• Korean Journal of Microbiology
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• v.52 no.1
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• pp.110-115
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• 2016

A small plasmid (pDK4) from the Antarctic marine organism Pseudoalteromonas sp. PAMC 21150, was purified, sequenced and analyzed. pDK4 was determined to be 3,480 bp in length with a G+C content of 41.64% and contains three open reading frames encoding a replication initiation protein (RepA), a conjugative mobilization protein (Mob) and a hypothetical protein. PCR-amplified pDK4 was cloned in high-copy pUC19 to yield the fusion vector pDOC153. The chloramphenicol resistance gene was inserted into pDOC153 to give an ampicillin and chloramphenicol-resistant, Pseudoalteromonas - Escherichia coli shuttle vector (7,216 bp; pDOC155). The TonB-dependent receptor (chi22718_IV ) and exochitinase (chi22718_III ) genes from Arctic marine P. issachenkonii PAMC 22718 were cloned into pDOC155 to produce pDOC158 and pDOC165, respectively. Both vector derivatives were transferred into plasmid-free Pseudoalteromonas sp. PAMC 22137 by the triparental mating method. PCR experiments showed that the genes were stably maintained both in Pseudoalteromonas sp. PAMC 22137 and E. coli $DH5{\alpha}$ cells, indicating the potential use of pDOC155 as a new gene transfer system into marine Pseudoalteromonas spp.

• Journal of Life Science
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• v.31 no.5
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• pp.496-501
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• 2021

In this study, the marine agar-degrading bacterium Pseudoalteromonas sp. JH-1 was isolated, and its growth and agarase properties were investigated. Seawater was collected from the offshore of the Yonggung Temple in Busan, and agar-degrading bacteria were isolated and cultured with marine agar medium. The bacterium Pseudoalteromonas sp. JH-1 was isolated through 16S rRNA gene sequencing. The extracellularly secreted enzyme was obtained from the culture broth of Pseudoalteromonas sp. JH-1 and was used to characterize its agarase. The extracellular agarase exhibited a maximum activity of 116.6 U/l at 50℃ and pH 6.0 of 20 mM Tris-HCl buffer. Relative activities were 31, 59, 94, 100, 45, and 31% at 20, 30, 40, 50, 60, and 70℃, respectively. Relative activities were 49, 85, 100, 86, 81, and 67% at pH 4, 5, 6, 7, 8, and 9, respectively. Residual activity was more than 85% after exposure at 20, 30, and 40℃ for 2 hr, and more than 82% after exposure at 50℃ for 2 hr. Zymogram analysis confirmed that Pseudoalteromonas sp. JH-1 produced at least two agarases of 55 and 97 kDa. As the products of α-agarase and β-agarase have antioxidation, antitumor, skin-whitening, macrophage activation, and prebiotic effects, further studies are needed on the agarase of Pseudoalteromonas sp. JH-1.

• Microbiology and Biotechnology Letters
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• v.39 no.4
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• pp.317-323
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• 2011

Strain A-3, an amylase-producing bacteria, was isolated from coastal seawater near Daecheon in the Republic of Korea. It was seen to possess a single polar flagella and grow well, on ASW-YP agar plates, at temperatures of between $20-37^{\circ}C$. However, it grew more slowly at the temperatures of $15^{\circ}C$ and $40^{\circ}C$. Similarly, it was observed to grow abundantly, in an Artificial Sea Water-Yeast extract-Peptone (ASW-YP) liquid medium, in a pH range of 6-9, but not grow at pHs of 4-5 and a pH of 10. Strain A-3 was noted as being close to Pseudoalteromonas phenolica O-$BC30^T$, Pseudoalteromonas luteoviolacea $NCIMB1893^T$, Pseudoalteromonas rubra $ATCC29570^T$, and Pseudoalteromonas byunsanensis $FR1199^T$, with 98.30%, 97.86%, 97.78%, and 97.25% similarities respectively, in its 16S rRNA sequence. A phylogenetic tree revealed that strain A-3 and P. phenolica O-$BC30^T$ belong to a clade. However, strain A-3 differed from P. phenolica O-$BC30^T$ in relation to a number of physiological characteristics. Strain A-3 exhibited no growth above 5% NaCl concentrations, no utilization of D-glucose, D-mannose, D-maltose, or D-melibose, and no lipase (C-14) activity. All of these properties strongly indicate that strain A-3 is distant from P. phenolica O-$BC30^T$ and thus led us to name it Pseudoalteromonas sp. A-3. Pseudoalteromonas sp. A-3 produces ${\alpha}$-amylase throughout growth. Maximal amylase activities of 144.48 U/mL and 149.20 U/mL were seen at pH 7.0 and $37^{\circ}C$, respectively. Pseudoalteromonas sp. A-3's high, stable production of ${\alpha}$-amylase in addition to its biochemical features, such as alkalitolerance, suggest that it is a good candidate for industrial applications.

• Journal of Microbiology and Biotechnology
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• v.19 no.3
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• pp.229-237
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• 2009

Among 12 marine bacterial strains from the China coast that exhibited interesting bioactivity (positive for both antimicrobial and cytotoxic activities), only four strains, namely, NJ6-3-1, NJ6-3-2, NB-6, and YTHM-17, had a KS domain or A domain when screened for PKS and NRPS genes using a PCR. Interestingly, two of these strains belonging to Pseudoalteromonas and associated with the marine sponge Hymeniacidon perleve were positive for both PKS and NRPS, whereas the other two strains of Pseudoalteromonas did not have a PKS or NRPS gene. A molecular phylogeny analysis and DGGE analysis of the Pseudoalteromonas sp. indicated that they had a specific affinity with the host marine sponge Hymeniacidon perleve. Furthermore, an analysis of a partial sequence of Pseudoalteromonas sp. NJ6-3-2 isolated from the marine sponge Hymeniacidon perleve obtained from genomic walking using a computational approach indicated a relatively complete PKS module including auxiliary domains (DH, KR, and Cy).

• Microbiology and Biotechnology Letters
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• v.43 no.2
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• pp.134-141
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• 2015

An agarolytic marine bacterium (H9) was isolated from the coastal seawater of the West Sea, South Korea. The isolate, H9, was gram-negative and rod-shaped with a smooth surface and polar flagellum. Cells grew at 20-30℃, between pH 5.0 and 9.0, and in ASW-YP (Artificial Sea Water-Yeast extract, Peptone) media containing 1-5% (w/v) NaCl. The G+C content was 41.56 mol%. The predominant isoprenoid quinone in strain H9 was ubiquinone-8. The major fatty acids (>10%) were C_16:1ω7c (34.3%), C_16:0 (23.72%), and C_18:1ω7c (13.64%). Based on 16S rRNA gene sequencing, and biochemical and chemotaxonomic characterization, the strain was designated as Pseudoalteromonas sp. H9 (=KCTC23887). In liquid culture supplemented with 0.2% agar, the cell density and agarase activity reached a maximum level of OD = 4.32 (48 h) and OD = 3.87 (24 h), respectively. The optimum pH and temperature for the extracellular crude agarases of H9 were 7.0 and 40℃, respectively. Thin-layer chromatography analysis of the agarase hydrolysis products revealed that the crude agarases hydrolyze agarose into neoagarotetraose and neoagarohexaose. Therefore, the new agar-degrading strain, H9, can be applicable for the production of valuable neoagarooligosaccharides and for the complete degradation of agar in bio-industries.

• Korean Journal of Environmental Agriculture
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• v.31 no.2
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• pp.175-184
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• 2012

BACKGROUND: The aim of this study was to isolate and identify algicidal bacterium that tends to kill the toxic dinoflagellate Alexandrium catenella, and to determine the algicidal activity. METHODS AND RESULTS: Among of four algicidal bacteria isolated in this study, NH-12 isolate was the strongest algicidal activity against A. catenella. NH-12 isolate was identified on the basis of biochemical characteristics and analysis of 16S rRNA gene sequences. The isolate showed 97.67% homology with Pseudoalteromonas prydzensis ACAM $620^T$ (U85855), and was designated Pseudoalteromonas sp. NH-12. The optimal culture conditions of this isolate were $25^{\circ}C$, initial pH 8.0, and 3.0% (w/v) NaCl concentration. The algicidal activity of NH-12 was significantly increased to maximum value in the late of logarithmic phase of bacterial culture. As a result of 'cell culture insert' experiment, NH-12 is assumed to produce secondary metabolites, as an indirect attacker. When 10% culture filtrate of NH-12 was applied to A. catenella, over 99% of algal cells were destroyed within 24 h. In addition, the killing effects were increased in dose and time dependent manners. CONCLUSION(S): Taken together, our results suggest that Pseudoalteromonas sp. NH-12 could be a candidate for controlling of toxic algal blooms.

• KSBB Journal
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• v.26 no.1
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• pp.57-61
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• 2011

The principal objective of this study was to determine the optimal conditions for the production of biosurfactant by the indigenous bacterium, Pseudoalteromonas sp. HK-3, originating from oil-spilled areas. The relationship between total biosurfactant production and the factors affecting biosurfactant production were evaluated by statistical analysis using SPSS software. The effects of various supplemental carbon sources (e.g., glucose, dextrose, mannitol, citrate, acetate) on the maximal production of biosurfactant by the test culture of Pseudoalteromonas sp. HK-3 was then evaluated. As a result, mannitol was found in this study to be the best supplemental carbon source for the production of biosurfactant. A spot inoculation of crude cultural liquid containing the HK-3 cells generated the largest clear zone, whereas only small clear zones appeared around the spots inoculated with either supernatant only or cell pellets following centrifugation. Our results demonstrated that the HK-3 test culture supplemented with 2% mannitol at an initial pH of 6 generated the maximal amount of biosurfactant within 72 h of incubation.

• Korean Journal of Microbiology
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• v.54 no.3
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• pp.305-307
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• 2018

The whole genome sequencing using PacBio RS II platform was performed for a marine bacterium Pseudoalteromonas donghaensis $HJ51^T$ isolated from East Sea of Korea. As a result, three assembled contigs consisting of a chromosome (size of 3,646,857 bp, and G + C content of 41.8%) and two plasmids (size of 842,855 bp and 244,204 bp, and G + C content of 41.3% and 40.4%, respectively) were obtained. The genome included 4,083 protein coding genes and 127 RNA genes. This result could be used for gene sources of biopolymers degradation and the development as a new host with secretion system similar to Escherichia coli.

• Korean Journal of Microbiology
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• v.54 no.3
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• pp.280-282
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• 2018

Pseudoalteromonas sp. meg-B1 belonging to Gammaproteobacteria was isolated from marine sediment in Jeju island. Here, we report the draft genome sequence of strain meg-B1 with a size of approximately 4.15 Mbp and a mean G + C content of 41.2%. The draft genome included 3,606 coding sequences, and 9 ribosomal RNA and 94 transfer RNA genes. In the draft genome, genes (e.g. choline dehydrogenase) involved in the accumulation of compatible solutes required for survival in marine environments have been identified.

• Korean Journal of Microbiology
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• v.45 no.4
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• pp.404-410
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• 2009

We investigated the physicochemical stabilities and biological activities of ethanol- extracted pigment from marine bacteria Pseudoalteromonas sp. Ju11-1 and Pseudoalteromonas sp. Ju14. The bacterial pigment of strain Ju11-1 was very stable at pH 5.0 below $25^{\circ}C$. The stability of the pigment showed higher stability in the presence of metal ions such as $Cu^{2+}$ and $Mg^{2+}$. The pigment has activity of free-radical scavenging ($IC_{50}$ $95.2{\mu}g$/ml) and the protective antioxidant effect ($ED_{50}$ $82.3{\mu}g$/ml) against DNA damage in human lymphocytes. The bacterial pigment of strain Ju14 was very stable at pH range between 4.0 and 8.0 below $40^{\circ}C$. In the presence of light, the pigment was also very stable, showing more than 90 percent of remaining absorbance during 14 days at $25^{\circ}C$. The stability of the pigment, when metal ions were present, showed higher stability in all examined metal ions except for $Fe^{2+}$, $Al^{3+}$, and $Cu^{2+}$, especially in the presence of $Na^+$. The pigment has activity of freeradical scavenging ($IC_{50}$ $208.6{\mu}g$/ml) and the protective antioxidant effect ($ED_{50}$ $ 96.4{\mu}g$/m) against DNA damage in human lymphocytes. The result indicates that the bacterial pigments from marine bacteria, Pseudoalteromonas sp. Ju11-1 and Pseudoalteromonas sp. Ju14 showed higher physicochemical stability and significant effects for reduction in oxidative DNA damage. Therefore, the results suggest that these bacterial pigments could be used as a natural colorant having the advantages of antioxidant.