• 미생물학회지
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• 제27권3호
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• pp.297-303
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• 1989

Attempts to isolate the naturally occurring ultra-violet resistant bacteria from environmental sources were made. The isolates, designated No.29, 100, and 107, among numbers of bacterial isolates revealed a remarkable resistance to UV ray, whose degree of resistance in dose/response kinetics was comparable to that of an endospore-former, Bacillus subtilis. In a range of 100-300 $Jm^{-2}$/min of UV irradiation, the isolates exhibited 500-1000 fold resistance compated with E. coli. The isolated appeared to possiss cell-bound pigment of organge or crimson-red. The isolate 29 is spherical in pairs or tetrads, whereas the isolates 100 and 107 are rod. All are Gram-gositive bacteria and seemed to be non-endospore-bearer. A number of biochemical studies pursued on the isolates suggested that they are quite different to each other. Electron microscopic examination and the physiological characters of the isolate 29 suggested that this UV resistant spherical bacterium might be one species of Deinococcus, probably Deinococus radiophilus. Since there is no documents on UV resistant, Gram-positive, non-sporeformer bacillus so far, the isolates 100 and 107 might be turned out as new kinds of UV resistant bacteria occurring in nature by further investigation.

• 미생물학회지
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• 제30권6호
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• pp.483-487
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• 1992

Among a few number of UV-resistant isolated form various environmental sources (10), we made a comparative physio-chemoanalytical study on one of spherical bacteria isolated from air dust, presumably Deinococcus sp. (CM strain 29) with an UV resistant bacterium, Deinococcus radiophilus ATCC 27603 as the reference strain. Our isolate of UV resistant coccus, Deinococcus sp. CM 29 and D. radiophilus ATCC 27603 showed more than 75% matching coefficient in metabolic activity of various substrates. The most predominant cellular fatty acid of both strains was palmitoleic acid (C 16 :1, cis 9), but the detail fatty acid profiles were slightly dissimilar to each other. Cell-bound arange pigment seemed to be an identical chemicals on spectrophotometric analysis. L-ornithine was detected as cell-wall amino acid in both strains. Galactose was detected as cell-wall sugar in D. radiophilus ATCC 27603, whereas glucose in Deinococcus sp. CM 29. G-C molar ratio of both strains was comparable, 63-65%.

• BMB Reports
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• 제36권3호
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• pp.282-287
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• 2003

The production of superoxide dismutase (SOD) varied in Deinococcus radiophilus, the UV resistant bacterium, depending upon different phases of growth, UV irradiation, and superoxide treatment. A gradual increase in total SOD activity occurred up to the stationary phases. The electrophoretic resolution of the SOD in cell extracts of D. radiophilus at each growth phase revealed the occurrence of MnSOD throughout the growth phases. The SOD profiles of D. radiophilus at the exponential phase received oxidative stress by the potassium superoxide treatment or UV irradiation also revealed the occurrence of a single SOD. However, these treatments caused an increase in SOD activity. The data strongly suggest that D. radiophilus has only one species of SOD as a constitutive enzyme, which seems to be a membrane-associated protein.

• Journal of Microbiology
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• 제45권4호
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• pp.318-325
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• 2007

Glucose 6-phosphate dehydrogenase (G6PDH, EC 1.1.1.49) in Deinococcus radiophilus, an extraordinarily UV-resistant bacterium, was investigated to gain insight into its resistance as it was shown to be involved in a scavenging system of superoxide $(O_2^{-1})$ and peroxide $(O_2^{-2})$ generated by UV and oxidative stresses. D. radiophilus possesses two G6PDH isoforms: G6PDH-1 and G6PDH-2, both showing dual coenzyme specificity for NAD and NADP. Both enzymes were detected throughout the growth phase; however, the substantial increase in G6PDH-1 observed at stationary phase or as the results of external oxidative stress indicates that this enzyme is inducible under stressful environmental conditions. The G6PDH-1 and G6PDH-2 were purified 122- and 44-fold (using NADP as cofactor), respectively. The purified G6PDH-1 and G6PDH-2 had the specific activity of 2,890 and 1,033 U/mg protein (using NADP as cofactor) and 3,078 and 1,076 U/mg protein (using NAD as cofactor), respectively. The isoforms also evidenced distinct structures; G6PDH-1 was a tetramer of 35 kDa subunits, whereas G6PDH-2 was a dimer of 60kDa subunits. The pIs of G6PDH-1 and G6PDH-2 were 6.4 and 5.7, respectively. Both G6PDH-1 and G6PDH-2 were inhibited by both ATP and oleic acid, but G6PDH-1 was found to be more susceptible to oleic acid than G6PDH-2. The profound inhibition of both enzymes by ${\beta}-naphthoquinone-4-sulfonic$ acid suggests the involvement of lysine at their active sites. $Cu^{2+}$ was a potent inhibitor to G6PDH-2, but a lesser degree to G6PDH-1. Both G6PDH-1 and G6PDH-2 showed an optimum activity at pH 8.0 and $30^{\circ}C$.

• Journal of Microbiology
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• 제33권3호
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• pp.239-243
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• 1995

Deinococcus radiophilus, an UV resistant bacterium seemed to contain three issoenzymes of catalase. Among them, the samllest and most abundant species in cell-free extract, catalase-3 which also exhibited peroxidase activity was purified to electrophoretic homogeneity (145-fold purification) by chromatographic procedures. Its molecular weight was 155 kDa composed of four 38 kDa subunits. The $K_{m}$ value of catalase-3 for H$\_$2/O$\_$2/ was approximately 0.5 mM. This enzyme showed a typical ferric heme spectrum with maximum absorption at 405 nm. Upon binding to cyanide, the 405 nm peak shifted to 420 nm. Catalase-3 was very sensitive to inhibitors of heme proteins, such as cyanide, azide and hydroxylamine. A ratio of A$\_$405/A$\_$28O/ was 0.5 Catalase-3 was active over a wide range of pH, between pH 7 and 10. The enzyme was rather heat-labile and partially sensitive to edthanol-chloroform treatment, but resistant to 3-amino-1, 2, 4-triazole. Catalase-3 of D. radiophilus, which is a bifunction catalatic peroxidatic enzyme seemed to share certain molecular properties with the typical catalase and the catalase-[roxidase along with its own unique features.

• Journal of Microbiology and Biotechnology
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• 제20권11호
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• pp.1513-1520
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• 2010

A bacterium capable of producing melanin pigment in the presence of L-tyrosine was isolated from a crop field soil sample and identified as Klebsiella sp. GSK based on morphological, biochemical, and 16S rDNA sequencing. The polymerization of this pigment occurs outside the cell wall, which has a granular structure as melanin ghosts. Chemical characterization of the pigment particles showed then to be acid resistant, alkali soluble, and insoluble in most of the organic solvents and water. The pigment got bleached when subjected to the action of oxidants as well as reductants. This pigment was precipitated with $FeCl_3$, ammoniacal silver nitrate, and potassium ferricynide. The pigment showed high absorbance in the UV region and decreased absorbance when shifted towards the visible region. The melanin pigment was further charecterized by FT-IR and EPR spectroscopies. A key enzyme, 4-hydroxyphenylacetic acid hydroxylase, that catalyzes the formation of melanin pigment by hydroxylation of L-tyrosine was detected in this bacterium. Inhibition studies with specific inhibitors, kojic acid and KCN, proved that melanin is synthesized by the DOPA-melanin pathway.

• Biodesign
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• 제6권4호
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• pp.96-99
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• 2018

The gram-positive bacterium Staphylococcus aureus is a common cause of abscesses, sinusitis and food poisoning. The emergence of antibiotic-resistant strains has caused significant clinical issues worldwide. The HslU-HslV complex was first identified as a prokaryotic homolog of eukaryotic proteasomes. HslU is an unfoldase that mediates the unfolding of the substrate proteins, and it works with the protease HslV in the complex. To date, the protein complex has been mostly studied in gram-negative bacteria. In this study, we report the purification and crystallization of the full-length HslU from S. aureus. The crystal diffracted X-rays to a $3.5{\AA}$ resolution, revealing that the crystals belong to space group $P2_1$, with unit cell parameters of a = 166.5, b = 189.6, $c=226.6{\AA}$, and ${\beta}=108.1^{\circ}$. We solved the phage problem by molecular replacement using the structure of HslU from Haemophilus influenzae as a search model. The cell content analysis with this molecular replacement solution revealed that 24 molecules are contained in the asymmetric unit. This structure provides insight into the structural and mechanistic difference of the HslUV complex of gram-positive bacteria.

• Molecules and Cells
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• 제43권8호
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• pp.694-704
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• 2020

HslUV is a bacterial heat shock protein complex consisting of the AAA+ ATPase component HslU and the protease component HslV. HslV is a threonine (Thr) protease employing the N-terminal Thr residue in the mature protein as the catalytic residue. To date, HslUV from Gram-negative bacteria has been extensively studied. However, the mechanisms of action and activation of HslUV from Gram-positive bacteria, which have an additional N-terminal sequence before the catalytic Thr residue, remain to be revealed. In this study, we determined the crystal structures of HslV from the Gram-positive bacterium Staphylococcus aureus with and without HslU in the crystallization conditions. The structural comparison suggested that a structural transition to the symmetric form of HslV was triggered by ATP-bound HslU. More importantly, the additional N-terminal sequence was cleaved in the presence of HslU and ATP, exposing the Thr9 residue at the N-terminus and activating the ATP-dependent protease activity. Further biochemical studies demonstrated that the exposed N-terminal Thr residue is critical for catalysis with binding to the symmetric HslU hexamer. Since eukaryotic proteasomes have a similar additional N-terminal sequence, our results will improve our understanding of the common molecular mechanisms for the activation of proteasomes.

• 한국미생물·생명공학회지
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• 제41권1호
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• pp.26-32
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• 2013

부탄올 용매에서 생존하는 부탄올 내성 미생물을 분리하였다. 분리된 미생물들의 세포성장은 부탄올 농도가 증가함에 따라 감소하였으며, 그 중에서 BRS02가 12.5 g/L에서 가장 높은 내성도를 나타내었다. 또한, UV를 이용하여 BRS02균의 변이를 유도하여 고농도 부탄올 내성균 BRS251을 개발하였다. 부탄올 생산 모델균주로 대장균과 함께 부탄올, 프로판올 및 펜탄올에 대한 내성도를 비교한 결과, 대장균은 7.5 g/L 부탄올과 20 g/L 프로판올, 2 g/L 펜탄올 농도까지 생육이 가능한 반편, BRS251은 더 고농도인 17.5 g/L 부탄올과 32.5 g/L 프로판올, 6 g/L 펜탄올 농도까지 생육이 가능하였다. 분리된 세균을 동정하기 위해서 그람염색 후 광학현미경으로 관찰한 결과 그람양성의 구균으로 확인이 되었으며, 6.5% NaCl에서 생육이 가능하였다. 생화학적 특성을 분석한 결과, arginine dihydrolase, ${\alpha}$-glucosidase, urease 효소활성을 가지고 있었으며, 호기적인 조건에서 D-galactose, Dmaltose, D-mannitol, D-mannose, methyl-${\beta}$-D-glucopyranoside, D-ribose, sucrose, D-trehalose를 탄소원으로 자화하여 산을 생성할 수 있었으며, bacitracin, vibriostatic agent O/129 및 optochin에 대한 항생제 내성을 나타내었다. 16S rRNA 유전자 서열을 결정하고 계통발생도 분석을 통해 BRS02는 최종적으로 Staphylococcus sp.임을 동정하였다.

• 한국균학회소식:학술대회논문집
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• 한국균학회 2018년도 춘계학술대회 및 임시총회
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• pp.33-33
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• 2018

Air-borne plant pathogenic fungus Fusarium graminearum and seed-borne plant pathogenic bacterium Burkholderia glumae are cause similar disease symptoms in rice heads. Here we showed that two pathogens frequently co-isolated in rice heads and F. graminearum is resistant to toxoflavin produced by B. glumae while other fungal genera are sensitive to the toxin. We have tried to clarify the resistant mechanism of F. graminearum against toxoflavin and the ecological reason of co-existence of the two pathogens in rice. We found that F. graminearum carries resistance to toxoflavin as accumulating lipid in fungal cells. Co-cultivation of two pathogens resulted in increased conidia and enhanced chemical attraction and attachment of the bacterial cells to the fungal conidia. Bacteria physically attached to fungal conidia, which protected bacterium cells from UV light and allowed disease dispersal. Chemotaxis analysis showed that bacterial cells moved toward the fungal exudation compared to a control. Even enhanced the production of phytotoxic trichothecene by the fungal under presence of toxoflavin and disease severity on rice heads was significantly increased by co-inoculation rather than single inoculation. This study suggested that the undisclosed potentiality of air-born infection of bacteria using the fungal spores for survival and dispersal.