• Korean Journal of Microbiology
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• v.30 no.6
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• pp.564-569
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• 1992

A green sulfur bacterium, Chlorobium limicola f. thiosulfatophilum NCIB 8327, was grown in modified Pfennig's medium including glu1amate as a nitrogen source. Glutamine synthetase was isolated through a series of ultracentrifugation. DEAE-Sepharose CL-6B ion exchange chromatography. Sephacryl S-300 gel permeation chromatography, and preparative HPLC. The recovery and purification fold of the enzyme were 2% and 46.3. respectively. The isolated enzyme was homogeneous on UV-Visible spectrum and polyacrylamide gel electrophoretogram. The relative molecular mass of the native enzyme was estimated to be 280,000 by gel permeation chromatography. The enzyme consisted of ten subunits with relative similar molecular mass. 30.000. which was estimated by SDS-polyacrylamide gel electrophoresis. The optimal temperature and pH of the enzyme were $30^{\circ}C$ and 7.0. Km values were 27.9 mM for L-glutamine and 0.92 mM for hydroxylamine-HCr. The enzyme activity was inhibited by alanine. glycine. and tryptophan considerably, but was not affected by asparagine, lysine. leucine. and valine.

• Korean Journal of Microbiology
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• v.43 no.4
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• pp.292-297
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• 2007

This study was performed to compare in vitro immune enhancing effects of polysaccharides extracted from cultivated mycelia of Agaricus blazei Murill. Carbohydrate contents of semi-purified polysaccharides were 85.6% and 95.3%, while ${\beta}$-glucan conents were 67.9% and 88.1% for intracellular and extracellular polysaccharide, respectively. Samples were adjusted to the same in their carbohydrate contents before efficacy tests. Both intracellular and extracellular polysaccharide increased nitric oxide (NO) synthesis of macrophage RAW 264.7 in dose dependent manner, and the maximum increase rate was 53.9 and 53.1% in intracellular and extraceltular polysaccharide, respectively. The polysaccharides also increased synthesis of cytokines such as interleukin (IL)-$1{\beta}$, IL-6 and tumor necrosis factor (TNF)-${\alpha}$ in RAW 264.7. For all the 3 cytokines, the increase rate of synthesis was much higher in extracellular polysaccharide compared to intracellular polysaccharide, especially at low concentration. Both polysaccarides increased the proliferation of splenocytes in vitro, intracellular polysaccharide showed increase in dose dependent manner while extraceltular polysaccharide showed increase untill medium concentration ($250\;{\mu}g/ml$). They did not show direct cytotoxicity against cancer cells such as B16F0 melanoma. As results, it was regarded that the both intracellular and extracellular polysaccharide from A. blazei showed immune enhancing effects in vitro, but the activity is higher in extracellular polysaccharide compared to intracellular polysaccharide.

• Korean Journal of Microbiology
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• v.43 no.4
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• pp.341-345
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• 2007

Fifty endophytic bacteria, which produced slime around the colonies, were isolated from Pueraria roots. In particular, HDN-14, TDG-3, and TNB-3 strains, which appeared to be high viscosity producers, were selected. These strains produced high levels of polysaccharides in Puerara root medium extract. The purified polysaccharide was digested with 1N HCI and analyzed by HPLC, with glucose ($45.6{\sim}63.1%$), maltose ($14.6{\sim}23.7%$), and fructose ($17.4{\sim}23.7%$) detected as constitutive sugars. When determined by the homology relationship of the 16S rDNA sequence with the relative taxa, the HDN-14 and TNB-3 strains were closely ($99.06{\sim}99.32%$) related to the Pseudomonas $koreensis^T$ and Pseudomonas $jessenii^T$, while TDG-3 were closely ($99.48{\sim}99.74%$) related to Pseudomonas $plecoglossicida^T$, Pseudomonas $mosselii^T$, and Pseudomonas $monteilii^T$. The major cellular Pseudomonas acids are $3OH-C_{10:0}$, $2OH-C_{12:0}$, $3OH-C_{12:0}$, and $3OH-C_{12:1}$, with these strains being further differentiated in species belonging to the genus Pseudomonas.

• Korean Journal of Microbiology
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• v.51 no.4
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• pp.364-373
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• 2015

As an effort to utilize alginate, 103 bacterial isolates that were positive for the alginate lyase activity were isolated from various clams and seawater samples collected in Incheon coastal area. Among them, 3 strains (M1-2-1, M6-1, and C8-15) were finally selected for further analysis based on their activities at higher levels than others. These isolates were all Gram-negative and rod shaped halophilic bacteria with motility. According to their physiological and biochemical properties as well as DNA sequence of their 16S rRNA genes, M1-2-1 and M6-1 were identified as a member of genus Pseudoalteromonas and C8-15 belonged to genus Vibrio. They exhibited the alginate degrading activity at the maximal level when they were cultured in APY broth for 6-8 h at $25^{\circ}C$. Both their growth and the enzyme activity were greatly enhanced when NaCl was added to the growth medium. The crude alginate lyases from the supernatants of the bacterial cultures showed the highest activity at $45^{\circ}C$ and pH 7.0-8.0. M1-2-1 and M6-1 produced 2.723 and 1.976 g/L of reducing sugar from alginate, respectively, suggesting that they have potential for commercial application.

• Korean Journal of Microbiology
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• v.51 no.4
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• pp.448-452
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• 2015

The bacterial strain that was isolated from chinese cabbage rhizosphere, showed inhibition of yeast growth. This strain was identified as Pseudomonas fluorescens BB2 by API 20NE test and 16S rRNA gene sequence analysis. P. fluorescens BB2 strain produced antibiotics against yeast as a secondary metabolite effectively when the culture was carried out in YM medium with 3% glucose at $20^{\circ}C$. The protein antibiotic of BB2 strain which was concentrated by ammonium sulfate precipitation and n-butanol extraction inhibited the growth of yeast with the minimal inhibitory concentration of $10{\mu}g/ml$ against Candida albicans KCTC 7965, and the growth of yeast was completely inhibited at $80{\mu}g/ml$. The hydrophilic fraction of n-butanol extraction inhibited the growth of Bacillus cereus ATCC 21366, showed orange halo on chrome azurol S plate, which means the fraction contained iron chelating siderophore. The results of crystal violet uptake through the cell membrane showed that membrane permeability was increased about 9% than control, when the concentration of hydrophobic antibiotic against yeast C. albicans was $60{\mu}g/ml$. As a result, the antibiotic produced by P. fluorescens BB2 against yeast Candida is considered antimicrobial peptide, and this is the first report in the genus Pseudomonas.

• Korean Journal of Microbiology
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• v.51 no.4
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• pp.427-434
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• 2015

Most of halophilic archaea are found in the various hypersaline environments including solar saltern, salt lake with very high salt concentration. The present study is about isolation and characterization of halphilic archaea from Gomso solar saltern known as a representative high salt environment in Korea. In order to isolate the halophilic archaea, we prepared and used high salt medium. Finally, total 7 strains obtained were tentatively identified based on comparative similarity analysis for 16S rRNA gene sequence and physiological traits. All halophilic archaea belonged to Haloruburm, Halogeometriucm, Halobacterium, and Haloarcula genera. These isolates were all Gram-staining negative, and growth was not observed using nitrate as an alternative electron acceptor under anaerobic conditions. In addition, all isolates required about 12-30% (w/v, NaCl) salt. This case study might provide basic information on microbial isolation technologies and related research in halophilic microorganisms from domestic halophilic environments, and contribute to obtaining useful indigenous halophilic archaea in a variety of extreme environmental conditions.

• Journal of Life Science
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• v.17 no.8 s.88
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• pp.1141-1146
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• 2007

This study investigated the effects on the biological activities of germinated brown rice soaked in mycelial culture broth of Phellinus linteus. The level of free amino acid was higher in the GBRP extract than those of BR and GBR. The major free amino acids were alanine, valine, isoleucine and methionine in both extracts. The level of ${\gamma}$-aminobutyric acid (GABA) was also increased significantly in the GBR and GBRP. Antioxidant activities of methanol extract of BR, GBR and GBRP were measured by using DPPH radical scavenging and SOD-like activity. Antioxidant activities showed the highest level of 83% and 76% when 100 mg/ml GBR and GBRP, respectively. Stimulation of the macrophages RAW264.7 cells with lipopolysaccharide (LPS) resulted in increased production of nitric oxide (NO) in the medium. However, the methanol extract of GBR and GBRP showed marked inhibition of NO synthesis in a does-dependant manner. These results showed that GBR and GBRP were significant role for activation of immune system in the pathogenesis of inflammatory diseases.

• Korean Journal of Microbiology
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• v.43 no.2
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• pp.106-110
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• 2007

The detection and distribution of Aeromonas in water supplies were investigated by using the USEPA Method 1605. Water samples were collected from the Han River, finished waters and tap waters supplied from Water Treatment Plants in Seoul monthly from July 2002 to December 2003. Aeromonas species in each water sample were quantified based on the development of yellow colonies on the surface of membrane filter using a selective medium (Ampicillin-Dextrin Agar with Vancomycin). The Quality Control (QC) for this study met the acceptance criteria of Method 1605. The concentrations of Aeromonas species in surface water samples ranged from $1.0{\times}10^{0}\;to\;9.8{\times}10^{3}\;CFU/ml$. Aeromonas species were found only in one tap water sample with concentration of 1 CFU/500 ml. No Aeromonas species were found in any finished water samples. Aeromonas species detected here were identified as A. salmonicida(51%), A. caviae(4.7%), A. schubertti(3.4%), A. sobria(3.8%), A. hydrophila(2.1%), and A. ichithiosmia(0.4%). A. salmonicida was the dominant species, which is of no significance to human health. Chlorine resistance of A. salmonicida was evaluated and as a result, 99.99% of A. salmonicida decreased after 30 seconds exposure at residual free chlorine 0.2 mg/L. These suggest that the waters supplied in Seoul may be safe against the pathogenic agent Aeromonas.

• Korean Journal of Microbiology
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• v.43 no.2
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• pp.77-82
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• 2007

In a homothallic ascomycete Aspergillus nidulans, sexual development is inhibited by various environmental stresses such as acetate medium, visible light and high osmolarity conditions. In order to study the genes involved in this stress-related regulatory network, we first attempted to isolate mutants that could develop cleistothecia even in the presence of any of those stresses including intensive visible light. More than 10,000 mutants were screened and 167 mutants were analyzed. Among them, 152 mutants underwent sexual development under the single stress condition of either high osmotic, high acetate or light condition but no sexual development in more than two stresses. Six mutants can produce cleistothecia under light or acetate stress but not in salt stress. Moreover, 6 mutants showed the ability to develop cleistothecia under the light but not under the acetate or osmo-stress. The mutants were revealed to have independent single gene mutation and grouped into different complementation groups (silA-F). The mutant alleles were all recessive to that of wild type. The light responsiveness of development implies the existence of delicate regulation process including reception and translocation of light signaling and determination of development.

• Korean Journal of Microbiology
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• v.45 no.2
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• pp.200-207
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• 2009

In this study, the characterization of purified erythritol 4-phosphate dehydrogenase, key enzyme of erythritol biosynthesis, produced by Penicillium sp. KJ81 was investigated. Optimum production conditions of erythritol 4-phosphate dehydrogenase was 1 vvm areration, 200 rpm agitation, at $37^{\circ}C$ for 8 days in the medium containing 30% sucrose, 0.5% yeast extract, 0.5% $(NH_4)_2SO_4$, 0.1% $KH_2PO_4$, and 0.05%$MgCl_2$. Erythritol 4-phosphate dehydrogenase was purified through ultrafiltration and preparative gel electrophoresis from cell extract of Penicillium sp. KJ81. This enzyme was especially active on erythrose 4-phosphate with 1.07 mM of Km value. It gave a single band on native polyacrylamide gel electrophoresis and an isoelectric point of 4.6. The enzyme had an optimal activity at pH 7.0 and $30^{\circ}C$. It was stable between pH 4.0 and 9.0, and also below $30^{\circ}C$. The enzyme activity was completely inhibited by 1mM $Cu^{2+}$ and 1 mM $Zn^{2+}$, but was not significantly affected by other cations tested. This enzyme was inactivated by treatment of tyrosine specific reagent, iodine and tryptophan specific reagent, N-bromosuccinimide. The substrate of the enzyme, erythrose 4-phosphate showed protective effect on the inactivation of the enzyme by both reagents. These results suggest that tryptophan and tyrosine residues are probably located at or near active site of the enzyme.