• Korean Journal of Environmental Biology
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• v.34 no.4
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• pp.304-313
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• 2016

One of the issues currently facing nuclear power plants is how to store spent nuclear waste materials which are contaminated with radionuclides such as $^{134}Cs$, $^{135}Cs$, and $^{137}Cs$. Bioremediation processes may offer a potent method of cleaning up radioactive cesium. However, there have only been limited reports on $Cs^+$ tolerant bacteria. In this study, we report the isolation and identification of $Cs^+$ tolerant bacteria in environmental soil and sediment. The resistant $Cs^+$ isolates were screened from enrichment cultures in R2A medium supplemented with 100 mM CsCl for 72 h, followed by microbial community analysis based on sequencing analysis from 16S rRNA gene clone libraries(NCBI's BlastN). The dominant Bacillus anthracis Roh-1 and B. cereus Roh-2 were successfully isolated from the cesium enrichment culture. Importantly, B. cereus Roh-2 is resistant to 30% more $Cs^+$ than is B. anthracis Roh-1 when treated with 50 mM CsCl. Growth experiments clearly demonstrated that the isolate had a higher tolerance to $Cs^+$. In addition, we investigated the adsorption of $0.2mg\;L^{-1}$ $Cs^+$ using B. anthracis Roh-1. The maximum $Cs^+$ biosorption capacity of B. anthracis Roh-1 was $2.01mg\;g^{-1}$ at pH 10. Thus, we show that $Cs^+$ tolerant bacterial isolates could be used for bioremediation of contaminated environments.

• Korean Journal of Organic Agriculture
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• v.25 no.4
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• pp.843-859
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• 2017

The JK-1 isolate which was the best producer of indole-3-acetic acid and carotenoid among the 388 strains isolated from 28 wetlands in Jeju, was identified to be Rhodopseudomonas palustirs belongs to a typical group of non sulfur purple bacteria based on 16S sRNA sequencing. This study investigated the effect of different cultural conditions of pH, temperature, agitation, light and aeration on growth, IAA and carotenoid production of photosynthetic bacterium JK-1 for optimization of IAA and carotenoid production. It was found that growth, IAA, carotenoid, and bacteriochlorophyll production with light (3,000~3,500 Lux) and agitation (100 rpm) showed better results than those with dark/static or dark/agitation (100 rpm) in anaerobic conditions. The optimal pH, temperature and agitation speed for cell growth were 7, $30^{\circ}C$, 150 rpm, for IAA production were 9, $30^{\circ}C$, 150rpm and for carotenoid production were 6, $25^{\circ}C$, 50 rpm, cultured for 72 h under anaerobic light, respectively. The growth and IAA production were high in aerobic culture compared with anaerocic culture, whereas carotenoid and bacteriochlorophyll content were decreased extremely in aerobic condition (0.5~1 vvm). Subsequently, the optimal culture conditions for JK-1 were selected with pH 7, $30^{\circ}C$ and 100 rpm under anaerobic light and the effect on plant growth was tested by pot assay. Inoculation of JK-1 with 3% (v/v) level caused increase in shoot and root dry weigh that varied from 20%~58% to 40%~28% in young radish in camparison to uninoculated treatment at 50 days of growth. The study suggests that the JK-1 isolate may serve as efficient biofertilizer inoculants to promote plant growth.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 1995.06b
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• pp.77-96
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• 1995

To obtain basic informations for the improvement of seed potato production in Korea, some etiological properties of potato virus Y(PVY) distributed in the major seed potato production area(Daekwanryeong) were characterized, and the nucleotide and amino acid sequences of the coat protein gene of the PVY strains isolated were analyzed. PVY strains in Daekwonryeong, an alpine area, were identified to be two strains, PVYo and PVYN by symptoms of indicator plants, and their distribution in potato fields was similar. Major symptom on potato varieties by PVY was grouped as either mosaic alone or mosaic accompanied with veinal necrosis in the lower leaves. The symptom occurrence of the two symptoms was similar with Irish Cobbler, but Superior showed a higher rate of mosaic symptom than the other. The PVY strain which was isolated from potato cv. Superior showing typical mosaic symptoms produced symptoms of PVY-O on the indicator plants of Chenopodium amaranticolor, Nicotiana tabacum cv. Xanthi nc and Physalis floridana, but no symptom o Capsicum annum cv. Ace. Moreover, results from the enzyme-linked immunosorbent assay with monoclonal and polyclonal antibodies showed that the isolated PVY reacts strongly with PYV-O antibodies but does not react specifically with PVY-T antibodies. The purified virus particles were flexious with a size of 730$\times$11nm. On the basis of the above characteristics, the strain was identified to be a PVY-O and named as of PVY-K strain. The flight of vector aphids was observed in late May, however, the first occurrence of infected plants was in mid June with the bait plants surrounded with PVY-infected potato plants and early July with the bait plants surrounded with PVY-free potato plants. PVY infection rates by counting symptoms on bait plants (White Burley) were 1.1% with the field surrounded with PVY-free potato plants and 13.7% the fields surrounded with PVY-infected potato plants, showing the effect of infection pressure. The propagated PVY-K strain on tobacco(N. sylvestris) was purified, and the RNA of the virus was extracted by the method of phenol extraction. The size of PVY-K RNA was measured to be 9, 500 nucleotides on agarose gel electrophoresis. The double-stranded cDNAs of PVY-K coat protein(CP) gene derived by the method of polymerase chain reaction were transformed into the competent cells of E. coli JM 109, and 2 clones(pYK6 and pYK17) among 11 clones were confirmed to contain the full-length cDNA. Purified plasmids from pYK17 were cut with Sph I and Xba I were deleted with exonuclease III and were used for sequencing analysis. The PVY-K CP gene was comprised of 801 nucleotides when counted from the clevage site of CAG(Gln)-GCA(Ala) to the stop codon of TGA and encoded 267 amino acids. The molecular weight of the encoded polypeptides was calculated to be 34, 630 daltons. The base composition of the CP gene was 33.3% of adenine, 25.2% of guanine, 20.1% of cytosine and 21.4% of uracil. The polypeptide encoded by PVY-K CP gene was comprised of 22 alanines, 20 threonines, 19 glutamic acids and 18 glycines in order. The homology of nucleotide sequence of PVY-K CP gene with those of PVY-O(Japan), PVY-T(Japan), PVY-TH(Japan), PVYN(the Netherlands), and PVYN(France) was represented as 97.3%, 88.9%, 89.3%, 89.6% and 98.5%, respectively. The amino acid sequence homology of the polypeptide encoded by PVY-K CP gene with those encoded by viruses was represented as 97.4%, 92.5%, 92.9%, 92.9%, and 98.5%, respectively.

• Journal of Life Science
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• v.30 no.2
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• pp.156-161
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• 2020

Since industrialization, the production and utilization of various chemicals has contributed to improving the quality of our lives, but the subsequent discharge of massive waste is inevitable, and environmental pollution is becoming more serious every day. Exposure to chemicals as a result of environmental pollution is having a negative effect on human health and the ecosystem, and cleaning up the polluted environment that can affect our lives is a very important issue. Toxic aromatic compounds have been detected frequently in soil, groundwater, and wastewater because of the extensive use of oil products, and phenol, which is used to produce synthetic resins, textiles, and dyes, is one of the major pollutants, along with insecticides and preservatives. Phenol can cause dyspnea, headache, vomiting, mutation, and carcinogenesis. Phenol-degrading bacterium DWB-1-8 was isolated from the activated sludge of textile wastewater; this strain was identified as Comamonas testosteroni by 16S rRNA gene sequencing. The optimal culture conditions for the cell growth and degradation of phenol were 0.7% K₂HPO₄, 0.6% NaH₂PO₄, 0.1% NH₄NO₃, 0.015% MgSO₄·7H₂O, 0.001% FeSO₄·7H₂O, an initial pH of 7, and a temperature of 30℃. The strain was also able to grow by using other toxic compounds, such as benzene, toluene, or xylene (BTX), as the sole source of carbon.

• Food Science and Preservation
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• v.24 no.8
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• pp.1168-1179
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• 2017

In this study, we tried to screen the Bacillus strain having safety probability by isolation of strains from traditional fermented food, measurement of probiotic properties, and the fermentative characteristics of Cheonggukjang. We isolated 400 Bacillus-like isolates from traditional fermented foods. Selected strains examined on the prevalent characteristic such as extracellular enzyme and antibacterial activities, and their safety probability was confirmed by biogenic amine productivity, hemolytic, and harmful substances and enzyme productivity. We selected the 5 strains by analysis of biogenic amine, antibacterial and B. cereus toxic associated gene. Five selected strains were examined on cell surface hydrophobicity, and bile and acid tolerance, and we selected the SRCM100730 as the final strain. SRCM100730 was confirmed B. amyloliquefaciens by 16S rRNA sequencing, and named the B. amyloloquefaciens SRCM100730 (KCCM11966P). Finally, we manufactured Cheonggukjang using SRCM100730 for confirmation of fermentation properties. Manufactured Cheonggukjang did not contain B. cereus, and showed that ${\gamma}$-PGA and extracellular enzyme activities were superior to commercial Chunggukjang. Amino nitrogen content was 544.02 mg% and 26 free amino acid were detected, and the bitterness-related amino acid content was lower than commercial Cheonggukjang. Especially, the amount of GABA was 3 fold higher than commercial Cheonggukjang. These results suggest that SRCM100730 have high availability in commercial probiotics market and fermented food industry.

• Korean Journal of Microbiology
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• v.42 no.3
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• pp.185-194
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• 2006

The aim of this study was to identify the bacteria isolated from endodontic lesions by cell culture and to determine the antimicrobial susceptibility of them against 8 antibiotics. The necrotic pulpal tissues were collected from 27 infected root canals, which were diagnosed as endodontic infection. Samples were collected aseptically from the infected pulpal tissue of the infected root canals using a barbed broach and a paper point. The cut barbed broaches and paper points were transferred to an eppendorf tube containing $500{\mu}l\;of\;1{\times}PBS$. The sample solution was briefly mixed and plated onto a BHI-agar plate containing 5% sheep blood. The agar plates were incubated in a $37^{\circ}C$ anaerobic chamber for 2 to 5 days. The bacteria grown on the agar plates were identified by comparison of 16S rRNA gene (rDNA) sequencing method at the species level. To test the sensitivity of the bacteria isolated from the infected root canals against 8 antibiotics, minimum inhibitory concentrations (MIC) were determined using broth dilution assay. The data showed that 101 bacterial strains were isolated and were identified. Streptococcus spp. (29.7%) and Actinomyces spp. (21.8%) were predominantly isolated. The 9 strains were excluded in antimicrobial susceptibility test because they were lost during the experiment or were not grown in broth culture. The percentage of bacteria susceptible for each antibiotic in this study was clindamycin, 87.0% (80 of 92); tetracycline, 75.0% (69 of 92); cefuroxime axetil, 75.0% (69 of 92); amoxicillin + clavulanic acid (5:1), 71.7% (66 of 92); penicillin G, 66.3% (61 of 92); erythromycin, 66.3% (61 of 92); amoxicillin, 44.6% (41 of 92); and ciprofloxacin, 31.5% (29 of 92). The susceptibility pattern of 8 antibiotics was dependent on the host of the bacteria strains rather than the kinds of bacterial species. These results indicate that antibiotic susceptibility test should be performed when antibiotics are needed for the treatment of infected root canals.

• Research in Plant Disease
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• v.25 no.1
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• pp.8-15
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• 2019

Plum pox virus (PPV) is a significant viral disease in Prunus spp. worldwide. A nationwide survey was started in Prunus spp. orchards, since PPV was first detected from peach in Korea, 2015. During 2016-2017, samples were collected from 30,333 trees in 1,985 orchards of stone fruits in 8 provinces and 4 cities, Korea and tested by RT-PCR using specific PPV primer set. As a result, 21 trees including peach (9 trees), Japanese apricot (4 trees), plum (1 tree), apricot (7 trees) in 10 orchards were infected and controlled by eradication program. Amplicons of the expected size (547 bp) were obtained from total RNA of seven peach trees in 2016, and directly sequenced. BLAST analysis revealed the highest nucleotide (NT) identity (99%) with a PPV D isolates (LC331298, LT600782) in Genbank. The seven isolates from shared nt sequence identities of 98 to 100% with one another. Phylogenetic analysis showed the isolates in peach clustered closely with the PPV-D isolates from Korea, Japan, USA, and Canada. This is, to our knowledge, the first report of the presence of PPV in Prunus spp. orchards in Korea, 2016-2017, we hope that our results and efforts will contribute to effective measures for eradication of PPV.

• 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.29 no.11
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• pp.1173-1178
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• 2019

The purpose of this study was to isolate an agar-degrading marine bacterium and characterize its agarase. Bacterium BK-1, from Gwanganri Beach at Busan, Korea, was isolated on Marine 2216 agar medium and identified as Agarivorans sp. BK-1 by 16S rRNA gene sequencing. The extracellular agarase, characterized after dialysis of culture broth, showed maximum activity at pH 6.0 and $50^{\circ}C$ in 20 mM Tris-HCl buffer. Relative activities at 20, 30, 40, 50, 60, and $70^{\circ}C$ were 67, 93, 97, 100, 58, and 52%, respectively. Relative activities at pH 5, 6, 7, and 8 were 59, 100, 95, and 91%, respectively. More than 90% of the activity remained after a 2 hr exposure to 20, 30, or $40^{\circ}C$; about 60% of the activity remained after a 2 hr exposure to $50^{\circ}C$. Almost all activity was lost after exposure to 60 or $70^{\circ}C$ for 30 min. Zymography revealed three agarases with molecular weights of 110, 90, and 55 kDa. Agarose was degraded to neoagarobiose (46.8%), neoagarotetraose (39.7%), and neoagarohexaose (13.5%), confirming the agarase of Agarivorans sp. BK-1 as a ${\beta}$-agarase. The neoagarooligosaccharides generated by this agarase could be used for moisturizing, bacterial growth inhibition, skin whitening, food treatments, cosmetics, and delaying starch degradation.

• Journal of Life Science
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• v.33 no.4
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• pp.357-362
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• 2023

This report looks at an agar-degrading marine bacterium and characterization of its agarase. Agar-degrading marine bacterium JS-1 was isolated with Marine agar 2216 media from seawater from the seashore of Sojuk-do, Changwon in Gyeongnam Province, Korea. The agar-degrading bacterium was named as Agarivorans sp. JS-1 by phylogenetic analysis based on 16S rRNA gene sequencing. The extracellular agarase was prepared from the culture media of Agarivorans sp. JS-1 and used for characterization. Relative activities at 20℃, 30℃, 35℃, 40℃, 45℃, 50℃, 55℃, and 60℃ were 70%, 74%, 78%, 83%, 87%, 100%, 74%, and 66%, respectively. Relative activities at pH 5, 6, 7, and 8 were 91%, 100%, 90%, and 89%, respectively. Its extracellular agarase showed maximum activity (207 units/l) at pH 6.0 and 50℃ in 20 mM Tris-HCl buffer. The residual activity after heat treatment at 20℃, 30℃, and 50℃ for 30 minutes was 90%, 70%, and 50% or more, respectively. After a 2-hour heat treatment at 20℃, 30℃, 35℃, 40℃, and 45℃, the residual activity was 80%, 68%, 65%, 63%, and 57%, respectively. At 50℃ and above, after heat treatment for 30 minutes, the residual activity was below 60%. Thin layer chromatography analysis suggested that Agarivorans sp. JS-1 produces extracellular β-agarases as they hydrolyze agarose to produce neoagarooligosaccharides such as neoagarohexaose (20.6%), neoagarotetraose (58.5%), and neoagarobiose (20.9%). Agarivorans sp. JS-1 and its thermotolerant β-agarase would be useful in the production of neoagarooligosaccharides, showing functional activity such as inhibition of bacterial growth and delay of starch degradation.