• International Journal of Industrial Entomology and Biomaterials
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• v.30 no.2
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• pp.64-74
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• 2015

Beetles Protaetia brevitarsis seulensis Kolbe (Coleoptera: Cetoniidae) and Allomyrina dichotoma Linn. (Coleoptera: Scarabaeidae) are widely used in traditional medicine, and the number of insect-rearing farms is increasing in South Korea. The purpose of this study was to establish a multiplex PCR-based assay for rapid simultaneous detection of multiple pathogens causing insect diseases. Six insect parasites such as fungi Beauveria bassiana (Bals.-Criv.) Vuill. (Hypocreales: Cordycipitaceae) and Metarhizium anisopliae (Metschn.) Sorokin (Hypocreales: Clavicipitaceae), bacteria Bacillus thuringiensis Berliner (Bacillales: Bacillaceae), Pseudomonas aeruginosa Migula (Pseudomonadales: Pseudomonadaceae), and Serratia marcescens Bizio (Enterobacteriales: Enterobacteriaceae), and Oryctes rhinoceros nudivirus were chosen based on the severity and incidence rate of insect diseases in South Korea. Pathogen-specific primers were designed and successfully applied for simultaneous detection of multiple infectious agents in farm-bred insects P. b. seulensis and A. dichotoma using multiplex PCR and high resolution capillary electrophoresis. Our results indicate that multiplex PCR is an effective and time-saving method for simultaneous detection of multiple infections in insects, and the QIAxcel capillary electrophoresis system is useful for quantitative evaluation of the individual impact of each infectious agent on the severity of insect disease. The approach designed in this study can be utilized for rapid and accurate diagnostics of infection in insect farms.

• Journal of Species Research
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• v.5 no.1
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• pp.166-178
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• 2016

An outcome of the study to discover indigenous prokaryotic species in Korea, a total of 26 bacterial species assigned to the classes Bacteroidetes and Firmicutes were isolated from diverse environmental samples collected from soil, tidal flat, freshwater, seawater, wetland, plant roots, and fermented foods. From the high 16S rRNA gene sequence similarity (>99.0%) and formation of a robust phylogenetic clade with the closest species, it was determined that each strain belonged to each independent and predefined bacterial species. There is no official report that these 26 species have been described in Korea; therefore 14 strains for the order Flavobacteriales and two strains for the order Cytophagales were assigned to the class Bacteroidetes, and 8 strains for the order Bacillales and 4 strains for the order Lactobacillales were assigned to the class Firmicutes are reported for new bacterial species found in Korea. Gram reaction, colony and cell morphology, basic biochemical characteristics, isolation source, and strain IDs are also described in the species description section.

• Fisheries and Aquatic Sciences
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• v.18 no.1
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• pp.1-6
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• 2015

In an effort to discover alternative antimicrobials against Listeria monocytogenes, several marine algae were screened. The methanolic extract of Ecklonia cava exhibited the highest antibacterial activity against L. monocytogenes, with the ethyl acetate (EtOAc) soluble fraction of E. cava methanolic extract having a MIC value of $256{\mu}g/mL$ and a MBC value of ${\mu}g/mL$. The MIC values of streptomycin in combination with the EtOAc fraction were markedly reduced up to 64-fold, suggesting that the antibacterial activity of the antibiotic was restored when combined with the EtOAc fraction. The interaction between streptomycin and the EtOAc fraction was assessed by fractional inhibitory concentration (FIC) indices. The combination of streptomycin and the EtOAc fraction against L. monocytogenes resulted in ${\Sigma}FIC_{min}$ range of 0.141 to 0.266 and ${\Sigma}FIC_{max}$ of 0.531 for all strains. The median ${\Sigma}FIC$ against L. monocytogenes strains ranged from 0.172 to 0.344. Thus, synergistic ranges of FIC <1 were observed for all combinations of streptomycin and the EtOAc fraction against L. monocytogenes strains. Indeed, the median ${\Sigma}FIC$ of streptomycin-EtOAc fraction ranged from 0.172 to 0.344, suggesting a marked synergy.

• Journal of Species Research
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• v.4 no.2
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• pp.127-136
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• 2015

As an outcome of the study on the bacterial species diversity in Korea, we report 24 unrecorded bacterial species of Korea belonging to the phylum Firmicutes. The unrecorded species excavated through this study were assigned to 12 different genera of 7 families, namely Bacillus, Halobacillus, Lysinibacillus and Thalassobacillus of Bacillaceae, Brevibacillus and Paenibacillus of Paenibacillaceae, Viridibacillus of Planococcaceae, Salinicoccus and Staphylococcus of Staphylococcaceae, Enterococcus of Enterococcaceae, Lactobacillus of Lactobacillaceae, and Lactococcus of Streptococcaceae, respectively. The bacterial isolates were obtained from various ecosystems in Korea. The isolates were identified based on 16S rRNA gene sequences, and those exhibiting at least 99% sequence similarity with known bacterial species but never reported in Korea were selected as unrecorded species. The selected isolates were subjected to further taxonomic characterization including the analysis of cell shape and fine structure using electron microscope, colony color and shapes, enzyme activities and carbon source utilization. The descriptive information on the 24 unrecorded species are provided.

• Journal of Species Research
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• v.7 no.1
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• pp.50-59
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• 2018

During the course of investigation of indigenous prokaryotic species in Korea, a total of 30 bacterial strains belonging to the phylum Firmicutes were isolated from diverse environmental sites such as soil, avian feces, wastewater treatment plants, fermented vegetables, seawater, algae, sea cucumber, octopus and tidal flat sediment. Phylogenetic analysis based on 16S rRNA gene sequences revealed that each strain showed high sequence similarity (${\geq}98.7%$) to the closest type strain and formed a robust phylogenetic clade with the most closely related species in the phylum Firmicutes. To date, there is no official record of these 30 species in Korea. Therefore, we report 26 species of 12 genera in the order Bacillales and 4 species of 4 genera in the order Lactobacillales which have not been reported in Korea. Morphological and biochemical characteristics, isolation sources and NIBR deposit numbers are described in the species descriptions.

• Microbiology and Biotechnology Letters
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• v.47 no.1
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• pp.125-131
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• 2019

We have isolated and identified 72 bacterial strains from four bentonite samples collected at the mining areas located in Gyeongsangbuk-do, Republic of Korea, and measured their hydrolytic enzyme (${\alpha}$-amylase, protease, and cellulase) activities to identify the isolates with industrial-use potential. Most of the isolates belonged to the Bacillaceae, with minor portions being from the Paenibacillaceae, Micrococcaceae, and Bacillales Family XII at the family level. Of the strains isolated, 33 had extracellular ${\alpha}$-amylase activity, 30 strains produced cellulase, and 35 strains produced protease. Strain MBLB1268, having the highest ${\alpha}$-amylase activity, was identified as Bacillus siamensis ($0.38{\pm}0.06U/ml$). Bacillus tequilensis MBLB1223, isolated from Byi33-b, showed the highest cellulase activity ($0.26{\pm} 0.04U/ml$), whereas Bacillus wiedmannii MBLB1197, isolated from Zdb130-b, exhibited the highest protease activity ($54.99{\pm}0.78U/ml$). These findings show that diverse bacteria of the Bacillaceae family adhere to and exist in bentonite and are potential sources of industrially useful hydrolytic enzymes.

• Journal of Species Research
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• v.10 no.2
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• pp.117-133
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• 2021

In 2019, 43 bacterial strains were isolated from food, soil, marine environments, human, and animals related sources from the Republic of Korea. Based on the analysis of 16S rRNA gene sequence, these isolates were allocated to the phyla Bacteroidetes and Firmicutes as unrecorded species in Korea. The 10 Bacteroidetes strains were classified into the families Bacteroidaceae, Chitinophagaceae, Cytophagaceae, Flavobacteriaceae, and Prolixibacteraceae (of the orders Bacteroidales, Chitinophagales, Cytophagales, Flavobacteriales, and Marinilabiliales, respectively). The 33 Firmicutes strains belonged to the families Bacillaceae, Paenibacillaceae, Planococcaceae, Staphylococcaceae, Clostridiaceae, Lachnospiraceae, Peptostreptococcaceae, Enterococcaceae, Lactobacillaceae, Leuconostocaceae, and Streptococcaceae (of the orders Bacillales, Clostridiales, and Lactobacillales). These unrecorded bacteria were determined based on taxonomic criterion (>98.7%; 16S rRNA gene sequence similarity). In addition, their phylogenetic affiliation, as well as cell and colony morphologies, staining reactions, and physiological and biochemical properties were investigated. Therefore, we report 43 isolates as unrecorded species, and described basic features, isolation source, and locations of these strains.

• Journal of Marine Bioscience and Biotechnology
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• v.16 no.1
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• pp.1-18
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• 2024

Coastal dunes must be conserved. Their native halophytes support coastal geography while their symbiotic microorganisms help vegetation thrive. The Goraebul coast has the largest, well-conserved dune system on the East Sea of the Korean Peninsula due to a climax mixed halophyte (C. soldanella, C. kobomugi, and E. mollis) vegetation support. This study identified rhizobacteria and their diversity in mixed halophyte communities unique to Goraebul. Five phyla, 12 genera, and 21 species were identified based on 16S rDNA sequences from 65 isolates. The phylum Bacillota, class Bacillota, order Bacillales, and family Bacillaceae were identified, with Bacillus as the dominant genus (46.15%). The richness and Shannon's diversity were higher at the species than at the genus level due to the dominance of Bacillus; however, various Bacillus species (7) were identified. Therefore, the climax mixed vegetation adapted to the Goraebul coast may exert natural selection pressure in favor of the common characteristics of Bacillus. However, despite this advantage, the Shannon equitability (0.86), Simpson (0.08), and Shannon diversity (2.79) indexes indicate a stable rhizosphere cluster and the climax mixed vegetation is affected by symbiotic relationships between healthy rhizosphere microbiota.

• Journal of Life Science
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• v.23 no.12
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• pp.1486-1494
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• 2013

A high protease-producing strain was isolated and identified from the digestive tract of octopus vulgaris by detecting a hydrolysis circle of protease and its activity. The strain was identified by morphology observation, biochemical experiments, and 16S rRNA sequence analysis. The protease obtained from the strain was purified by a three-step process involving ammonium sulfate precipitation, carboxy methyl-cellulose (CM-52) cation-exchange chromatography, and DEAE-Sephadex A50 anion-exchange chromatography. The properties of protease were characterized as well. The strain Bacillus sp. QDV-3, which produced the highest activity of protease, was isolated. On the basis of the phenotypic and biochemical characterization and 16S rRNA gene-sequencing studies, the isolate was identified as follows: domain: Bacteria; phylum: Firmicutes; class: Bacilli; order: Bacillales; family: Bacillaceae; and genus: Bacillus. The isolate was shown to have a 99.2% similarity with Bacillus flexus. A high active protease designated as QDV-E, with a molecular weight of 61.6 kDa, was obtained. The enzyme was found to be active in the pH range of 9.0-9.5 and its optimum temperature was $40^{\circ}C$. The protease activity retained more than 96% at the temperature of $50^{\circ}C$ for 60 min. Phenylmethylsulfonyl fluoride (PMSF) inhibited the enzyme activity, thus confirming that this protease isolated from Bacillus sp. QDV-3 is an alkaline serine protease. Metal ions, $Mn^{2+}$ and $Mg^{2+}$, were determined to enhance the protease activity, whereas $Ba^{2+}$, $Zn^{2+}$, and $Cu^{2+}$ were found to inactivate the enzyme.

• Journal of Life Science
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• v.34 no.4
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• pp.254-263
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• 2024

Korean jang is a food made using fermented soybeans, and the typical products include gochujang (GO), doenjang (DO), cheonggukjang (CH), and ganjang (GA). In this study, 16S rRNA metagenome analysis was performed on a total of 200 types of GO, DO, CH, and GA using next-generation sequencing to analyze the microbial community of fermented soybean foods and compare taxonomic (biomarker) differences. Alpha diversity analysis showed that in the CHAO index, the species richness index tended to be significantly higher compared to the DO and GA groups (p<0.001). The results of the microbial distribution analysis of the GO, DO, CH, and GA products showed that at the order level, Bacillales was the most abundant in the GO, DO, and CH groups, but Lactobacillales was most abundant in the GA group. Linear discriminant analysis effect (LEfSe) analysis was used to identify biomarkers at the family and species levels. Leuconostocaceae, Thermoactinomycetaceae, Bacillaceae, and Enterococcaceae appeared as biomarkers at the family level, and Bacillus subtilis, Kroppenstedtia sanguinis, Bacillus licheniformis, and Tetragenococcus halophilus appeared at the species level. Permutational multivariate analysis of variance (PERMANOVA) analysis showed that there was a significant difference in the microbial community structure of the GO, DO, CH, and GA groups (p=0.001), and the microbial community structure of the GA group showed the greatest difference. This study clarified the correlation between the characteristics of Korean fermented foods and microbial community distribution, enhancing knowledge of microorganisms participating in the fermentation process. These results could be leveraged to improve the quality of fermented soybean foods.