• Proceedings of the Korean Society of Life Science Conference
• /
• 2007.10a
• /
• pp.109.1-109.1
• /
• 2007

See Full Text

• Journal of Practical Agriculture & Fisheries Research
• /
• v.23 no.1
• /
• pp.81-87
• /
• 2021

Olive flounders which were cultured in commercial fish farm showed abnormal swimming behavior in November 2020. Then, gradual mortality was observed in the fish farm. During the diagnosis, bacterial strain KNCFKW-PN1 was isolated from the kidney of the dead fish. Based on the sequence of gyrase B subunit gene, KNCFKW-PN1 was proved to be Pseudoalteromonas nigrifaciens, showing 99.59% nucleotide identity with that of P. nigrifaciens LMG 2227^T. According to the result of antibiotic susceptibility test, P. nigrifaciens KNCFKW-PN1 showed intermediate resistance to ciprofloxacin, and was resistant to ampicillin, cefepime, cefotaxime, ceftazidime, and amikacin. This is the first report of the isolation of multiple-antibiotic-resistant P. nigrifaciens from olive flounder.

• Korean Journal of Microbiology
• /
• v.40 no.4
• /
• pp.302-306
• /
• 2004

A total of forty strains of pigment-producing halophilic bacteria were isolated from the solar saltern and coastal seawater in Korea. The diversity of those bacteria were determined on the basis of PCR-RFLP and 16S rDNA sequences. The isolated strains were clssified into nine genera: Pseudoalteromonas, Photobacterium, Vibrio, Halobacillus, Bacillus, Paracoccus, Salinicoccus, Tenacilbaculum, and Flavobacterium. While more than $80\%$ of the pigment-producing halophilic bacteria isolated from the coastal seawater were classified as gram-negative Pseudolateromonas, most of the strains isolated from the solar saltern were classified into gram-positive Halo­bacillus. The other strain was KK7, which may be identified as novel species belonging to the genus, Salini­coccus.

• Journal of Life Science
• /
• v.22 no.12
• /
• pp.1724-1728
• /
• 2012

A marine bacterial strain that degraded fucoidan from Ecklonia cava was isolated from seawater. The crude fucoidanase of this strain efficiently degraded fucoidan at pH 8 and $50^{\circ}C$. The crude enzyme hydrolyzed 7.1% (w/w) fucoidan within 24 hrs from an 1% (w/v) fucoidan solution and produced oligosaccharides by endo-type hydrolysis as the reaction products. The results of 16S rRNA gene sequence analysis and biochemical tests permitted a tentative identification of strain SB 1493 as a Pseudoalteromonas species.

• Microbiology and Biotechnology Letters
• /
• v.35 no.1
• /
• pp.11-16
• /
• 2007

The arylsulfatase gene (astA, 984 bp ORF) from Pseudoalteromonas carrageenovora genome was amplified by PCR and subcloned into the pHCE-IA vector, in which the hyper consitutive expression (HCE) promoter from the D-amino acid aminotransferase (D-AAT) gene of Geobacillus toevii was employed. The transformant cell, Escherichia coli BL21 (DE3)/pHCE-AST, on LB agar plate containig 4-methylumbelliferyl sulfate, showed an intense fluorescence at 360 nm, indicating that 4-methylumbelliferone was liberated by desulfatate activity. When BL21 (DE3)/pHCE-AST was grown on LB media containing 0.4% glucose or 0.4% glycerol, the arylsulfatase activity was higher at glycerol rather than at glucose. On 2% glycerol medium, the arylsulfatase activity reached 15.0 unit/ml, which was 2.6-fold higher expression level than that with 1% glycerol. The DNA ladder in agarose prepared from agar by this recombinant enzyme revealed similar resolution and migration patterns with a commercial agarose. This results suggests that arylsulfatase overexpressed in E. coli could be applicable to the economic production of electrophoretic-grade agarose.

• Korean Journal of Microbiology
• /
• v.51 no.4
• /
• pp.364-373
• /
• 2015

As an effort to utilize alginate, 103 bacterial isolates that were positive for the alginate lyase activity were isolated from various clams and seawater samples collected in Incheon coastal area. Among them, 3 strains (M1-2-1, M6-1, and C8-15) were finally selected for further analysis based on their activities at higher levels than others. These isolates were all Gram-negative and rod shaped halophilic bacteria with motility. According to their physiological and biochemical properties as well as DNA sequence of their 16S rRNA genes, M1-2-1 and M6-1 were identified as a member of genus Pseudoalteromonas and C8-15 belonged to genus Vibrio. They exhibited the alginate degrading activity at the maximal level when they were cultured in APY broth for 6-8 h at $25^{\circ}C$. Both their growth and the enzyme activity were greatly enhanced when NaCl was added to the growth medium. The crude alginate lyases from the supernatants of the bacterial cultures showed the highest activity at $45^{\circ}C$ and pH 7.0-8.0. M1-2-1 and M6-1 produced 2.723 and 1.976 g/L of reducing sugar from alginate, respectively, suggesting that they have potential for commercial application.

• Korean Chemical Engineering Research
• /
• v.49 no.6
• /
• pp.840-845
• /
• 2011

Degradation products of agarose are biologically active and thus used as an ingredient in pharmaceuticals or functional cosmetics. Furthermore, it has been strongly considered as a substrate for bio-ethanol fermentation. Recently, we isolated new agarase-producing microorganism, Pseudoalteromonas sp. from south sea of Korea. In this study, we aimed to separate and purify the agarase from culture broth of this strain. Separation of agarase was performed by ion- exchange chromatography on DEAE-Sepharose resin. Equilibrium pH and volume ratio of resin to the amount of protein were optimized for the efficient adsorption of protein. 410 ${\mu}g$ of protein was completely adsorbed to 3 mL of resin at pH 7.5. The total amount of eluted protein increased as NaCl concentration increased to 400 mM at isocratic elution. Agarase was separated by linear gradient elution of NaCl (0~1,000 mM). Three major protein peaks were observed and the presence or absence of agarase in these eluted proteins was measured by Lugol's staining technique. Only six eluted protein fractions showed strong agarase activity.

• Microbiology and Biotechnology Letters
• /
• v.40 no.1
• /
• pp.1-9
• /
• 2012

This study's aim was to isolate microorganisms producing agarase with a high activity, with possible applications in improving the performance of the pretreatment processes for bioethanol production. Marine algaes were collected from the south coast of Korea, from which three kinds of microorganisms were isolated. After a 4-day culture of these strains at $25^{\circ}C$, crude enzymes were obtained from culture supernatant or cell-free extract by ammonium sulfate precipitation and membrane dialysis. Agarase activity was observed in these crude enzymes. Notably higher specific activity was observed in the crude enzyme obtained from the culture supernatant rather than that from the cell-free extract. This indicates that a secreted enzyme has a much greater activity than a cellular enzyme. Crude enzymes from the GNUM08122 strain were inferred to have ${\alpha}$-agarase activity because release of p-nitrophenol was observed, possibly due to the cleavage of p-nitrophenyl-${\alpha}$-D-galactopyranoside. The 16S rRNA sequence of GNUM08122 showed a close relationship to Pseudoalteromonas issachenkonii KMM 3549 (99.8%) and Pseudoalteromonas tetraodonis IMA 14160 (99.7%), which led us to assign it to the genus Pseudoalteromonas. Biochemical and physiological study revealed that this strain can grow well at $40^{\circ}C$ under a wide range of pH (pH 4~8) in high-salt conditions (10% NaCl).

• 한국생물공학회:학술대회논문집
• /
• 2003.04a
• /
• pp.571-575
• /
• 2003

A marine aerobic Gram-negative bacterium, Pseudoalteromonas carrageenovora, has been blown to hydrolyze carrageenans, the sulfated galactans of red algae, and to desulfate oligo kappa-carrageenans. Recently, the gene encoding arylsulfatase (aryl-sulfate sulfohydrolase, E.C.3.1.6.1) of A. carrageenovora was cloned and the nucleotide sequence was reported. Enzymatic hydrolysis of sulfate groups in agaropectin simplifies the process of agarose preparation. In order to overproduce the enzyme, the arylsulfatase gene (astA, 984 bp ORF) from P. carrageenovora genome was amplified by PCR and subcloned into the pET21a vector. When the constructed plasmid pAST-A1 was introduced into E, coli BL21(DE3), the transformant on LB plate containing IPTG showed the hydrolyzing activity for p-nitrophenyl sulfate. Most of arylsulfatase activity was found in the cell lysate, but at $50\;{\sim}\;5000\;{\mu}M$ IPTG concentration the activity was found both in the culture supernatant and the cell lysate. The molecular weight of the recombinant enzyme was estimated to be 34 kDa by SDS-PAGE.

• Ocean and Polar Research
• /
• v.27 no.2
• /
• pp.215-219
• /
• 2005

We isolated and identified three protease-producing bacteria that had inhabited the region around the Korean Arctic Research Station Dasan located at Ny-Alesund, Svalbard, Norway $(79^{\circ}N,\;12^{\circ}E)$. Biofilms were collected from the surface of a floating pier and from dead brown algae in a tide pool near the seashore. The biofilm samples were transported to the Korea Polar Research Institute (KOPRI) under frozen conditions, diluted in sterilized seawater, and cultured on Zobell agar plates with 1% skim milk at $10^{\circ}C$. Three clear zone forming colonies were selected as protease-producing bacteria. Phylogenetic analysis based on 16S rDNA sequences showed that these three stains shared high sequence similarities with Pseudoalteromonas elyakovii, Exiguobacterium oxidotofewm Pseudomonas jessenii, respectively. We expect these Arctic bacteria may be used to develop new varieties of protease that are active at low temperatures.