• Journal of Microbiology
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• 제44권6호
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• pp.694-698
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• 2006

Several marine bacterial strains were isolated from Undaria pinnatifida (Miyok in Korean). Sixty-six strains were isolated on R2A agar media at $10^{\circ}C$ and identified by a phylogenetic analysis of the 16S rRNA gene sequences. They were grouped into 10 different sequence types based on the initial sequence analysis of the 5' domain of the gene (approximately 500 bp). Full sequences of 16S rRNA gene, were obtained from one strain in each sequence type and the species-affiliation was determined using phylogenetic and sequence similarity analyses. The results of the analyses indicated that they were closely related to Psychrobacter aquimaris, P. celer, P. nivimaris, P. pulmonis, Psychromonas arctica or Bacillus psychrodurans. These bacteria are marine or psychrotrophic bacteria. Because the sporophytes of U. pinnatifida are cultured on the costal area during winter, the U. pinnatifida-associated bacteria appeared to grow at low temperatures. U. pinnatifida sporophytes can be a good source for the isolation of psychrotrophic bacteria.

• Bulletin of the Korean Chemical Society
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• 제32권7호
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• pp.2405-2409
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• 2011

The psychrophilic bacteria Psychromonas arctica survives at subzero temperatures by having adapted several protective mechanisms against freezing and oxidative stresses. Many reactive oxygen species are likely generated in P. arctica as a result of reduced metabolic turnover rates. A previous study identified the pasod gene for superoxide dismutase from P. arctica using a series of PCR amplifications. Here, upon cloning into a His-tag fused plasmid, the sod gene from P. arctica (pasod) was successfully expressed by IPTG induction. His-tagged PaSOD was subsequently purified by $Ni^{2+}$-NTA affinity chromatography. The purified PaSOD exhibited a higher SOD activity than that of Escherichia coli (EcSOD) at all temperatures. The difference in activity between PaSOD and EcSOD becomes even more significant at 4$^{\circ}C$, indicating that PaSOD plays a functional role in the cold adaptation of P. arctica in the Arctic.

• Bulletin of the Korean Chemical Society
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• 제32권6호
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• pp.1925-1930
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• 2011

Psychrophilic bacteria have acquired cold-resistance in order to protect themselves against freezing temperatures, which would otherwise be lethal. DnaK/DnaJ/GrpE systems are molecular chaperones which facilitate proper folding of newly synthesized proteins. Efficient folding processes are of great importance especially in a cold environment, such as the Arctic. In order to understand the protection mechanisms of psychrophilic bacteria against cold temperatures, we have explored a genome of KOPRI22215, tentatively identified as Psychromonas arctica, whose genome sequence has not yet been discovered. With an aim of searching for a coding gene of DnaK from KOPRI22215, we have applied a series of polymerase chain reactions (PCR) with homologous primers designed from other Psychromonas species and LA PCR in vitro cloning. 1917 bp complete coding sequence of dnaK from KOPRI22215 was identified including upstream promoter sites. Recombinant plasmids to overexpress PaDnaK along with EcDnaK (DnaK of E. coli) were then constructed in pAED4 vector and the pET-based system to induce PaDnaK expression by IPTG. Characterization assays of expressed PaDnaK were carried out by measuring survival rates upon 4 day incubation at 4 $^{\circ}C$: a refolding assay as molecular chaperone, and ATPase assay for functional activity. Taking account of all the data together, we conclude that PaDnaK was identified, successfully expressed, and found to be more efficient in providing cold-resistance for bacterial cells.

• Bulletin of the Korean Chemical Society
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• 제31권4호
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• pp.887-890
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• 2010

The family of ClpB protein is a molecular chaperone which protects cellular proteins from being aggregated upon exposure to severe environmental stresses in association with DnaK/DanJ/GrpE in the ATP-dependent manner. In a psychrophilic bacterium which survives at a subzero temperature, any functional role of cold-active ClpB protein can be rather crucial. In order to identify a ClpB encoding gene from a cold-adapted bacterium whose genome sequence has not been fully discovered, we have employed a series of PCR technologies, including a gradient PCR with homologous primers, an inverse PCR and a cassette PCR. The full sequence of PaclpB gene was successfully identified and compared with those of other psychrophilic species. We have further cloned the gene in E.coli expression systems and were able to induce PaClpB protein expression by IPTG, which help us understand a molecular mechanism for survival against extremely cold environments.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2007년도 International Meeting of the Microbiological Society of Korea
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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.

• Journal of Species Research
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• 제12권2호
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• pp.165-173
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• 2023

Bacterial communities inhabiting islands play a vital role in the functioning and formation of a unique, isolated ecosystem. Nevertheless, there has been a lack of systematic research on the indigenous microbiological resources of the islands in Korea. To excavate microbial resources for further studies on the metabolism and biotechnological potential, a standard dilution plating was applied to coastal seawater samples collected from islands along the west coast of the Korean Peninsula, including Deokjeokdo, Baengnyeongdo, and Daebudo in 2022. A total of 2,007 bacterial strains were isolated from the samples as single colonies and identified using 16S rRNA gene sequence analyses. A total of 20 strains, with ≥98.7% 16S rRNA gene sequence similarity to bacterial species having validly published names but not reported in Korea, were designated as unrecorded bacterial species in Korea. The unrecorded bacterial strains were phylogenetically diverse and belonged to four phyla, five classes, 12 orders, 17 families, and 18 genera. The unreported species were assigned to Algimonas, Amylibacter, Notoacmeibacter, Roseibium, and Terasakiella of the class Alphaproteobacteria; Alteromonas, Congregibacter, Marinagarivorans, Marinicella, Oceanospirillum, Psychromonas, Thalassotalea, Umboniibacter, and Vibrio of the class Gammaproteobacteria; Lutibacter and Owenweeksia of the class Flavobacteriia; Paenibacillus of the class Bacilli; and Pelagicoccus of the class Opitutae. The taxonomic characteristics of the unreported species, including morphology, biochemistry, and phylogenetic position are provided in detail.