Limnovirga soli gen. nov., sp. nov., isolated from river sediment.

A cream-coloured, Gram-stain-negative, rod-shaped bacterium, designated strain KSC-6T, was isolated from soil sampled at the Gapcheon River watershed in Daejeon, Republic of Korea. The organism does not require NaCl for growth and grows at pH 6.0-8.0 (optimum, pH 7.0) and 10-37 °C (optimum, 25 °C). Phylogenetic trees based on the 16S rRNA gene sequences reveal that strain KSC-6T belongs to the family Chitinophagaceae within the order Chitinophagales and is most closely related to Panacibacter ginsenosidivorans Gsoil 1550T (95.9% similarity). The genomic DNA G+C content was 38.9 mol%. The major cellular fatty acids (>8 %) of strain KCS-6T were iso-C15:0, iso-C15 : 1 G and iso-C17 : 0 3-OH. The predominant respiratory quinone was menaquinone 7 and the predominant polar lipids were phosphatidylethanolamine, five unidentified aminolipids and two unidentified lipids. Based on genome analyses, low digital DNA-DNA hybridization, average nucleotide identity and average amino acid identity values with closely related genera, and differential chemotaxonomic and physiological properties, we suggest that strain KCS-6T represents a novel species in a new genus in the family Chitinophagaceae, for which the name Limnovirga soli gen. nov., sp. nov. (type strain KCS-6T=KCCM 43337T=NBRC 114336T) is proposed.

Paucibacter aquatile sp. nov. isolated from freshwater of the Nakdong River, Republic of Korea.

A Gram-negative, aerobic, motile, and rod-shaped bacterial strain designated CR182T was isolated from freshwater of the Nakdong River, Republic of Korea. Optimal growth conditions for this novel strain were found to be: 25-30 °C, pH 6.5-8.5, and 3% (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequence indicates that the strain CR182T belongs to type strains of genus Paucibacter. Strain CR182T showed 98.0% 16S rRNA gene sequence similarity with Paucibacter oligotrophus CHU3T and formed a robust phylogenetic clade with this species. The average nucleotide identity value between strain CR182T and P. oligotrophus CHU3T was 78.4% and the genome-to-genome distance was 22.2% on average. The genomic DNA G+C content calculated from the genome sequence was 66.3 mol%. Predominant cellular fatty acids of strain CR182T were summed feature 3 (C16:1 ω7c and/or C16:1 ω6c) (31.2%) and C16:0 (16.0%). Its major respiratory quinine was ubiquinone Q-8. Its polar lipids consisted of diphosphatidylglycerol, phosphatidylethanolamine, and two unidentified phospholipids. Its genomic DNA G+C content was 66.3%. Based on data obtained from this polyphasic taxonomic study, strain CR182T represents a novel species belonging to genus Paucibacter, for which a name of P. aquatile sp. nov. is proposed. The type strain is CR182T (= KCCM 90284T = NBRC 113032T).

Halostella salina gen. nov., sp. nov., an extremely halophilic archaeon isolated from solar salt.

A novel halophilic archaeon designated strain CBA1114T was isolated from solar salt in the Republic of Korea. Strain CBA1114T, cells of which were coccoid and Gram-stain-negative, grew in the presence of 15-30 % (w/v) NaCl (optimum, 20 %) and at 20-50 °C (optimum, 40 °C) and pH 7.0-9.0 (optimum, pH 8.0). Strain CBA1114T required Mg2+ for growth. Strain CBA1114T had three 16S rRNA genes, rrnA, rrnB and rrnC; levels of similarity between the sequences were 99.7-99.9 %. The 16S rRNA gene sequence of strain CBA1114T showed 91.7 % similarity to that of Haloterrigena thermotolerans PR5T. In multilocus sequence analysis (MLSA), five housekeeping genes, atpB, EF-2, radA, rpoB' and secY, were found to be closely related to those of the members of the genera Halorientalis(89.7 % similarity of the atpB gene sequence), Halomicroarcula(91.9 %, EF-2), Haloterrigena(85.4 %, radA), Natronoarchaeum(89.2 %, rpoB') and Natrinema(75.7 %, secY). A phylogenetic tree generated from the results of MLSA of the five housekeeping genes showed that strain CBA1114T was closely related to species of the genus Halorientalis in the family Halobacteriaceae. The major polar lipids were identified as phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and unidentified lipids. The G+C content of the genomic DNA of strain CBA1114T was 68.1 mol%. According to the results of phylogenetic, phenotypic and chemotaxonomic analyses, we designate strain CBA1114T (=JCM 30111T=KCTC 4206T) as the type strain of Halostella salina gen. nov., sp. nov., a novel species of a new genus within the family Halobacteriaceae.

Halolamina sediminis sp. nov., an extremely halophilic archaeon isolated from solar salt.

An extremely halophilic archaeal strain, halo-7T, was isolated from brine sediment of the Gomso solar saltern, Republic of Korea. Cells of strain halo-7T were pleomorphic, stained Gram-negative, lysed in distilled water and formed red-pigmented colonies. Strain halo-7T grew in the range of 25-45°C (optimum 37-40°C), pH 6.5-9.5 (optimum pH 7.0-8.0), 15-30% (w/v) NaCl (optimum, 20-25%), and 0.05-0.5 M MgCl2 (optimum 0.1-0.3 M). The minimal NaCl concentration to prevent cell lysis of strain halo-7T was 10% (w/v). The major polar lipids of the isolate were phosphatidylglycerol, phosphatidylglycerolphosphate methyl ester, an unidentified phospholipid, an unidentified lipid, and two unidentified glycolipids. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain halo-7T is closely related to the members of the genus Halolamina, Halolamina salina WSY15-H3T (98.7% 16S rRNA gene sequence similarity), Halolamina pelagica TBN21T (98.2%) and Halolamina rubra CBA1107T (97.4%). The genomic DNA G+C content determined for strain halo-7T (68.0 mol%) was slightly higher than those of H. salina JCM 18549T and H. rubra CBA1107T. DNA-DNA hybridization values between strain halo-7T and reference strains were < 25%. Based on phenotypic, chemotaxonomic and phylogenetic properties, we describe a novel species of the genus Halolamina, represented by strain halo-7T, for which we propose the name Halolamina sediminis sp. nov. The type strain is halo-7T ( = JCM 30187T = CECT 8739T).

Croceitalea litorea sp. nov., isolated from seashore sand.

Strain CBA3205T is a Gram-stain-negative, non-motile and rod-shaped bacterium that was isolated from the seashore sand of Jeju Island in South Korea. Based on the phylogenetic analysis, the most closed related species was Croceitalea eckloniae DOKDO 025T, with 94.8 % sequence similarity for the 16S rRNA gene. Strain CBA3205T was observed to grow optimally at 25-30 °C and at pH 8.5 in the presence of 2-3 % (w/v) NaCl. The major fatty acids of strain CBA3205T were iso-C15 : 0, iso-C15 : 1 G, and iso-C17 : 0 3-OH. The major respiratory quinone was MK-6 and the major polar lipids were two unidentified amino-group-containing phospholipids and an unidentified polar lipid. The G+C content of the genomic DNA of strain CBA3205T was 62.5 mol%. Based on the phenotypic, genotypic and phylogenetic analyses, strain CBA3205T was considered to be a novel species belonging to the genus Croceitalea within the family Flavobacteriaceae, for which the name Croceitalea litorea sp. nov. is proposed. The type strain is CBA3205T ( = KACC 17669T = JCM 19531T).

Vulcanisaeta thermophila sp. nov., a hyperthermophilic and acidophilic crenarchaeon isolated from solfataric soil.

An anaerobic, rod-shaped, hyperthermophilic and acidophilic crenarchaeon, designated strain CBA1501(T), was isolated from solfataric soil of the Mayon volcano in the Republic of the Philippines. Phylogenetic analysis showed that strain CBA1501(T) is affiliated with the genus Vulcanisaeta in the phylum Crenarchaeota. DNA sequence similarities between the 16S rRNA gene of strain CBA1501(T) and those of Vulcanisaeta distributa IC-017(T) and Vulcanisaeta souniana IC-059(T) were 98.5 and 97.4 %, respectively. Strain CBA1501(T) grew between 75-90 °C, over a pH range of 4.0-6.0 and in the presence of 0-1.0 % (w/v) NaCl, with optimal growth occurring at 85 °C, pH 5.0, and with 0 % (w/v) NaCl. Fumarate, malate, oxidized glutathione, sulfur and thiosulfate were used as final electron acceptors, but FeCl3, nitrate and sulfate were not. The DNA G+C content of strain CBA1501(T) was 43.1 mol%. On the basis of polyphasic taxonomic analysis, strain CBA1501(T) represents a novel species of the genus Vulcanisaeta in the phylum Crenarchaeota, for which we propose the name Vulcanisaeta thermophila sp. nov. The type strain is CBA1501(T) ( = ATCC BAA-2415(T) = JCM 17228(T)).

Draconibacterium filum sp. nov., a new species of the genus of Draconibacterium from sediment of the east coast of the Korean Peninsula.

A Gram-stain negative, long rod shaped, facultatively anaerobic bacterium, designated strain F2(T), was isolated from coastal sediment of the Korean Peninsula. Strain F2(T) was found to grow at 10-40 °C (optimum 30 °C), at pH 6.0-8.5 (optimum pH 7.5) and at 0.0-8.0 % (w/v) NaCl (optimum 3.0 %). Phylogenetic analysis of the 16S rRNA gene sequence showed that strain F2(T) is closely related to Draconibacterium orientale FH5(T) (with 97.9 % 16S rRNA gene similarity) of the family Prolixibacteraceae of the phylum Bacteroidetes. The major isoprenoid quinone was identified as MK-7 and the main fatty acids as iso-C15:0 (24.1 %), anteiso-C15:0 (15.4 %), C16:0 (10.7 %), iso-C17:0 3-OH (7.6 %) and iso-C16:0 3-OH (5.9 %). The major polar lipids were identified as phosphatidylethanolamine, phosphatidylinositol and four unidentified polar lipids. The genomic DNA G+C content of strain F2(T) was determined to be 44.7 mol% and the DNA-DNA relatedness of strain F2(T) with D. orientale DSM 25947(T) was 34.6 ± 4.3 %. Nitrate reduction capability and cell morphology of strain F2(T) are distinct from those of the closest relative, D. orientale DSM 25947(T). Based on these properties, we propose strain F2(T) represents a novel species within the genus Draconibacterium, with the name Draconibacterium filum sp. nov. The type strain of D. filum is F2(T) (=KCTC 32486(T) = JCM 19986(T)).

Bizionia psychrotolerans sp. nov., a psychrophilic bacterium isolated from the intestine of a sea cucumber (Apostichopus japonicus).

A novel Gram-negative, non-flagellated, non-gliding, rod-shaped bacterial strain, designated PB-M7(T), was isolated from the intestine of a sea cucumber collected from Pohang, South Korea. Growth was observed at 4-30 °C (optimum, 25 °C), pH 6.0-9.0 (optimum, pH 7.0-8.0), and with 2.0-6.0% (w/v) NaCl (optimum, 2.0%). In a phylogenetic analysis based on 16S rRNA gene sequences, strain PB-M7(T) was found to belong to the genus Bizionia and to be most closely related to Bizionia echini KMM 6177(T) (99.0% 16S rRNA gene sequence similarity), Bizionia hallyeonensis T-y7(T) (97.9%), Bizionia algoritergicola APA-1(T) (97.5%), Bizionia argentinensis JUB59(T) (97.5%) and Bizionia myxarmorum ADA-4(T) (97.1%). The predominant fatty acids of strain PB-M7(T) were identified as iso-C(15:0) (22.2%), iso-C(15:1) G (10.8%), iso-C(17:0) 3-OH (16.7%) and summed feature 3 (C(16:1) ω7c and/or C(16:1) ω6c; 11.2%). The major respiratory quinone was identified as menaquinone-6 (MK-6). The polar lipid profile of strain PB-M7(T) was found to contain phosphatidylethanolamine, an unidentified phospholipid, three unidentified aminolipids and three unidentified lipids. The DNA G + C content of strain PB-M7(T) was determined to be 33.4 mol% and the mean DNA-DNA relatedness values with the type strains of B. echini, B. hallyeonensis, B. algoritergicola, B. argentinensis, and B. myxarmorum were 52.9, 48.5, 46.5, 37.1 and 26.6%, respectively. Based on the data presented, strain PB-M7(T) represents a novel species of the genus Bizionia, for which the name Bizionia psychrotolerans sp. nov. is proposed. The type strain of B. psychrotolerans is PB-M7(T) (= KCCM 43042 (T) = JCM 19924 (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)).

Halolamina rubra sp. nov., a haloarchaeon isolated from non-purified solar salt.

Two Gram-stain negative, rod-shaped and motile extreme halophiles, designated CBA1107(T) and CBA1108, were isolated from non-purified solar salt. Based on the phylogenetic analysis, strains CBA1107(T) and CBA1108 were shown to belong to the genus Halolamina, with similarities for the 16S rRNA gene sequences between strains CBA1107(T) and Halolamina pelagica TBN21(T) , Halolamina salina WSY15-H3(T) and Halolamina salifodinae WSY15-H1(T) of 98.3, 97.6 and 97.3 %, respectively; the similarities for the rpoB' gene sequences between the same strains were 96.0, 95.3 and 94.6 %, respectively. The colonies of both strains were observed to be red pigmented on growth medium. Strain CBA1107(T) was observed to grow at 20-50 °C, in the presence of 15-30 % NaCl, at pH 6.0-9.0, and with 0.005-0.5 M Mg(2+). The cells of both strains lysed in distilled water. The DNA-DNA hybridization experiments showed that strain CBA1107(T) shared 97 % relatedness with CBA1108 and <50 % relatedness with H. pelagica JCM 16809(T), H. salina JCM 18549(T) and H. salifodinae JCM 18548(T). The genomic DNA G+C content of strain CBA1107(T) was determined to be 65.1 mol%. The major polar lipids of the two strains were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and glycolipids including sulfated mannosyl glucosyl diether and mannosyl glucosyl diether. Based on the polyphasic taxonomic analyses, the strains are considered to represent a new taxon for which the name Halolamina rubra sp. nov. is proposed, with the type strain CBA1107(T) (=CECT 8421(T) =JCM 19436(T)).

Halococcus sediminicola sp. nov., an extremely halophilic archaeon isolated from a marine sediment.

A novel, red-pigmented and coccoid haloarchaeon, designated strain CBA1101(T), was isolated from a marine sediment. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain CBA1101(T) is most closely related to the genus Halococcus in the family Halobacteriaceae. Strain CBA1101(T) had a highest 16S rRNA gene sequence similarity of 98.4 % with Halococcus dombrowskii DSM 14522(T), followed by 93.7-98.3 % with sequences of other type strains in the genus Halococcus. The RNA polymerase subunit B' gene sequence similarity of strain CBA1101(T) with that of Halococcus qingdaonensis JCM 13587(T) is 89.5 % and lower with those of other members of the genus Halococcus. Strain CBA1101(T) was observed to grow at 25-40 °C, pH 6.0-9.0 and in the presence of 15-30 % (w/v) NaCl, with optimal growth at 35-40 °C, pH 7.0 and with 20 % NaCl. The cells of strain CBA1101(T) are Gram-negative and did not lyse in distilled water. The major polar lipids were identified as phosphatidylglyerol, phosphatidylglycerol phosphate methyl ester, sulfated diglycosyl diether, unidentified phospholipids and unidentified glycolipids. The genomic DNA G+C content was determined 66.0 mol%. The DNA-DNA hybridization experiment showed that there was less than 40 % relatedness between strain CBA1101(T) and the reference species in the genus Halococcus. Based on this polyphasic taxonomic analysis, strain CBA1101(T) is considered to represent a new species in the genus Halococcus, for which the name Halococcus sediminicola sp. nov. is proposed. The type strain is CBA1101(T) (=JCM 18965(T) = CECT 8275(T)).

Halapricum salinum gen. nov., sp. nov., an extremely halophilic archaeon isolated from non-purified solar salt.

A halophilic archaeal strain, designated CBA1105(T), was isolated from non-purified solar salt. Strain CBA1105(T) was found to have three 16S rRNA genes, rrnA, rrnB and rrnC; similarities between the 16S rRNA gene sequences were 99.5-99.7 %. The phylogenetic analysis of the 16S rRNA gene sequences showed that strain CBA1105(T) forms a distinct clade with the strains of the closely related genera, Halorientalis and Halorhabdus, with similarities of 94.2 % and 93.9-94.2 %, respectively. Multilocus sequence analysis confirmed that strain CBA1105(T) is closely related to the genus Halorhabdus or Halorientalis. Growth of the strain was observed in 15-30 % NaCl (w/v; optimum 20 %), at 30-45 °C (optimum 37 °C) and pH 7.0-8.0 (optimum pH 7.0) and with 0-0.5 M MgCl2·6H2O (optimum 0.05-0.2 M). The cells of the strain were observed to be Gram-stain negative and pleomorphic with coccoid or ovoid-shape. The cells lysed in distilled water. Tweens 20, 40 and 80 were found to be hydrolysed but starch, casein and gelatine were not. The cells were unable to reduce nitrate under aerobic conditions. Assays for indole formation and urease activity were negative and no growth was observed under anaerobic conditions. Cells were found to be able to utilize L-glutamate, D-glucose, L-maltose, D-mannose and sucrose as sole carbon sources. The polar lipids were identified as phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, unidentified glycolipids and an unidentified phospholipid. The G+C content of strain CBA1105(T) was determined to be 66.0 mol%. The phenotypic, chemotaxonomic and phylogenetic properties suggest that the strain represents a novel species of a novel genus within the family Halobacteriaceae, for which the name Halapricum salinum is proposed with CBA1105(T) (= KCTC 4202(T) = JCM 19729(T)) as the type strain.

Halorubrum halophilum sp. nov., an extremely halophilic archaeon isolated from a salt-fermented seafood.

A novel, red-pigmented, pleomorphic and short rod-shaped haloarchaeon, designated B8(T), was isolated from a salt-fermented seafood. Strain B8(T) was found to be able to grow at 20-45 °C, in the presence of 15-30 % (w/v) NaCl and at pH 7.0-9.0. The optimum requirements were found to be a temperature range of 35-40 °C, pH 8.0 and the presence of 25 % NaCl. The cells of strain B8(T) were observed to be Gram-staining negative and lysed in distilled water. Anaerobic growth did not occur in the presence of nitrate, L-arginine, dimethyl sulfoxide or trimethylamine N-oxide. The catalase and oxidase activities were found to be positive and nitrate was reduced in aerobic conditions. Tween 20, 40 and 80 were found to be hydrolyzed, whereas casein, gelatin and starch were not hydrolyzed. Indole or H2S was not formed and urease activity was not detected. A phylogenetic analysis based on the 16S rRNA gene sequences indicated that strain B8(T) is most closely related to members of the genus Halorubrum in the family Halobacteriaceae. Strain B8(T) was found to have three 16S rRNA genes, rrnA, rrnB and rrnC; similarities between the 16S rRNA gene sequences are 99.0-99.8 %. Strain B8(T) shared 99.0 % 16S rRNA gene sequence similarity with Halorubrum (Hrr.) lipolyticum JCM 13559(T) and Hrr. saccharovorum DSM 1137(T), 98.8 % with Hrr. kocurii JCM 14978(T), 98.3 % with Hrr. lacusprofundi DSM 5036(T), 98.0 % with Hrr. arcis JCM 13916(T), 97.7 % with Hrr. aidingense JCM 13560(T) and 97.0 % with Hrr. aquaticum JCM 14031(T), as well as 93.7-96.5 % with other type strains in the genus Halorubrum. The RNA polymerase subunit B' gene sequence similarity of strain B8(T) with Hrr. kocurii JCM 14978(T) is 97.2 % and lower with other members of the genus Halorubrum. DNA-DNA hybridization experiments showed that strain B8(T) shared equal or lower than 50 % relatedness with reference species in the genus Halorubrum. The genomic DNA G+C content of strain B8(T) was determined to be 64.6 mol%. The major isoprenoid quinone of strain B8(T) was identified as menaquinone-8 and the major polar lipids as phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate, sulfated mannosyl glucosyl diether and an unidentified phospholipid. Based on this polyphasic taxonomic study, strain B8(T) is considered to represent a new species in the genus Halorubrum, for which the name Hrr. halophilum sp. nov. is proposed. The type strain is B8(T) (=JCM 18963(T) = CECT 8278(T)).

Ferrimonas pelagia sp. nov., isolated from seawater.

A Gram-stain-negative bacterium, designated strain CBA4601(T), was isolated from a seawater sample obtained off the coast of Jeju Island, Korea. The organism grew in the presence of 0-4% (w/v) NaCl and at 20-35 °C and pH 7.0-9.0, with optimal growth in 2% NaCl, and at 25 °C and pH 8.0. Phylogenetic trees based on 16S rRNA gene sequences showed that strain CBA4601(T) was related to the genus Ferrimonas within the class Gammaproteobacteria. 16S rRNA gene sequence similarity between strain CBA4601(T) and Ferrimonas marina A4D-4(T), the most closely related species, was 96.9%. The G+C content of the genomic DNA from strain CBA4601(T) was 54.2 mol%, and the isoprenoid quinones menaquinone 7 (MK-7), ubiquinone 7 (Q-7) and ubiquinone 8 (Q-8) were detected. The major fatty acids were C(17:1)ω8c, C(18:1)ω9c and C(16:0), and the major polar lipids were phosphatidylethanolamine, phosphatidylglycerol and an unidentified ninhydrin-positive phospholipid. On the basis of this taxonomic study using a polyphasic approach, strain CBA4601(T) represents a novel species of the genus Ferrimonas, for which the name Ferrimonas pelagia sp. nov. is proposed. The type strain is CBA4601(T) ( =KACC 16695(T) =KCTC 32029(T) =JCM 18401(T)).

Gillisia marina sp. nov., from seashore sand, and emended description of the genus Gillisia.

A Gram-staining-negative, strictly aerobic, rod-shaped bacterium, designated CBA3202(T), was isolated from seashore sand on Jeju Island, Republic of Korea. Based on the 16S rRNA gene sequence analysis, strain CBA3202(T) was allocated to the genus Gillisia (family Flavobacteriaceae) and was most closely related to the type strain of Gillisia mitskevichiae (99.0 % 16S rRNA gene sequence similarity). Optimal growth occurred at 25 °C and with 3 % NaCl. The only isoprenoid quinone was menaquinone-6 (MK-6), the predominant fatty acids were C16 : 0, iso-C15 : 1 G, iso-C16 : 0 and summed feature 3 (comprising C16 : 1ω6c and/or C16 : 1ω7c), and the DNA G+C content was 34.9 mol%. The polar lipids were phosphatidylethanolamine, two unidentified aminolipids and several unidentified polar lipids. Based on phylogenetic inference and phenotypic data, we conclude that strain CBA3202(T) represents a novel species of the genus Gillisia, for which the name Gillisia marina sp. nov. is proposed. The type strain is CBA3202(T) ( = KACC 16693(T) = KCTC 32030(T) = JCM 18402(T)). An emended description of the genus Gillisia is also provided.

Methanomethylovorans uponensis sp. nov., a methylotrophic methanogen isolated from wetland sediment.

A novel mesophilic, methylotrophic, methanogenic archaeon, designated strain EK1(T), was enriched and isolated from wetland sediment. Phylogenetic analysis showed that strain EK1(T) was affiliated with the genus Methanomethylovorans within the family Methanosarcinaceae, and shared the highest 16S rRNA and methyl-coenzyme M reductase alpha-subunit gene sequence similarity with the type strain of Methanomethylovorans hollandica (98.8 and 92.6 %, respectively). The cells of strain EK1(T) were observed to be Gram-negative, non-motile and irregular cocci that did not lyse in 0.1 % (w/v) sodium dodecyl sulfate. Methanol, mono-, di- and trimethylamine, dimethyl sulfide and methanethiol were found to be used as catabolic and methanogenic substrates, whereas H2/CO2, formate, 2-propanol and acetate were not. Growth was observed at 25-40 °C (optimum, 37 °C), at pH 5.5-7.5 (optimum, pH 6.0-6.5) and in the presence of 0-0.1 M NaCl (optimum, 0 M). Growth and methane production rates were stimulated in the presence of H2/CO2 although methane production and growth yields were not significantly affected; acetate, formate, 2-propanol and CO/CO2/N2 did not affect methane production. CoCl2 (0.6-2.0 µM) and FeCl2 (25 mg/l) stimulated growth, while yeast extract and peptone did not. The DNA-DNA hybridization experiment revealed a relatedness of <20 % between EK1(T) and the type strains of the genus Methanomethylovorans. The DNA G+C content of strain EK1(T) was determined to be 39.2 mol%. Based on the polyphasic taxonomic study, strain EK1(T) represents a novel species belonging to the genus Methanomethylovorans, for which the name Methanomethylovorans uponensis sp. nov. is proposed. The type strain is strain EK1(T)(=NBRC 109636(T) = KCTC 4119(T) = JCM 19217(T)).

Natronomonas gomsonensis sp. nov., isolated from a solar saltern.

A halophilic archaeal strain, SA3(T), was isolated from sediment of a solar saltern in Gomso Bay, Republic of Korea. Cells of strain SA3(T) were observed to be coccoid-shaped, to lyse in distilled water, Gram stain-negative and to form red-pigmented colonies. Strain SA3(T) was found to require at least 18 % (w/v) NaCl for growth. Optimal growth was observed at 24 % (w/v) NaCl and 6 % (w/v) MgCl2. The optimum pH and temperature for growth were determined to be pH 7.0 and 40 °C, respectively, while the strain was found to grow within pH and temperature ranges of 5.5-8.0 and 20-45 °C, respectively. The polar lipids were determined to consist of phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, unidentified phosphoglycolipids and unidentified phospholipids. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain SA3(T) was most closely related to the members of the genus Natronomonas, Natronomonas moolapensis JCM 14361(T) (95.2 %) and Natronomonas pharaonis JCM 8858(T) (95.1 %). The genomic DNA G+C content (61.8 mol%) determined for strain SA3(T) was slightly lower than those of N. moolapensis JCM 14361(T) (63.4 mol%) and N. pharaonis JCM 8858(T) (64.3 mol%). DNA-DNA hybridization values between N. moolapensis JCM 14361(T) and N. pharaonis JCM 8858(T) and strain SA3(T) were <20 %. Based on phenotypic, chemotaxonomic and phylogenetic properties, we describe a new species of the genus Natronomonas, represented by strain SA3(T) (=JCM 17867(T) = KCTC 4088(T)), for which we propose the name Natronomonas gomsonensis sp. nov.

Mychonastes sp. 246 Suppresses Human Pancreatic Cancer Cell Growth via IGFBP3-PI3K-mTOR Signaling.

Previously, we confirmed that Mychonastes sp. 246 methanolic extract (ME) markedly reduced the viability of BxPC-3 human pancreatic cancer cells. However, the underlying mechanism ME remained unclear. Hence, we attempted to elucidate the anticancer effect of ME on BxPC-3 human pancreatic cancer cells. First, we investigated the components of ME and their cytotoxicity in normal cells. Then, we confirmed the G1 phase arrest mediated growth inhibitory effect of ME using a cell counting assay and cell cycle analysis. Moreover, we found that the migration-inhibitory effect of ME using a Transwell migration assay. Through RNA sequencing, Gene Ontology-based network analysis, and western blotting, we explored the intracellular mechanisms of ME in BxPC-3 cells. ME modulated the intracellular energy metabolism-related pathway by altering the mRNA levels of IGFBP3 and PPARGC1A in BxPC-3 cells and reduced PI3K and mTOR phosphorylation by upregulating IGFBP3 and 4E-BP1 expression. Finally, we verified that ME reduced the growth of three-dimensional (3D) pancreatic cancer spheroids. Our study demonstrates that ME suppresses pancreatic cancer proliferation through the IGFBP3-PI3K-mTOR signaling pathway. This is the first study on the anticancer effect of the ME against pancreatic cancer, suggesting therapeutic possibilities and the underlying mechanism of ME action.

Improvement of Lutein and Zeaxanthin Production in Mychonastes sp. 247 by Optimizing Light Intensity and Culture Salinity Conditions.

In this study, we sought to improve lutein and zeaxanthin production in Mychonastes sp. 247 and investigated the effect of environmental factors on lutein and zeaxanthin productivity in Mychonastes sp. The basic medium selection and N:P ratio were adjusted to maximize cell growth in one-stage culture, and lutein and zeaxanthin production conditions were optimized using a central composite design for two-stage culture. The maximum lutein production was observed at a light intensity of 60 µE/m2/s and salinity of 0.49%, and the maximum zeaxanthin production was observed at a light intensity of 532 µE/m2/s and salinity of 0.78%. Lutein and zeaxanthin production in the optimized medium increased by up to 2 and 2.6 folds, respectively, compared to that in the basic medium. Based on these results, we concluded that the optimal conditions for lutein and zeaxanthin production are different and that optimization of light intensity and culture salinity conditions may help increase carotenoid production. This study presents a useful and potential strategy for optimizing microalgal culture conditions to improve the productivity of lutein and zeaxanthin, which has applications in the functional food field.

Draft genome sequence of Gillisia sp. strain CBA3202, a novel member of the genus Gillisia, which belongs to the family Flavobacteriaceae.

Gillisia sp. strain CBA3202, which belongs to the family Flavobacteriaceae, was isolated from sand of the seashore on Jeju Island, Republic of Korea. The draft genome of Gillisia sp. CBA3202 contains 2,981,404 bp with a G+C content of 34.9%. This is the second genome sequence of the Gillisia strains.