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Protocol for efficient recovery of high-quality DNA from microbiome of marine invertebrates
Yeong-Jun Park, Jae Kyu Lim, Yeon-Ju Lee, Kae Kyoung Kwon
J. Microbiol. 2025;63(9):e2507003.   Published online September 30, 2025
DOI: https://doi.org/10.71150/jm.2507003
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AbstractAbstract PDF

Marine organisms often form symbiotic relationships with various microorganisms to adapt and thrive in harsh environments. These symbiotic microbes contribute to host survival by providing nutrition, modulating the hosts’ immune system, and supporting overall physiological stability. Advances in high-throughput sequencing technologies have enabled a deeper understanding of the structure and function of symbiotic microbial communities, as well as host-microbe interactions. Notably, symbiotic bacteria associated with marine invertebrates such as corals and sponges are recognized as a potential source of useful bioactive compounds, including antibiotics and enzymes. However, obtaining high-quality microbial DNA from host tissues still remains a technical challenge due to the presence of unknown substances. This study focuses on optimizing sample preparation and DNA extraction procedures and additional purification to improve the recovery of microbial DNA while minimizing host DNA contamination. Comparison between several methods was conducted using sponge samples to evaluate DNA quality and microbial recovery. A sample designated as 2110BU-001 was collected from the east coast of the Republic of Korea and used for culture-independent microbial cell isolation. Total bacterial DNA was extracted by using a manual Phenol-Chloroform protocol and three commercial kits. DNA extracted using the standard manual method showed both the highest yield and the largest fragment size. However, PCR (Polymerase chain reaction) test showed that quality of manually extracted DNA was not enough for sequencing. Therefore, the quality of DNA was improved through additional purification steps. Briefly, host eukaryotic cells were removed by mechanical process and almost only bacterial DNA was successfully obtained by combination of manual extraction method and further purification processes. The established protocol was successfully introduced to extraction of metagenomic DNA from mussel and jellyfish microbiomes, indicating that it can be widely applied to various marine organisms.

Journal Articles
Flavivirga spongiicola sp. nov. and Flavivirga abyssicola sp. nov., Isolated from Marine Environments
Sung-Hyun Yang , Mi-Jeong Park , Hyun-Myung Oh , Yeong-Jun Park , Kae Kyoung Kwon
J. Microbiol. 2024;62(1):11-19.   Published online February 6, 2024
DOI: https://doi.org/10.1007/s12275-023-00102-z
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AbstractAbstract PDF
Two novel Gram-stain-negative, strictly-aerobic, rod-shaped (1.2 ± 3.4 μm × 0.3 ± 0.7 μm), and non-motile marine bacterial species, designated MEBiC05379T and MEBiC07777T, were isolated from a marine sponge Pseudaxinella sp. in Gangneung City and deep-sea sediments of the Ulleung basin in the East Sea of Korea, respectively. The 16S rRNA gene sequence analysis revealed high levels of similarities between these strains and members of the genus Flavivirga (97.0–98.4% sequence identities). Both novel strains revealed as mesophilic, neutrophilic in pH and slightly halophilic. Similar to those of other Flavivirga members, the primary cellular fatty acids of both strains were iso-C15:0, iso-C15:1 G, iso-C15:03-OH, and iso-C17:0 3-OH, with MEBiC05379T and MEBiC07777T containing relatively higher proportions of C12: 0 and summed feature 3 ( C16:1ω7c and/or C16: 1ω6c). In both taxa, the major isoprenoid quinone was MK-6. The DNA G + C contents of MEBiC05379T and MEBiC07777T genomes were 32.62 and 32.46 mol%, respectively. Compared to other members of Flavivirga, both strains exhibited similar DNA G + C ratio and fatty acids pattern, yet enzyme expression and carbon sources utilization pattern were different. Genomes of the genus Flavivirga showed enzyme preferences to fucoidan and sulfated galactans. Considering the monophyly rule, AAI values delineate the genus Flavivirga from adjacent genera calculated to be 76.0–78.7%. Based on the phenotypic, genomic and biochemical data, strains for MEBiC05379T and MEBiC07777T thus represent two novel species in the genus Flavivirga, for which the names Flavivirga spongiicola sp. nov. ( MEBiC05379T [= KCTC 92527 T = JCM 16662 T]), and Flavivirga abyssicola sp. nov. ( MEBiC07777T [= KCTC 92563 T = JCM 36477 T]) are proposed.

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  • Phycobium rhodophyticola gen. nov., sp. nov. and Aliiphycobium algicola gen. nov., sp. nov., isolated from the phycosphere of marine red algae
    Jeong Min Kim, Woonhee Baek, Byeong Jun Choi, Hülya Bayburt, Jae Kyeong Lee, Sung Chul Lee, Che Ok Jeon
    Journal of Microbiology.2025; 63(6): e2503014.     CrossRef
  • Rubrivirga aquatilis sp. nov. and Rubrivirga halophila sp. nov., isolated from Korean coastal surface seawater
    Jisoo Han, Yeonjung Lim, Mirae Kim, Jang-Cheon Cho
    Journal of Microbiology.2025; 63(8): e2504017.     CrossRef
  • Rhodobacteraceae are Prevalent and Ecologically Crucial Bacterial Members in Marine Biofloc Aquaculture
    Meora Rajeev, Jang-Cheon Cho
    Journal of Microbiology.2024; 62(11): 985.     CrossRef
  • Validation List no. 220. Valid publication of new names and new combinations effectively published outside the IJSEM
    Aharon Oren, Markus Göker
    International Journal of Systematic and Evolutionary Microbiology .2024;[Epub]     CrossRef
  • Optimization of Culture Medium for the Production of an Exopolysaccharide (p-CY02) with Cryoprotective Activity by Pseudoalteromonas sp. RosPo-2 from the Antarctic Sea
    Pilsung Kang, Sung Jin Kim, Ha Ju Park, Il Chan Kim, Se Jong Han, Joung Han Yim
    Journal of Microbiology and Biotechnology.2024; 34(5): 1135.     CrossRef
Description of Fervidibacillus gen. nov. with Two Species, Fervidibacillus albus sp. nov., and Fervidibacillus halotolerans sp. nov., Isolated from Tidal Flat Sediments and Emendation of Misclassificed Taxa in the Genus Caldibacillus
Sung&# , Mi&# , Hyun&# , Kae Kyoung Kwon
J. Microbiol. 2023;61(2):175-187.   Published online February 17, 2023
DOI: https://doi.org/10.1007/s12275-023-00022-y
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AbstractAbstract PDF
Two Gram-stain-positive, motile, endospore-forming, facultatively anaerobic strains, designated MEBiC13591T and MEBiC13594T, were isolated from tidal flat sediment of the Incheon City on the west coast of Korea. Growth of both novel strains was observed at pH 5–9 (optimum, pH 7–7.5), and in 0–8% NaCl (optimum, 2% for MEBiC13591T and 3% for MEBiC13594T). Strains MEBiC13591T and MEBiC13594T grew optimally at 50 °C, (37.5–56.1 °C) and 44 °C (20.7–50.7 °C), respectively. The main cellular fatty acids of strain MEBiC13591T were iso-C15: 0, anteiso-C15: 0, iso-C16: 0, iso-C17: 0 and anteiso-C17: 0, while those for strain MEBiC13594T were C14: 0, iso-C14: 0, iso-C15: 0, anteiso-C15: 0 and C16: 0. In both taxa, the major isoprenoid was MK-7. The genomic DNA G + C contents were 34.1 and 37.0 mol% for MEBiC13591T and MEBiC13594T, respectively. A 16S rRNA gene sequence analysis revealed that the novel strains showed high similarity with members of the genera Aeribacillus (95.0%) and Caldibacillus (93.5–94.5%); however, showed lower than 90% with Caldibacillus debilis TfT. Phylogenetic and Phylogenomic analysis revealed that two novel strains comprised distinct phyletic line with members formerly assigned to Caldibacillus. Based on genomic indices, such as AAI and ANI, members formerly affiliated with Caldibacillus and Bacillus as well as the novel strains should be classified into five independent genera. Based on the phenotypic, genomic and biochemical data, strains MEBiC13591T and MEBiC13594T represent two novel species in the novel genus, for which the names Fervidibacillus albus gen. nov., sp. nov. ( MEBiC13591T [= KCCM 43317T = KCTC 43181T = JCM 33662T = MCCC 1K04565T]), and Fervidibacillus halotolerans sp. nov. ( MEBiC13594T [= KCTC 43182T = JCM 34001T]) are proposed. Three additional genera Caldifermentibacillus, Palidibacillus, and Perspicuibacillus are also proposed by reclassification of the several species with valid names that formerly affiliated with the genera Caldibacillus.

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  • Rubrivirga aquatilis sp. nov. and Rubrivirga halophila sp. nov., isolated from Korean coastal surface seawater
    Jisoo Han, Yeonjung Lim, Mirae Kim, Jang-Cheon Cho
    Journal of Microbiology.2025; 63(8): e2504017.     CrossRef
  • An update on novel taxa and revised taxonomic status of bacteria isolated from domestic companion and agricultural animals described in 2023
    Sara D. Lawhon, Claire R. Burbick, Trinity Krueger, Elena Ruiz-Reyes, Erik Munson, Romney M. Humphries
    Journal of Clinical Microbiology.2024;[Epub]     CrossRef
  • Congregibacter variabilis sp. nov. and Congregibacter brevis sp. nov. Within the OM60/NOR5 Clade, Isolated from Seawater, and Emended Description of the Genus Congregibacter
    Hyeonsu Tak, Miri S. Park, Hyerim Cho, Yeonjung Lim, Jang-Cheon Cho
    Journal of Microbiology.2024; 62(9): 739.     CrossRef
  • Validation List no. 213. Valid publication of new names and new combinations effectively published outside the IJSEM
    Aharon Oren, Markus Göker
    International Journal of Systematic and Evolutionary Microbiology .2023;[Epub]     CrossRef
Constantimarinum furrinae gen. nov., sp. nov., a marine bacterium isolated from saline volcanic rock aquifer (lava seawater) at Jeju Island, Republic of Korea
Sung-Hyun Yang , Hyun-Myung Oh , Mi-Jeong Park , Dongil Jang , Kae Kyoung Kwon
J. Microbiol. 2022;60(1):11-17.   Published online December 29, 2021
DOI: https://doi.org/10.1007/s12275-022-1468-6
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AbstractAbstract PDF
A Gram-stain-negative, aerobic, rod-shaped (0.3–0.5 × 1.0– 1.9 μm), non-motile marine bacterium designated as ALE3EIT was isolated from a saline volcanic rock aquifer (lava seawater) on Jeju Island, Republic of Korea. The 16S rRNA gene sequence analysis revealed that strain ALE3EIT showed high similarity to ‘Altibacter lentus’ JLT2010T (97.2%), followed by Marixanthomonas ophiurae KMM 3046T (94.5%). Growth was observed at 10–41°C (optimum, 30°C), at pH 6.0–8.5 (optimum, pH 7.5) and at 0.5–8% (optimum, 4.0%) NaCl. The predominant cellular fatty acids were iso-C15:0 (23.5%), iso-C16:0 (10.2%), iso-C16:0 3OH (10.5%), and iso-C17:0 3OH (16.8%). The DNA G + C contents was 40.4 mol%. The major respiratory quinone was MK-6. The major polar lipids were determined to be phosphatidylethanolamine, two unidentified glycolipids, and two unidentified aminolipids. Several phenotypic characteristics such as production of acetoin, activities of arginine dihydrolase and acid phosphatase, and utilization pattern of carbon sources differentiate strain ALE3EIT from ‘A. lentus’ JLT2010T. Activities of the lipase, trypsin, α- chymotrypsin and gelatinase and utilization pattern of carbon sources differentiate strain ALE3EIT from M. ophiurae KMM 3046T. The genome of strain ALE3EIT is 3.0 Mbp long and its ANI and AAI values against ‘A. lentus’ JLT2010T were 76.58 and 72.76, respectively, however, AAI values against members in other genera were lower than 72%. The phylogenomic tree inferred by PhyloPhlAn clearly differentiated the strain ALE3EIT together with strain JLT2010T from other genera in the Falvobacteriaceae. This polyphasic taxonomic data indicates that strain ALE3EIT should be identified as a novel species in the genus ‘Altibacter’, however, the name has not been validated. Therefore, the strain is classified as a novel genus and is proposed as Constantimarinum furrinae gen. nov., sp. nov. The type strain is ALE3EIT (= KCCM 43303T = JCM 33022T).

Citations

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  • Mesohalobacter salilacus sp. nov., a novel halotolerant, flexirubin-type pigment-producing bacterium of the family Flavobacteriaceae
    Shuai-Bo Han, Yang-Huan Yu, Xiao-Long He, Jin-Jia Liang, Min Wu, Can Chen, Xue-Wei Xu
    International Journal of Systematic and Evolutionary Microbiology .2025;[Epub]     CrossRef
  • Comparison of the anti-inflammatory effects of carotenoids-rich product from microalgal mass-cultured between natural seawater and magma seawater
    Jayeon Cheon, Eun-A Kim, Nalae Kang, Taeho Kim, Lei Wang, Soo-Jin Heo, Seon-Heui Cha
    Biocatalysis and Agricultural Biotechnology.2024; 62: 103420.     CrossRef
  • Validation List no. 212. Valid publication of new names and new combinations effectively published outside the IJSEM
    Aharon Oren, Markus Göker
    International Journal of Systematic and Evolutionary Microbiology .2023;[Epub]     CrossRef
Thermococcus indicus sp. nov., a Fe(III)-reducing hyperthermophilic archaeon isolated from the Onnuri Vent Field of the Central Indian Ocean ridge
Jae Kyu Lim , Yun Jae Kim , Jhung-Ahn Yang , Teddy Namirimu , Sung-Hyun Yang , Mi-Jeong Park , Yong Min Kwon , Hyun Sook Lee , Sung Gyun Kang , Jung-Hyun Lee , Kae Kyoung Kwon
J. Microbiol. 2020;58(4):260-267.   Published online April 1, 2020
DOI: https://doi.org/10.1007/s12275-020-9424-9
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AbstractAbstract PDF
A strictly anaerobic, dissimilatory Fe(III)-reducing hyperthermophilic archaeon, designated as strain IOH1T, was isolated from a new deep-sea hydrothermal vent (Onnuri Vent Field) area in the Central Indian Ocean ridge. Strain IOH1T showed > 99% 16S rRNA gene sequence similarity with Thermococcus celericrescens TS2T (99.4%) and T. siculi DSM 12349T (99.2%). Additional three species T. barossii SHCK-94T (99.0%), T. celer Vu13T (98.8%), and T. piezophilus (98.6%) showed > 98.6% of 16S rRNA gene sequence similarity, however, the maximum OrthoANI value is 89.8% for the genome of T. celericrescens TS2T. Strain IOH1T cells are coccoid, 1.2–1.8 μm in diameter, and motile by flagella. Growth was at 70–82°C (optimum 80°C), pH 5.4–8.0 (optimum pH 6.0) with 2–4% (optimum 3%) NaCl. Growth of strain IOH1T was enhanced by starch, pyruvate, D(+)-maltose and maltodextrin as a carbon sources, and elemental sulfur as an electron acceptor; clearly different from those of related species T. celecrescens DSM 17994T and T. siculi DSM 12349T. Strain IOH1T, T. celercrescence DSM 17994T, and T. siculi DSM 12349T reduced soluble Fe(III)-citrate present in the medium, whereas the amount of total cellular proteins increased with the concomitant accumulation of Fe(II). We determined a circular chromosome of 2,234 kb with an extra-chromosomal archaeal plasmid, pTI1, of 7.7 kb and predicted 2,425 genes. The DNA G + C content was 54.9 mol%. Based on physiological properties, phylogenetic, and genome analysis, we proposed that strain IOH1T (= KCTC 15844T = JCM 39077T) is assigned to a new species in the genus Thermococcus and named Thermococcus indicus sp. nov.

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  • Fluid chemistry evolution in deep-sea hydrothermal environments: Unraveling mineral-fluid-microorganism interactions through continuous culture experiment
    Lise Artigue, Valérie Chavagnac, Christine Destrigneville, David François, Françoise Lesongeur, Anne Godfroy
    Deep Sea Research Part I: Oceanographic Research Papers.2025; 218: 104456.     CrossRef
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    Solveig I. Bühring, Stefanie Böhnke-Brandt, Alexander Diehl, Martha Gledhill, Laura Haffert, Charlotte Kleint, Andrea Koschinsky, Sinikka Lennartz, Mirjam Perner, Sylvia G. Sander, Christoph Völker, Ying Ye
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    Yuan Yuan, Yongqin Liu
    Earth Science-Journal of China University of Geosciences.2025; 50(3): 887.     CrossRef
  • Macrobenthic communities in the polymetallic nodule field, Indian Ocean, based on multicore and box core analysis
    Santosh Gaikwad, Sabyasachi Sautya, Samir Damare, Maria Brenda Luzia Mascarenhas-Pereira, Vijayshree Gawas, Jayesh Patil, Mandar Nanajkar, Sadiq Bukhari
    Frontiers in Marine Science.2024;[Epub]     CrossRef
  • Thermococcus argininiproducens sp. nov., an arginine biosynthesis archaeal species isolated from the Central Indian Ocean ridge
    Yeong-Jun Park, Jae Kyu Lim, Yun Jae Kim, Sung-Hyun Yang, Hyun Sook Lee, Sung Gyun Kang, Jung-Hyun Lee, Youngik Yang, Kae Kyoung Kwon
    International Journal of Systematic and Evolutionary Microbiology .2023;[Epub]     CrossRef
  • Spatial comparison and temporal evolution of two marine iron-rich microbial mats from the Lucky Strike Hydrothermal Field, related to environmental variations
    Aina Astorch-Cardona, Mathilde Guerre, Alain Dolla, Valérie Chavagnac, Céline Rommevaux
    Frontiers in Marine Science.2023;[Epub]     CrossRef
  • Formate and hydrogen in hydrothermal vents and their use by extremely thermophilic methanogens and heterotrophs
    James F. Holden, Harita Sistu
    Frontiers in Microbiology.2023;[Epub]     CrossRef
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    WeiShu ZHAO, Xiang XIAO
    SCIENTIA SINICA Vitae.2022;[Epub]     CrossRef
  • Limitations of microbial iron reduction under extreme conditions
    Sophie L Nixon, Emily Bonsall, Charles S Cockell
    FEMS Microbiology Reviews.2022;[Epub]     CrossRef
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    Aharon Oren, George M. Garrity
    International Journal of Systematic and Evolutionary Microbiology .2021;[Epub]     CrossRef
  • Thermococcus aciditolerans sp. nov., a piezotolerant, hyperthermophilic archaeon isolated from a deep-sea hydrothermal vent chimney in the Southwest Indian Ridge
    Xue-Gong Li, Hong-Zhi Tang, Wei-Jia Zhang, Xiao-Qing Qi, Zhi-Guo Qu, Jun Xu, Long-Fei Wu
    International Journal of Systematic and Evolutionary Microbiology.2021;[Epub]     CrossRef
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Paradesulfovibrio onnuriensis gen. nov., sp. nov., a chemolithoautotrophic sulfate-reducing bacterium isolated from the Onnuri vent field of the Indian Ocean and reclassification of Desulfovibrio senegalensis as Paradesulfovibrio senegalensis comb. nov.
Yun Jae Kim , Jhung-Ahn Yang , Jae Kyu Lim , Mi-Jeong Park , Sung-Hyun Yang , Hyun Sook Lee , Sung Gyun Kang , Jung-Hyun Lee , Kae Kyoung Kwon
J. Microbiol. 2020;58(4):252-259.   Published online February 27, 2020
DOI: https://doi.org/10.1007/s12275-020-9376-0
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AbstractAbstract PDF
An anaerobic, rod-shaped, mesophilic, chemolithoautotrophic, sulfate-reducing bacterial strain IOR2T was isolated from a newly found deep-sea hydrothermal vent (OVF, Onnuri Vent Field) area in the central Indian Ocean ridge (11°24􍿁88􍿂􀁇 S 66°25􍿁42􍿂􀁇E, 2021 m water depth). The 16S rRNA gene sequence analysis revealed that the strain IOR2T was most closely related to Desulfovibrio senegalensis BLaC1T (96.7%). However, it showed low similarity with the members of the family Desulfovibrionaceae, such as Desulfovibrio tunisiensis RB22T (94.0%), D. brasiliensis LVform1T (93.9%), D. halophilus DSM 5663T (93.7%), and Pseudodesulfovibrio aespoeensis Aspo-2T (93.2%). The strain IOR2T could grow at 23– 42°C (optimum 37°C), pH 5.0–8.0 (optimum pH 7.0) and with 0.5–6.5% (optimum 3.0%) NaCl. The strain could use lactate, pyruvate, H2, and glycerol as electron donors and sulfate, thiosulfate, and sulfite as electron acceptors. The major fatty acids of the strain IOR2T were iso-C15:0, iso-C17:0, anteiso- C15:0, and summed feature 9 (C16:0 methyl/iso-C17:1ω9c). Both the strains IOR2T and BLaC1T could grow with CO2 and H2 as the sole sources of carbon and energy, respectively. Genomic evidence for the Wood-Ljungdahl pathway in both the strains reflects chemolithoautotrophic growth. The DNA G + C content of the strain IOR2T and BLaC1T was 58.1–60.5 mol%. Based on the results of the phylogenetic and physiologic studies, Paradesulfovibrio onnuriensis gen. nov., sp. nov. with the type strain IOR2T (= KCTC 15845T = MCCC 1K04559T) was proposed to be a member of the family Desulfovibrionaceae. We have also proposed the reclassification of D. senegalensis as Paradesulfovibrio senegalensis comb. nov.

Citations

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  • Sulfate-Reducing Bacteria Isolated from an Oil Field in Kazakhstan and a Description of Pseudodesulfovibrio karagichevae sp. nov.
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Isolation, cultivation, and genome analysis of proteorhodopsincontaining SAR116-clade strain Candidatus Puniceispirillum marinum IMCC1322
Junhak Lee , Kae Kyoung Kwon , Seung-Il Lim , Jaeho Song , Ah Reum Choi , Sung-Hyun Yang , Kwang-Hwan Jung , Jung-Hyun Lee , Sung Gyun Kang , Hyun-Myung Oh , Jang-Cheon Cho
J. Microbiol. 2019;57(8):676-687.   Published online June 14, 2019
DOI: https://doi.org/10.1007/s12275-019-9001-2
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AbstractAbstract PDF
Strain IMCC1322 was isolated from a surface water sample from the East Sea of Korea. Based on 16S rRNA analysis, IMCC1322 was found to belong to the OCS28 sub-clade of SAR116. The cells appeared as short vibrioids in logarithmicphase culture, and elongated spirals during incubation with mitomycin or in aged culture. Growth characteristics of strain IMCC1322 were further evaluated based on genomic information; proteorhodopsin (PR), carbon monoxide dehydrogenase, and dimethylsulfoniopropionate (DMSP)-utilizing enzymes. IMCC1322 PR was characterized as a functional retinylidene protein that acts as a light-driven proton pump in the cytoplasmic membrane. However, the PR-dependent phototrophic potential of strain IMCC1322 was only observed under CO-inhibited and nutrient-limited culture conditions. A DMSP-enhanced growth response was observed in addition to cultures grown on C1 compounds like methanol, formate, and methane sulfonate. Strain IMCC1322 cultivation analysis revealed biogeochemical processes characteristic of the SAR116 group, a dominant member of the microbial community in euphotic regions of the ocean. The polyphasic taxonomy of strain IMCC1322 is given as Candidatus Puniceispirillum marinum, and was confirmed by chemotaxonomic tests, in addition to 16S rRNA phylogeny and cultivation analyses.

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    Hyun-Myung Oh, Ji Hyen Lee, Ahyoung Choi, Sung-Hyun Yang, Gyung-Hoon Shin, Sung Gyun Kang, Jang-Cheon Cho, Hak Jun Kim, Kae-Kyoung Kwon
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    Shan-Hui Li, Jaeho Song, Ilnam Kang, Juchan Hwang, Jang-Cheon Cho
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    Suhyun Kim, Miri S. Park, Jaeho Song, Ilnam Kang, Jang-Cheon Cho
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    Michael W. Henson, V. Celeste Lanclos, David M. Pitre, Jessica Lee Weckhorst, Anna M. Lucchesi, Chuankai Cheng, Ben Temperton, J. Cameron Thrash, Robert M. Kelly
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Comparative genome analysis of the Flavobacteriales bacterium strain UJ101, isolated from the gut of Atergatis reticulatus
Jhung-Ahn Yang , Sung-Hyun Yang , Junghee Kim , Kae Kyoung Kwon , Hyun-Myung Oh
J. Microbiol. 2017;55(7):583-591.   Published online June 30, 2017
DOI: https://doi.org/10.1007/s12275-017-7172-2
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AbstractAbstract PDF
Here we report the comparative genomic analysis of strain UJ101 with 15 strains from the family Flavobacteriaceae, using the CGExplorer program. Flavobacteriales bacterium strain UJ101 was isolated from a xanthid crab, Atergatis reticulatus, from the East Sea near Korea. The complete genome of strain UJ101 is a 3,074,209 bp, single, circular chromosome with 30.74% GC content. While the UJ101 genome contains a number of annotated genes for many metabolic pathways, such as the Embden–Meyerhof pathway, the pentose phosphate pathway, the tricarboxylic acid (TCA) cycle, and the glyoxylate cycle, genes for the Entner-Douddoroff pathway are not found in the UJ101 genome. Overall, carbon fixation processes were absent but nitrate reduction and denitrification pathways were conserved. The UJ101 genome was compared to genomes from other marine animals (three invertebrate strains and 5 fish strains) and other marine animal- derived genera. Notable results by genome comparisons showed that UJ101 is capable of denitrification and nitrate reduction, and that biotin-thiamine pathway participation varies among marine bacteria; fish-dwelling bacteria, freeliving bacteria, invertebrate-dwelling bacteria, and strain UJ101. Pan-genome analysis of the 16 strains in this study included 7,220 non-redundant genes that covered 62% of the pan-genome. A core-genome of 994 genes was present and consisted of 8% of the genes from the pan-genome. Strain UJ101 is a symbiotic hetero-organotroph isolated from xanthid crab, and is a metabolic generalist with nitrate-reducing abilities but without the ability to synthesize biotin. There is a general tendency of UJ101 and some fish pathogens to prefer thiamine-dependent glycolysis to gluconeogenesis. Biotin and thiamine auxotrophy or prototrophy may be used as important markers in microbial community studies.

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Proteomic characterization of the outer membrane vesicle of the halophilic marine bacterium Novosphingobium pentaromativorans US6-1
Sung Ho Yun , Sang-Yeop Lee , Chi-Won Choi , Hayoung Lee , Hyun-Joo Ro , Sangmi Jun , Yong Min Kwon , Kae Kyoung Kwon , Sang-Jin Kim , Gun-Hwa Kim , Seung Il Kim
J. Microbiol. 2017;55(1):56-62.   Published online December 30, 2016
DOI: https://doi.org/10.1007/s12275-017-6581-6
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AbstractAbstract PDF
Novosphingobium pentaromativorans US6-1 is a Gram-negative halophilic marine bacterium able to utilize several polycyclic aromatic hydrocarbons such as phenanthrene, pyrene, and benzo[a]pyrene. In this study, using transmission electron microscopy, we confirmed that N. pentaromativorans US6-1 produces outer membrane vesicles (OMVs). N. pentaromativorans OMVs (hereafter OMVNovo) are spherical in shape, and the average diameter of OMVNovo is 25–70 nm. Proteomic analysis revealed that outer membrane proteins and periplasmic proteins of N. pentaromativorans are the major protein components of OMVNovo. Comparative proteomic analysis with the membrane-associated protein fraction and correlation analysis demonstrated that the outer membrane proteins of OMVNovo originated from the membrane- associated protein fraction. To the best of our knowledge, this study is the first to characterize OMV purified from halophilic marine bacteria.

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Metagenomic analysis reveals the contribution of anaerobic methanotroph-1b in the oxidation of methane at the Ulleung Basin, East Sea of Korea
Jin-Woo Lee , Kae Kyoung Kwon , Jang-Jun Bahk , Dong-Hun Lee , Hyun Sook Lee , Sung Gyun Kang , Jung-Hyun Lee
J. Microbiol. 2016;54(12):814-822.   Published online November 26, 2016
DOI: https://doi.org/10.1007/s12275-016-6379-y
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AbstractAbstract PDF
We have previously identified a sulfate methane transition zone (SMTZ) within the methane hydrate-bearing sediment in the Ulleung Basin, East Sea of Korea, and the presence of ANME-1b group in the sediment has been shown by phylogenetic analysis of a 16S rRNA gene. Herein, we describe taxonomic and functional profiling in the SMTZ sample by metagenomic analysis, comparing with that of surface sediment. Metagenomic sequences of 115 Mbp and 252 Mbp were obtained from SMTZ and surface sediments, respectively. The taxonomic profiling using BLASTX against the SEED within MG-RAST showed the prevalence of methanogens (19.1%), such as Methanosarcinales (12.0%) and Methanomicrobiales (4.1%) predominated within the SMTZ metagenome. A number of 185,200 SMTZ reads (38.9%) and 438,484 surface reads (62.5%) were assigned to functional categories, and methanogenesis-related reads were statistically significantly overrepresented in the SMTZ metagenome. However, the mapping analysis of metagenome reads to the reference genomes, most of the sequences of the SMTZ metagenome were mapped to ANME-1 draft genomes, rather than those of methanogens. Furthermore, the two copies of the methyl-coenzyme M reductase gene (mcrA) segments of the SMTZ metagenome were clustered with ANME-1b in the phylogenetic cluster. These results indicate that ANME- 1b reads were miss-annotated to methanogens due to limitation of database. Many of key genes necessary for reverse methanogenesis were present in the SMTZ metagenome, except for N5,N10-methenyl-H4MPT reductase (mer) and CoBCoM heterodisulfide reductase subunits D and E (hdrDE). These data suggest that the ANME-1b represents the primary player the anaerobic methane oxidation in the SMTZ, of the methane hydrate-bearing sediment at the Ulleung Basin, East Sea of Korea.

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Research Support, Non-U.S. Gov't
Abyssisolibacter fermentans gen. nov. sp. nov., isolated from deep sub-seafloor sediment
Wonduck Kim , Jung-Hyun Lee , Kae Kyoung Kwon
J. Microbiol. 2016;54(5):347-352.   Published online April 20, 2016
DOI: https://doi.org/10.1007/s12275-016-6048-1
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AbstractAbstract PDF
A Gram-staining-negative, thin rod-shaped, anaerobic bacterium designated MCWD3T was isolated from sediment of the deep sea in Ulleung Basin, East Sea, Korea. The ranges of temperature, pH and NaCl for growth of this strain were 15– 40°C (optimum 29°C), 5.0–10.0 (optimum pH 6.5), and 1–5%, respectively. The major fatty acids were iso-C15:0 (30%) and iso-C15:0 dimethyl acetal (17%). The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, and unidentified aminophospholipids, phospholipids, and aminolipids. The fermentation product from yeast extract was acetate. Phylogenetic analysis based on 16S rRNA genes indicated that the isolate was related to Sporosalibacterium faouarense (92.8% sequence identity), Clostridiisalibacter paucivorans (92.6%), and Brassicibacter mesophilus (92.4%). However, the isolate was differentiated from these genera by both physiological and chemotaxonomical properties. On the basis of a polyphasic taxonomic analysis, we propose that MCWD3T represents a novel taxon with the name Abyssisolibacter fermentans gen. nov. sp. nov.

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  • Validation List no. 220. Valid publication of new names and new combinations effectively published outside the IJSEM
    Aharon Oren, Markus Göker
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    Sung-Hyun Yang, Mi-Jeong Park, Hyun-Myung Oh, Kae Kyoung Kwon
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    Neak Muhammad, Tra T.H. Nguyen, Yong-Jae Lee, Jaeho Ko, Forbes Avila, Song-Gun Kim
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    Teddy Namirimu, Jihyun Yu, Jhung-Ahn Yang, Sung-Hyun Yang, Yun Jae Kim, Kae Kyoung Kwon
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Optimal Protocol for Enumeration of Attached Bacteria on Glass Slides
Hyun Sang Lee , Kae Kyoung Kwon , Jong Ho Lee , Hong Kum Lee
J. Microbiol. 1999;37(4):263-266.
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AbstractAbstract PDF
In examining bacterial growth on glass surfaces immersed in sea water, we found serious differences between enumeration methods. Therefore, we compared various methods and found sonication and direct count methods were superior to other methods. Since the direct count method was not suitable for long-term investigation, we chose the sonication method and confirmed that sonication periods 8 times for 30 seconds was optimal for the detachment of bacteria from glass surfaces.
Morphological Diversity of Marine Microorganisms on Different Isolation Media
Shin Hye Park , Kae Kyoung Kwon , Deuk-Soo Lee , Hong Kum Lee
J. Microbiol. 2002;40(2):161-165.
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AbstractAbstract PDF
Isolation frequency of microorganisms from marine sources was examined with different media and samples collected from the coastal area of Cheju Island. From sea water samples, about 1% of microorganisms from the total number of bacteria were recovered. Microorganisms were cultured at the much lower frequency of 10^-4 ?0^-6 from other marine sources, such as sediment, sponges and corals. The frequency of duplicated isolation was examined with 140 morphologically different colonies isolated on different media. Fourteen percent of them exhibited the same morphology on two different media. The duplication frequency of the isolates among three different media was 33%.
Isolation and Identification of Biofilm-Forming Marine Bacteria on Glass Surfaces in Dae-Ho Dike, Korea
Kae Kyoung Kwon , Hyun Sang Lee , Sung-Young Jung , Joung-Han Yim , Jung-Hyun Lee , Hong Kum Lee
J. Microbiol. 2002;40(4):260-266.
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AbstractAbstract PDF
Bacterial strains were isolated from biofilms formed on glass slides submerged in seawater in Dae-Ho Dike. Eight strains showing fast attaching ability were selected and identified. Their exopolysaccharide(EPS)-producing ability and EPS properties were characterized. Based on Microlog System, 4 among the 8 strains were identified as Micrococcus luteus and the rest were Bacillus thuringiensis, Bacillus megaterium, Staphylococcus saprophyticus and Agrobacterium vitis. A. vitis was reidentified as Sulfitobacter pontiacus based on 16S rDNA sequence data. The amount of water-soluble EPS produced by the 8 strains ranged from 0.114 to 1.329 g·l^-1 and the productivity was negatively correlated with the cell biomass. The molecular weight of the produced EPS ranged from 0.38 to 25.19x 10^4 Da. Glucose and galactose were ubiquitous sugar components. Mannose, ribose, and xylose were also major sugar components. The molecular weight and composition of the EPS showed strain-specific variation.

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