• Toxicological Research
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• 제14권3호
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• pp.435-441
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• 1998

Alginase$Alginase^{ⓡ}$ (Arginine esterase) is one of the snake venoms which is mainly consisted of arginine esterase and acts as a thrombus -forming inhibitor/thrombus-lysin. These present studies were performed to investigate of the acute and subacute toxicity of the Alginase$Alginase^{ⓡ}$ in rats. In acute toxicity study, rats were single administered intravenously with dosages of 0.001, 0.01. 0.1, 1 and 10U/kg B.W. and examined the number of death, clinical sign, body weight and pathological change for 7days after administration of Alginase$Alginase^{ⓡ}$. At maximum dose level (10U/kg B.W.), Alginase$Alginase^{ⓡ}$ induced symptoms of shock with cyanosis and dyspnea. But these symptoms dissappeared after 30~50 minutes and we could not find any other toxic effect in rats. Therefore, $LD_{50}$ Value of Alginase was over 10U/kg B.W. in rats. In four-week intravenous toxicity study of Alginase$Alginase^{ⓡ}$, rats were administered intravenously seven days per week for 28 days, with dosages of 0, 0.0125, 0.125 and 1.25U/kg B.W./day, respectively. Alginase$Alginase^{ⓡ}$ did not caused any death and showed any clinical signs in rats. No significant Alginase$Alginase^{ⓡ}$ -related changes were found in feed uptake, water consumption, hematology, serum biochemistry, urinalysis, ocular examination, organ weight and histopathological examination. From the results, Alginase$Alginase^{ⓡ}$ seems not to have any toxic effect in rats when it were given daily intravenous injections below the dosage 1.25U/kg B.W./day for four weeks.

• 생명과학회지
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• 제13권5호
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• pp.718-722
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• 2003

다양한 종류의 미역으로부터 알긴산 분해 활성을 가지는 해양 미생물인 Vibrio sp. AEBL-211을 분리하였다. 우선, 알긴산 고체배지에서의 halo 크기가 크고 환원당 생성량이 가장 많은 해양 미생물을 선발하고, 생화학 및 영양적인 특성 분석 결과 등을 바탕으로 Vibro 속으로 동정하였다. DE 52-cellulose 및 Sephacryl G-200를 이용한 음이온 교환 및 gel permeation chromatography를 통해 부분 정제된 alginase 효소액을 얻었다. 부분 정제된 alginase 효소를 이용하여 guluronic acid에 대한 mannuronic acid의 비율이 다른 sodium alginate 시료에 대한 분해능을 평가한 결과, guluronic acid의 상대적 양이 많은 알긴산에 대한 활성이 상대적으로 우수하여 Vibrio sp. AEBL-211 유래의 alginase는 guluronic acid가 많은 G-rich block에 대한 분해활성이 높은 특성을 가지는 것으로 판단되었다.

• 한국미생물·생명공학회지
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• 제23권4호
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• pp.432-438
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• 1995

The intracellular alginase from Vibrio sp. AL-145 was purified by ion chromatography on DEAE-Cellulose column, Q-Sepharose column, and gel filtration on Sephadex G-100 column. The optimum pH and temperature for the activity of the purified intracellular enzyme were 8.0 and 37$\circ$C, respectively. The enzyme was stable at the pH range of 7.5-8.5, and at 30$\circ$C for 30 min. The molecular weight of the intracellular enzyme was estimated to be about 23, 000 daltons by SDS-polyacrylamide gel electrophoresis. NaCl was required for enzyme activity and the optimum concentration was 0.5 M. The activity of intracellular enzyme was inhibited by Co$^{2+}$, Hg$^{2+}$, Zn$^{2+}$, 0-phenanthroline, $\rho$-CMB, EDTA and iodoacetate, and stimulated by Ca$^{2+}$, L-cysteine and 2-mercaptoethanol. This enzyme was an alginase specifically degrading alginic acid.

• 한국식품영양과학회지
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• 제35권2호
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• pp.231-237
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• 2006

알긴산을 선택적으로 분해하는 Bacillus licheniformis AL-577 균주가 생산하는 균체 외 효소를 정제하고 특성을 밝혔다. CM-Cellulose, DEAE-Sepharose 및 Gel 크로마토 그래프 등의 순서대로 정제하여 정제도가 약 98배 정도인 효소를 얻었다. 정제효소의 활성 최적 pH 및 온도는 5.0 및 $35^{\circ}C$이었고, pH 5.5이하와 pH 9.5이상의 반응조건에서는 불안정하였으며, $20^{\circ}C$이상의 온도에서는 불활성화가 쉽게 일어나는 효소였다. 얻어진 효소를 SDS-PAGE로 분자량을 측정한 결과 25,500 Da으로 추정되었다. NaCl 0.2 M 농도에서 최대의 활성을 나타내었고, 무첨가시에도 활성은 약간 나타내었다. $Cu^{2+},\;Fe^{2+},\;Mg^{2+},\;Zn^{2+}$ 등과 같은 2가 금속이 온에 의해서는 활성이 현저히 억제되었고, $K^+,\;Li^+$ 이온에 의해서 활성이 촉진됨을 알 수 있었다. 화학약품인 dithiothreitol 와 O-phenanthroline의 첨가로 인하여 약간의 활성 증가를 가져 왔으며 반면 EDTA, L-cysteine는 현저하게 감소하였다. 이 효소는 알긴산에만 특이적으로 작용함으로써 본 효소는 alginase 또는 alginate lyase인 것으로 판단되었다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제11권5호
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• pp.382-390
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• 2006

Various microorganisms were isolated from the surface waters and sediments of eutrophic lakes and reservoirs in Korea to enable an investigation of bacteria having algal lytic activities against Anabaena flos-aquae when water blooming occurs and to study enzyme profiles of algal lytic bacteria. Two bacterial strains, AFK-07 and AFK-13, were cultured, characterized and identified as Acinetobacter johnsonii and Sinorhizobium sp., respectively. The A. johnsonii AFK-07 exhibited a high level of degradatory activities against A. flos-aquae, and produced alginase, caseinase, lipase, fucodian hydrolase, and laminarinase. Moreover, many kinds of glycosidase, such as ${\beta}-galactosidase,\;{\beta}-glucosidase,\;{\beta}-glucosaminidase,\;and\; {\beta}-xylosidase$, which hydrolyzed ${\beta}-O-glycosidic$ bonds, were found in cell-free extracts of A. johnsonii AFK-07. Other glycosidases such as ${\alpha}-galactosidase,\;{\alpha}-N-Ac-galactosidase,\;{\alpha}-mannosidase,\; and\;{\alpha}-L-fucosidase$, which cleave ${\alpha}-O-glycosidic$ bonds, were not identified in AFK-07. In the Sinorhizobium sp. AFK-13, the enzymes alginase, amylase, proteinase (caseinase and gelatinase), carboxymethyl-cellulase (CMCase), laminarinase, and lipase were notable. No glycosidase was produced in the AFK-13 strain. Therefore, the enzyme system of A. johnsonii AFK-07 had a more complex mechanism in place to degrade the cyanobacteria cell walls than did the enzyme system of Sinorhizobium sp. AFK-13. The polysaccharides or the peptidoglycans of A. flos-aquae may be hydrolyzed and metabolized to a range of easily utilized monosaccharides or other low molecular weight organic substances by strain AFK-07 of. A. johnsonii, while the products of polysaccharide degradation or peptidoglycans were more likely to be utilized by Sinorhizobium sp. AFK-13. These bacterial interactions may offer an alternative effective approach to controlling the water choking effects of summer blooms affecting our lakes and reservoirs.

• 생태와환경
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• 제36권2호통권103호
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• pp.108-116
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• 2003

부영양화 현상을 나타내는 석촌호수와 팔당호의 표층수와 저니로부터 178개의 균주를 분리한 후, Anabaena cylindrica lawn 상에서 plaque를 형성하는 9개의 균주를 선별하였으며 이들 중에서 남조류 생장 억제 능력이 가장 우수한 AK-07를 선발하였다. AK-07의 특성과 16S rDNA의 염기 서열 분석을 기초로 하여 유연관계를 조사한 결과, 형태적, 생리적 생화학적 특징들은 Acinetobacter속의 특성들과 유사하였으며, 165 rDNA의 염기 서열 분석한 결과는 Acinetobacter johnsonii와 99.5%의 유사성을 나타내어, Acinetobacter johnsonii AK-07로 명명하였다. 남조류 분해 특성을 조사하기 위해서, AK-07를 A. cylindrica와 혼합 배양시 접종 2일 후에 남조류의 분해가 관찰되었고, 접종 10일 후에는 남조류가 완전히 사멸하였으며, AK-07의 세포 수는 $8\;{\times}\;10^8\;cfu\;ml^{-1}$까지 증가하였다. 그러나 배양 상등액을 A. cylindrica와 혼합 배양 하였을 때에는 남조류의 분해는 관찰 되지 않았다. 따라서 AK-07는 남조류를 직접 접촉하여 분해하는 것으로 사료되어, AK-07에 세포에 존재하는 효소의 활성을 조사한 결과 Pretense와 glycanases중에서 ${\beta}$-Xylosidase의 활성이 가장 높았으며, Alginase, Laminarinase, Lipase, ${\beta}$-Galactosidase 및 ${\beta}$-Glucosidase의 활성도 높은 수준으로 관찰되었다. A. johnsonii AK-07은 A. cylindrica의 polysaccharides나 peptidoglycans를 monosaccharides이나 저분자 유기물로 분해하는 것으로 여겨진다.

• 환경생물
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• 제22권1호
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• pp.184-191
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• 2004

To investigate bacteria with algalytic activities against Anabaena cylindrica when water blooming occurs and to study enzyme profiles associated with alga-lytic activity, various bacterial strains were isolated from surface waters and sediments in eutrophic lakes or reservoirs in Korea. Among 178 isolates, only nine isolates exhibited lytic abilities against A cylindrica on the agar plates, and then the isolate AK-13 was selected as the strongest in lysing the cyanobacterium A. cytindrica. The strain AK-13 was characterized and identified as Sinorhizobium sp. based on fatty acid methyl ether profiles and 16S rDNA sequence. According to the results of the enzyme assays, in the strain An-13 of Sinorhizobium sp., alginase, amylase, proteinase (caseinase and gelatinase), carboxymethyl-cellulase (CMCase), laminarinase, and lipase was produced, namely CMCase, laminarinase and protease were highly active. None of glycosidase was produced. Therefore, enzyme systems of Sinorhizobium sp. AK-13 were very complex to degrade cell walls of A. cylindrica. The peptidoglycans of A. cylindrica mat be hydrolyzed and metabolized to a range of easily utilizable monosaccharides or other low molecular weight organic substances by Sinorhizobium sp. AK-13.

• 한국균학회소식:학술대회논문집
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• 한국균학회 2016년도 춘계학술대회 및 임시총회
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• pp.11-11
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• 2016

Penicillium species are commonly isolated from various outdoor and indoor environments, including marine environments such as sponges, algae and sand. Penicillium is especially important because numerous bioactive compounds have been isolated. Penicillium was the most common species in intertidal zone in Korea, however the diversity and ecological roles of Penicillium in intertidal zone are not clarified. We explored diversity and ecological roles of marine-derived Penicillium from tidal flat and sea sand in Korea. The diversity of marine-derived Penicillium from Korea was investigated using both culture-dependent and culture-independent approach by ${\beta}$-tubulin sequence. In addition, we evaluated optimal temperature, halo-tolerance, and enzyme activity of Penicillium strains, such as extracellular alginase, endoglucanase, ${\beta}$-glucosidase, and protease. For culture-dependent approach, a total of 182 strains of 62 Penicillium species were isolated, with 53 species being identified. The most common species was Penicillium oxalicum, followed by P. crustosum, P. brasilianum, P. koreense, and P. griseofulvum. Species richness and composition were not significantly different by season, substrates, and seaside. For culture-independent approach using Illumina sequencing, 73 OTUSs were detected. The most frequently observed species was P. antarcticum, followed by P. koreense, P. crustosum, and P. brevicompactum. Diversity of Penicillium was higher during winter season than during summer season and in western sea than in southern sea, respectively. Community structure was significantly different by season and sea side. 52 species were detected by both methods. Unique species were isolated from each of methods - 10 from culture methods and 21 from Illumina sequencing. Furthermore, salinity adaption of the Penicillium varied depending on species. Many Penicillium species showed endoglucanase, ${\beta}$-glucosidase, and protease activity. Some species including P. paneum and P. javanicum degraded the polycyclic aromatic hydrocarbons. Thus, our results demonstrate that intertidal zone in Korea harbors diverse Penicillium community and marine-derived Penicillium play important ecological roles as decomposers of organic material in marine environments.

• 생명과학회지
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• 제22권6호
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• pp.751-759
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• 2012

본 연구에서는 기장 해안의 해수에서 분리한 해양 미생물로부터 해조류의 난소화성 복합다당류를 분해하는 균주를 분리하고 동정하여 Microbulbifer sp. SC092로 명명하였다. Microbulbifer sp. SC092 미생물에 의하여 생성된 조효소의 기질 특이성 분석으로 agar 및 alginic acid를 분해하는 효소활성이 각각 146.2%로 agarase와 alginase 효소활성이 높은 것으로 나타났다. 감태 효소분해산물의 항산화력은 DPPH radical 소거능과 SOD 활성으로 측정하였으며, DPPH radical 소거능과 SOD 활성은 각각 84.1, 89.6%로 나타났다. 감태 효소분해산물의 혈당 강하능은 ${\alpha}$-glucosidase 활성억제 효과를 측정하였으며 ${\alpha}$-glucosidase 활성은 58.7%로 나타났다. 감태 효소분해산물의 항염증 소재로써 이용 가능성은 인체의 위의 pH와 비슷한 조건에서 아질산염소거능 측정으로 확인하였는데, 0.5 mg/ml 농도에서 56.3%를 나타내어 positive control로 사용한 Vit C 보다 높은 활성을 보였다. 알코올분해능은 ADH와 ALDH 활성을 측정한 결과는 각각 123.3, 215.2%로 나타나 감태 효소분해산물의 우수한 숙취해소 효능을 보였다. 감태 효소분해산물의 미백 및 주름개선기능은 tyrosinase 및 elastase 저해효과로 분석하였다. 감태 효소분해산물의 농도가 증가함에 따라 미백 및 주름개선기능은 농도 의존적으로 증가하는 경향을 나타내었으며, 분해산물의 농도 2.5 mg/ml 농도에서 각각 42.2%와 73.1%로 높은 저해 효능을 나타내었다. 이러한 결과는 효소 추출법이 새로운 기능성 소재를 발굴하는데 효과적인 추출법인 것으로 기대된다.