• Journal of Life Science
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• v.30 no.1
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• pp.32-39
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• 2020

Rhododendron mucronulatum, native to Korea, Mongolia, Russia and parts of northern China, is known not only for its medicinal properties but also as a tourist attraction. One of the most famous tourist destinations with R. mucronulatum is in Biseul Mountain County Park, Daegu, Korea. To investigate the relationship between R. mucronulatum and microbiome communities in the surrounding soil, three sites within the park were chosen for sampling in February and August. The soil samples were then passed through a pyrosequencing process for analysis of the bacterial communities, and a total of 404,899 sequencing reads were obtained. Between 2,349 and 4,736 operational taxonomic units (OTUs) were observed across the three sampling zones and two seasons; samples from the park entrance showed a higher number of OTUs than the other two sites, and samples from August had more OTUs than those from February. The sample from the second observation site displayed the fewest OTUs, particularly in February. According to Chao1 and Shannon indices, samples from the park entrance in August demonstrated the highest degree of species richness and diversity. Studying the bacterial communities across the six samples identified the common population as comprising 287 genera, 45 of which are only present in Biseul Mountain County Park and are expected to participate in the colonization of R. mucronulatum.

• Applied Biological Chemistry
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• v.50 no.4
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• pp.262-267
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• 2007

To access the natural product antibiotics produced by uncultured microorganisms, six cosmid libraries of DNA extracted directly from soil samples (environmental DNA, eDNA) were constructed and screened for the production of antibacterial active molecules. Of the approximately 60,000 clones screened, one antibacterial clone (YS92B) was detected. Ethyl acetate extracts of clone YS92B showed antibacterial activity against various pathogenic bacteria (Listeria monocytogenes, Bacillus subtilis, Pseudomonas syringae, Xanthomonas campestris pv. oryzae, Staphylococcus epidemis). Active constituents from cultures of YS92B were isolated and purified using a bioassay-guided fractionation against B. subtilis through a series of procedures (ethyl acetate extraction, Sephadex LH20 column chromatography, High Performance Liquid Chromatography). NMR (Nuclear Magnetic Resonance) spectral analysis of a major antibacterial active YS92B-VII indicated that it is a lauric acid linked to tyrosine. This report describes the characterization of antibacterially active long chain N-acyl derivatives of tyrosine that are produced by eDNA clones hosted in Escherichia coli from Korean soils.

• Microbiology and Biotechnology Letters
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• v.36 no.4
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• pp.260-267
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• 2008

The choice of expression vector is very important for industrial production of proteins. Therefore, the systematic mining of promoters over a wider range of genetic resource and/or host is required. We previously reported a novel bidirectional reporting system (pBGR) for the isolation of promoters from metagenome and screened useful promoters that functioned constitutively in E. coli under general culture conditions. Among them, three promoter sequences including each upstream region were amplified by PCR and used to construct new expression vectors. To facilitate subcloning, a multi-cloning site was incorporated into the downstream region of the revere primer sequence. At these sites, GFP, esterase and $\beta$-glucosidase were subcloned and analyzed the constitutive expression ability of new promoter in terms of protein solubility and expression level. As a result, these vectors expressed the proteins constitutively to a level of $2{\sim}3%$ of the total cell protein in soluble fraction (>80 %). This study suggested that excavation of metagenomic promoters for construction of expression vector in a certain strain could provide a way for the development of the expression systems.

• Journal of Microbiology and Biotechnology
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• v.29 no.12
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• pp.1947-1956
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• 2019

The gut microbiome influences the health and well-being of dogs. However, little is known about the impact of breed on the fecal microbiome composition in dogs. Therefore, we aimed to investigate the differences in the fecal microbiome in three breeds of dog fed and housed under the same conditions, namely eight Maltese (8.0 ± 0.1 years), eight Miniature Schnauzer (8.0 ± 0.0 years), and nine Poodle dogs (8.0 ± 0.0 years). Fresh fecal samples were collected from the dogs and used to extract metagenomic DNA. The composition of the fecal microbiome was evaluated by 16S rRNA gene amplicon sequencing on the MiSeq platform. A total of 840,501 sequences were obtained from the 25 fecal samples and classified as Firmicutes (32.3-97.3% of the total sequences), Bacteroidetes (0.1-62.6%), Actinobacteria (0.2-14.7%), Fusobacteria (0.0-5.7%), and Proteobacteria (0.0-5.1%). The relative abundance of Firmicutes was significantly lower in the Maltese dog breed than that in the other two breeds, while that of Fusobacteria was significantly higher in the Maltese than in the Miniature Schnauzer breed. At the genus level, the relative abundance of Streptococcus, Fusobacterium, Turicibacter, Succinivibrio, and Anaerobiospirillum differed significantly among the three dog breeds. These genera had no correlation with age, diet, sex, body weight, vaccination history, or parasite protection history. Within a breed, some of these genera had a correlation with at least one blood chemistry value. This study indicates that the composition of the fecal microbiome in dogs is affected by breed.

• Microbiology and Biotechnology Letters
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• v.46 no.2
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• pp.102-110
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• 2018

A bacterial strain capable of metabolizing myo-inositol (MI) and converting to other substances was isolated from soil of orchard. The isolate, named YB-46, was grown on minimal medium supplemented with MI as the sole carbon source and was presumed to belonging to genus Enterobacter according to the 16S rDNA sequence. Escherichia coli transformant converting MI into unknown metabolites was selected from a metagenomic library prepared with fosmid pCC1FOS vector. Plasmid was isolated from the transformant, and the inserted gene was partially sequenced. From the nucleotide sequence, an iolG gene was identified to encode myo-inositol dehydrogenase (IolG) consisting of 336 amino residues. The IolG showed amino acid sequence similarity of about 50% with IolG of Enterobacter aerogenes and Bacillus subtilis. The His-tagged IolG (HtIolG) fused with hexahistidine at C-terminus was produced and purified from cell extract of recombinant E. coli. The purified HtIolG showed maximal activity at $45^{\circ}C$ and pH 10.5 with the highest activity for MI and D-glucose, and more than 90% of maximal activity for D-chiro-inositol, D-mannitol and D-xylose. $K_m$ and $V_{max}$ values of the HtIolG for MI were 1.83 mM and $0.724{\mu}mol/min/mg$ under the optimal reaction condition, respectively. The activity of HtIolG was increased 1.7 folds by $Zn^{2+}$, but was significantly inhibited by $Co^{2+}$ and SDS.

• Tuberculosis and Respiratory Diseases
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• v.79 no.3
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• pp.165-178
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• 2016

Background: Although recent metagenomic approaches have characterized the distinguished microbial compositions in airways of asthmatics, these results did not reach a consensus due to the small sample size, non-standardization of specimens and medication status. We conducted a metagenomics approach by using terminal restriction fragment length polymorphism (T-RFLP) analysis of the induced whole sputum representing both the cellular and fluid phases in a relative large number of steroid $na{\ddot{i}}ve$ asthmatics. Methods: Induced whole sputum samples obtained from 36 healthy subjects and 89 steroid-$na{\ddot{i}}ve$ asthma patients were analyzed through T-RFLP analysis. Results: In contrast to previous reports about microbiota in the asthmatic airways, the diversity of microbial composition was not significantly different between the controls and asthma patients (p=0.937). In an analysis of similarities, the global R-value showed a statistically significant difference but a very low separation (0.148, p=0.002). The dissimilarity in the bacterial communities between groups was 28.74%, and operational taxonomic units (OTUs) contributing to this difference were as follows: OTU 789 (Lachnospiraceae), 517 (Comamonadaceae, Acetobacteraceae, and Chloroplast), 633 (Prevotella), 645 (Actinobacteria and Propionibacterium acnes), 607 (Lactobacillus buchneri, Lactobacillus otakiensis, Lactobacillus sunkii, and Rhodobacteraceae), and 661 (Acinetobacter, Pseudomonas, and Leptotrichiaceae), and they were significantly more prevalent in the sputum of asthma patients than in the sputum of the controls. Conclusion: Before starting anti-asthmatic treatment, the microbiota in the whole sputum of patients with asthma showed a marginal difference from the microbiota in the whole sputum of the controls.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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• v.17 no.3
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• pp.160-180
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• 2012

We have studied the eukaryotic plankton species diversity to compare the community structure of fresh and brackish waters in the lower reaches of the Nakdong River using metagenomic methods. We constructed 18S rDNA clone libraries of total DNAs extracted from environmental water samples collected at Mulgeum (MG100929, fresh) and Eulsukdo bridge (ES, brackish). Through the steps of colony PCR, PCR-RFLP, sequencing and similarity analysis, we discovered the diverse species composition of eukaryotic plankton. Total 338 clones (170 at MG100929 and 168 at ES) were analyzed, and then we found 74 phylotypes (49 for MG100929 and 25 for ES). From the phylogenetic analysis, we confirmed various eukaryotic plankton of broad range of taxonomic groups, including Stramenopiles, Cryptophyta, Viridiplantae, Alveolata, Rhizaria, Metazoa, and Fungi. We also found several unreported species in Korea and candidates of new taxonomic entities at levels higher than genus. Especially, the cryptic species diversity including unreported phylotypes of Pirsonia (Stramenopiles) and Perkinsea (Alveolata) suggests that the molecular monitoring method can produce new informative biological data in monitoring the changes in the Nakdong River Mouth ecosystem.

• Korean Journal of Microbiology
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• v.52 no.3
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• pp.365-374
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• 2016

In high-latitude regions, temperature has risen ($0.6^{\circ}C$ per decade) and this leads to the increase in microbial degradability against soil organic carbon (SOC). Furthermore, the decomposed SOC is converted into green-house gases ($CO_2$ and $CH_4$) and their release could further increase the rate of climate change. Thus, understanding the microbial diversity and their functions linked with SOC degradation in soil-thawing model is necessary. In this study, we divided tundra soil from Council, Alaska into two depth regions (30-40 cm and 50-60 cm of depth, designated as SPF and PF, respectively) and incubated that for 108 days at $0^{\circ}C$. A total of 111,804 reads were obtained through a pyrosequencing-based metagenomic study during the microcosm experiments, and 574-1,128 of bacterial operational taxonomic units (OTUs) and 30-57 of archaeal OTUs were observed. Taxonomic analysis showed that the distribution of bacterial taxa was significantly different between two samples. In detail, the relative abundance of phyla Actinobacteria and Firmicutes largely increased in SPF and PF soil, respectively, while phyla Crenarchaeota was increased in both soil samples. Weight measurement and gel permeation chromatography of the SOC extracts demonstrated that polymerization of humic acids, main component of SOC, occurred during the microcosm experiments. Taken together our results indicate that these bacterial and archaeal phyla could play a key function in SOC degradation and utilization in cold tundra soil.

• Korean Journal of Food Science and Technology
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• v.52 no.3
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• pp.304-309
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• 2020

Different types of storage containers, such as polypropylene (PP), stainless steel (STS), and ceramic were used for kimchi storage at 0℃ in a refrigerator, and the characteristics were compared for 32 days. The pH of kimchi samples in PP and STS containers reached 4.59 and 4.53, respectively at day 16, while a pH of 4.92 could be observed in ceramic containers. This trend persisted until day 32. Titratable acidities of the PP and STS container contents reached 0.83 and 0.82%, respectively, on day 16, while it reached 0.73% in the case of the ceramic container contents. The viable cell counts of lactic acid bacteria in kimchi samples in PP, STS, and ceramic containers fluctuated and finally reached 4.87, 5.44, and 5.35 Log CFU/g, respectively. Weissella koreensis occupied a large portion of the kimchi sample of the ceramic container on day 20 based on the metagenomic analysis. Taken together, ceramic container might be desirable for the storage of kimchi in low temperature refrigerators.

• Journal of Life Science
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• v.28 no.10
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• pp.1244-1253
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• 2018

Rumen microbiome consists of a wide variety of microorganisms, such as bacteria, archaea, protozoa, fungi, and viruses, that are in a symbiotic relationship in a strict anaerobic environment in the rumen. These rumen microbiome, a vital maker, play a significant role in feed fermentation within the rumen and produce different volatile fatty acids (VFAs). VFAs are essential for energy metabolism and protein synthesis of the host animal, even though emission of methane gas after feed fermentation is considered a negative indicator of loss of dietary energy of the host animal. To improve rumen microbial efficiency, a variety of approaches, such as feed formulation, the addition of natural feed additives, dietary feed-microbes, etc., have taken to increase ruminant performance. Recently with the application of high-throughput sequencing or next-generation sequencing technologies, especially for metagenomics and metatranscriptomics of rumen microbiomes, our understanding of rumen microbial diversity and function has significantly increased. The metaproteome and metabolome provide deeper insights into the complicated microbial network of the rumen ecosystem and its response to different ruminant diets to improve efficiency in animal production. This review summarized some recent advances of rumen microbiome techniques, especially "meta-omics," viz. metagenomic, metatranscriptomic, metaproteomic, and metabolomic techniques to increase feed fermentation and utilization in ruminants.