• Title/Summary/Keyword: hemolysin

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Cloning of a Hemolytic Mosquitocidal Delta-endotoxin Gene (cyt) of Bacillus thuringiensis 73E10-2 (serotype 10) into Bacillus subtilis and Characterization of the cyt Gene Product

  • Kim, Kwang-Hyeon;Ohba, Michio;Kim, Byung-Woo
    • Journal of Microbiology and Biotechnology
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    • v.6 no.5
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    • pp.326-330
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    • 1996
  • To illustrate whether a hemolysin in $\delta$-endotoxins of Bacillus thuringiensis strain 73E10-2 and subsp. israelensis had immunological identity, a cyt gene of the strain 73E10-2 which encodes a hemolysin was cloned to B. subtilis (transformant 2753). The transformant 2753 containing cyt gene produced the hemolysin which lysed sheep erythrocytes after treatment of proteinase K. The hemolysin was proved also to be toxic against mosquito larvae (Aedes aegypti). The molecular weight of the hemolysin produced from the transformant 2753 was determined to be about 25 kDa by SDS-PAGE and immunoblot. The hemolysin in $\delta$-endotoxin of subsp. israelensis and subsp. kyushensis did not react on immunoblot using polyclonal anti-$\delta$-endotoxin of the strain 73E10-2, but 70-140 kDa mosquitocidal toxins in $\delta$-endotoxin of subsp. kyushuensis reacted.

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병원성 비브리오균의 용혈독소와 단백분해효소에 관한 연구

  • 박미연;김현진;장동석
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2002.10a
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    • pp.177-178
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    • 2002
  • 병윈성 비브리오균의 병원인자에는 hemolysin, protease, phospholipase A2, siderophore 외에도 콜레라균 만이 생산하는 cholera toxin 등이 있다. 이 중에서도 대부분의 병원성 비브리오균에서 생성되는 대표적인 인자는 hemolysin과 protease로 알려져 있다. Hemolysin은 혈액을 분해하는 독소로서 병원성 비브리오균의 분리ㆍ동정에 널리 이용 되고 있다. Hemolysin은 균의 배양초기에서 부터 서서히 생성되기 시작하여 대수증식기 말에 최대의 활성을 나타내며 안정기에 접어들면서 활성이 급격히 감소되는 것으로 보고되고 있다 (Kim et al., 1997). (중략)

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Identification of hemolysin as one of the important virulent factors in vibrio anguillarum V7

  • Choe, Young-Chool;Jeong, Gajin
    • Journal of Microbiology
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    • v.33 no.4
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    • pp.283-288
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    • 1995
  • We have identified hemolysin rendering virulency of Vibrio anguilarum grown at 23.deg.C which was evaluated on human RBCs. Hemolysin itself was separated as a single band on non-denaturing gel electrophoresis. Vibrio hemolysin was destroyed by trypsin and proteinase K and was heat labile. Optimal pH for activity was around pH 6 while pI of the molecule was recognized as 5.7, with relative distance (R$\sub$f/) on non-denaturing gel was 0.7. Addition of EDTA and FeCI$\sub$3/ drew the possibility that the production of hemolysin was mainly induced to overcome iron deficiency inside host animalsd infection.

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Evidence That Temporally Alternative Expression of the Vibrio vulnificus Elastase Prevents Proteolytic Inactivation of Hemolysin

  • Rhee, Jee-Eun;Lee, Jeong-Hyun;Jeong, Hye-Sook;Park, U-Ryung;Lee, Dong-Ha;Woo, Gun-Jo;Miyoshi, Shin-Ichi;Choi, Sang-Ho
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.1021-1026
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    • 2003
  • Numerous secreted and cell-associated virulence factors have been proposed to account for the fulminating and destructive nature of Vibrio vulnificus infections. Among the putative virulence factors are an elastase, elastolytic protease, and a cytolytic hemolysin. Effects of the elastase on the hemolysin were assessed by evaluating changes of hemolytic activities either in the presence or absence of the protease. Although hemolytic activity in the culture supernatant was lowered by the purified elastase added in vitro, the cellular level of hemolytic activity was unaffected by the mutation of vvpE encoding the elastase. Growth kinetic studies revealed that hemolysin reached its maximum level in the exponential phase of growth, and the elastase appeared at the onset of the stationary phase. These results have provided insight into the regulation of virulence factors: temporally coordinate regulation of virulence factors is essential for the overall success of the pathogen during pathogenesis.

Purification of Hemolysin from Vibrio anguillarum Isolated from Fish (어류분리 Vibrio anguillarum 용혈소의 정제)

  • 김영희
    • Journal of Life Science
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    • v.8 no.5
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    • pp.598-603
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    • 1998
  • A marine microbe, Vibrio anguillarum was isolated from fish and studied for its concerning pathogenic substance of hemolysin. Purification of hemolysin was achieved by the procedure of ammonium sulfate precipitation from cul-ture filtrate, DEAE-cellulose chromatography, and G-200 gel filtration with 36 fold of purification and 2.3% yield. The molecular weight of the purified hemolysin was 38,000 dalton by SDS-PAGE. The purified hemolysin was stable at pH 6-9, below 45$^{\circ}C$, and up to 1% of NaCl, respectively. $Ca^{2+}, Cu^{2+}, Zn^{2+}, Fe^{2+}$ inhibited the hemolytic activity whereas EDTA and $Mg^{2+}$ did not.

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Virulence of Environmental Urease-Positive and Kanagawa Phenomenon-Negative Vibrio parahaemolyticus

  • Park, Mi-Yeon
    • Journal of Microbiology and Biotechnology
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    • v.14 no.2
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    • pp.330-336
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    • 2004
  • Fifty-two pathogenic Vibrio parahaemolyticus strains were isolated from the environments of Busan and Yeosu, Korea. Forty-three of these strains showed protease activities, whereas 4 strains showed $\alpha / \beta$ hemolysin activities and 6 strains had urease activities. Their pathogenic factors were not overlapping except one strain, which had both protease and hemolysin activities. The 6 urease-positive strains (V. parahaemolyticus YKB4, YKB14, S25, YFB20, YFO21, and YFO22) showed the same biochemical characteristics as a reference strain [V. parahaemolyticus KCTC 2471 (urease-negative)], except for urease production. The 6 urease-positive strains showed different urease activities in their culture supernatant during the growth. The urease activity of S25 increased sharply at the late exponential phase, and was the highest at the initial stationary phase and was kept until the late stationary phase. The other 5 isolates, except C25, showed urease activities at the mid-stationary phase and increased steadily until the late stationary phase, when the urease activity was maximal. To compare the degree of virulence of V. parahaemolyticus with different pathogenic factors, hemolysin, protease, or urease-positive strains were injected into groups of 10 each of ICR mice (7- to l0-week-old males). The lethal rates of urease-positive V. parahaemolyticus, YKB14, YKB4, and S25, were significantly high, being 50, 70, and 80%, respectively. Protease-positive V. parahaemolyticus strains FM39 and FM50 showed 40% and 60% of lethal rate, respectively. Hemolysin-positive V. parahaemolyticus strains S34 and S72 had no mortality, similar to nonpathogenic V. parahaemolyticus FM12.

Vibrio fluvialis의 세포외 Hemolysin(용혈소)에 관한 특성

  • 허문수;송춘복;이제희;여인규;이미헌
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2000.05a
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    • pp.455-456
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    • 2000
  • 병원성 Vibrio spp등의 추정되는 병원성 인자들로서는 Hemolysin(용혈소) Protease, Phospholipase와 Siderophores등 여러 가지가 있으며 이 가운데 Hemolysin 은 공통인자로 중요시되어 왔다. V. fluvialis는 자연환경에서도 분리가능 하였지만, 임상에서도 분리되었다. 본 연구에서는 병원성 비브리오속인 V. fluvialis의 용혈소를 분리정제하여 그 물리화학적 특성을 확인하였다. (중략)

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Purification and Characterization of the Extracellular Hemolysin of Vibrio furnissii (Vibrio furnissii로부터 세포외 용혈소의 정제 및 특성)

  • HEO Moon-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.34 no.5
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    • pp.502-508
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    • 2001
  • The extracellualr hemolysin produced by pathogenic Vibrio furnissii was isolated and purified by ammonium sulfate precipitation and followed by DEAE-Sephadex A-50 (1st and 2nd) ion-exchange and Sephadex G-100 gel permeation chromatography. The molecular weight of the purified hemolysin was 61 kDa. The enzyme activity was inactivated by heating at 60 for 5 min, and inhibited by additions of $Cu^{2+},\;Zn^{2+}$ and a high concentration of cholesterol. Lysis of human erythrocytes by the enzyme was temperature dependent, and optimal temperature of the enzyme was $37^{\circ}C$. The purified hemolysin was unstable at pH 6.0, 6.5 and 10.0. The optimal concentration of human erythrocytes on hemolysins was about $1.5\%$. The purified hemolysin was active for erythrocytes from three animal species including mouse, rabbit and rat, and was the most active against rabbit erythrocytes. B blood group of human erythrocytes also showed a high sensitivity to the enzyme.

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Sclareol Protects Staphylococcus aureus-Induced Lung Cell Injury via Inhibiting Alpha-Hemolysin Expression

  • Ouyang, Ping;Sun, Mao;He, Xuewen;Wang, Kaiyu;Yin, Zhongqiong;Fu, Hualin;Li, Yinglun;Geng, Yi;Shu, Gang;He, Changliang;Liang, Xiaoxia;Lai, Weiming;Li, Lixia;Zou, Yunfeng;Song, Xu;Yin, Lizi
    • Journal of Microbiology and Biotechnology
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    • v.27 no.1
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    • pp.19-25
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    • 2017
  • Staphylococcus aureus (S. aureus) is a common gram-positive bacterium that causes serious infections in humans and animals. With the continuous emergence of methicillin-resistant S. aureus (MRSA) strains, antibiotics have limited efficacy in treating MRSA infections. Accordingly, novel agents that act on new targets are desperately needed to combat these infections. S. aureus alpha-hemolysin plays an indispensable role in its pathogenicity. In this study, we demonstrate that sclareol, a fragrant chemical compound found in clary sage, can prominently decrease alpha-hemolysin secretion in S. aureus strain USA300 at sub-inhibitory concentrations. Hemolysis assays, western-blotting, and RT-PCR were used to detect the production of alpha-hemolysin in the culture supernatant. When USA300 was co-cultured with A549 epithelial cells, sclareol could protect the A549 cells at a final concentration of $8{\mu}g/ml$. The protective capability of sclareol against the USA300-mediated injury of A549 cells was further shown by cytotoxicity assays and live/dead analysis. In conclusion, sclareol was shown to inhibit the production of S. aureus alpha-hemolysin. Sclareol has potential for development as a new agent to treat S. aureus infections.