In-Tae Cha

Metabolic versatility of toluene-degrading, iron-reducing bacteria in tidal flat sediment, characterized by stable isotope probing-based metagenomic analysis

DNA stable isotope probing and metagenomic sequencing were used to assess the metabolic potential of iron-reducing bacteria involved in anaerobic aromatic hydrocarbon degradation in oil spill-affected tidal flats. In a microcosm experiment, (13) C-toluene was degraded with the simultaneous reduction of Fe(III)-NTA, which was also verified by quasi-stoichiometric (13) C-CO2 release. The metabolic potential of the dominant member affiliated with the genus Desulfuromonas in the heavy DNA fraction was inferred using assembled scaffolds (designated TF genome, 4.40 Mbp with 58.8 GC mol%), which were obtained by Illumina sequencing. The gene clusters with peripheral pathways for toluene and benzoate conversion possessed the features of strict and facultative anaerobes. In addition to the class II-type benzoyl-CoA reductase (Bam) of strict anaerobes, the class I-type (Bcr) of facultative anaerobes was encoded. Genes related to the utilization of various anaerobic electron acceptors, including iron, nitrate (to ammonia), sulfur and fumarate, were identified. Furthermore, genes encoding terminal oxidases (caa3 , cbb3 and bd) and a diverse array of genes for oxidative stress responses were detected in the TF genome. This metabolic versatility may be an adaptation to the fluctuating availability of electron acceptors and donors in tidal flats.

Roseovarius halotolerans sp. nov., isolated from deep seawater.

A Gram-reaction-negative, non-motile, aerobic bacterium, designated HJ50(T), was isolated from deep seawater of the East Sea, South Korea. Cells were ovoid to rod-shaped (0.5-0.8x1.3-3.0 mum), often with unequal ends, suggesting a budding mode of reproduction. The strain had an absolute requirement for sea salts and tolerated up to 20 % (w/v) sea salts. Propionate, dl-lactate, 2-ketogluconate, 3-hydroxybutyrate and rhamnose were used as growth substrates, but not mannitol, salicin, 4-hydroxybenzoate or acetate. The major fatty acid was summed feature 7 (C(18 : 1)omega7c/omega9t/omega12t) and the DNA G+C content was 59.0+/-0.1 mol%. Phylogenetic analyses based on 16S rRNA gene sequences showed that this strain was affiliated with the genus Roseovarius. Similarities between the 16S rRNA gene sequence of strain HJ50(T) (1430 nt) and those of type strains of members of the genus Roseovarius were 94.1-96.3 %. DNA-DNA relatedness values between strain HJ50(T) and the type strains of members of the genus Roseovarius were low (1.3-24.6 %). Physiological and biochemical differences support assignment of strain HJ50(T) to the genus Roseovarius as a representative of a novel species. The name Roseovarius halotolerans sp. nov. is proposed, with HJ50(T) (=KCTC 22224(T) =LMG 24468(T)) as the type strain.

Unveiling abundance and distribution of planktonic Bacteria and Archaea in a polynya in Amundsen Sea, Antarctica.

Polynyas, areas of open water surrounded by sea ice, are sites of intense primary production and ecological hotspots in the Antarctic Ocean. This study determined the spatial variation in communities of prokaryotes in a polynya in the Amundsen Sea using 454 pyrosequencing technology, and the results were compared with biotic and abiotic environmental factors. The bacterial abundance was correlated with that of phytoplankton, Phaeocystis spp. and diatoms. A cluster analysis indicated that the bacterial communities in the surface waters of the polynya were distinct from those under the sea ice. Overall, two bacterial clades, Polaribacter (20-64%) and uncultivated Oceanospirillaceae (7-34%), dominated the surface water in the polynya while the Pelagibacter clade was abundant at all depths (7-42%). The archaeal communities were not as diverse as the bacterial communities in the polynya, and marine group I was dominant (> 80%). Canonical correspondence analysis indicated that the oceanographic properties facilitated the development of distinct prokaryotic assemblages in the polynya. This analysis of the diversity and composition of the psychrophilic prokaryotes associated with high phytoplankton production provides new insights into the roles of prokaryotes in biogeochemical cycles in high-latitude polynyas.

Molecular analysis of spatial variation of iron-reducing bacteria in riverine alluvial aquifers of the Mankyeong River.

Alluvial aquifers are one of the mainwater resources in many countries. Iron reduction in alluvial aquifers is often a major anaerobic process involved in bioremediation or causing problems, including the release of As trapped in Fe(III) oxide. We investigated the distribution of potential iron-reducing bacteria (IRB) in riverine alluvial aquifers (B1, B3, and B6 sites) at the Mankyeong River, Republic of Korea. Inactive iron reduction zones, the diversity and abundance of IRB can be examined using a clone library and quantitative PCR analysis of 16S rRNA genes. Geobacter spp. are potential IRB in the iron-reducing zone at the B6 (9 m) site, where high Fe(II) and arsenic (As) concentrations were observed. At the B3 (16 m) site, where low iron reduction activity was predicted, a dominant clone (10.6%) was 99% identical in 16S rRNA gene sequence with Rhodoferax ferrireducens. Although a major clone belonging to Clostridium spp. was found, possible IRB candidates could not be unambiguously determined at the B1 (18 m) site. Acanonical correspondence analysis demonstrated that, among potential IRB, only the Geobacteraceae were well correlated with Fe(II) and As concentrations. Our results indicate high environmental heterogeneity, and thus high spatial variability, in thedistribution of potential IRB in the riverine alluvial aquifersnear the Mankyeong River.

Salinisphaera orenii sp. nov., isolated from a solar saltern

A taxonomic study was performed on two isolates, designated strains MK-B5(T) and MK-B7, isolated from sediment of a solar saltern pond in Gomso Bay, Republic of Korea. Comparative 16S rRNA gene sequence analysis showed that strains MK-B5(T) and MK-B7 belong to the Gammaproteobacteria and are related most closely to Salinisphaera shabanensis JCM 11575(T) ( = E1L3A(T)) (96.3 and 96.5% similarity, respectively), Salinisphaera dokdonensis KCCM 90064(T) ( = CL-ES53(T)) (95.6 and 95.6%) and Salinisphaera hydrothermalis JCM 115514(T) ( = EPR70(T)) (95.1 and 95.3%). The level of 16S rRNA gene sequence similarity between strains MK-B5(T) and MK-B7 was 99.8%. The G+C contents of their genomic DNAs were 63.4 and 63.6 mol%, respectively, and the major respiratory quinone was ubiquinone-8. DNA-DNA relatedness between strains MK-B5(T) and MK-B7 was 98%, indicating that the two isolates represent a single species. However, the level of DNA-DNA relatedness between the two isolates and S. shabanensis E1L3A(T) (26.4-30.8%) indicates that they represent a novel species. Strains MK-B5(T) and MK-B7 possessed C(14:0), C(16:0) and C(19:0)ω8c cyclo as major fatty acids. The two isolates were Gram-stain-negative, strictly aerobic, short rod-shaped and motile. They grew at 10-40 °C (optimum, 35-37 °C), at pH 5.0-8.5 (optimum, 7.0-7.5) and with 5-25% (w/v) NaCl (optimum, 15% NaCl). On the basis of phenotypic and phylogenetic analyses, strains MK-B5(T) and MK-B7 are thus considered to represent a novel species of the genus Salinisphaera, for which the name Salinisphaera orenii sp. nov. is proposed. The type strain is MK-B5(T) ( = KCTC 23198(T) = JCM 17073(T)).

Phylogenetic analysis of microalgae based on highly abundant proteins using mass spectrometry

The blooms of toxic phototrophic microorganisms, such as microalgae and cyanobacteria, which are typically found in freshwater and marine environments, are becoming more frequent and problematic in aquatic systems. Due to accumulation of toxic algae, harmful algal blooms (HABs) exert negative effects on aquatic systems. Therefore, rapid detection of harmful microalgae is important for monitoring the occurrence of HABs. Mass spectrometry-based methods have become sensitive, specific techniques for the identification and characterization of microorganisms. Matrix-assisted laser desorption/ionization (MALDI) with time-of-flight (TOF) mass spectrometry (MS) allows us to measure a unique molecular fingerprint of highly abundant proteins in a microorganism and has been used for the rapid, accurate identification of bacteria and fungi in clinical microbiology. Here, we tested the specificity of MALDI-TOF MS using microalgal strains (Heterocapsa, Alexandrium, Nannochloropsis, Chaetoceros, Chlorella, and Dunaliella spp.). Our research suggested that this method was comparable in terms of the rapid identification of microalgea to conventional methods based on genetic information and morphology. Thus, this efficient mass spectrometry-based technique may have applications in the rapid identification of harmful microorganisms from aquatic environmental samples.

Occurrence of viable, red-pigmented haloarchaea in the plumage of captive flamingoes

Flamingoes (Phoenicopterus spp.) whose plumage displays elegant colors, inhabit warm regions close to the ocean throughout the world. The pink or reddish color of their plumage originates from carotenoids ingested from carotenoid-abundant food sources, since flamingoes are unable to synthesize these compounds de novo. In this study, viable red-colored archaeal strains classified as extremely halophilic archaea (i.e., haloarchaea) and belonging to the genera Halococcus and Halogeometricum were isolated from the plumage of flamingoes in captivity. Detailed analysis for haloarchaeal community structure in flamingo feathers based on metagenomic data identified several haloarchaeal genera and unclassified sequences of the class Halobacteria at the genus level. Carotenoid pigment analyses showed that a bacterioruberin precursor carotenoid in haloarchaea was identical to one of the pigments found in flamingo plumage. To the best of our knowledge, this is the first report of viable extremophilic archaea in avian plumage, thus contributing to our understanding of the ecology of haloarchaea. The potential influence of haloarchaea as an environmental factor determining avian plumage coloration should be investigated in further studies.

Enhanced Production of Gamma-Aminobutyric Acid by Optimizing Culture Conditions of Lactobacillus brevis HYE1 Isolated from Kimchi, a Korean Fermented Food.

This study evaluated the effects of culture conditions, including carbon and nitrogen sources, L-monosodium glutamate (MSG), and initial pH, on gamma-aminobutyric acid (GABA) production by Lactobacillus brevis HYE1 isolated from kimchi, a Korean traditional fermented food. L. brevis HYE1 was screened by the production analysis of GABA and genetic analysis of the glutamate decarboxylase gene, resulting in 14.64 mM GABA after 48 h of cultivation in MRS medium containing 1% (w/v) MSG. In order to increase GABA production by L. brevis HYE1, the effects of carbon and nitrogen sources on GABA production were preliminarily investigated via one-factor-at-a-time optimization strategy. As the results, 2% maltose and 3% tryptone were determined to produce 17.93 mM GABA in modified MRS medium with 1% (w/v) MSG. In addition, the optimal MSG concentration and initial pH were determined to be 1% and 5.0, respectively, resulting in production of 18.97 mM GABA. Thereafter, response surface methodology (RSM) was applied to determine the optimal conditions of the above four factors. The results indicate that pH was the most significant factor for GABA production. The optimal culture conditions for maximum GABA production were also determined to be 2.14% (w/v) maltose, 4.01% (w/v) tryptone, 2.38% (w/v) MSG, and an initial pH of 4.74. In these conditions, GABA production by L. brevis HYE1 was predicted to be 21.44 mM using the RSM model. The experiment was performed under these optimized conditions, resulting in GABA production of 18.76 mM. These results show that the predicted and experimental values of GABA production are in good agreement.

Marinoscillum luteum sp. nov., isolated from marine sediment.

A novel strain, designated SJP7(T), was isolated from sediment of the Tofua Arc of the Tonga Trench. The 16S rRNA gene sequence of the isolate showed the highest similarity to that of Marinoscillum pacificum KCCM 42325(T) (95.9%). Phylogenetic analysis revealed that the isolate formed a distinct phyletic line with Marinoscillum pacificum KCCM 42325(T) and Marinoscillum furvescens LMG 13023(T) within the family Cytophagaceae. Cells of strain SJP7(T) were Gram-stain-negative and appeared as long rods that were motile by gliding. Growth was observed at 15-40 °C (optimum, 30 °C), at pH 5.5-9.0 (optimum, pH 7.5-8.0) and in the presence of 0.5-7.0% (w/v) NaCl (optimum, 2.5-3%). The major respiratory quinone was MK-7. The dominant fatty acids were summed feature 3 (comprising C(16:1)ω7c and/or C(16:1)ω6c), iso-C(15:0) and C(16:1)ω5c. The DNA G+C content was 43.5 mol%. These properties support the affiliation of strain SJP7(T) with the genus Marinoscillum. Further phenotypic differentiation of strain SJP7(T) from other species of the genus Marinoscillum was indicated by the results of physiological and biochemical tests. On the basis of evidence from our polyphasic taxonomic study, strain SJP7(T) represents a novel species of the genus Marinoscillum, for which the name Marinoscillum luteum sp. nov. is proposed. The type strain of Marinoscillum luteum is SJP7(T) ( =KCTC 23939(T) =NCAIM B02491(T)).

Reichenbachiella faecimaris sp. nov., isolated from a tidal flat, and emended descriptions of the genus Reichenbachiella and Reichenbachiella agariperforans.

A taxonomic study was carried out on two bacterial strains, PCP11(T) and PCP104, isolated from a tidal flat of the Yellow Sea, Korea. Comparative 16S rRNA gene sequence studies showed that these strains belonged to the family Cytophagaceae, phylum Bacteroidetes. Strains PCP11(T) and PCP104 shared 99.4 % sequence similarity and were related most closely to Reichenbachiella agariperforans KMM 3525(T) (95.8 and 96.0 % sequence similarity, respectively). Members of the genera Fulvivirga, Roseivirga, Fabibacter and Marinoscillum were the next closest relatives of the new isolates, with sequence similarities ≤ 91 %. The two isolates were Gram-staining-negative, strictly aerobic, gliding bacteria. They grew in the presence of 1-5 % NaCl, at pH 5.5-8.5 and at 4-35 °C. Strains PCP11(T) and PCP104 shared a number of physiological and biochemical properties with Reichenbachiella agariperforans KMM 3525(T), but they differed from this strain in the hydrolysis of biopolymers and in the production of carotenoid and flexirubin-type pigments. Both strains possessed iso-C(15 : 0), summed feature 4 (C(16 : 1)ω7c and/or iso-C(15 : 0) 2-OH) and C(15 : 0) as major cellular fatty acids. The major respiratory quinone was menaquinone 7 (MK-7). The G+C contents of the genomic DNA of strains PCP11(T) and PCP104 were 39.6 and 41.9 mol%, respectively. On the basis of phenotypic data and phylogenetic inference, it is proposed that the two isolates represent a novel species, Reichenbachiella faecimaris sp. nov., with strain PCP11(T) ( = KACC 14523(T)  = JCM 16588(T)) as the type strain. Emended descriptions of the genus Reichenbachiella and Reichenbachiella agariperforans are also proposed.