• 한국생물공학회:학술대회논문집
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• 2005.10a
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• pp.343-346
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• 2005

The isolated strain, SC2-1 was Gram-positive, catalase positive, facultatively anaerobic, oxidase negative, motile and small rods. The strain utilized sucrose, dextrose, fructose, mannitol and maltose as a sole carbon and energy source and sodium chloride required for the bacteria growth. The radical scavenging activity of the culture supernatants was determined by DPPH (1,1-diphenyl-2-picrylhydrazyl) method. This bacterium was identified based on cellular fatty acids analysis and 16S rDNA sequencing then named Exiguobacterium sp. SC2-1. The optimum culture conditions for production of antioxidant were $25^{\circ}C,$ pH 7.8 and NaCl concentration were 4%. The modified optimal medium compositions were maltose 2.5% (w/v), yeast extract 1.5% (w/v) and $KH_2PO_4$ 0.05% (w/v). Free radical scavenging activity of under optimal culture conditions were 93%.

• Journal of Marine Bioscience and Biotechnology
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• v.9 no.2
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• pp.49-57
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• 2017

We isolated marine bacterium, isolate DG-2 which produces the antibiotics against MRSA (methicillin-resistant Staphylococcus aureus). This isolate DG-2 was examined by its morphological, biochemical properties, and 16S rRNA sequencing analysis. And then, isolate DG-2 was identified to the genus Streptomyces. Therefore, this isolate was designated as Streptomyces sp. DG-2. Streptomyces sp. DG-2 grew relatively well at $25^{\circ}C$, pH 7.0, and NaCl 1.0%. For the pre-purification of the bioactive compounds, DG-2 was fermented in 30 L PPES-II medium, and the culture filtrates of DG-2 was extracted by ethyl acetate. The ethyl acetate extract of DG-2 showed the significant anti-MRSA and antibacterial activities.

• Journal of Microbiology and Biotechnology
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• v.14 no.1
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• pp.220-223
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• 2004

Club root caused by Plasmodiophora brassicae is found in crucifers. Among the over hundreds of Streptomyces isolated from soil in Korea. One strain showed prominent activity against P. brassicae. The strain was identified based on 16S rDNA sequencing and the morphology by a method of scanning electron microscopy. An active compound in the fermented broth obtained from the strain was separated. Even though the complete assignments of the compound remain for future work, the results regarding the isolation and characterization of the strain with a certain activity against P. brassicae are shown in this paper.

• Tuberculosis and Respiratory Diseases
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• v.50 no.1
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• pp.94-105
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• 2001

Background : Examining the biological susceptibility of Mycobacterium tuberculosis to pyrazinamide (PZA) in vitro is very difficult as PZA is inactive under normal culture conditions. The biological susceptibility test, an enzyme assay for Pzase activity, and a genetic test for pncA gene mutations, were performed in order to predict PZA resistance. Methods : 28 cultured clinical isolates of Mycobacterium tuberculosis were tested. The biological susceptibility was performed by the absolute concentration method using Lowenstein-Jensen media. The PZase activity was tested by means of Wayne's method. A 710-bp region includes the entire open reading frame of pncA was amplified and sequenced. Results : All six strains with positive PZase activity exhibited no pncA mutations with one strain showing a false resistance in the biological susceptibility test. Among the 22 strains with no PZase activity, 21 exhibited showed pncA mutations. In the biological susceptibility test, 20 strains were resistant, and one was susceptible, and the other flied to test. The mutation types varied with ten missense, one silent and one nonsense mutation 1 slipped-strand mispairing, and 6 frameshift mutations. Three strains had an adenine to guanine mutation at position -11 upstream of the start codon. Conclusion : The mutation at the pncA promotor region is frequent at -11 upstream position. Automatic sequencing of pncA is a useful tool for rapid and accurate detection of PZA resistant M. tuberculosis, and for demonstrating the epidemiological relatedness of the PZA resistant M. tuberculosis strains.

• Korean Journal of Clinical Laboratory Science
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• v.38 no.3
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• pp.196-202
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• 2006

Treatment of hepatitis B virus (HBV) with lamivudine is effective in suppressing virus replication and results in reduced inflammatory activity. However the most troublesome problem of lamivudine treatment is the emergence of lamivudine-resistant strains with amino acid substitution in the YMDD motif of DNA polymerase gene during the treatment. The aim of this study was to determine the mutation of YMDD motif (codon 552) and codon 528 in chronic HBV patients with lamivudine therapy using PCR-direct sequencing and to investigate the relationship between lamivudine mediated HBV mutation and HBeAg. HBV DNA was extracted from serum samples of HBV patients and amplified by nested PCR with two sets of primer pairs selected in HBV DNA polymerase gene. Amplified PCR product was analyzed by 2% agarose gel electrophoresis and direct sequencing. HBV mutation was detected in 124 out of 207 samples (60%). Single mutation was 50.8% for M552I, 43.5% for M552V, 5.7% for M552I/V and the L528M mutation was 67.0%. Double mutation was 43.6% for M552V/L528M, 33.1% for M552I/L528(wild type), 17.7% for M552I/L528M and 5.6% for M552I/V/L528M. Serine mutation at YMDD motif (M552S) was not found and the L528M mutation frequently accompanied M552V type. In this study, the typical difference of frequencies for HBV mutation depending on HBeAg was not found. Moreover, the PCR-direct sequencing method used in this study might be a powerful tool for the mutation study in clinical reference laboratories with high volume.

• Microbiology and Biotechnology Letters
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• v.42 no.3
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• pp.293-296
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• 2014

This study was conducted to investigate the effect of culture temperature on the growth rate and protease activity of Antarctic microorganisms. The Antarctic microorganisms PAMC 25641, 25614, 25719 and 25617 were obtained from the Polar and Alpine Microbial Collection (PAMC) at the Korea Polar Research Institute. These microorganisms were confirmed for the excretion of protease on a plate with skim milk. The identification of microorganisms was carried out using the 16S rDNA sequencing method. PAMC 25641 showed the highest protease activity among the subjects tested, and PAMC 25617 exhibited the highest growth rate. The growth rates of the microorganisms were not affected by temperature, except for PAMC 25617. However, protease activities were increased for all strains in a temperature dependent fashion. These results suggest the possible application of Antarctic microorganisms for the efficient production of low temperature proteases.

• Journal of Microbiology and Biotechnology
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• v.34 no.7
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• pp.1452-1463
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• 2024

Fungi generate different metabolites some of which are intrinsically bioactive and could therefore serve as templates for drug development. In the current study, six endophytic fungi namely Aspergillus flavus, Aspergillus tubigenesis, Aspergillus oryzae, Penicillium oxalicum, Aspergillus niger, and Aspergillus brasiliensis were isolated and identified from the medicinal plant, Silybum marianum. These endophytic fungi were identified through intra transcribed sequence (ITS) gene sequencing. The bioactive potentials of fungal extracts were investigated using several bioassays such as antibacterial activity by well-diffusion, MIC, MBC, anti-biofilm, antioxidant, and haemolysis. The Pseudomonas aeruginosa PAO1 was used to determine the antibiofilm activity. The ethyl acetate extract of Aspergillus flavus showed strong to moderate efficacy against Staphylococcus aureus, Escherichia coli, P. aeruginosa, and Bacillus spizizenii. Aspergillus flavus and Aspergillus brasiliensis exhibited significant antibiofilm activity with IC₅₀ at 4.02 and 3.63 mg/ml, while A. flavus exhibited maximum antioxidant activity of 50.8%. Based on HPLC, LC-MS, and NMR experiments kojic acid (1) and carbamic acid (methylene-4, 1-phenylene) bis-dimethyl ester (2) were identified from A. flavus. Kojic acid exhibited DPPH free radical scavenging activity with an IC₅₀ value of 99.3 ㎍/ml and moderate activity against ovarian teratocarcinoma (CH1), colon carcinoma (SW480), and non-small cell lung cancer (A549) cell lines. These findings suggest that endophytic fungi are able to produce promising bioactive compounds which deserve further investigation.

• Environmental Sciences Bulletin of The Korean Environmental Sciences Society
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• v.1 no.2
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• pp.125-136
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• 1997

Digestion of a municipal wastewater sludge by the anaerobic sequencing batch reactor(ASBR) was investigated to evaluate the performance of the ASBR process at a critical condition of high-solids-content feed. The reactors were operated at an HRT of 10 days with an equivalent loading rate of 0.8-1.5 gVS/L/d at $35^{\circ}C.$ The main conclusions drawn from this study were as follows: 1. Digestion of a municipal wastewater sludge was possible using the ASBR in spite of high concentration of settleable solids in the sludge. The ASBRS with 3- and 4-day cycle period showed almost identical high digestion performances. 2. No adverse effect on digestion stability was observed in the ASBRS in spite of withdrawal and replenishment of $30\%\;or\;40\%$ of liquid contents. A conventional anaerobic digester could be easily converted to the ASBR without any stability problem. 3. Flotation thickening occurred in thicken step of the ASBRS throughout steady state, and floating bed volume at the end of thicken period occupied about $70\%$ of the working volume of the reactor. Efficiency of flotation thickening in the ASBRS could be comparable to that of additional gravity thickening of a completely mixed digester. 4. Solids were accumulated rapidly in the ASBR during start-up period. Solids concentrations in the ASBRS were 2.6 times higher than that in the completely mixed control reactor at steady state. Dehydrogenase activity had a strong correlation with the solids concentration. Dehydrogenase activity of the digested sludge in the ASBR was 2.9 times higher than that of the sludge in the control reactor, and about 25 times higher than that of the subnatant in the ASBR. 5. Remarkable increase in equivalent gas production of $52\%$ was observed at the ASBRS compared with the control reactor in spite of similar Quality of clarified effluent from the ASBRS and control reactor. The increase in gas production from the ASBRS was believed to be combined results of accumulation of microorganisms, higher driving force applied, and additional long-term degradation of organics continuously accumulated.

• Journal of Applied Biological Chemistry
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• v.66
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• pp.394-403
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• 2023

In the pursuit of sustainable and environmentally-friendly agricultural practices, we conducted an extensive study on the rhizosphere bacteria inhabiting soils that have been devoid of chemical fertilizers for an extended period exceeding 40 years. Through this investigation, we isolated a total of 80 species of plant growth-promoting rhizosphere bacteria and assessed their potential to enhance plant growth. Among these isolates, Burkholderia cepacia CD2 displayed remarkable plant growth-promoting activity, making it an optimal candidate for further analysis. Burkholderia cepacia CD2 exhibited a range of beneficial characteristics conducive to plant growth, including phosphate solubilization, siderophore production, denitrification, nitrate utilization, and urease activity. These attributes are well-known to positively influence the growth and development of plants. To validate the taxonomic classification of the strain, 16S rRNA gene sequencing confirmed its placement within the Burkholderia genus, providing further insights into its phylogenetic relationship. To delve deeper into the potential mechanisms underlying its plant growth-promoting properties, we sought to confirm the presence of specific genes associated with plant growth promotion in CD2. To achieve this, whole genome sequencing (WGS) was performed by Plasmidsaurus Inc. (USA) utilizing Oxford Nanopore technology (Abingdon, UK). The WGS analysis of the genome of CD2 revealed the existence of a subsystem function, which is thought to be a pivotal factor contributing to improved plant growth. Based on these findings, it can be concluded that Burkholderia cepacia CD2 has the potential to serve as a microbial fertilizer, offering a sustainable alternative to chemical fertilizers.

• Journal of Life Science
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• v.12 no.4
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• pp.496-503
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• 2002

To develop a yeast strain of Pichia pastoris producing an antibacterial peptide, we have attempted the expression and secretion of an insect defensin. The nucleotide sequences corresponding to mature defensin were chemically synthesized by 6 oligomers, assembled in vitro and the synthesized gene was identified by nucleotide sequencing. The prepro sequence of yeast mating factor $\alpha$1 and the defensin gene were recombined into a Pichia expression vector, pPIC9K. The resulting plasmid, pPIDE, was transformed into P. pastoris GSl15 and transformants selected on histidine-deficient minimal plates were tested for antibacterial activity against Micrococcus luteus. Four strains with different antibacterial activity were selected for further analysis. Southern hybridization and RT-PCR verified the defensin gene was maintained and transcribed in a host. Four strains were cultivated in YPD broth for 96 hours to compare cell growth and antibacterial activity, They showed no difference in cell growth, however, each strain showed different antibacterial activity pattern with culture time. The maximal activity was about 550 AU/ $m\ell$.