18795^T) is proposed.     

Chryseobacterium yeoncheonense sp. nov., with ginsenoside converting activity isolated from soil of a ginseng field

A Gram-staining negative, aerobic, non-motile, non-flagellate, yellow-pigmented, rod-shaped bacterial strain, designated strain DCY67^T, was isolated from ginseng field in Republic of Korea. Strain DCY67^T contained β-glucosidase activity which converts ginsenoside Rb1 to compound K. Optimum growth of DCY67^T occurred at 30 °C and pH 6.0–6.5. Analysis of the 16S rRNA gene sequences revealed that strain DCY67^T belonged to the family Flavobacteriaceae and was most closely related to Chryseobacterium ginsenosidimutans THG 15^T (97.5 %). The genomic DNA G+C content was 36.1 mol%. The predominant quinones were MK-6 (90.9 %) and MK-7 (9.15 %). The major fatty acids were iso-C_15:0, summed feature 3 (containing C_16:1 ω7c and/or C_16:1 ω6c) and iso-C_17:0 3-OH. On the basis of these phenotypic, genotypic and chemotaxonomic studies, strain DCY67^T represents a novel species of the genus Chryseobacterium, for which, name Chryseobacterium yeoncheonense sp. nov. proposed the type strain is DCY67^T (=KCTC 32090^T = JCM 18516^T).     

Halobellus rufus sp. nov., an extremely halophilic archaeon isolated from non-purified solar salt

A halophilic archaeon, designed strain CBA1103^T, was isolated from non-purified solar salt. The cells of strain CBA1103^T were observed to be Gram-stain negative and pleomorphic, and the colonies appear red. Strain CBA1103^T was observed to grow between 20 and 55 °C (optimum 37 °C), and in NaCl concentrations of 10–30 % (w/v) (optimum 15 %) with 0–0.5 M MgSO₄·7H₂O (optimum 0.1 M) and at pH 6.0–9.0 (optimum pH 7.0). Additionally, the cells lyse in distilled water. The major polar lipids of strain CBA1103^T are phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and two glycolipids chromatographically identical to sulfated mannosyl glucosyl diether and manosyl glucosyl diether. Strain CBA1103^T is shown to belong to the Halobellus genus and exhibits similarity to related taxa; the 16S rRNA gene sequence similarity between strain CBA1103^T and Halobellus rarus 18362^T, Hbs. limi 16811^T, Hbs. litoreus JCM 17118^T, Hbs. inordinatus YC20^T, Hbs. clavatus TNN18^T and Hbs. salinus CSW2.24.4^T is 97.3, 96.5, 96.5, 94.5, 94.5 and 93.7 %, respectively. The RNA polymerase subunit B gene sequence of strain CBA1103^T shows 93.7 % similarity with the sequence of Hbs. litoreus JCM 17118^T; the similarity is lower with sequences from the type strains of other species of Halobellus. The genomic DNA G+C content of strain CBA1103^T was determined to be 67.0 mol% a value which is in the range of the genomic DNA G+C content of members of the genus Halobellus (61.5–69.2 mol%). These results suggest that strain CBA1103^T should be considered to represent a new taxon for which the name Halobellus rufus sp. nov. is proposed, with the type strain CBA1103^T (=CECT 8423^T =JCM 19434^T).     

Rothia marina sp. nov., isolated from an intertidal sediment of the South China Sea

A novel non-sporulating, non-motile, catalase-positive, oxidase-negative, facultatively anaerobic, Gram-positive coccus, designated strain JSM 078151^T, was isolated from an intertidal sediment sample collected from Naozhou Island in the South China Sea, China. Growth was found to occur in the presence of 0–15 % (w/v) NaCl (optimum 0.5–3 % (w/v) NaCl), at pH 6.5–10.5 (optimum pH 7.0–8.0) and at 5–35 °C (optimum 25–30 °C). The peptidoglycan type was determined to be A3a, containing lysine, glutamic acid and alanine. The major cellular fatty acid identified was anteiso-C_15:0 and the predominant menaquinones are MK-7 and MK-8. The polar lipids were found to consist of diphosphatidylglycerol, phosphatidylglycerol, glycolipid and one unidentified phospholipid. The genomic DNA G+C content of strain JSM 078151^T was determined to be 55.2 mol%. A phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain JSM 078151^T should be assigned to the genus Rothia, and was most closely related to Rothia nasimurium CCUG 35957^T (98.3 % sequence similarity), followed by Rothia amarae J18^T (97.5 %) and Rothia terrae L-143^T (97.3 %). A combination of phylogenetic analysis, DNA–DNA relatedness values, phenotypic characteristics and chemotaxonomic data supports the suggestion that strain JSM 078151^T represents a novel species of the genus Rothia, for which the name Rothia marina sp. nov. is proposed. The type strain is JSM 078151^T (= DSM 21080^T = KCTC 19432^T).     

Microlunatus cavernae sp. nov., a novel actinobacterium isolated from Alu ancient cave, Yunnan, south-west China

A Gram-positive, coccoid, non-endospore-forming actinobacterium, designated YIM C01117^T, was isolated from a soil sample collected from Alu ancient cave, Yunnan province, south-west China. Based on the 16S rRNA gene sequence analysis, strain YIM C01117^T was shown to belong to the genus Microlunatus, with highest sequence similarity of 97.4 % to Microlunatus soli DSM 21800^T. The whole genomic DNA relatedness as shown by the DNA–DNA hybridization study between YIM C01117^T and M. soli DSM 21800^T had a low value (47 ± 2 %). Strain YIM C01117^T was determined to contain LL-diaminopimelic acid with Gly, Glu and Ala amino acids (A3γ′ type) in the cell wall. Whole-cell hydrolysates were found to contain glucose, galactose, mannose and ribose. The major polar lipids were determined to be phosphatidylglycerol and diphosphatidylglycerol. The predominant menaquinone system present is MK-9(H₄), while the major fatty acids were identified to be anteiso-C_15:0 (24.1 %), iso-C_16:0 (22.3 %) and iso-C_15:0 (11.4 %). The G+C content of the genomic DNA was determined to be 65.9 mol%. The chemotaxonomic and genotypic data support the affiliation of the strain YIM C01117^T to the genus Microlunatus. The results of physiological and biochemical tests allow strain YIM C01117^T to be differentiated phenotypically from recognized Microlunatus species. Strain YIM C01117^T is therefore considered to represent a novel species of the genus Microlunatus, for which the name Microlunatus cavernae sp. nov. is proposed. The type strain is YIM C01117^T (= DSM 26248^T = JCM 18536^T).     

Sphingomonas ginsenosidivorax sp. nov., with the ability to transform ginsenosides

A Gram-negative, strictly aerobic, non-motile, non-spore-forming and rod-shaped bacterial strain designated KHI67^T was isolated from sediment of the Gapcheon River in South Korea and its taxonomic position was investigated by using a polyphasic approach. Strain KHI67^T was observed to grow optimally at 25–30 °C and at pH 7.0 on nutrient and R2A agar. On the basis of 16S rRNA gene sequence similarity, strain KHI67^T was shown to belong to the family Sphingomonadaceae and was related to Sphingomonas faeni MA-olki^T (97.6 % sequence similarity), Sphingomonas aerolata NW12^T (97.5 %) and Sphingomonas aurantiaca MA101b^T (97.3 %). The G + C content of the genomic DNA was determined to be 65.6 %. The major ubiquinone was found to be Q-10, the major polyamine was identified as homospermidine and the major fatty acids identified were summed feature 8 (comprising C_18:1 ω7c/ω6c; 37.0 %), C_16:0 (13.0 %), summed feature 3 (comprising C_16:1 ω7c/C_16:1 ω6c; 12.8 %) and C_14:0 2OH (9.3 %). DNA and chemotaxonomic data supported the affiliation of strain KHI67^T to the genus Sphingomonas. The DNA–DNA relatedness values between strain KHI67^T and its closest phylogenetic neighbours were below 15 %. Strain KHI67^T could be differentiated genotypically and phenotypically from the recognised species of the genus Sphingomonas. The isolate therefore represents a novel species, for which the name Sphingomonas ginsenosidivorax sp. nov. is proposed, with the type strain KHI67^T (=KACC 14951^T = JCM 17076^T = LMG 25801^T).     

Sabulilitoribacter multivorans gen. nov., sp. nov., a polysaccharide-degrading bacterium of the family Flavobacteriaceae isolated from seashore sand

A Gram-negative, aerobic, non-flagellated, non-gliding and rod-shaped bacterial strain, designated M-M16^T, was isolated from seashore sand around a seaweed farm on the South Sea, South Korea, and its taxonomic position was investigated by using a polyphasic study. Strain M-M16^T grew optimally at 30 °C, at pH 7.0–8.0 and in the presence of 2 % (w/v) NaCl. Strain M-M16^T exhibited the highest 16S rRNA gene sequence similarity values to the type strains of Gaetbulibacter lutimaris (96.5 %) and Flaviramulus basaltis (95.8 %). Neighbour-joining and maximum-parsimony phylogenetic trees based on 16S rRNA gene sequences revealed that strain M-M16^T clustered with the type strains of Gaetbulibacter species and F. basaltis. Strain M-M16^T contained MK-6 as the predominant menaquinone and iso-C_15:1 G, iso-C_15:0 and iso-C_17:0 3-OH as the major fatty acids. The major polar lipids detected in strain M-M16^T were phosphatidylethanolamine and one unidentified lipid. The DNA G+C content of strain M-M16^T was 37.4 mol%. The phylogenetic and chemotaxonomic data and other phenotypic properties revealed that strain M-M16^T represents a novel genus and species within the family Flavobacteriaceae, for which the name Sabulilitoribacter multivorans gen. nov., sp. nov. is proposed. The type strain of S. multivorans is M-M16^T (= KCTC 32326^T = CCUG 63831^T).     

Bacillus bingmayongensis sp. nov., isolated from the pit soil of Emperor Qin’s Terra-cotta warriors in China

A Bacillus-like isolate, strain FJAT-13831^T, isolated from the No. 1 pit soil of Emperor Qin’s Terra-cotta Warriors in Xi’an City, China, was studied to determine its taxonomic status. Dominant fatty acids of this organism included iso-C_15:0, iso-C_17:0, C_16:0, iso-C_13:0, anteiso-C_15:0, and iso-C_17:1ω5c. Comparative 16S rRNA gene sequence analysis confirmed the affiliation of this isolate to the genus Bacillus and indicated that it was closely related to Bacillus pseudomycoides DSM 12442^T (99.72 % similarity). A phylogenetic analysis of the gyrB gene sequence similarities exhibited independent clustering of the isolate FJAT-13831^T and showed 93.8 % (<95 %) sequence similarity with its closest phylogenetic neighbour B. pseudomycoides DSM 12442^T. Separate standing of the strain FJAT-13831^T was supported by a whole genome-based phylogenetic analysis with an average nucleotide identity value of 91.47 (<95 %) between isolate FJAT-13831^T and B. pseudomycoides DSM 12442^T and was consistent with the results of DNA–DNA hybridization (69.1 % relatedness). These findings support the conclusion that the isolate FJAT-13831^T represents a novel species, for which the name Bacillus bingmayongensis sp. nov. is proposed. The type strain is FJAT-13831^T (= CGMCC 1.12043^T = DSM 25427^T).     

Pontibacter humi sp. nov., Isolated from Mountain Soil

A Gram-negative, aerobic, non-motile, rod-shaped bacterial strain, designated as SWU8^T, was isolated from a mountain soil collected in Seoul Women’s University campus at South Korea. Phylogenic analysis, using 16S rRNA gene sequence of the new isolate, showed that strain SWU8^T belongs to the genus Pontibacter. The highest sequence similarities were 96.2 % with Pontibacter saemangeumensis GCM0142^T, 95.5 % with Pontibacter toksunensis ZLD-7^T, 95.3 % with Pontibacter roseus DSM 17521^T, and 95.1 % with Pontibacter odishensis JC130^T. Chemotaxonomic data showed that the most abundant fatty acids were summed feature 4 (comprising iso-C_17:1 I/anteiso-C_17:1 B; 26.9 %), iso-C_15:0 (25.6 %), and iso-C_17:0 3OH (10.6 %), and major polar lipid was phosphatidylethanolamine. The DNA G+C content of strain SWU8^T was 48.5 mol%. Together, the phenotypic, phylogenetic, and chemotaxonomic data supported that strain SWU8^T presents a novel species of the genus Pontibacter, for which the name Pontibacter humi sp. nov. is proposed. The type strain is SWU8^T (=KEMC 9004-131^T = JCM 19178^T).