• Journal of Microbiology
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• 제45권6호
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• pp.510-514
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• 2007

Twenty-five bacterial strains that secrete mucous materials were isolated from sediment obtained from King George Island, Antarctica. Seven of these strains proved capable of producing cryoprotective exopolysaccharides. The strain KOPRI 21653 was selected for the further study of an anti-ice-nucleating polysaccharide (ANP), which originated from a polar region. KOPRI 21653 was identified as Pseudoalteromonas arctica as the result of 16S rRNA analysis. The exopolysaccharide, P-21653, was purified completely from the KOPRI 21653 cell culture via column chromatography and protease treatment. The principal sugar components of P-21653 were determined to be galactose and glucose, at a ratio of 1:1.5, via GC-MS analysis. The cryoprotective activity of P-21653 was characterized via an E. coli viability test. In the presence of 0.1% (w/v) P-21653, the survival ratio of E. coli cells was as high as 82.6% over three repeated freeze-thaw cycles. The survival ratio decreased drastically to 71.5 and 48.1 %, respectively, in five and seven repeated cycle conditions; however, the survival ratios were greater over three (96.6-92.1%) to seven (100.5-91.6%) freeze-thaw cycles in the presence of 0.5 and 1.0% (w/v) P-21653. In addition, at much lower concentrations (0.1-1.0%), P-21653 resulted in survival ratios (83.1-98.4%) similar to those of two commercially available cryoprotectants ($V_{EG}$ plus X-1000, 92.9% and $V_{M3}$, 95.3%), which were utilized at the recommended concentrations (90%). The biochemical characteristics of exopolysaccharide P-21653 reflect that this compound may be developed as a useful cryoprotectant for use in medical applications and in the food industry.

• Journal of Microbiology and Biotechnology
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• 제30권8호
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• pp.1195-1206
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• 2020

Beringraja pulchra, Cham-hong-eo in Korean, is a mottled skate which is belonging to the cartilaginous fish. Although this species is economically valuable in South Korea as an alkaline-fermented food, there are few microbial studies on such fermentation. Here, we analyzed microbial changes and pH before, during, and after fermentation and examined the effect of inoculation by a skin microbiota mixture on the skate fermentation (control vs. treatment). To analyze microbial community, the V4 regions of bacterial 16S rRNA genes from the skates were amplified, sequenced and analyzed. During the skate fermentation, pH and total number of marine bacteria increased in both groups, while microbial diversity decreased after fermentation. Pseudomonas, which was predominant in the initial skate, declined by fermentation (Day 0: 11.39 ± 5.52%; Day 20: 0.61 ± 0.9%), while the abundance of Pseudoalteromonas increased dramatically (Day 0: 1.42 ± 0.41%; Day 20: 64.92 ± 24.15%). From our co-occurrence analysis, the Pseudoalteromonas was positively correlated with Aerococcaceae (r = 0.638) and Moraxella (r = 0.474), which also increased with fermentation, and negatively correlated with Pseudomonas (r = -0.847) during fermentation. There are no critically significant differences between control and treatment. These results revealed that the alkaline fermentation of skates dramatically changed the microbiota, but the initial inoculation by a skin microbiota mixture didn't show critical changes in the final microbial community. Our results extended understanding of microbial interactions and provided the new insights of microbial changes during alkaline fermentation.

• 생명과학회지
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• 제24권10호
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• pp.1110-1117
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• 2014

갯벌은 영양염 순환과 오염물질 제거 등 환경적으로 가지는 의미가 크다. 이에 갯벌에서 아밀라아제, 셀룰라아제, 프로테아제, 리파아제를 생성하여 유기물을 분해하는 190균주를 순수분리하고 효소 활성 실험을 통해 그 중 활성이 좋은 균주를 선택하였으며 16S rRNA유전자 분석을 통해 Pseudoalteromonas sp. IC35 (KE804087)로 동정하였다. 또 무기황(S0)을 에너지원으로 하여 최소배지에서 생존 가능한 황산화 세균을 31균주를 순수 분리하고 sulfate ion을 측정하여 황산화능력이 뛰어난 세균을 16S rRNA 분석하여 Halothiobacillus neapolitanus IC_S22 (KE804088)로 동정하였다. 이렇게 분리한 균주들이 환경에서 활성을 가지는 지를 측정하기 위해 유기물 분해에 대해(대조군 M1, 접종군 M2)와 황산화능력에 대해(대조군 M3, 접종군 M4)의 microcosam 반응기를 각각 구축하였다. Pseudoalteromonas sp. IC35를 접종한 M1에서 비접종군인 M2에 비해 유기물 분해 효율이 증가되었으며 microcosm 반응기 내의 생물량도 증가됨을 확인하였다. H. neapolitanus IC_S22을 접종한 M3에서 비접종군인 M4에 비해 환원성 황($S_0$)을 산화하는 능력이 상승되었으며 real time PCR을 이용하여 pilot 내에서 세균의 생물량이 증가함에 따라 초기 sulfate ion의 양도 증가함을 확인하였다. 이번 연구를 통해 대사활성이 우수한 균주를 부가적으로 환경에 접종함으로써 기존 세균 군집에 의한 환경정화 능력을 증가 시키는 것으로 사료된다.

• 생명과학회지
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• 제19권8호
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• pp.1132-1138
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• 2009

해양세균 Pseudoalteromonas psicida TA20의 ethanol 색소 추출물에 대한 안정성과 기능성을 검토한 결과, 본 해양세균 색소는 pH 4.0에서 pH 8.0의 조건과 $40^{\circ}C$ 이하에서 매우 안정하였으며, $25^{\circ}C$ 14일간 90% 이상의 잔존율을 나타내어 빛에 대한 안정성이 매우 우수한 것으로 나타났다. 금속이온의 경우 $Al^{3+}$과 $Cu^{2+}$를 제외한 실험된 모든 금속이온에 대하여 매우 안정하였으며 특히, 다른 색소추출물들에 비해 $Fe^{2+}$에 대한 안정성이 높은 것으로 나타났다. 해양세균 색소추출물에 대한 생리활성 실험 결과, free radical 소거 활성 (3495 ${\mu}g/ml$)이 나타났으며, 색소농도 10 ${\mu}g/ml$일 때 44%의 인체세포 DNA 손상 회복능이 있음을 보여 항산화능이 우수함을 알 수 있었다. 또한 주요 식품유해세균에 대하여 항균활성을 나타내었다. 따라서 본 해양세균 색소는 항산화능과 항균활성을 갖는 식품 색소로의 적용을 검토할 수 있을 것으로 기대된다.

• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2006년도 제47회 학술심포지움 및 추계국제학술대회
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• pp.102.2-102.2
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• 2006

• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2003년도 제40회 국제학술심포지움
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• pp.102-102
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• 2003

• Journal of Microbiology and Biotechnology
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• 제14권5호
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• pp.952-958
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• 2004

Of 23 psychrotrophic bacteria isolated from the west Pacific deep-sea sediments, 19 were assigned to the $\gamma$-Proteobacteria, 3 to the <$\beta$-Proteobacteria, and 1 to the Gram-positive bacteria, as determined by their 16S rDNA sequences. Ten psychrotrophs, affiliated to the Psychrobacter, Pseudoalteromonas, and Pseudomonas genera in the $\gamma$-Proteobacteria group, were screened for lipolytic bacteria. The majority of the lipolytic isolates had growth temperatures between 4-$30^\circ{C}$, and all of them were neutrophilic, aerobic, or facultatively anaerobic, and some were able to produce multiple kinds of ectohydrolytic enzymes. The deep-sea strains Psychrobacter sp. wp37 and Pseudoalteromonas sp. wp27 were chosen for further lipase production analysis. Both strains had the highest lipase production when grown at 10 to $20^\circ{C}$; their highest lipase production occurred at the late-exponential growth stage; and the majority of the enzymes were excreted to the outside of the cells. Lipases from both strains had the same optimal reaction temperature and pH (20-$30^\circ{C}$, pH 7-8) and could retain about 60% of their highest activity at $4^\circ{C}$. Furthermore, SDS-PAGE and an in-gel activity test showed that they had the same high molecular mass of about 85 kDa.

• KSBB Journal
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• 제29권4호
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• pp.278-284
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• 2014

We present microfluidic method to rapidly analyze the effect of temperature on the change of morphologies of Antarctic bacteria (Pseudoalteromonas sp., Shewanella vesiculosa, Shewanella sp., and Cellulophaga sp.). The microfluidic device is able to generate stable temperature gradient from 7 to$40^{\circ}C$ and dramatically reduce the number of experiments, experimental cost and labor, and amount of sample. Based on this approach, we found that specific bacteria transforming morphology into filament or elongated body strongly depends on cultivation temperature. Interestingly, we found that the morphologies of Pseudoalteromonas sp., Shewanella vesiculosa, Shewanella sp., and Cellulophaga sp. are elongated at below $25^{\circ}C$, above $20^{\circ}C$, above $15^{\circ}C$ and above $35^{\circ}C$, respectively. We envision the microfluidic device is a useful approach to analyze biological events with a high throughput manner.

• 한국동물위생학회지
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• 제37권3호
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• pp.203-212
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• 2014

Frog culture industry is not yet familiar but has much potential. Generally, in farm, the population density is higher than that of in nature and frog farm is not the exception. But when population density is high, it can easily leads to stressful condition, poor sanitation. When a disease occur, it is a primary factor that makes the population more susceptible and the results more grave. Because of severe Rhabditoidea- helminth infection and subsequent bacterial septicemia, 50~70% of the total population had been died in a farm in Jeong-sun in Gangwon-do and Chungju in Chungcheongbuk-do from late June, 2012 to September, 2012. Diseased frogs showed ruptured lung, bloody ascites, liver discoloration, myocardium weakness, congested kidney, microcytic anemia and so on. Enterobacteriacea, Citrobacter.sp, Cupriavidus metallidurans, Acinetobacter.sp were isolated as major bacterium that had caused septicemia in frogs. Among isolated bacterium, Cupriavidus metallidurans, Ewingella americana, Shewanella aquimarina and Pseudoalteromonas sp. have not reported as potential pathogens in frogs before. It is a good example that severe helminth infection in frogs can lead to secondary infection of bacteria.

• 한국미생물·생명공학회지
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• 제37권4호
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• pp.355-360
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• 2009

P. carageenovora 유래 arylsulfatase 유전자(astA)의 효모표면발현 plasmid pCTAST(7.1 kb)를 구축하여 S. cerevisiae EBY100에 형질전환하였고, 선별된 형질전환체들을 YPDG 배지에서 배양했을 때 4-methylumbelliferyl sulfate를 분해하여 형광을 보였다. 이는 효모에서 arylsulfatase가 활성형으로 생산되었음을 의미하였다. S. cerevisiae EBY100/pCTAST를 플라스크 배양했을 때 arylsulfatase 활성은 galactose가 완전히 소모된 배양 48시간째 최대 활성인 1.2 unit/mL로 나타났으며 배양 상등액에서는 활성이 나타나지 않았다. 효모 S. cerevisiae의 세포표면에서 발현된 재조합 arylsulfatase로 제조된 agarose를 agar와 시판용 agarose와 함께 ${\lambda}DNA$ HindIII marker를 사용하여 DNA 전기영동 성능을 비교 실험한 결과, agar보다는 재조합 arylsulfatase 처리로 제조한 agarose가 이동성이나 분리능에서 우수하였으며, 시판용 agarose와 비교하여 이동성이나 분리능이 유사한 결과를 확인할 수 있었다. 본 연구의 결과, 효모 S. cerevisiae의 세포표면에서 발현된 재조합 arylsulfatase 효소를 이용하여 한천으로부터 전기영동용 고순도 친환경적인 agarose 생물 생산 공정에 적용 가능함을 알 수 있었다.