• 제목/요약/키워드: Extracellular

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Streptomyces subrutilus P5가 생산하는 철 함유 superoxide dismutase의 분비 (Secretion of the iron containing superoxide dismutase of Streptomyces subrutilus P5)

  • 박재승;김재헌
    • 미생물학회지
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    • 제51권2호
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    • pp.108-114
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    • 2015
  • 본 연구에서는 Streptomyces subrutilus P5의 생장과 세포내 외 철 함유 superoxide dismutase 활성을 비교 분석하여 철함유 superoxide dismutase의 분비 시점을 확인하고 분자 수준에서 이 효소의 분비에 관여하는 유전정보를 확인하고자 하였다. Streptomyces subrutilus P5의 균체 생장은 건체 중량을 측정하여 결정하였다. Glucose는 log phase에서 급격히 소모되어 24시간 후에 이르러 완전히 고갈되었다. 세포내의 철 함유 superoxide dismutase는 배양 후 3시간에 나타나며 세포외 철 함유 superoxide dismutase는 배양 후 7.5시간부터 나타난다. 따라서 superoxide dismutase는 용균에 의해서가 아니라 능동적인 분비기작에 의해서 세포 외로 분비된 것으로 추측할 수 있다. Streptomyces subrutilus P5의 sodF에는 signal peptide 유전정보가 존재하지 않았다. 그러나 sodF의 상류지역에서 다른 세균의 type III 분비단백질 유전자와 유사한 type III 분비상자가 발견되었다. Streptomyces 균주에서 type III 분비단백질이 존재할 가능성이 있음을 처음으로 제시하였다.

단백질 분해효소를 분비하는 Micrococcus sp. HJ19의 분리 및 특성 (Isolation and Charcterization of Micrococcus sp. HJ-19 Secreting Extracellular Protease)

  • 차인태;오용식;노동현
    • 미생물학회지
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    • 제43권3호
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    • pp.222-226
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    • 2007
  • 단백질 분해효소는 아미노산간에 존재하는 peptide결합을 절단하는 효소로 산업적으로 많이 사용된다. 이러한 단백질 분해효소의 새로운 공급원을 찾기 위하여 체외 단백질 분해효소를 생산하는 세균을 둥해 심층수로부터 집식배양으로 분리하였다. 이들 중 빠른 성장과 skim milk를 첨가한 배지에서 가장 큰 투명환을 보여준 균주를 HJ19로 명명하였다. 분리균의 형태학적, 생리생화학적 특성과 16S rRNA gene의 염기서열을 조사한 결과 Micrococcus sp.에 속하는 것으로 나타났다. 분리된 Micrococcus sp. HJ19는 $10^{\circ}C$에서는 자라지 못하였으며 $20^{\circ}C$부터 생육과 효소활성을 보여주었다. 최적 생육 온도는 $37^{\circ}C$였으며, 효소 활성은 $30^{\circ}C$에서 약 480 unit/ml의 활성을 보여 주었다.

유산균이 생성하는 세포외 다당류의 최적 생성조건에 관한 연구 (Study on the Optimum Cultural Conditions for the production of Extracellular Polysaccharide by Lactic Acid Bacteria)

  • 강국희;정후길
    • 한국식품위생안전성학회지
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    • 제5권1호
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    • pp.27-34
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    • 1990
  • 유산균이 생성하는 다당류에 관한 연구의 일환으로서 발효유 제품의 종균으로 널리 사용되고 있는 Str. themophilus, Lb. bulgaricus, Lb. casei 등을 10% 환원멸균 탈지유에 배양하면서 다당류 생성에 따른, 우유 배양액 점도의 경시적 변화 및 다당류의 최적 생성 조건에 대하여 조사하였다. 1. 시험균 중에서 Str. thermophilus 510이 가장 높은 점도를 나타냈으며 $41^{\circ}C$ 배양 5일째에 5,000 CP로서 다른균에 비해서 1,000 CP 이상 높았다. 2. 인공합성배지를 사용하여 흡광도를 측정한 결과 $41^{\circ}C$에서의 흡광도가 가장 높았으며 다당류 합성을 위한 탄소원으로서는 Sucrose의 이용성이 1~5% 농도에서 가장 높았다. 3. 다당류 합성효소의 금속염에 대한 영향을 살펴본 결과 $Mg^{++}$의 효과가 인정되었다.

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Antiapoptotic Effect of Aurintricarboxylic Acid; Extracellular Action versus Inhibition of Cytosolic Protein Tyrosine Phosphatases

  • Lee, Dong-Yoon;Kim, Mee-Kyung;Kim, Mi-Jeong;Bhattarai, Bharatraj;Kafle, Bhooshan;Lee, Keun-Hyeung;Kang, Jae-Seung;Cho, Hyeong-Jin
    • Bulletin of the Korean Chemical Society
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    • 제29권2호
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    • pp.342-346
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    • 2008
  • Aurintricarboxylic acid (ATA) prevents apoptosis in a wide range of cell types, including PC12 cells. ATA is known to increase the phosphorylation level of IGF-1 receptor (IGF-1R) and downstream signaling proteins. ATA can translocate across the plasma membrane of PC12 cells and inhibit protein tyrosine phosphatases (PTPs) and, therefore, it is not clear whether ATA exerted its antiapoptotic effect through activation of IGF-1R or by inhibition of cytosolic PTPs. When PC12 cells, deprived of serum, were treated with Fab fragment of anti-IGF-1R antibody to prevent the binding of ATA to the extracellular domain of IGF-1R, ATA was found to penetrate into the cytosolic space of the cells. Under these conditions, the survival-promoting effects of ATA were abolished, and the increase of phosphorylation and characteristic cleavage of IGF-1R were not observed. These results indicate that the antiapoptotic effect of ATA in PC12 cells is due to the binding of ATA to the extracellular domain of IGF-1R and subsequent activation of the IGF-1R, not inhibition of cytosolic PTP(s).

Zinc Enhances Neutrophil Extracellular Trap Formation of Porcine Peripheral Blood Polymorphonuclear Cells through Tumor Necrosis Factor-Alpha from Peripheral Blood Mononuclear Cells

  • Heo, Ju-Haeng;Kim, Hakhyun;Kang, Byeong-Teck;Yang, Mhan-Pyo
    • 한국임상수의학회지
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    • 제37권5호
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    • pp.249-254
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    • 2020
  • Neutrophil extracellular trap (NET) formation is an immune response for the invasion of microbes. The purpose of this study is to examine the effect of zinc on NET formation of porcine peripheral blood polymorphonuclear cells (PMNs). The NET formation of PMNs was measured by fluorescence microplate reader. The production of tumor necrosis factor (TNF)-α in the culture supernatants from zinc-treated peripheral blood mononuclear cells (PBMCs) was measured by enzyme-linked immunosorbent assay (ELISA). Zinc itself did not have no effect on NET formation. However, the NET formation of PMNs was increased by culture supernatants from PBMCs treated with zinc. Also, the NET formation of PMNs was increased by recombinant porcine (rp) TNF-α. The production of TNF-α in PBMCs culture supernatants was shown to increase upon zinc treatments. These NET formations of PMNs increased by either culture supernatant from PBMCs treated with zinc or rpTNF-α were inhibited by treatment of anti-rpTNF-α polyclonal antibody (pAb). These results suggested that zinc has an immunostimulating effect on the NET formation of PMNs, which is mediated by TNF-α released from zinc-treated PBMCs. Therefore, zinc may play an important role for NET formation in the defense of porcine inflammatory diseases.

차가버섯의 균사체 및 세포외다당체의 생산조건과 면역활성 (Optimun Conditions for Production of Mycelia and Extracellular Polysaccharide from Inonotus obliquus and Their Immunomodulating Acitivities)

  • 박희성;신동일;정일경;양병근
    • 생명과학회지
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    • 제19권11호
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    • pp.1617-1622
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    • 2009
  • 차가버섯의 균사체 생장과 세포외 다당체 생산을 위한 최적의 배양조건을 조사하고, 세포내 외 다당체의 항보체 활성과 대식세포의 lysosomal enzyme 활성을 관찰하였다. 균사체 생장과 세포외 다당체 생산을 위한 최적 pH, 온도, 및 교반속도는 각각 5.5, $25^{\circ}C$, 150 rpm으로 나타났으며, 배양기간은 11일째 최대 균사체 생장(10.89 g/l)과 세포외다당체 생산(1.25 g/l)을 보였다. 항보체 활성은 세포내 외 다당체의 처리 농도가 높을수록 활성이 증가함을 보였고, 대식세포의 lysosomal enzyme 활성은 $100{\mu}g/ml$ 농도에서 음성대조군에 비해 세포내 다당체는 약 2.2배, 세포외 다당체는 약 2배 정도 증가하였다.

Enzymatic Characteristics of an Extracellular Agarase of Cytophaga sp. KY-1 and Molecular Cloning of the Agarase gene

  • Kim, Young-Ho;Kim, Youn-Sook;Lee, Jae-Ran;Lee, Eun-Kyung
    • Journal of Microbiology and Biotechnology
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    • 제3권1호
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    • pp.31-38
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    • 1993
  • A bacterial strain KY-l isolated from sewage was able to produce an extracellular agarase(agarose 4-glycanohydrolase. EC 3.2.1.81). The strain KY-1 was identified as Cytophaga fermentans subsp. agarovorans based on its morphological and physiological characteristics. The agarase was purified by ammonium sulfate precipitation followed by DEAE-Sephadex A-50. Bio-Gel P-100. and CM-Cellulose column chromatography. The molecular weight of the purified enzyme was 24 kDa by SDS-polyacrylamide gel electrophoresis. The optimum temperature and pH for the enzyme activity were 30^{circ}C and 7.5, respectively. The enzyme activity was significantly inhibited in the presence of 0.1 mM $HgCl_2$. whereas it was elevated 3 times by $MnSO_4$ at 1 mM concentration. The Km value and Vmax were 16.67 mg/ml and 3.77 unit/ml.min. The agarase gene was cloned into Escherichia coli MC1061 using the plasmid vector pBR322. A 1.4 Kb DNA fragment of PstI-digested chromosomal DNA of C. fermentans KY-l was inserted into the PstI site of pBR322. expressed in the E. coli. and up to 60% of the total enzyme was extracellularly secreted. Enzymatic properties of the extracellular agarases produced by both the transformant and the donor were very similar in terms of optimal pH and temperature.

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다양한 조류종들의 세포외 유기물질에서의 염소 소독부산물 생성 특성 (Characteristics of Formation of Chlorination Disinfection By-Products in Extracellular Organic Matter of Various Algal Species)

  • 손희종;박홍기;황영도;정종문;김상구
    • 한국환경과학회지
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    • 제24권4호
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    • pp.541-547
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    • 2015
  • Formation of disinfection by-products (DBPs) including trihalomethans (THMs) and haloacetic acids (HAAs) from chlorination of six different species (Chlorella vulgaris, Scenedesmus sp., Anabaena cylindrical, Microcystis aeruginosa, Asterionella formosa and Aulacoseira sp.) of algal extracellular organic matter (EOM). The EOM characteristics evaluation of six algal species reaching at the stationary phase in the growth curve showed most of its SUVA254 showed below 1 and this means hydrophilic organic matter is much higher than hydrophobic organic matter. Chloroform formation potential (CFFP), dichloroacetic acid formation potential (DCAAFP) and trichloroacetic acid formation potential (TCAAFP) were mainly composed of THMFP and HAAFP in the EOM of various algal species. In the case of THMFP/DOC and HAAFP/DOC values, EOM of blue-green algae has appeared highest and EOM of green algae and diatom in order. THMFP/DOC was higher than HAAFP/DOC in EOM of blue-green algae. In comparison of formation potential by unit DOC composed of HAAFP in algal species EOM, DCAAFP/DOC was 1.5 times to 7.5 time higher than TCAAFP/DOC in the EOM of blue-green algae, while DCAAFP/DOC was found to be relatively high compared to TCAAFP/DOC in the EOM of green algae and diatom.

태생 및 생후 구순.구개열에 나타나는 조직변성에 대한 성장인자와 세포외 기질 단백의 면역조직화학적 연구 (IMMUNOHISTOCHEMICAL DETECTION OF GROWTH FACTORS AND EXTRACELLULAR MATRIX PROTEINS IN THE DEGENERATING TISSUES OF PRE-AND POSTNATAL HUMAN CLEFT LIP AND PALATE)

  • 민봉기;이석근;박영욱
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제28권6호
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    • pp.421-433
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    • 2002
  • In order to elucidate the pathogenesis of cleft lip and palate, first of all, it is necessary to understand the developmental mechanisms of growth factors and extracellular matrix proteins in the tissues of cleft lip and palate. We have performed immunohistochemical studies on human cleft lip and palate tissues to elucidate the pathogenetic implications of cleft lip and palate. 16 specimens from postnatal human cleft lip and palate subjects and 17 specimens from autopsy of prenatal human cleft lip and palate were fixed in 10% buffered formalin, embedded in paraffin. The sections were routinely stained by hematoxylin and eosin, also stained by PAS, and followed by immunohistochemical stainings using the antiseras of growth factors and extracellular matrix proteins such as PCNA, S-100, c-erb-B2, MMP-3, MMP-10, HSP-70, transglutaninase-C, E-cadherin, VEGF, vWF. Both the prenatal and postnatal specimens of cleft lip and palate showed dysplastic proliferation of the basal cell layer, increased infiltration of melanocytes into mucosal epithelium, sebaceous gland hyperplasia ingrowing into the muscular tissue of lip and palate, and fatty infiltration into the submucosal deep connective tissue. The strong reactions of MMP-3 and HSP-70 were detected in the tissues of cleft lip and palate, especially increased in degenerating muscle bundles, while the immunostainings of PCNA and c-erb-B2 were weakly positive in the tissues of cleft lip and palate. These data suggest that the retrogressive tissue degeneration around the cleft areas persistently exist during the prenatal and postnatal period after cleft formation, and the sebaceous gland hyperplasia and fatty infiltration with the intense expression of MMP-3 and HSP-70 is closely related to the muscular degeneration around the cleft area.

세포외 기질 물질의 물리·화학적 영향에 따른 심근세포(HL-1)의 성장 연구 (Physical and Chemical Effects of Extracellular Matrix on the Growth of Cardiomyocytes (HL-1))

  • 홍윤미;최성균;김선민
    • 대한기계학회논문집B
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    • 제35권11호
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    • pp.1229-1235
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    • 2011
  • 본 연구에서는 세포의 성장속도, 배양표면 부착, 체내 특이성을 유지에 중요한 요소인 세포외 기질 물질에 따른 심근세포주(HL-1 cell)의 성장, 생존률 및 표현형 등을 분석했다. 서로 다른 5 가지의 세포외 기질 환경을 조성하여 세포 성장에 미치는 영향에 대하여 분석했다. 세포외 기질 물질이 처리된 배양 표면의 물리적인 형태는 원자현미경을 통하여 분석했으며, 세포의 표면부착, 성장 및 증식과 생존률 및 표현형은 역상형광현미경 및 면역염색법을 통해 분석했다. 본 연구를 통하여 서로 다른 세포외 기질 물질이 세포의 부착 및 성장 속도에 영향을 미치며, 심근세포의 특이성을 나타내는 단백질의 형성에도 차이를 보이는 것을 확인했다.