• Research in Plant Disease
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• v.21 no.4
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• pp.280-289
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• 2015

Maize (Zea mays L.) is an economically important crop in worldwide. While the consumption of the maize is steadily increasing, the yield is decreasing due to continuous mono-cultivation and infection of soil-borne fungal pathogens such as Fusarium species. Recently, stalk rot disease in maize, caused by F. subglutinans and F. temperatum has been reported in Korea. In this study, we isolated bacterial isolates in rhizosphere soil of maize and subsequently tested for antagonistic activities against F. subglutinans and F. temperatum. A total of 1,357 bacterial strains were isolated from rhizosphere. Among them three bacterial isolates (GC02, GC07, GC08) were selected, based on antagonistic effects against Fusarium species. The isolates GC02 and GC07 were most efficient in inhibiting the mycelium growth of the pathogens. The three isolates GC02, GC07 and GC08 were identified as Bacillus methylotrophicus, B. amyloliquefaciens and B. thuringiensis using 16S rRNA sequence analysis, respectively. GC02 and GC07 bacterial suspensions were able to suppress over 80% conidial germination of the pathogens. GC02, GC07 and GC08 were capable of producing large quantities of protease enzymes, whereas the isolates GC07 and GC08 produced cellulase enzymes. The isolates GC02 and GC07 were more efficient in phosphate solubilization and siderophore production than GC08. Analysis of disease suppression revealed that GC07 was most effective in suppressing the disease development of stalk rot. It was also found that B. methylotrophicus GC02 and B. amyloliquefaciens GC07 have an ability to inhibit the growth of other plant pathogenic fungi. This study indicated B. methylotrophicus GC02 and B. amyloliquefaciens GC07 has potential for being used for the development of a biological control agent.

• Korean Journal of Microbiology
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• v.45 no.2
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• pp.119-125
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• 2009

To understand the ecological function of heterotrophic bacterial community in water column of large freshwater lakes in the permafrost zone, we investigated the structure and function of bacterial community in Lake Khuvsgul, Mongolia. Species composition of overall bacterial community was analyzed by denaturing gradient gel electrophoresis (DGGE) of 16S rRNA gene fragments, and bacteria that can be cultured at 10oC were isolated and characterized. Based on the depth profile of environmental parameters, thermocline and chemocline were recognized at the 5~10 m zone of the water column. The stratified DGGE profile indicated that the discontinuity of water properties might influence the structure of bacterial community: band profiles in the 0~5 m zone were diverse with large change by depth, but the profile was relatively stable at the $\geq$10 m zone, with predominance of the band identified as Acidovorax facilis. Bacterial cultures were screened for protease, cellulase, amylase and lipase activity, and 23 isolates were selected for high activity of the hydrolytic enzymes. The isolates were identified based on their 16S rRNA gene sequences. In the surface water (zero meter depth), Acidovorax defluvii and Sphingobacterium faecium with high cellulase activity were present. Flavobacterium succinicans, Mycoplana bullata and A. facilis were stably predominant isolates at 2 m, 5 m, and $\geq$10 m depths, respectively. F. succinicans isolates showed high protease activity while M. bullata isolates showed moderate levels of protease and celluase activity. A. facilis isolates showed either cellulase or lipase activity, exclusively to each other. According to the profile of growth rates of the isolates in the temperature range of $0\sim42^{\circ}C$, the surface-zone (0~5 m) isolates were facultative psychrophiles while isolates from $\geq$10 m depth were typical mesophiles. This stratification is believed to be due to stratified availability of organic materials to the bacterial decomposers. In the water column below the chemoline, the environment is extremely oligotrophic so that the trait of rapid growth in low temperature might not be demanded by deep-lake decomposers. The stratified distribution of community composition and decomposer activity in Lake Khuvsgul implies that ecological functions of bacterial community in lakes of cold region are sharply divided by water column stratification.

• Journal of Life Science
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• v.27 no.2
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• pp.202-210
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• 2017

As an effort to find a potential biopreservative, we isolated bacterial strains producing bacteriocin from fermented foods. A strain was finally selected and characteristics of the bacteriocin were investigated. The selected strain was identified as Bacillus subtilis E9-1 based on the 16S rRNA gene analysis. The culture supernatant of B. subtilis E9-1 showed antimicrobial activity against Gram-positive bacteria. Subtilisin A, ${\alpha}$-chymotrypsin, trypsin and proteinase K inactivated the antimicrobial activity, which means its proteinaceous nature, a bacteriocin. The bacteriocin activity was fully retained at the pH range from 2.0 to 8.0 and stable at up to $100^{\circ}C$ for 60 min. Solvents such as ethanol, isopropanol and methanol had no effect on the antimicrobial activity at the concentration of 100% but acetone and acetonitrile reduced the activity at up to 100% concentration. Cell growth of four indicator strains was dramatically decreased in dose-dependent manner. Listeria monocytogenes was the most sensitive, but Enterococcus faecium was the most resistant. Bacillus cereus and Staphylococcus aureus showed the medium sensitivity. The bacteriocin showed its antimicrobial activity against B. cereus and L. monocytogenes via bactericidal action. The number of viable cells of L. monocytogenes started to reduce after addition of bacteriocin to the minced beef. The bacteriocin was purified through acetone concentration, gel filtration chromatography and RP-HPLC. The whole purification step led to a 6.82 fold increase in the specific activity and 6% yield of bacteriocin activity. The molecular weight of the purified bacteriocin was determined to be 3.3 kDa by MALDI-TOF/TOF mass spectrometry.

• Korean Journal of Microbiology
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• v.54 no.4
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• pp.384-397
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• 2018

The aim of this study is to screen the strains of Bacillus spp. possessing safety, probiotic activity, and so on, which can be utilized as probiotic resource for using the feed and supplement food of companion animal. About 300 isolates were isolated from traditional Korean sauces, four isolates that did not have or produce the six kinds of B. cereus type vomiting and diarrhea toxin genes, ${\beta}$-hemolytic, and three kinds of carcinogenic enzymes were selected. Antibiotic gene retention, cell surface hydrophobicity, antibiotic sensitivity, and glucose utilization were analyzed for four isolates, and finally SRCM 100731 was selected. SRCM 100731 was named as Bacillus amyloliquefaciens SRCM 100731 16S rRNA sequencing analysis, and carried out optimization of cell growth for industrial applications such as pet food and feed. The effects of 14 different components on cell growth were investigated and three significant positive factors, molasses, sodium chloride, and potassium chloride were selected as the main factors based on a Plackett-Burman design. In order to find out optimal concentration on each constituent, we carried out central composite design. The predicted optimized concentrations were 7% molasses, 1.1% sodium chloride, 0.5% potassium chloride. Finally, an overall about 7-fold increase in dry cell weight yield ($12.6625{\pm}0.0658g/L$) was achieved using the optimized medium compared with the non-optimized medium ($1.8273{\pm}0.0214g/L$). This research is expected to be highly utilized in the growing pet industry by establishing optimal cultivation conditions for industrial application as well as screening Bacillus amyloliquefaciens SRCM 100731 as probiotic resource for companion animal.

• 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.

• Journal of Microbiology and Biotechnology
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• v.27 no.10
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• pp.1744-1752
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• 2017

Myeolchi jeotgals (MJs) were prepared with purified salt (PS), solar salt aged for 1 year (SS), and bamboo salt (BS) melted 3 times at 10% and 20% (w/w) concentrations, and fermented for 28 weeks at $15^{\circ}C$. BS MJ showed higher pH and lower titratable acidities than the other samples because of the alkalinity of bamboo salt. Lactic acid bacteria counts increased until 4-6 weeks and then decreased gradually, and were not detected after 20 weeks from MJs with 10% salt. Yeast counts of PS MJs were higher than those of BS and SS MJs. Bacilli were detected in relatively higher numbers throughout the 28 weeks, like marine bacteria, but archae were detected in lower numbers during the first 10 weeks. When 16S rRNA genes were amplified from total DNA from PS MJ (10% salt) at 12 weeks, Tetragenococcus halophilus was the major species. However, Staphylococcus epidermidis was the dominant species for BS MJ at the same time point. In SS MJ, T. halophilus was the dominant species and S. epidermidis was the next dominant species. BS and SS MJs showed higher amino-type nitrogen, ammonia-type nitrogen, and volatile basic nitrogen contents than PS MJs. SS and BS were better than PS for the production of high-quality MJs.

• Korean Journal of Food Science and Technology
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• v.50 no.4
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• pp.400-406
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• 2018

Takju and yakju are traditional Korean alcoholic beverages that are prepared by fermentation of glutinous rice with nuruk, a cereal starter containing various bacteria, fungi, and yeast. In this study, physicochemical and microbial properties of a total of 12 commercial takju and yakju samples were analyzed; their pH, sweetness, and alcohol content were varied, depending on the type of alcohol, from pH 3.64-4.8, $5.1-24.8^{\circ}Bx$, and 4.6-18.5%, respectively. Microbial communities were analyzed with 16S rRNA amplicon sequencing using MiSeq system. At the phylum level, Firmicutes (86.2%) was the most dominant, followed by Proteobacteria (8.08%), Actinobacteria (2.56%), and Cyanobacteria (3.13%). Lactic acid bacteria, including Lactobacillus, Lactococcus, Leuconostoc, and Weissella were also frequently detected. Among eukaryotes, Saccharomyces cerevisiae was the most dominant in these samples.

• Journal of Soil and Groundwater Environment
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• v.20 no.5
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• pp.47-51
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• 2015

Biocatalytic degradation of total petroleum hydrocarbons (TPHs) in contaminated soil by hemoglobin and hydrogen peroxide is an effective soil remediation method. This study used a laboratory soil reactor experiment to evaluate the effectiveness of a nonspecific biocatalytic reaction with hemoglobin and H₂O₂ for treating TPH-contaminated soil. We also quantified changes in the soil microbial community using real-time PCR analysis during the experimental treatment. The results show that the measured rate constant for the reaction with added hemoglobin was 0.051/day, about 3.5 times higher than the constant for the reaction with only H₂O₂ (0.014/day). After four weeks of treatment, 76% of the initial soil TPH concentration was removed with hemoglobin and hydrogen peroxide treatment. The removal of initial soil TPH concentration was 26% when only hydrogen peroxide was used. The soil microbial community, based on 16S rRNA gene copy number, was higher (7.1 × 10⁶ copy number/g of bacteria, and 7.4 × 10⁵ copy number/g of Archaea, respectively) in the hemoglobin catalyzed treatment. Our results show that TPH treatment in contaminated soil using hemoglobin catalyzed oxidation led to the enhanced removal effectiveness and was non-toxic to the native soil microbial community in the initial soil.

• The Plant Pathology Journal
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• v.24 no.2
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• pp.207-210
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• 2008

In March of 2003, tumors (galls) were observed on ginseng seedling roots in ginseng seedbeds at Yeoju, Gyeonggi province, Korea. Symptoms were spherical or galls with about 0.5-1.0cm in diameter formed on the upper through middle parts of the primary roots. Bacterial isolates obtained from the root galls were Gram-negative, rod-shaped with peritrichous flagella, aerobic, not forming yellow or orange colonies on nutrient glucose agar, yeast extract-dextrose $CaCO_3$ agar and nutrient-broth yeast extract agar, non-fluorescent on King's B agar, and non-spore forming, which were identical to characteristics of the genus Agrobacterium. They were identified as Agrobacterium tumefaciens with 0.732-0.993 similarities in 100% probability by the Biolog analyses. The 16S rRNA gene partial sequences of the six isolates tested (Genbank Accession EF486308-EF486313) were 100% homologous to those of other A. tumefaciens strains (GenBank accession AF501343, AY701900, AY701898, AY701899). The above results confirmed that this bacterium is A. tumefaciens. Pathogenicity of the bacteria was proved by the inoculation test on carrot root discs and tomato seedlings. This is the first description of A. tumefaciens causing root gall in ginseng seedling. The disease occurred locally and sparsely, but considering its appearances in seedbeds suggests that the ginseng root gall may become a threat to ginseng in Korea.

• Journal of Microbiology and Biotechnology
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• v.26 no.10
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• pp.1736-1745
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• 2016

The interactions of microbiota in the gut play an important role in promoting or maintaining the health of hosts. In this study, in order to investigate and compare the effects of dietary supplementation with Lactobacillus pentosus HC-2 (HC-2), Enterococcus faecium NRW-2, or the bacteria-free supernatant of a HC-2 culture on the bacterial composition of Litopenaeus vannamei, Illumina sequencing of the V1-V2 region of the 16S rRNA gene was used. The results showed that unique species exclusively existed in specific dietary groups, and the abundance of Actinobacteria was significantly increased in the intestinal bacterial community of shrimp fed with the bacteria-free supernatant of an HC-2 culture compared with the control. In addition, the histology of intestines of the shrimp from the four dietary groups was also described, but no obvious improvements in the intestinal histology were observed. The findings in this work will help to promote the understanding of the roles of intestinal bacteria in shrimps when fed with probiotics or probiotic supernatant.