• 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 Species Research
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• 제7권1호
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• pp.9-12
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• 2018

In 2016, as part of a larger effort to discover indigenous prokaryotic species in Korea, we isolated the family Deinococcaceae and Planctomycetaceae as unrecorded bacterial species. From the high 16S rRNA gene sequence similarity (>98.5%) and formation of a robust phylogenetic clade with known species, it was determined that each strain was a distinct bacterial species. There are no official reports that these two species have been described in Korea; therefore, the bacterial strains of Deinococcus and Blastopirellula are described for the first time in Korea. Gram reaction, colony and cell morphology, basic biochemical characteristics, and isolation sources are also described in the species description section.

• 생명과학회지
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• 제28권3호
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• pp.361-368
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• 2018

본 연구는 제주연안에서 채집한 옥돔(Branchiostegus japonicus)의 장내기관으로부터 장내미생물의 분리하여 다양성을 조사하였다. 1차로 1.5% BHIA, MA, TSA 및 R2A agar 배지상 순수분리한 결과, 1.5% BHIA에서 가장 많은 colony개수를 나타냈다. 호기성, 혐기배양은 평균 $1.7{\times}10^6CFU/g^{-1}$, $1.1{\times}10^5CFU/g^{-1}$로 나타났으며 총 147개의 순수 colony가 분리되었다. 16S rDNA염기서열 분석결과 58속 74종으로 나타났으며 기본균주와 95-100%의 유사도를 나타내었다. 크게 5문(Phylum)으로 나뉘었으며, 주요 계통군으로 Proteobacteria 문이 50%로 Moraxellaceae, Rhodobacteraceae, Shewanellae, Halomondaceae, Enterobacteriaceae, Vibrionaceae, Hahellaceae, Pseudomonadaceae, Erythrobacteraceae 총 9과 35속 35종으로 우점도가 제일 높게 나타났다. Actinobacteria문은 24%, Microbacteriaceae, Intrasporangiaceae, Dietziaceae, Dermabacteraceae, Dermacoccaceae, Nocardiodaceae, Brevibacteriaceae, Propionobacteriacea 총 8과 11속 17종, Frimicutes문은 16%, Bacillaceae, Staphylcoccaceae, Planococcaceae, Streptococcaceae, Paenibacillaceae, Clostridiaceae 총 6과 8속 17종, bacteroidetes문은 6%, Cyclobacteriaceae, Flavobacteriaceae 총 2과 3속 4종을, Deinococcus-thermus문은 4%로 Deinococcaceae 단일 1과 1속 1종으로 나타났다.

• 미생물학회지
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• 제47권1호
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• pp.66-73
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• 2011

각 지역에서 수집한 무당벌레(JK, CK, CJ)의 소화기관을 채취하여 장내세균의 밀도를 조사한 결과, 호기배양의 경우 $6.0{\times}10^4$ CFU/gut, 혐기배양 결과 $8.0{\times}10^6$ CFU/gut로 계수되었다. 호기적 조건에서 배양된 세균 집락은 총 7가지 형태로 분류되었으며, 혐기적 조건에서 배양된 집락은 총 3가지 형태로 유사한 특징을 나타내었다. 무당벌레 각 소화기관으로부터 호기성세균 34균주와 혐기성세균 82균주, 총 116균주의 장내세균을 순수분리하였다. 호기성 세균 34균주를 대상으로 16S rRNA 유전자 염기서열을 해석한 결과, ${\alpha}$-Proteobacteria (3균주), ${\gamma}$-Proteobacteria (2 균주), Firmicutes (24 균주), Actinobacteria (4 균주) 그리고 Deinococcus-Thermus (1균주) 계통군으로 분류되었다. Firmicutes 계통군의 Bacillus thuringiensis와 Staphylococcus 속의 다양한 종은 JK, CK 및 CI 무당벌레 소화기관에서 모두 공통적으로 분리되었다. 형태적으로 유사한 혐기세균의 16S rRNA-ARDRA 패턴양상을 분석하여 유사도 70%에서 비교한 결과, 17개 ARDRA group으로 분류되었다. 각 ARDRA group에 속하는 대표 혐기성세균의 16S rRNA 유전자 염기서열을 해석한 결과, 무당벌레 소화기관에서 분리된 모든 혐기성 장내세균은 ${\gamma}$-Proteobacteria 계통군에 속하는 것으로 나타났으며 Hafnia alvei, Enterobacter ludwigii, Enterobacter kobei, Pseudomonas oryzihabitans 그리고 Pseudomonas koreensis와 높은 유연관계를 갖는 것으로 확인되었다. 무당벌레 소화기관으로부터 분리된 전체 장내세균의 약 70%가 ${\gamma}$-Proteobacteria 계통군에 속하였으며, 23%가 Firmicutes 계통군으로 무당벌레 소화기관 내 주요 계통군임이 확인되었다.

• 생명과학회지
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• 제28권12호
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• pp.1477-1488
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• 2018

본 연구는 제주 연안에서 채집한 오분자기(Sulculus diversicolor supertexta)를 구성하는 미생물 군집의 다양성을 알아보기 위하여 근육, 장, 생식소 각 부위별로 조사하였다. 배지로 1차 순수 분리한 결과 근육은 MA, 장 1% BHIA, 생식소 1% TSA에서 각각 최대 군락 계수가 나타났다. 16S rRNA sequence로 표준 균주와 비교 유사도 분석 결과 총 190개의 순수 colony가 분리되었다. NBLAST program 분석 결과 크게 5문 25과 39속 71종으로 나타났다. 표준 균주와 상동성은 91-100%를 나타냈다. 오분자기 내 미생물 군집은 크게 Probacteria (Gamma-proteobacteria, Alpha-proteobacteria) 48%, Actinobacteria 32.5%, Firmicutes 16.9%, Bacteroide 1.3%, Deinococcus-thermus 1.3%로 나타났다. 근육, 장, 생식소 모든 부위에서 Moraxellaceae과 Psychrobacter cibarius가 우점하였다. 근육, 장, 생식소 모든 부위에서 Alteromonadaceae, Enterobacteriaceae, Pasturellaceae, Moraxellaceae, Rhodobacteraceae, Geminicoccaceae, Dietziaceae, Intrasporangiaceae, Microbacteriaceae, Micrococcaceae, Micromonosporaceae, Streptomycetaceae, Aerococcaceae, Bacillaceae, Paenibacillaceae, Planococcaceae, Staphylcoccaceae가 공통적으로 분리되었으며, 장에서 Flavobacteriaceae, Corynebacteriaceae, Yesiniaceae, Vibrionaceae, Hahellaceae, Pseudomonadaceae가 추가 분리되었다. 분리 균주로부터 인체 유해 질병 세균에 대한 항균활성 모니터링 결과 Sterptomyces albus (96%)가 4균주 모두 항균활성을 보였고 Agrococcus baldri (99%), Psychrobacter nivimaris (99%)가 E. coli, E. aerogens에 대한 항균활성을 나타냈다. 그 외 상동성이 낮은 일부 균주가 분리되어 신균주 실험을 비롯한 항균활성물질 정제 등 추가 실험이 필요한 것으로 사료된다. 본 실험은 오분자기 미생물 군집의 다양성과 유전학적 자원을 확보하는데 의의를 두며, 응용 미생물의 개발 가능성에 있어 기초 자료를 제공하고자 하였다.

• Journal of Species Research
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• 제6권2호
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• pp.163-170
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• 2017

As a part of the research program 'Survey of freshwater organisms and specimen collection', freshwater samples were collected from Lakes Soyang and Chungju in 2016. Hundreds of bacterial strains were isolated from the samples and were identified based on 16S rRNA gene sequences. Among the bacterial isolates, strains showing higher than 98.7% sequence similarity with validly published bacterial species not reported in Korea were selected as unrecorded bacterial species. Based on 16S rRNA gene sequence similarity, 17 strains were identified as unrecorded bacterial species in Korea. The 17 bacterial strains were phylogenetically diverse and belonged to four phyla, seven classes, 13 orders, 14 families, and 16 genera. At generic level, the unreported species were affiliated with Caulobacter, Paracoccus, and Mesorhizobium of the class Alphaproteobacteria, Deefgea, Undibacterium, Chitinimonas, Inhella, and Sphaerotilus of the class Betaproteobacteria, Vibrio and Cellvibrio of the class Gammaproteobacteria, Sanguibacter and Clavibacter of the phylum Actinobacteria, Lactococcus of the phylum Firmicutes, Deinococcus of the class Deinococci, and Chryseobacterium and Flavobacterium of the phylum Bacteroidetes. The unreported species were further characterized by examining Gram reaction, colony and cell morphology, biochemical properties, and phylogenetic position. The detailed description of the 17 unreported species are also provided.

• 생명과학회지
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• 제6권1호
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• pp.56-65
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• 1996

10%의 sucrose를 첨가한 홍차추출물에 tea fungus를 접종하여 (30^{\circ.)$에서 정치배양하여 발효하였다. 14일 동안의 발효에 의해 배양액의 전표면에 7~8mm의 두꺼운 피막이 생겼으며, 배양액의 pH가 2.5 부근으로 저하되었다. 발효 과정 중 아래쪽의 배양액에서는 효모(Saccharomyces cerevisiae and Eeniella sp.)와 여러종류의 세균(Bacillus subtilis, Kurthia zopfli, Gluconobacter oxydans와 Deinococcus sp.)이 분리되었다. 반면에 피막 부위에서는 Acetobacter aceti의 단일균주가 분리되었으며 이 세균은 통상의 Acetobacter와는 달리 점질상의 덩어리로 성장하였다. 발효음료는 달고 새콤한 맛과 약간의 달콤한 과일향을 나타내었다. 이상의 결과로부터 tea fungus에 의한 홍차발효는 다양한 미생물이 공동으로 작용하여 진행되는 symbiotic acetate발효로서, 발효음료는 생물학적으로 안전할 뿐만아니라 적당한 발효조건에 의해 좋은 향미를 갖춘 유망한 음료로 판단되었다.

• 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.

• Molecules and Cells
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• 제39권6호
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• pp.460-467
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• 2016

Bacteriophytochromes are phytochrome-like light-sensing photoreceptors that use biliverdin as a chromophore. To study the biochemical properties of the Deinococcus radiodurans bacteriophytochrome (DrBphP) protein, two anti-DrBphP mouse monoclonal antibodies (2B8 and 3H7) were generated. Their specific epitopes were identified in our previous report. We present here fine epitope mapping of these two antibodies by using truncation and substitution of original epitope sequences in order to identify minimized epitope peptides. The previously reported original epitope sequences for 2B8 and 3H7 were truncated from both sides. Our analysis showed that the minimal peptide sequence lengths for 2B8 and 3H7 antibodies were nine amino acids (RDPLPFFPP) and six amino acids (PGEIEE), respectively. We further characterized these peptides in order to investigate their reactivity after single deletion and single substitution of the original peptides. We found that single-substituted 2B8 epitope (RDPLPAFPP) and dual-substituted 3H7 epitope (PGEIAD) showed significantly increased reactivity. These two antibodies with high reactivity for the short modified peptide sequences are valueble for developing new peptide tags for protein research.

• Journal of Species Research
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• 제10권2호
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• pp.99-116
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• 2021

In 2018, as a subset study to discover indigenous prokaryotic species in Korea, a total of 18 unreported bacterial strains were discovered. From the high 16S rRNA gene sequence similarity (>98.8%) and formation of a robust phylogenetic clade, it was determined that each strain belonged an independent and predefined bacterial species. There were no official report that these 18 species were previously described in Korea; therefore, one strain of Williamsia, one strain of Rhodococcus, three strains of Microbacterium, three strains of Agromyces, one strain of Arthrobacter, one strain of Paeniglutamicibacter, one strain of Pseudarthrobacter, one strain of Nocardioides, one strain of Fibrella, one strain of Hymenobacter, one strain of Deinococcus, two strains of Fictibacillus, and one strain of Paenibacillus are described as unreported bacterial species in Korea. Gram reaction, basic biochemical characteristics, and colony and cell morphologies are described in the species description section.