• Title/Summary/Keyword: toxin production

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Cultural and Physiological Conditions for T-2 Toxin Production by Fusarium sp. (Fusarium 균주의 배양 조건 및 생리적 조건에 따른 T-2 toxin의 생성 조건)

  • 홍성희;양규환
    • Korean Journal of Microbiology
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    • v.36 no.2
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    • pp.91-96
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    • 2000
  • The cultural and physiological conditions for the T-2 toxin [4,15-diacetoxy-8-(3-mety1butyloxy)-12,13- epoxy-trichothec-9-en-3-01, $C_{24}H_{30}O_9$] production by Fusarium spp. were studied. Thin layer chromatography (TLC) assay and the microbiological assay uslng Rhodotomla rubra were used to quantitate tbe T- 2 toxin. Among the four strains of Fusarium spp., F tn'cinctum NRRL 3299 was best for T-2 toxin production. In solid culture, white com grit medium was best for T-2 toxm production. Temperature played a critical role in the production of T-2 toxin. T-2 toxin production was favored by long duration of low-temperature incubation. The growth and toxin production were relatively high on galactose, fructose, glucose, and sucrose media, when each was used as a sole carbon source, and relatively low on sorbitol, glycerol, and lactose media. For nitrogen sources, $NH_4^(+) and NO_3^{-}were used well as a sole nitrogen source, but $NO_2^-$ was not used. Initial pH and speed of shaker also affected the production of T-2 toxin. From temperature shifting experiment, it is clear that T-2 toxin metabolic pathway is regulated by temperature-dependent enzyme depression or enzyme induction system.

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Production and partial purification of Staphylococcus aureus alpha toxin

  • Park, Hee-myung;Oh, Tae-ho;Han, Hong-ryul
    • Korean Journal of Veterinary Research
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    • v.39 no.5
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    • pp.1028-1032
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    • 1999
  • Alpha toxin of S aureus has cytolytic activity respectively. This antigen has been received the most attention since it is a major virulence factor in pathogenesis of staphylococcal mastitis. Thus, alpha toxin has been focused as potential candidate of vaccine to minimize mastitis in cows. The purpose of this study was to develop a simple, efficient production and purification methods of sufficient amount of alpha toxin antigen from S aureus. Alpha toxin production measured by hemolytic activity was the highest at 18 hrs postinoculation in yeast extract culture medium supplemented with thiamine, nicotinic acid and casamino acid. Alpha toxin was purified by ammonium sulfate precipitation (65%) and ultrafiltration. Molecular weight of the toxin was 33 kDa in the analysis with SDS-PAGE. Conclusionally, when alpha toxin was included in the vaccine, the optimal harvest time of alpha toxin was at 18 hrs after inoculation in yeast extract medium supplemented with thiamine and nicotinic acid.

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Optimization of Culture Conditions for the Production of Diphtheria Toxin (디프테리아 toxin 생산을 위한 발효조건 최적화)

  • Cho, Min;Ryu, Yeon-Woo
    • KSBB Journal
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    • v.14 no.2
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    • pp.241-247
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    • 1999
  • Experimental studies were carried out to optimize the culture conditions of Corynebacterium diphtheriae for the production of diphtheria toxin. A new media which does not contain any meat digest products was selected. The main ingredient of new medium was enzymatic digests of casein known as NZ-Case. In fermenter experiments, the toxin production was increased with the increase of cell growth. The optimum initial pH of media, air flow rate and agitation speed were 7.0, 0.22, vvm and 400 rpm, respectively. The contents of iron and calcium-phosphate precipitate were important for maximal cell growth and toxin production. The optimum concentration of iron was 0.3 mg/L and calcium-phosphate precipitate could serve in gradual supply of iron to maintain the optimal culture condition which is required for enhanced yield of toxin production. In potency test, the potency of toxoid from fermentor culture was higher than that from static culture. When diphtheria toxin is produced by fermentor culture, it is possible to produce higher levels of toxin and better toxoid quality in terms of safety, yield, productivity and immunity.

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Effects of Staurosporine and Genistein on Superoxide and HOCl Production in C5a- or PMA-activated Neutrophils (Staurosporine과 Genistein이 C5a 또는 PMA에 의하여 활성화된 호중구에서의 Superoxide와 HOCl 생성에 나타내는 영향)

  • Yun Young-Chul;Kang Hee-Jeong;Shin Yong-Kyoo;Lee Chung-Soo
    • The Korean Journal of Pharmacology
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    • v.31 no.1 s.57
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    • pp.115-122
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    • 1995
  • Effects of staurosporine, genistein and pertussis toxin on superoxide and HOCl production in C5a- or PMA-activated neutrophils were investigated. A C5a-induced superoxide and $H_2O_2$ production was inhibited by staurosporine, genistein and pertussis toxin. The stimulatory effect of PMA was inhibited by staurosporine but was not affected by pertussis toxin, whereas it was further promoted by genistein. Staurosporine and genistein inhibited superoxide production by sodium fluoride, but pertussis toxin did not affect it. PMA-induced $H_2O_2$ production was inhibited by staurosporine but was not affected by pertussis toxin. Genistein did not show a stimulatory effect on PMA-induced $H_2O_2$ production. Staurosporine and pertussis toxin inhibited HOCl production by C5a- or PMA, whereas genistein stimulated it. C5a-or PMA-induced myeloperoxidase release was inhibited by genistein, in this response the effect of pertussis toxin was not detected. Staurosporine did not affect the stimulatory effect of PMA on the release. Myeloperoxidase activity was markedly increased by genistein but was not affected by staurosporine and pertussis toxin. These results indicate that the respiratory burst of neutrophils may be regulated by protein kinase C and protein tyrosine kinase. Superoxide production induced by the direct activation of protein kinase C might be affected by protein tyrosine kinase oppositely. Genistein probably pro-motes HOCl production by activating myeloperoxidase.

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Effects of Polyphosphates on the Growth and T-2 Toxin Production of Fusarium sporotrichioides M-1-1 (인산염이 Fusarium sporotrichioides M-1-1 성장과 T-2 toxin 생성에 미치는 영향)

  • 장덕화;송재영;김일환
    • Journal of Food Hygiene and Safety
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    • v.10 no.4
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    • pp.199-204
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    • 1995
  • The antifungal effects of polyposphates on the growth and T-2 toxin production of Fusarium sporotrichioides M-1-1 were investigated. The growth of the strain was significantly inhibited in the potatoes dextrose agar medium treated with 1.5% polyphosphates or more. When we checked T-2 toxin by the indirect competitive ELISA, the strain produced 11.25 ug/ml and 10.90 ug/ml levels of T-2 toxin rice and corn containing 50% moisture contents, respectively. However, T-2 toxin was little detected in rice medium and corn medium with 1.5% polyphosphates addition for short(14 days) and prolonged incubation time(45 days). We also observed the destruction of cell wall and outflow of cell ingredients with 1% polyphosphates treatment to the strain. Therefore, moisture and polyphosphates greatly effected on the growth and T-2 toxin production of the strain.

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Modulation of Interleukin Production in Anthrax Lethal Toxin-treated Macrophages by Melatonin and Dehydroepiandrosterone

  • Shin, Sung-Ho;Hur, Gyeung-Haeng;Yeon, Kyu-Baek;Kim, Yun-Bae;Park, Kyung-Jin;Park, Young-Min;Lee, Woo-Sung;Cho, Bong-Huey;Kim, Won-Yong;Chung, Sang-In;Choi, Chul-Soon
    • BMB Reports
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    • v.33 no.6
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    • pp.463-468
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    • 2000
  • Anthrax lethal toxin, which consists of two separate protein, protective antigen (83 KDa) and lethal factor (85 KDa) is responsible for major symptoms and death from systemic infection of Bacillus anthracis. High concentrations of this toxin are cytolytic to macrophages, whereas sublytic concentrations of lethal toxin induce these cells to produce interleukin $1{\beta}$ ($IL-1{\beta}$). It is proposed that melatonin and dehydroepiandrosterone (DHEA) may play an important role in modifying immune dysfunction. In this study, we investigated whether or not melatonin and DHEA could prevent $IL-1{\beta}$ production that is induced by anthrax lethal toxin in mouse peritoneal macrophages. Treatment of melatonin or DHEA alone, as well as together, prevented the production of $IL-1{\beta}$ caused by anthrax lethal toxin. We found that melatonin at a concentration of $10^{-6}-10^{-7}$ M inhibits $IL-1{\beta}$ production induced by anthrax lethal toxin. As expect, treatment of DHEA at a concentration $10^{-6}-10^{-7}$ M also suppressed production of $IL-1{\beta}$ by lethal toxin stimulated macrophages. The results of these studies suggest that melatonin and DHEA, immunomodulators, may have an important role in reducing the increase of cytokine production in anthrax lethal toxin-treated macrophages.

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EFFECT OF FEEDING A LATHYRUS TOXIN ON EGG WEIGHT, EGG PRODUCTION AND EGG MALFORMITY IN LAYING CHICKEN

  • Chowdhury, S.D.
    • Asian-Australasian Journal of Animal Sciences
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    • v.4 no.3
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    • pp.275-280
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    • 1991
  • Beta-aminopropionitrile (BAPN)-a Lathyrus toxin was fed to laying chicken to investigate effects on egg weight, production and malformity. Two feeding trials were conducted where the levels of BAPN varied from 0.1 to 0.6 g per kg diet. The responses with regard to egg weight, egg production and the incidence of malformed eggs due to varying concentrations of the toxin were recorded and the results on weekly changes in these important variables were evaluated by plotting graphs. The results showed that there were increasing trends in egg weight and egg shell abnormalities and a decreasing trend in egg production as the dietary level of BAPN was increased indicating that the responses of layers were dose dependent. The effects on egg weight, egg production and egg malformity disappeared following withdrawal of the toxin from the diets.

Stimulatory Effects of cyclic AMP on Vitellogenin Induction by Estradiol-17$\beta$ in the Primary Culture of Hepatocytes in the Rainbow Trout Oncorhynchus mykiss

  • Yeo In-Kyu
    • Fisheries and Aquatic Sciences
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    • v.1 no.2
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    • pp.153-158
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    • 1998
  • Effects of cyclic (c) AMP and G-protein related reagents (3-isobutyl-l-methyxanthine (IBMX), Forskolin (FSK), cholera toxin (CTX), and pertussis toxin (PTX≫ on estradiol-17$\beta$ induced vitellogenin (VTG) induction were examined in primary hepatocyte cultures in rainbow trout Oncorhynchus mykiss. The addition of IBMX, FSK, or CTX to the incubation medium markedly increased VTG production, while PTX was not effective in stimulating the production. It is well known that cAMP regulates phosphorylation and dephosphorylation through mediation of protein kinase A. These results suggest that VTG production is highly dependent on cAMP state in hepatocytes because of its highly phosphorylated nature.

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Production of Hepatotoxin by the Cyanobacterium Scytonema sp. Strain BT 23

  • Ashok, Kumar;Singh, D.P.;Tyagi, M.B.;Kumar, Arvind;Prasuna, E.G.;Thakur, J.K.
    • Journal of Microbiology and Biotechnology
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    • v.10 no.3
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    • pp.375-380
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    • 2000
  • The preliminary screening of several cyanobacteria, using mice bioassay, reveale the production of a hepatotoxin by the cyanobacterium Scytonema sp. strain BT 23 isolated from soil. An intraperitoneal injection of the crude toxin (LD50 56 mg/kg body wt) from this strain caused the death of the mice within 40 min, and the anmals showed slinical signs of mice within 40 min, and the animals showed clinical signs of hepatotoxicity. The toxin was purified and partially characterized. The active fraction appears to be nonpolar in nature and shows absorption peaks at 240 and 285 nm. The purified toxin had an LD50 of TEX>$100<\mu\textrm{g}/kg$ body wt and the test mice died within 40 min of toxin administration. The toxin-treated mice showed a 1.65-fold increase in liver weight at 40 min and the liver color chnged to dark red due to intrahepatic hemorrhage and pooling of blood. Furthermore, the administration of the toxin to test mice induced a 2.58, 2.63, and 2.30-fold increse in the activity of the serum enzymes alanine aminotransferase, lactate dehydrogenase, and alkaline phosphatase, respectively. Further experiments with the 14C-labeled toxin revealed a maximum accumulation of the toxin in the liver. The clinical symptoms in the mice were similar to those produced by microcystin-L.R. These results suggest that hepatotoxins may also be produced in non bloom-forming planktonic cyanobacteria.

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Amino acids at N- and C-termini are required for the efficient production and folding of a cytolytic γ-endotoxin from Bacillus thuringiensis

  • Thammachat, Siriya;Pathaichindachote, Wanwarang;Krittanai, Chartchai;Promdonkoy, Boonhiang
    • BMB Reports
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    • v.41 no.11
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    • pp.820-825
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    • 2008
  • Bacillus thuringiensis Cyt2Aa toxin is a mosquito-larvicidal and cytolytic $\delta$-endotoxin, which is synthesized as a protoxin and forms crystalline inclusions within the cell. These inclusions are solubilized under alkaline conditions and are activated by proteases within the larval gut. In order to assess the functions of the N-and C-terminal regions of the protoxin, several N- and C-terminal truncated forms of Cyt2Aa were constructed. It was determined that amino acid removal at the N-terminal, which disrupts the $\beta$1 structure, might critically influence toxin production and inclusion formation. The deletion of 22 amino acids from the C-terminus reduced the production and solubility of the toxin. However, the removal of more than 22 amino acids from the C-terminus or the addition of a bulky group to this region could result in the inability of the protein to adopt the proper folding. These findings directly demonstrated the critical roles of N- and C-terminal amino acids on the production and folding of the B. thuringiensis cytolytic $\delta$-endotoxin.