• Journal of Marine Bioscience and Biotechnology
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• v.7 no.2
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• pp.42-51
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• 2015

The community structure of cultivable bacteria associated with the marine sponge, Petrosia corticata, collected from Jeju Island in summer (September) of 2012 and winter (January) of 2013, were compared by the PCR-ARDRA method. Bacterial strains were cultured for 4 days at $26^{\circ}C$ on Zobell medium and marine agar medium. After PCR amplification of 16S rRNA gene of individual strains, the restriction enzymes MspI and HaeIII were used to make restriction patterns. As a result, 24 ARDRA patterns from the summer sponge and 20 ARDRA patterns from the winter sponge were obtained. The sequencing result of 1-3 selected strains from each pattern showed over 98% similarities with the known sequences from the public database. At the phylum level, the bacterial community structures of both sponges (summer and winter) were identical qualitatively and composed of 4 phyla : Proteobacteria, Actinobacteria, Bacteroidetes, and Firmicutes. Alphaproteobacteria accounted for 42.5% of total in summer sponge and 25.2% in winter, decreasing in the winter sample. Gammaproteobacteria accounted for 27.5% of total in summer sponge and 35.2% in winter, increasing in the winter sample. At the genus and species level, summer sponge had more diverse bacterial communities than winter sponge. Actinobacteria, Bacteroidetes, and Firmicutes increased in the winter sample.

• Korean Journal of Microbiology
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• v.45 no.2
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• pp.170-176
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• 2009

Culture-independent 16S rDNA-DGGE profiling and phylogenetic analysis were used to examine the predominant bacterial communities associated with the two sponges, Dictyonella sp. and Spirastrella abata from Jeju island. The culture-independent approach involved extraction of total bacterial DNA, PCR amplification of the 16S ribosomal DNA using primer pair 341f-GC and 518r, and separation of the amplicons on a denaturing gradient gel. Denaturing gradient gel electrophoresis banding patterns indicated 8 and 7 bands from the two sponge species, Dictyonella sp. and Spirastrella abata, respectively. There were not common major bands in two different sponges. Comparative sequence analysis of variable DGGE bands revealed from 93% to 98% similarity to the known published sequences. The dominant bacterial group of Dictyonella sp. belonged to uncultured Gammaproteobacteria, while, that of Spirastrella abata belonged to uncultured Alphaproeobacteria and Firmicutes. DGGE analysis indicated predominant communities of the sponge-associated bacteria differ in the two sponges from the same geographical location. This result revealed that bacterial community profiles of the sponges were host species-specific.

• Korean Journal of Microbiology
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• v.45 no.2
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• pp.155-162
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• 2009

A cultivation-based approach was employed to compare the culturable bacterial diversity associated with two phylogenetically closely related marine sponges, Spirastrella abata and Spirastrella panis, which have geologically overlapping distribution patterns. The bacteria associated with sponge were cultivated using MA medium supplemented with 3% sponge extracts. Community structures of the culturable bacteria of the two sponge species were analyzed with PCR-RFLP (restriction fragment length polymorphism) based on 16S rDNA sequences. The RFLP fingerprinting of 16S rDNA digested with HaeIII and MspI, revealed 24 independent RFLP types, in which 1-5 representative strains from each type were partially sequenced. The sequence analysis showed >98.4% similarity to known bacterial species in public databases. Overall, the microbial populations of two sponges investigated were found to be the members of the classes; Alphaproteobacteria, Gammaproteobacteria, Firmicutes, and Actinobacteria. The Alphaproteobacteria were predominant in the bacterial communities of the two sponges. Gammaproteobacteria represented 38.5% of bacterial community in S. abata. Whereas only 1.6% of this class was present in S. panis. Bacillus species were dominat in S. panis. Bacillus species were found to be 44.3% of bacterial species in S. panis, while they were only 9.7% in S. abata. It is interesting to note that Planococcus maritimus (8.1%, phylum Firmicutes) and Psychrobacter nivimaris (28.9%, phylum Gammaproteobacteria) were found only in S. abata. This result revealed that profiles of bacterial communities from the sponges with a close phylogenetic relationship were highly species-specific.

• Korean Journal of Microbiology
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• v.46 no.3
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• pp.255-261
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• 2010

The bacterial community structure associated with two marine sponges, Hyrtios sp. 604 and Callyspongia sp. 612 collected from the South Pacific Ocean were analyzed by 16S rDNA-denaturing gradient gel electrophoresis (DGGE). The phylogenetic analysis showed that the bacterial community associated with Hyrtios sp. 604 contained diverse bacterial groups such as Chloroflexi, Firmicutes, Cyanobacteria, Alphaproteobacteria, Gammaproteobacteria, Actinobacteria, and Acidobacteria. Callyspongia sp. 612 harbored Chloroflexi, Cyanobacteria, Alphaproteobacteria, and Gammaproteobacteria. Hyrtios sp. 604 belonging to genus Hyrtios known to produce natural products showed greater bacterial diversity than Callyspongia sp. 612. Phylum Actinobacteria was shown to be one of dominant bacterial groups in Hyrtios sp. 604. Although the same phyla of bacteria were found in both sponge species, the spongeassociated predominant bacterial groups differed between the two sponges with different chemical characteristics from the same geographical location. Uncultured bacteria represented over 90% of the bacteria diversity present in all bacterial communities of the sponges.

• Korean Journal of Microbiology
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• v.46 no.2
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• pp.177-182
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• 2010

The bacterial community structure of two marine sponges, Spirastrella abata and Cinachyrella sp. collected from Jeju Island, in April 2009, was analyzed by 16S rDNA-denaturing gradient gel electrophoresis (DGGE). DGGE banding patterns indicated 8 and 7 bands for Spirastrella abata and Cinachyrella sp., respectively. Comparative sequence analysis of variable DGGE bands revealed from 92% to 100% similarity to the known published sequences. The bacterial groups associated with Spirastrella abata were Alphaproteobacteria and Deltaproteobacteria. The bacterial community of Cinachyrella sp. consisted of Alphaproteobacteria, Gammaproteobacteria, and Actinobacteria. Alphaproteobacteria was common and predominant in both the sponge species. Deltaproteobacteria was found only in Spirastrella abata while Actinobacteria and Gammaproteobacteria were found only in Cinachyrella sp. The results revealed that though the common bacterial group was found in both the sponges, the bacterial community profiles differed between the two sponge species obtained from the same geographical location.

• Journal of Marine Bioscience and Biotechnology
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• v.6 no.2
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• pp.76-83
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• 2014

Bacteria associated with marine sponges seemed to be concerned in halogenation/dehalogenation process of natural compounds. In the present study, the effect of hexabromobenzene (HBB) on the community structure of bacteria associated with a marine sponge Axinella sp. from Chuuk State under anaerobic condition was investigated. Regardless of 100 ppm HBB, most of detected microorganisms displayed high similarity with clones reported from coral or sponges. Amongst, Desulfovibrio marinisediminis like clones were dominant. Clones affiliated with Lentisphaerae and Fusibacter paucivorans (Clostridia) were detected at the conditions without HBB but clones affiliated with Vallitalea guaymasensis (Clostridia) increased its proportion with HBB. From these results and previous reports clones affiliated with D. marinisediminis and V. guaymasensis seemed to be concerned in halogenation/dehalogenation process.

• Korean Journal of Microbiology
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• v.46 no.4
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• pp.366-374
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• 2010

Culture-dependent RFLP and culture-independent DGGE were employed to investigate the bacterial community associated with the marine sponge Spirastrella abata. A total of 164 bacterial strains associated with the sponge were cultivated using Zobell and Natural sea salt media. PCR amplicons of the 16S rDNA from the bacterial strains were digested with the restriction enzymes HaeIII and MspI, and then assigned into different groups according to their restriction patterns. The 16S rDNA sequences derived from RFLP patterns showed more than 95% similarities compared with known bacterial species, and the isolates belonged to four phyla, Proteobacteria (Alphaproteobacteria, Gammaproteobacteria), Actinobacteria, Firmicutes, and Bacteriodetes, of which Alphaproteobacteria was dominant. DGGE fingerprinting of 16S rDNAs amplified from the sponge- derived total gDNA showed five major DGGE bands, and their sequences showed more than 96% similarities compared with available sequences. The sequences derived from DGGE bands revealed high similarity with the uncultured bacterial clones. DGGE revealed that bacterial community consisted of four phyla, including Proteobacteria (Alphaproteobacteria, Gammaproteobacteria), Actinobacteria, Spirochetes, and Chloroflexi. Alphaproteobacteria, Gammaproteobacteria, and Actinobacteria were commonly found in bacteria associated with S. abata by both RFLP and DGGE methods; however, overall bacterial community in the sponge differed depending on the analysis methods.

• Natural Product Sciences
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• v.20 no.3
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• pp.202-205
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• 2014

Three compounds including 7,7-bis(3-indolyl)-p-cresol (1), cyclo-(S-Pro-R-Leu) (2) and cyclo-(S-Pro-R-Val) (3) were isolated from the strain of Bacillus megaterium LC derived from the marine sponge Haliclona oculata. All the isolated compounds showed antimicrobial activity at MIC values ranging from 0.005 to $5{\mu}g/mL$ against Gram-negative bacteria Vibrio vulnificus and V. parahaemolyticus, gram-positive bacteria Bacillus cereus and Micrococcus luteus, and the dermatophyte Trichophyton mentagrophytes. The results suggested that these compounds might have potential to be developed as agents treating dermatosis and controlling vibriosis in aquaculture.

• Korean Journal of Microbiology
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• v.48 no.4
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• pp.275-283
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• 2012

Culture-dependent RFLP and culture-independent DGGE were employed to investigate the bacterial community associated with the marine sponge Asteropus simplex collected from Jeju Island. A total of 120 bacterial strains associated with the sponge were cultivated using modified Zobell and MA media. PCR amplicons of the 16S rDNA from the bacterial strains were digested with the restriction enzymes HaeIII and MspI, and then assigned into different groups according to their restriction patterns. The 16S rDNA sequences derived from RFLP patterns showed more than 94% similarities compared with known bacterial species, and the isolates belonged to five phyla, Alphaproteobacteria, Gammaproteobacteria Actinobacteria, Bacteroidetes, and Firmicutes, of which Gammaproteobacteria was dominant. DGGE fingerprinting of 16S rDNAs amplified from the sponge-derived total gDNA showed 12 DGGE bands, and their sequences showed more than 90% similarities compared with available sequences. The sequences derived from DGGE bands revealed high similarity with the uncultured bacterial clones. DGGE revealed that bacterial community consisted of seven phyla, including Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Deltaproteobacteria, Actinobacteira, Chloroflexi, and Nitrospira. Alphaproteobacteria, Gammaproteobacteria, and Actinobacteria were commonly found in bacteria associated with A. simplex by both RFLP and DGGE methods, however, overall bacterial community in the sponge differed depending on the analysis methods. Sponge showed more various bacterial community structures in culture-independent method than in culture-dependent method.

• 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.