• 제목/요약/키워드: proteinase K

검색결과 321건 처리시간 0.028초

Antimicrobial Substance against Escherichia coli O157:H7 Produced by Lactobacillus amylovorus ME1

  • Jung, Byung-Moon;Woo, Suk-Gyu;Chung, Kun-Sub
    • Food Science and Biotechnology
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    • 제17권3호
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    • pp.679-682
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    • 2008
  • A lactic acid bacterium producing an antimicrobial substance against Escherichia coli O157:H7 was isolated from raw milk and identified as Lactobacillus amylovorus ME-1. In addition to E. coli O157 :H7, the antimicrobial substance also inhibited the growth of Bacillus cereus, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, Staphylococcus aureus, Streptococcus agalactiae, Streptococcus pyrogenes, and Yersinia enterocolitica. The antimicrobial substance was stable at pH 2-12 and $121^{\circ}C$ for 15 min and insensitive to proteinase K, protease, amylase, and catalase. Purification of the antimicrobial substance was conducted through methanol and acetonitrile/ethylacetate extraction, ultrafiltration with a 500 Da cutoff, thin layer chromatography (TLC) with silicagel 60, and high performance liquid chromatography (HPLC) with a $C_{18}$ reverse phase column. The ${\lambda}_{max}$ of the purified antimicrobial substance was determined as 192 nm by ultra violet (UV) scanning, while the molecular weight was estimated as 453 Da based on the mass spectrum. Accordingly, the current results suggest that the antimicrobial substance from the L. amylovorus ME-1 was not a bacteriocin, but rather a new non-proteinaceous substance distinct from acidophilin, acidolin, diacetyl, and reuterin.

Blockade of STAT3 in T Cells Inhibits Germinal Center Reactions against Intranasal Allergens

  • Choi, Garam;Chung, Yeonseok
    • Biomolecules & Therapeutics
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    • 제24권3호
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    • pp.244-251
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    • 2016
  • Understanding the developmental mechanisms of humoral immunity against intranasal antigens is essential for the development of therapeutic approaches against air-borne pathogens as well as allergen-induced pulmonary inflammation. Follicular helper T (Tfh) cells expressing CXCR5 are required for humoral immunity by providing IL-21 and ICOS costimulation to activated B cells. However, the regulation of Tfh cell responses against intranasal antigens remains unclear. Here, we found that the generation of Tfh cells and germinal center B cells in the bronchial lymph node against intranasal proteinase antigens was independent of $TGF-{\beta}$. In contrast, administration of STAT3 inhibitor STA-21 suppressed the generation of Tfh cells and germinal center B cells. Compared with wild-type OT-II T cells, STAT3-deficient OT-II T cells transferred into recipients lacking T cells not only showed significantly reduced frequency Tfh cells, but also induced diminished IgG as well as IgE specific for the intranasal antigens. Cotransfer study of wild-type OT-II and STAT3-deficient OT-II T cells revealed that the latter failed to differentiate into Tfh cells. These findings demonstrate that T cell-intrinsic STAT3 is required for the generation of Tfh cells to intranasal antigens and that targeting STAT3 might be an effective approach to ameliorate antibody-mediated pathology in the lung.

만성폐쇄성폐질환의 병태생리 (Pathophysiology of Chronic Obstructive Pulmonary Disease)

  • 김현국;이상도
    • Tuberculosis and Respiratory Diseases
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    • 제59권1호
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    • pp.5-13
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    • 2005
  • COPD는 비가역적인 기류제한을 특징으로 하는 질병의 상태이다. 이러한 기류제한은 유해입자나 가스등에 대한 폐의 비정상적인 염증반응과 관련되어 있고 자연경과 중 지속적으로 진행하는 양상을 띤다. 비정상적인 염증반응 및 단백분해효소와 항단백분해효소간의 불균형과 산화 스트레스 등의 기전에 의해 점액의 과분비, 섬모의 기능장애, 소기도의 섬유화와 협착, 폐실질의 파괴, 폐혈관손상 등이 발생한다. 이러한 병리학적 변화에 의해 기류제한이 발생하고 가스교환 장애, 폐고혈압, 폐성심, 전신적인 염증이나 골격근의 기능장애 등이 유발된다. COPD 환자의 일반적인 증상인 기침, 객담, 호흡곤란 등은 이러한 병태생리학적 변화로 설명할 수 있다.

Enzyme Profiles of Alga-Lytic Bacterial Strain AK-13 Related with Elimination of Cyanobacterium Anabaena cylindrica

  • Kim, Jeong-Dong;Han, Myung-Soo
    • 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.

Inhibitory effect of bacteriocin-producing lactic acid bacteria against histamine-forming bacteria isolated from Myeolchi-jeot

  • Lim, Eun-Seo
    • Fisheries and Aquatic Sciences
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    • 제19권10호
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    • pp.42.1-42.10
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    • 2016
  • The objectives of this study were to identify the histamine-forming bacteria and bacteriocin- producing lactic acid bacteria (LAB) isolated from Myeolchi-jeot according to sequence analysis of the 16S rRNA gene, to evaluate the inhibitory effects of the bacteriocin on the growth and histamine accumulation of histamine-forming bacteria, and to assess the physico-chemical properties of the bacteriocin. Based on 16S rRNA gene sequences, histamine-forming bacteria were identified as Bacillus licheniformis MCH01, Serratia marcescens MCH02, Staphylococcus xylosus MCH03, Aeromonas hydrophila MCH04, and Morganella morganii MCH05. The five LAB strains identified as Pediococcus acidilactici MCL11, Leuconostoc mesenteroides MCL12, Enterococcus faecium MCL13, Lactobacillus sakei MCL14, and Lactobacillus acidophilus MCL15 were found to produce an antibacterial compound with inhibitory activity against the tested histamine-producing bacteria. The inhibitory activity of these bacteriocins obtained from the five LAB remained stable after incubation at pH 4.0-8.0 and heating for 10 min at $80^{\circ}C$; however, the bacteriocin activity was destroyed after treatment with papain, pepsin, proteinase K, ${\alpha}$-chymotrypsin, or trypsin. Meanwhile, these bacteriocins produced by the tested LAB strains also exhibited histamine-degradation ability. Therefore, these antimicrobial substances may play a role in inhibiting histamine formation in the fermented fish products and preventing seafood-related food-borne disease caused by bacterially generated histamine.

뽕나무버섯으로부터 Fibrinolytic enzyme의 정제 및 특성 연구 (Purification and Characterization of Fibrinolytic Enzyme from Armillariella mellea)

  • 김준호;김양선
    • 한국균학회지
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    • 제26권4호통권87호
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    • pp.583-588
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    • 1998
  • 뽕나무버섯 추출물에서 fibrinolytic enzyme을 분리 정제하였다. 이 효소의 분자량은 약 19800Da 이고 pH 7.0에서 최고의 활성을 보여주는 neutral protease로 $55^{\circ}C$에서 최대 활성을 나타냈다. EDTA에 의해 활성이 저해되는 것으로 보아 metalloprotease로 추정되며 $Hg^{+2}$의 영향을 받아 활성이 상실되었다. N-terminal amino acid 분석 결과 15번째까지의 아미노산 잔기 순서는 X-X-T-N-G-X-T-X-S-R-Q-T-T-L-V 이었으며 지금까지 알려진 것과는 다른 새로운 효소이다.

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Acanthoic acid blocks production of pro-inflammatory mediators by inhibiting the ERK activation in trypsin-stimulated human leukemic mast cells

  • Kang, Ok-Hwa;Tae, Jin;Choi, Yeon-A;Kwon, Dong-Yeul;Kim, Yun-Kyung;Cai, Xing-Fu;Kim, Young-Ho;Bae, Ki-Hwan;Lee, Young-Mi
    • 대한약학회:학술대회논문집
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    • 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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    • pp.88.3-89
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    • 2003
  • Acanthoic acid (AA) is a pimaradiene diterpene isolated from the Korean medicinal plant, Acanthopanax koreanum (Araliaceae), which has been traditionally used as a tonic and sedative as well as in the treatment of rheumatism and diabetes in korea. Proteinase-activated receptor-2 (PAR-2) agonist trypsin plays a role in inflammation, and human leukemic mast cells (HMC-l) express PAR-2. In the present study, the effect of acanthoic acid on production of tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) and tryptase in trypsin-stimulated HMC-1 was examined. (omitted)

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Identification of Receptor-like Protein for Fructose-1,6-bisphosphatase on Yeast Vacuolar Membrane

  • Ko, Je-Sang
    • BMB Reports
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    • 제33권6호
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    • pp.448-453
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    • 2000
  • In yeast the key gluconeogenic enzyme, fructose-1,6-bisphosphatase (FBPase), is selectively targeted from the cytosol to the lysosome (vacuole) for degradation when glucose starved cells are replenished with glucose. The pathway for glucose induced FBPase degradation is unknown. To identify the receptor-mediated degradation pathway of FBPase, we investigated the presence of the FBPase receptor on the vacuolar membrane by cell fractionation experiments and binding assay using vid mutant (vacuolar import and degradation), which is defective in the glucose-induced degradation of FBPase. FBPase sedimented in the pellets from vid24-1 mutant after centrifugation at $15,000{\times}g$ for 15 min, suggesting that FBPase is associated with subcellular structures. Cell fractionation experiments revealed that FBPase is preferentially associated with the vacuole, but not with other organelles in vid24-1. FBPase enriched fractions that cofractionated with the vacuole were sensitive to proteinase K digestion, indicating that FBPase is peripherally associated with the vacuole. We developed an assay for the binding of FBPase to the vacuole. The assay revealed that FBPase bound to the vacuole with a Kd of $2.3{\times}10^6M$. The binding was saturable and specific. These results suggest that a receptor for FBPase degradation exists on the vacuolar membrane. It implies the existence of the receptor-mediated degradation pathway of FBPase by the lysosome.

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Caveolin-1 inhibits membrane-type 1 matrix metalloproteinase activity

  • Kim, Hye-Nan;Chung, Hye-Shin
    • BMB Reports
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    • 제41권12호
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    • pp.858-862
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    • 2008
  • Membrane-type 1 matrix metalloproteinase (MT1-MMP) is a zinc-dependent proteinase found in cholesterol-rich lipid rafts on the plasma membrane. MT1-MMP hydrolyzes extracellular matrix (ECM) proteins, activates pro-matrix metalloproteinase-2 (proMMP-2) and plays an important role in ECM remodeling, cancer cell migration and metastasis. The role of caveolin-1, an integral protein of caveolae, in the activation of MT1-MMP remains largely unknown. Here, we show that the expression of caveolin-1 attenuates the activation of proMMP-2, reduces proteolytic cleavage of ECM and inhibits cell migration. We utilized the cytoplasmic tail domain deletion (${\Delta}CT$) or the E240A mutant of MT1-MMP. Co-expression of caveolin-1 with the wild-type or the ${\Delta}CT$ MT1-MMP decreased the proMMP-2 activation and inhibited collagen degradation and cell migration. Caveolin-1 had no effect on the catalytically inert E240A MT1-MMP. Our findings suggest that caveolin-1 is essential in the down-regulation of MT1-MMP activity by promoting internalization from the cell surface.

신이 에탄올 추출물의 PAR2-유발 부종에 대한 항염증 활성 (Anti-inflammatory Activity of the Ethanol Extract from Magnoliae Flos on PAR2-mediated Edema)

  • 임종필;박영서
    • 한국약용작물학회지
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    • 제13권6호
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    • pp.245-249
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    • 2005
  • 신이는 전통적으로 비염 등의 치료에 사용되어 왔는데, 신이 에탄올 추출물은 그람 양성균인 Staphylococcus aureus 그람 음성균인 Pseudomonas fluorescens균 및 곰팡이균인 Aspergillus niger에 대하여 모두 항균활성을 보였으며, proteinase로 활성화된 receptor-2에 의한 흰쥐 뒷발바닥 부종모델에서 신이 에탄올 추출물을 경구투여한 경우 typsin이나 $tc-NH_2$로 유발된 발바닥 부종, 혈관투과성, myoloperoxidase 활성도에서 유의성 있는 억제율을 보여 항염증 활성이 뚜렷함을 나타냈다.