• Korean Journal of Microbiology
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• v.41 no.1
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• pp.74-80
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• 2005

For the reseach of the natural marine antioxidant, an antioxidant-producing bacterium was isolated from seawater in Jeju costal area. The isolated strain SC2-1 was Gram-positive, catalase positive, oxidase negative, motile and small rods. The strain utilized sucrose, dextrose, fructose, mannitol and maltose as a sole carbon and energy source and NaCl required for growth. The radical scavenging activity of the culture supernatant was detemined by DPPH method. This bacterium was identified based on morphological, biochemical characteristics and 16S rDNA sequence analysis, and then was named Exiguobacterium sp. SC2-1. The optimum conditions of culture for production antioxidnat were $25^{\circ}C$, pH 6-8 and $4\%$ NaCl. The stain showed the highest activity and growth cultured in medium which added $1\%$ maltose. Hydroxyl radical activity of the supernatant of Exiguobacterium sp. SC2-1 was $73\%$. The SOD activity of the culture supernatant was estimated about $35\%$.

• Journal of Microbiology and Biotechnology
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• v.29 no.2
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• pp.235-243
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• 2019

A special eosinophilic agarase exo-type ${\beta}$-agarase gene, AgaDL6, was cloned from a marine agar-degrading bacterium, Flammeovirga sp. HQM9. The gene comprised 1,383-bp nucleotides encoding a putative agarase AgaDL6 of 461 amino acids with a calculated molecular mass of 52.8 kDa. Sequence analysis revealed a ${\beta}$-agarase domain that belongs to the glycoside hydrolase family (GH) 16 and a carbohydrate-binding module (CBM_4_9) unique to agarases. AgaDL6 was heterologously expressed in Escherichia coli BL21 (DE3). Enzyme activity analysis of the purified protein showed that the optimal temperature and pH of AgaDL6 were $50^{\circ}C$ and 3.0, respectively. AgaDL6 showed thermal stability by retaining more than 98% of activity after incubation for 2 h at $50^{\circ}C$, a feature quite different from other agarases. AgaDL6 also exhibited outstanding acid stability, retaining 100% of activity after incubation for 24 h at pH 2.0 to 5.0, a property distinct from other agarases. This is the first agarase characterized to have such high acid stability. In addition, we observed no obvious stimulation or inhibition of AgaDL6 in the presence of various metal ions and denaturants. AgaDL6 is an exo-type ${\beta}$-1,4 agarase that cleaved agarose into neoagarotetraose and neoagarohexaose as the final products. These characteristics make AgaDL6 a potentially valuable enzyme in the cosmetic, food, and pharmaceutical industries.

• Membrane and Water Treatment
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• v.10 no.6
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• pp.395-404
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• 2019

The influence of chlorine on marine bacterial communities was examined in this study. A non-chlorine-adapted marine bacterial community (NCAM) and a chlorine-adapted bacterial community (CAM, bacterial community treated with $0.2mg-Cl_2/L$ chlorine) were cultivated for 1 month. A distinct difference was observed between the NCAM and CAM, which shared only eight operational taxonomic units (OTUs), corresponding to 13.1% of the total number of identified OTUs. This result suggested that chlorine was responsible for the changes in the marine bacterial communities. Kordiimonas aquimaris was found to be a chlorine-resistant marine bacterium. The effect of intermittent chlorination on the two marine biofilm communities formed on the reverse osmosis (RO) membrane surface was investigated using various chlorine concentrations (0, 0.2, 0.4, 0.6 and 0.8 mg $Cl_2/L$). Although the average number of adherent marine bacteria on the RO membrane over a period of 7 weeks decreased with increasing chlorine concentration, disinfection efficiencies showed substantial fluctuations throughout the experiment. This is due to chlorine depletion that occurs during intermittent chlorination. These results suggest that intermittent chlorination is not an effective disinfection strategy to control biofilm formation.

• Journal of the Korean Society of Food Science and Nutrition
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• v.25 no.2
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• pp.294-300
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• 1996

우리나라 남해 연안 해역으로부터 적색색소를 생산하는 균주를 분리하여 동정한 결과 Vibrio sp.로 판명되었다. 본 균주는 성장과정 중에 적색색소를 생산하여 세포내에 축적시키며 이때의 적색색소 생산량은 배양 후 24시간 이후부터 최고치에 도달하였으며, 배지중의 첨가물로서 0~2%의 NaCl, 1% fructose, 0.3%의 $(NH_{4})_{2}SO_{4}$를 첨가되었을 때 높은 생산량을 보였다. 한편 배양하여 얻어진 균체로부터 methanol로 추출한 적색색소는 UV-VIS spectrophotometer로 분석한 결과 최대 흡수파장이 531nm이였으며, 현재까지 널리 알려진 anthocyan계열의 색소와 동일한 흡수파장을 지니는 것으로 알 수 있었다. 또한 methanol 추출 색소를 TLC와 HPLC로 분리.정제하여 GC/MS로 분석한 결과 분자량 281과 236인 2종류의 물질이 검출되었으며, 281의 분자량을 가지는 물질의 경우 anthocyanin의 기본 구조에 OH기가 5개 존재하는 cyanidin으로 추정되었다. 따라서 본 실험에서 사용한 해양 유래 Vibrio sp.가 생산하는 적색색소는 cyanidin을 주성분으로 하는 anthocyan 계열의 색소이이 확인되었다. 유전공학적 기법을 이용한 균주 개량과 대량생산 체제가 구축된다면, 현재 널리 사용되고 있는 화학 합성색소를 대체할 수 있는 천연색소로서, 식품, 의약품, 화장품, 화학, 염료 공업 등에 사용될 수 있으리라 사료된다.

• Journal of Life Science
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• v.12 no.6
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• pp.759-764
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• 2002

A halophilic bacterium for the production of the immunostimulant was isolated from domestic marine, it was identified as Burkholderia sp. IS-203. The optimal conditions for the production of the immunostimulant were 1 % dextrose and 1 % yeast extract in artificial sea water for carbon and nitrogen sources, respectively. The initial pH and growth temperature for the prodution were 8.0 and $30^{\circ}C$ under the presence of oxygen, respectively.

• Biomolecules & Therapeutics
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• v.16 no.4
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• pp.356-363
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• 2008

A bacterial strain with good antibacterial activities against Staphylococus aureus and Escherichia coli was isolated from a marine sponge Stelleta sp., and it was identified as a Psychrobacter sp. by comparative 16S rDNA sequence analysis. In our search for bioactive secondary metabolites from this psychrophillic and halotolerent bacterium, sixteen cyclic dipeptides (1-16) were isolated and their structures were identified on the basis of NMR analysis. In the test of the compounds for the protective effect against Vibrio vulnificusinduced cytotoxicity in human intestinal epithelial cells, cyclo-(L-Pro-L-Phe) (5) exhibited significant protective effect. Compounds 2, 6, and 11, which contain D-amino acid, were first isolated from bacteria.

• Proceedings of the Microbiological Society of Korea Conference
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• 2008.05a
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• pp.163-163
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• 2008

Methylophaga aminisulfidivorans $MP^T$, a restricted facultative marine methylotrophic bacterium, was able to utilize methanol as a sole carbon and energy source, and possessed a methanol dehydrogenase (MDH) that is a key enzyme in the process of methanol oxidation. During purification of MDH, three types of MDH (MDH I, II, and III) were obtained in the cell free extracts from $MP^T$ cells grown on methanol. When analyzed by SDS-PAGE and ESI-FT ICR MS, MDH I was confirmed to consist of two subunits and with molecular masses of ~66 and ~10 kDa, respectively, in a form of ${\alpha}_2{\beta}_2$. While MDH II and MDH III contained an additional ~30 kDa protein, designated ${\gamma}$, in a form of ${\alpha}_2{\beta}_2{\gamma}$ and ${\alpha}_2{\beta}_2{\gamma}_2$, respectively. MDH III showed 1.5.2.0 times higher activity than MDH II, while MDH I remained the lowest activity. Based on these observations and experimental data, it seems that the original MDH conformation is ${\alpha}_2{\beta}_2{\gamma}2$ within $MP^T$ growing on methanol, and subunit ${\gamma}$ keeps MDH in an active form, and/or makes MDH easily bind to the substrate, methanol.

• Fisheries and Aquatic Sciences
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• v.12 no.1
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• pp.54-59
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• 2009

Carotenoids such as $\beta$-carotene and astaxanthin are used as food colorants, animal feed supplements and for nutritional and cosmetic purposes. In a previous study, an astaxanthin biosynthesis gene cluster was isolated from the marine bacterium, Paracoccus haeundaensis. Geranylgeranyl pyrophosphate (GGPP) synthase (CrtE), encoded by the ortE gene, catalyzes the formation of GGPP from farnesyl pyrophosphate (FPP), which is an essential enzyme for the biosynthesis of carotenoids in early steps. In order to study the biochemical and enzymatic characteristics of this important enzyme, a large quantity of purified GGPP synthase is required. To overproduce GGPP synthase, the crtE gene was subcloned into a pET-44a(+) expression vector and transformed into the Escherichia coli BL21(DE3) codon plus cell. Transformants harboring the crtE gene were cultured and the crtE gene was over-expressed. The expressed protein was purified to homogeneity by affinity chromatography and applied to study its biochemical properties and molecular characteristics.

• Journal of Marine Bioscience and Biotechnology
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• v.2 no.4
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• pp.263-266
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• 2007

Vibrio vulnificusis a causative agent of serious diseases in humans resulting from the contact of wound with seawater or consumption of raw seafood. Several studies aimed at detecting V. vulnificus have targeted vvh as a representative virulence toxin gene belonging to the bacterium. In this study, we targeted the rpoS gene, a general stress regulator, to detect V. vulnificus. PCR specificity was identified by amplification of 8 V. vulnificus templates and by the loss of a PCR product with 36 non-V. vulnificus strains. The PCR assay had the 273-bp fragment and the sensitivity of 10 pg DNA from V. vulnificus. SYBR Green I-based real-time PCR assay targeting the rpoS gene showed a melting temperature of approximately $84^{\circ}C$ for V. vulnificus strains. The minimum level of detection by real-time PCR was 2 pg of purified genomic DNA, or $10^3$ V. vulnificus cells from pure cultured broth and $10^3$ cells in 1g of oyster tissue homogenates. These data indicate that real-time PCR is a sensitive, species-specific, and rapid method for detecting this bacterium using the rpoS gene in pure cultures and in infected oyster tissues.

• Proceedings of the Microbiological Society of Korea Conference
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• 2007.05a
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• pp.80-82
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• 2007

We have isolated numerous cold deep-sea adapted microorganisms (piezophilic, formerly referred to as "barophilic" bacteria) using deep-sea research submersibles. Many of the isolates are novel psychrophilic bacteria, and we have identified several new piezophilic species, i.e., Photobacterium profundum, Shewanella violacea, Moritella japonica, Moritella yayanosii, Psychromonas kaikoi, and Colwellia piezophila. These piezophiles are involving to five genera in gamma-Proteobacteria subgroup and produce significant amounts of unsaturated fatty acids in their cell membrane fractions to maintain the membrane fluidity in cold and high-pressure environments. Piezophilic microorganisms have been identified in many deep-sea bottoms of many of the world oceans. Therefore, these microbes are well distributed on our planet. One of the isolated deep-sea piezophiles, Shewanella violacea strain DSS12 is a psychrophilic, moderately piezophilic bacterium from a sediment sample collected at the Ryukyu Trench (depth: 5,110 m), which grows optimally at 30 MPa and $8^{\circ}C$ but also grows at atmospheric pressure (0.1 MPa) and $8^{\circ}C$. We have examined this strain to elucidate the molecular basis for gene regulation at different pressure conditions because this strain is useful as a model bacterium for comparing the various features of bacterial physiology under pressure conditions. In addition, we completed the sequencing of the entire genome of this piezophilic bacterium and we expect that many biotechnologically useful genes will be identified from the genome information.