• 미생물학회지
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• 제31권2호
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• pp.141-147
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• 1993

백색 부후균인 plaurotus ostreatus 의 4가지의 균주로부터 각각 10.2 kb 와 7.2 kb (NFFA 2), 두 종류의 10.2 kb (NFFA 4001) 11.2 kb (NFFA 4501), 10.2kb 와 11.2 kb(KFCC 11635) 크기의 미토콘드리아 플라스미드들을 분리해 내었다. NFFA 2 의 변종인 NFFA 2m1 과 NFFA 2m2 에서는 이들 플라스미드가 관찰되지 않았다. 분별 원심분리에 의해 얻은 미토콘드리아에서 핵산을 추출하여 agarose gel 에서 전기영동시키면 플라스미드가 관찰되지 않았으나 proteinase K 를 처리하고 핵산을 추출하여 전기영동한 결과 이들 플라스미드가 관찰되었는에, 이는 플라스미드상에 단백질인 결합되어 있음을 시사한다. Proteinase K 를 처리한 플라스미드 DNA 와 exonuclease 를 반응시킨 결과, 이들 플라스미드들은 5'말단에 단백질이 결합된 성형 이중가닥 DNA의 구조를 가진 것을 확인하였다. 각 플라스미드들의 상호관계를 조사하기 위하여 Southern hybridization 을 수행한 결과 최소 3가지 종류의 플라스미드들로 분류할 수 있었다. 이중 한 그룹 (group I) 은 모든 P. ostreatus 균주들에서 공통적으로 발견되었다. Pleurotus 속의 5가지 다른 종(P. cornucopiae, P. florida, P. pulmonarius, P. sajor-cuja, P. spodoleucus) 의 균주들로부터 미토콘드리아 플라스미드들을 분리하였다. 이들은 한균주당 1-4 개 까지의 플라스미드를 가지며, 플라스미드의 크기는 7.2 kb-14kb 범위에 있었다. P. ostreatus 의 NFFA 2 의 10.2 kb(group I) 플라스미드와 hybridization 을 수행한 결과 P. cornucopiae ASI 2011을 제외한 다른 모든 균주들이 유사한 염기서열의 플라스미드를 갖고 있음을 알았다.

• 한국토양비료학회지
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• 제15권4호
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• pp.241-250
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• 1982

발아초기 녹두의 부위별 alkaline inorganic PPase의 활성변화 및 잎 부위에서 부분 정제하여 얻은 효소를 이용하여 효소적 성질을 조사한 결과를 요약하면 다음과 같다. 1. 잎 부위가 타 부위보다 약 2~4배의 더 높은 활성을 나타냈으며 발아가 진행됨에 따라 잎과 뿌리 그리고 지엽에서는 효소 활성이 초기에 증가하다가 점차 감소하는 경향을 보인 반면, 상배축에서는 초기부터 계속 감소하였으며 하배축에서는 계속 증가하는 경향을 보였다. 2. 정제 과정동안 23.9%의 수율로 86배가 정제되었으며 전기 영동상의 Rm value는 0.35였으며 homogenenity는 아니었고, Km value는 0.89mM로 나타났다. 3. 이 효소는 $Mg^{2+}$에 대해 대단히 Specific하였으며 $Cu^{2+}$와 $Fe^{2+}$도 $Mg^{2+}$에 비해 각각 56%, 55%의 activating effect를 나타냈다. 그러나, $Cu^{2+}$, $Zn^{2+}$, $Mn^{2+}$, $Co^{2+}$와 $Ni^{2+}$은 이 효소에 대해 저해체로 작용하였다. 4. 이 효소는 pH 8-9와 $50^{\circ}C$에서 최대의 활성을 보였으며 열에 대해서도 상당히 안정하였다.

• Journal of Microbiology and Biotechnology
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• 제14권5호
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• pp.985-990
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• 2004

Levans produced from Microbacterium laevaniformans were isolated, characterized, and fractionated by molecular weight. TLC, HPLC, and GC-MS analyses of the exopolysaccharide showed that it was a fructan-type polymer and was composed of (2,6)- and (2,1)-glycosidic linkages. $^{13}C$-NMR analysis proved that the polysaccharide was mainly a $\beta$-(2,6)-linked levan-type polysaccharide. To investigate the cytotoxicity of the acetone-precipitated levan fractions such as M1, M2, and M3, HepG2, P388D1, U937, SNU-1, and SNUC2A cell lines were screened. Among the cell lines tested, the cytotoxicity of M1- M3 fractions were detected from only SNU-1 and HepG2 cells at the dosage level of $100-800\mu\textrm{g}ml$. The M2 fraction M_r$, 80,000) at 400 $mu{g/ml}$ had the greatest cell growth inhibition (84.6%) on SNU-1, while the M1 $(M_r$, 50,000) at $800\mu\textrm{g}ml$ showed the greatest (46.32%) on HepG2. To obtain more uniform M_r$ fractions of levan, the levan was further fractionated from S1 $(M_r$ 1,000,000) to S5 $(M_r$ 10,000) using gel permeation chromatography. Again, the S1-S5 fractions had strong cytotoxicity on SNU-1 and HepG2 cell lines. The greatest inhibition effects of S4 $(M_r$ 80,000) on SNU-1 and S5 $(M_r$ 10,000) on HepG2 were shown to be 49.5% and 73.0%, respectively. The cytotoxicity of the levan fractions was more effective on SNU-1 than on HepG2. Although the relationship between the Mw and the cytotoxicity was not clear, smaller $M_r$, fractions of levan showed greater growth inhibition effect on the cancer cell lines in general. Therefore, it was indicated that a specific Mw class of levan is responsible for the effective cytotoxicity.

• Journal of Microbiology and Biotechnology
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• 제19권12호
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• pp.1656-1664
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• 2009

Bioaugmentation of bioreactors focuses on the removal of numerous organics, with little attention typically paid to the maintenance of high and stable nitrite accumulation in partial nitrification. In this study, a bioaugmented membrane bioreactor (MBR) inoculated with enriched ammonia-oxidizing bacteria (AOB) was developed, and the effects of dissolved oxygen (DO) and temperature on the stability of partial nitrification and microbial community structure, in particular on the nitrifying community, were evaluated. The results showed that DO and temperature played the most important roles in the stability of partial nitrification in the bioaugmented MBR. The optimal operation conditions were found at 2-3 mgDO/l and $30^{\circ}C$, achieving 95% ammonia oxidization efficiency and nitrite ratio ($NO_2^-/{NO_x}^-$) of 0.95. High DO (5-6 mg/l) and low temperature ($20^{\circ}C$) had negative impacts on nitrite accumulation, leading to nitrite ratio drop to 0.6. However, the nitrite ratio achieved in the bioaugmented MBR was higher than that in most previous literatures. Denaturing gradient gel electrophoresis (DGGE) and fluorescence in situ hybridization (FISH) were used to provide an insight into the microbial community. It showed that Nitrosomonas-like species as the only detected AOB remained predominant in the bioaugmented MBR all the time, and coexisted with numerous heterotrophic bacteria. The heterotrophic bacteria responsible for mineralizing soluble microbial products (SMP) produced by nitrifiers belonged to the Cytophaga-Flavobacterium-Bacteroides (CFB) group, and $\alpha$-, $\beta$-, and $\gamma$- Proteobacteria. The fraction of AOB ranging from 77% to 54% was much higher than that of nitrite-oxidizing bacteria (0.4-0.9%), which might be the primary cause for the high and stable nitrite accumulation in the bioaugmented MBR.

• 미생물학회지
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• 제19권3호
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• pp.142-152
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• 1981

One hundred and thirteen healed pulmonary tuberculosis patients and 11 patients with other underlying diseases were studied for evidence of pulmonary fungal infection because of persisting hemoptysis or chronic cough. Rediological, mycological and serological investigations revealed that 54 out of 124 patients were evidently infected with one or more species of fungi. A. fumigatus was isolated from 4 out of 70 patients whose sera did not react with antigens from this fungus, while it was isolated from 43 out of 47 serological reactors to this fungus. Chest radiography showed a distinct fungus ball in a cyst of one patient and in a preformed cavity in the lung of 17 healed tuberculosis patients and two other patients. The latter two patients were infected with A.flavus. Two patients, who were under the long period of immunosuppressive therapy, apparently succumbed to invasive aspergillosia due to A.fumigatus. A single or dual infection with A. flavus, A. nidulans, A.nidulans var. latus, C. albicans, and P. boydii were noticed in some patients without mycetomal shadow on chest radiographs. Young mycelial extract (ME) of A.fumigatus detected antibody in 95.8 percent of the sera from patients infected with this fungus, while it was isolated from 43 out of 47 serological reactors to this fungus. Chest radiography showed a distinct fungus ball in a cyst of one patient and in a performed cavity in the lung of 17 healed tuberculosis patients and two other patients. The latter two patients were infected with A. flavus. Two patients, who were under the long period of immunosuppressive therapy, apparently succumbed to invasive aspergillosis due to A.fumigatus. A single or dual infection with A. flavus, A. nidulans, A. niduans var. latus, C. albicans, and P. boydii were noticed in some patients without mycetomal shadow on chest radiographs. Young mycelial extract (ME) of A.fumigatus detected antibody in 95.8 percent of the sera from patients infected with this fungus, while the commercial culture filtrate antigen (GL) yielded 78.7 per cent positive result. Culture filtrate antigen, however, was comparable with ME. There was no single antigen with which all the serum specimens reacted. Fractionation of ME resulted in a loss of some activity although it excluded substances that reacted with C-reactive protein in a loss of some activity although it excluded substances that reacted with C-reactive protein. Most reactive and specific precipitinogens distributed in the fraction (FB) which was precipitable at 75 percent saturation with ammonium sulfate and eluted in a second peak in order from gel-filtration and which contained mostly proteinic components. Glycoproteins or polysaccharides rich fractions (FA and ASI) were relatively less effective in detecting antibody. Demonstration of antibody in the serum from patients using a battery of fungal antigens and of etiologically related fungi from clinical specimens are very useful laboratory procedures for the diagnosis of pulmonary fungal infection which is a common complication of tuberculosis.

• 생명과학회지
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• 제13권5호
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• pp.668-674
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• 2003

홍어를 발효하여 발효기간에 따른 항고혈압 효과를 조사하고 그에 따른 항고혈압성 물질을 분리하기 위해 GPC system을 사용하여 항고혈압 물질을 조 분리하였다. 항고혈압성 물질을 농축하기 위해 홍어 내장 및 가식부 열수 추출물을 대량 포집 하였다. 그리고 그에 따른 ACE 억제효과를 검색하였다. 즉, 홍어 가식 부의 ACE 저해 작용은 2% 첨가시 29%로, 다소 낮으나 상시 섭취될 수 있는 식품이란 측면에서 볼 때 그 유용성이 기대된다고 할 수 있으며 홍어 내장 열수 추출물의 ACE 저해 효과는 시료 2% 첨가의 경우 발효 0일째에 71.0%로 가장 높게 나타났다. 발효가 진행되면서 ACE 저해 효과는 감소하는 것으로 나타났다. 이러한 ACE 억제효과를 나타내는 성분을 추정하기 위하여 홍어 내장 열수 추출물의 일반성분과 원소분석을 실시한 결과 일반성분 중 순단백질이 58.7%로 가장 높게 나타났으며, 원소분석 결과 C, H, O, N의 성분비가 당류라기 보다는 peptide계인 것으로 나타났다. 또한, 질소화합물의 분석결과 ACE를 억제하는 기능성 peptide의 성분인 Tyr, Phe, Val, His 등이 가식부 보다 많아서 ACE를 억제하는 peptide를 함유하리라 예상되었다. ACE억제 물질을 조 분리하기 위하여 Sephadex G-25 column chromatography에 의해 분자량별로 분획한 결과, 분획물 들의 ACE 저해 효과는 분획물 B(111-160)의 농도 0.2%에서 67.8%로 가장 높은 ACE저해 효과를 나타내었다. 이상의 결과로 미루어 보아 홍어 내장 열수 추출물의 ACE 저해인자는 가열에 대하여 안정한 비교적 저분자의 peptide와 같은 물질이라고 추정할 수 있었다.

• 대한수의학회지
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• 제29권4호
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• pp.445-455
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• 1989

The proteins of the ruminant erythrocyte membranes were analysed by polyacrylamide gel electrophoresis in sodium dodecyl sulfate, and their relations to the slow erythrocyte sedimentation rate(ESR) of the ruminants were investigated by treating the erythrocytes with proteinases such as trypsin, chymotrypsin and pronase, and glycosidases such as neuraminidase and galactosidase. Protein content in the erythrocyte membrane was $2.85{\pm}0.28$ in human, $3.60{\pm}0.41$ in Korean cattle, $3.71{\pm}0.36$ in Holstein, $4.13{\pm}0.83$ in Korean native goat and $3.94{\pm}0.56mg/ml$ in sheep, showing higher in ruminant animals than in human(p<0.01). Although the general protein profiles of the ruminant erythrocyte membranes were almost similar to that of human, all the ruminant erythrocyte membranes showed one additional protein band, called band-Q in the previous report on proteins of bovine erythrocyte membrane, which migrated electrophoretically to the mid position between band-2 and band-3 in human erythrocyte membranes. The glycoprotein profiles of ruminant erythrocyte membranes revealed by periodic acid Schiff(PAS) stain showed a marked difference from that of human. The PAS-1(glycophorin) and PAS-2(sialoglycogrotein) present in human erythrocyte membranes were almost absent from the ruminant animals. Instead, a strong PAS-positive band near the origin of the electrophorograms, which was named as PAS-B in the previous report on proteins of bovine erythrocyte membranes, was shown in the ruminant animals except sheep. In addition, the erythrocyte membranes of Korean native goat and sheep showed a moderate PAS-negative band near the tracking dye of the electrophorograms, which was named as PAS-G in this study. In the erythrocyte treated with the enzymes, the migration of each protein fracture of erythrocyte membranes in response to each enzyme was diverse according to different species or breed of ruminant animals. Among others, band-Q present in ruminants was slightly or moderately decreased by trypsin-, chymotrypsin-, and pronase- treatments of the erythrocytes, but not only in sheep. It was particularly noticeable that PAS-B, a fraction of glycoprotein, present in ruminants except sheep, was better digested by proteinases than by glycosidases, showing remarkable increase(p<0.01) of the ESR in accord with complete digestion(disappearance) of the PAS-B band by pronase, trypsin or chymotrypsin treatment of erythrocytes. In sheep, there was almost no any response to the various enzymes in general protein and glycoprotein profiles of the erythrocyte membranes except PAS-G, which was markedly decreased by pronase treatment of the erythrocytes. Nevertheless, the ESRs were accelerated in erythrocytes treated with pronase, trypsin, chymotrypsin and neuraminidase. Erythrocyte osmotic fragility was increased in erythrocytes treated with only pronase among five enzymes in all the human and ruminant animals used in this study.

• 한국산학기술학회논문지
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• 제17권2호
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• pp.528-535
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• 2016

Ethyl acrylate(EA) 혹은 lauryl acrylate(LA)와 hydroxy ethyl acrylate(HEA)를 용액중합법에 의해 공중합하여 2종의 친유성 아크릴 전구중합체를 얻은 다음, 여기에 가교제로서 hexamethylene diisocyanate(HDI)를 첨가 배합하여, 폴리프로필렌 velvet 편직물에 통상의 pad-dry-cure방법으로 수지가공을 실시하여 유흡착재를 제조하였다. 가교 아크릴 수지(EA-HEA-HDI)의 겔화도는 전체 acrylate에 대한 HEA의 사용량과 가교제인 HDI의 농도가 증가할수록 증가하였다. 얻어진 유흡착재의 수지 부착율에 따른 흡유능과 유보유율은 시험유로서 n-decane, toluene, 대두유, 윤활유, 벙커C유를 사용하여 평가 하였고 대두유를 제외하고 사용된 모든 오일에 걸쳐 흡유능에 미치는 최적의 부착율은 약 6%로 나타났다. 한편 유보유율은 부착율의 증가에 따라 증가하였고 더욱이 고비중, 고점도 오일의 경우에서 전반적으로 보다 증가된 값을 나타내었다. 또한 흡유능은 LA-HEA-HDI로 처리된 유흡착재가 EA-HEA-HDI의 그것에 비하여 높았으며, 아크릴 수지의 측쇄 알킬기의 길이가 증가할수록 보다 높은 흡유능을 나타내었다.

• 대한수의학회지
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• 제29권1호
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• pp.13-20
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• 1989

The proteins of the bovine erythrocyte membrane were analyzed by polyacrylamide gel electrophoresis in sodium dodecyl sulfate, and their relations to the slow sedimentation rate of bovine erythrocytes were investigated by treating the erythrocytes with trypsin. The erythrocyte sedimentation rates of bovine erythrocytes from Holstein and Korean native cattle were very slow compared with the human one (1/7 as slow as the human one) as reported previously. However, when human and Holstein erythrocytes were treated with trypsin (0.2 and 0.5 mg/ml) for 1 hour at ${37^{\circ}C}$, their sedimentation rates were markedly accelerated while the sedimentation rate of Korean native cattle's erythrocytes were not affected. Although the general protein profiles of the bovine erythrocyte membranes were almost similar to that of human, bovine erythrocyte membranes showed one additional protein band, called band Q in this study, which migrated electrophoretically to the mid-position between band 2 and band 3 in human erythrocyte membranes. Treatment of Holstein and human erythrocytes with trypsin caused a decrease or disapperance of the band Q from the erythrocyte membrane. Although the band Q in Korean native cattle's erythroyte membrane was decreased by trypsin treatment of the erythrocytes, the magnitude of the decrement was not so pronounced as in the case of human and Holstein erythrocytes. The glycoprotein profiles of the bovine erythrocyte membranes revealed by periodic acid-Schiff stain showed a marked difference from that of human. The PAS-1 (glycophorin) and PAS-2 (sialoglycoprotein) present in human erythrocyte membrane were almost absent from the bovine erythrocyte membranes. Instead, the bovine erythrocyte membranes showed a strong PAS-positive band near the origin of the electrophorograms, which is named as PAS-B in this study. The PAS-B band was disappered completely by the trypsin treatment of Holstein erythrocytes whereas the PAS-B band in Korean native cattle's erythrocyte membrane still remained after the trypsin treatment. The trypsin treatment of Korean native cattle's erythrocytes, however, led to the appearance of small molecular weight peptides, indicating that the high molecular weight glycoproteins were degraded by trypsin as in human and Holstein ones. These results suggest that the slow sedimentation rate of bovine erythrocytes is due in part to the presence of band Q protein fraction and PAS-B glycoprotein in the bovine erythrocytes.

• 생명과학회지
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• 제22권8호
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• pp.1092-1098
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• 2012

본 연구에서는 우방자를 70% 에탄올로 추출하여 얻어진 추출물을 n-hexane, methylene chloride, ethyl acetate로 순차용매 분획하였다. 각 분획물에 대해 activity-guided isolation을 수행하여 활성물질의 분리 정제를 실시하였다. 활성을 나타내는 물질은 silica gel chromatography (230 mesh), sephadex LH 20, recrystallization method를 이용하여 분리하였다. 각 화합물의 화학구조는 NMR 스펙트럼 데이터 해석하였고 arctiin, arctigenin, diarctigenin, matairesinol 으로 동정하였다. 이들을 human dermal fibroblast HS68 세포에 처리하여 얻어진 상등액은 ELISA kit를 활용하여 procollagen type I 생합성과 MMP-1 저해활성을 측정하였다. 측정결과 procollagen type I 생합성과 MMP-1 저해활성 결과 모두 arctiin이 가장 우수한 결과를 보였다. 이와 같은 결과를 통해 우방자에서 분리한 리그난 화합물들을 이용하여 주름개선 소재로 개발할 수 있을 것으로 사료 된다.