• Korean Journal of Food Science and Technology
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• v.23 no.3
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• pp.366-369
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• 1991

In order to elucidate texturization mechanism of extrudated protein, egg white lysozyme was heated under high pressure conditions, and its solubility and changes of molecular weight were investigated. Under high pressure conditions of $100,\;300\;and\;600\;kg/cm^2$, solubility decreased gradually with increasing temperature in the samples heated at $70,\;120\;and\;150^{\circ}C$ and decreased notably with increasing pressure at $200^{\circ}C$. Polymerization was found in the samples heated at $150\;and\;200^{\circ}C$ while a band which located below monomer(low-molecular) could be recognized. Molecular weight of the low-molecular was estimated to be about $6,000{\sim}9,000$ and no smaller peptide was recognized. The polymerization may have occured by disulfide crosslinking in the samples heated at $120^{\circ}C$ but other crosslinking may have played a role in those at $150\;and\;200^{\circ}C$.

• Journal of fish pathology
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• v.12 no.1
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• pp.16-23
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• 1999

The immunomodulatory effects of levamisole (LMS) were evaluated in leucocytes of eels in vitro. Proliferation of lymhocytes treated with T-cell mitogen (Con A or PHA) was markedly inhibited by LMS in a dose dependent manner. B cell mitogen (LPS), in contrast, slightly increased the proliferaion. On the other hand, production of MIF and MAF when treated with Con A was increased in a dose-dependent way. NK cell activities were somewhat increased when LMS was pretreated and this augmentation was due to an increase in binding capacity of effector-target cell, but not due to the target cell lytic activity of effector cells. Phagocytic activity, superoxide anion formation, hydrogen peroxide formation and lysozyme activity of leucocytes were enhanced by LMS in a dose related-manner. These results suggest that LMS might modulate the immmune responses by activation of cytokine production and by augmentation of leukocyte activity but not by increment of immunocompetent cell numbers.

• Microbiology and Biotechnology Letters
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• v.17 no.5
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• pp.412-420
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• 1989

To know how the ribosomes involved in secretory protein synthesis were attached to the cytoplasmic membrane in Bacillus amyloliquefaciens, the cells were treated with puromycin combinated with magnesium at the logarithmic phase, and the variation of cell-bound and extracellular $\alpha$-amylase activity was assayed for determining the $\alpha$-amylase translocation blocking through the cytoplasmic membrane. In the abnormal $\alpha$-amylase producing mutant in which the C-terminal of the $\alpha$-amylase structure was deleted, B. umytotiquefaciens CH10-2, the $\alpha$-amylase was translocated normally through the cytoplasmic membranes, and the translocation blocking by puromycin was revealed to have a similar pattern as that in the wild type. This means that the C-terminal part of the enzyme structure may not have a signal for secretion. The cell death of the logarithmic phase cells in both strains was not affected much under 20$\mu\textrm{g}$/$m\ell$ of puromycin, however, the $\alpha$-amylase translocation was blocked markedly under less than 10$\mu\textrm{g}$/$m\ell$ of the puromycin concentration. The blocking of the enzyme secretion by puromycin may be due to the detachment of the ribosomes from cytoplasmic membranes by disturbing the nascent polypeptide synthesis. Further evidence for confirming this was that the detachment was increased in 50 mM of magnesium ion because the extracellular $\alpha$-amylase activity was decreased more under this condition. If the cells were treated with trypsin combinated with Iysozyme, the extracellular $\alpha$-amylase activity from the cultured medium was reduced markedly, however, the activity from the cells treated with trypsin only was not reduced. This means that the nascent polypeptides protruding from the cytoplasmic membrane were sensitive to the trypsin digestion, whereas the matured ones were not. Therefore, the protruding polypeptides from the cytoplasmic membranes may be truncated by trypsin before forming their final tertiary structures by folding in the cell wall layer.

• Journal of Life Science
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• v.13 no.1
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• pp.90-98
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• 2003

An allium rhizobacterium Serratia plymuthica AL-1 was previously selected as a biocontrol agent of allium white rot. The chitinase from S. plymuthica AL-1 produced in medium containing colloidal chitin was purified by ammonium sulfate precipitation (40~70%), affinity adsorption, column chromatography on DEAE-sephadex A-50 and sephadex C-200 gel filtration. The enzyme was purified 10.8-fold with a yield of 7.3% from the starting culture broth. The purified chtinase gave a single band on sodium dodecyl sulfate polyacrylamide gel electrophoresis, it's molecular weight was estimated to be 55 kDa. The optimum pH and temperature of the purified enzyme were pH 5.5 and $55^{\circ}C$, respectively and it is stable up to $50^{\circ}C$ and maintains around 90% of its activity for 60min. The enzyme were activated by $Ca^{2+}$, $Mn^{2+}$ and $Mg^{2+}$ and inhibited by $Cu^{2+}$, SDS, $\rho$-CMB, MIA, respectively. The purified chitinase showed broad spectrum of antifungal activities against plant pathogenic fungi Sclerotium cepivoruin, Alternana alternnta, Colletotrichum glceosporioidrs, Phoma sp., Sclerotinia sclerotiorum, Stemphylium solani, Fusarium oxysporium f. sp. niveum but rarely inhibited Phytophthora capsici and Pythium ultimum.. The purified chitinase from S. plymuthica AL-1 caused swelling, lysis, deceleration and degradation of the hyphal tips of S. sczerotiorum causing allium white rot. It suggest that S. prymuthica AL-1 chitinase play an important part in the bifunctional chitinase / lysozyme activity.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.39 no.6
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• pp.460-465
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• 2006

An 8-week feeding trial was conducted to investigate the effects of dietary supplementation with probiotics as a feed additive for Juvenile olive flounder (Paralichthys olivaceus). Three experimental diets supplemented with Bacillus polyfermenticus (BP), Bacillus licheniformis (BL), or Bacillus polyfermenticus plus Saccharomyces cerevisiae, (BP+SC) at $1.0{\times}10^7CFU/kg$ diet on a dry-matter basis were prepared. The basal diet was used as a control. After the 8-week feeding trial, the respiratory burst activity (NBT assay) of fish fed the BP + SC diet was significantly higher than that of fish fed the control diet. Fish fed the BP, BL and BP + SC diets had significantly lower cumulative mortality than did fish fed the control diet after the third day of the challenge test (P<0.05). However, there were no significant differences among fish fed the experimental diets in weight gain, feed efficiency, protein efficiency ratio, hematosomatic index, condition factor, survival rate, or Iysozyme activity. Results could suggest that dietary B. polyfermenticus, B. licheniformis, and B. polyfermenticus +S. cerevisiae enhance nonspecific immunity and disease resistance in juvenile olive flounder.

• The Korean Journal of Cytopathology
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• v.6 no.2
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• pp.106-115
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• 1995

Reactive humsn mesothelial cells were examined by immunocytochemical stain with intermediate filaments (cytokeratin [CK1, CK7, CK8, CK18, CD19), vimentin, desmin, actin), epithelial membrane antigen, carcinoembryonic antigen (CEA), MHC class II antigen (HLA-DR), LeuM-1 (CD15), $\alpha1-antitrypsin$(ACT), $\alpha1-antichymotrypsin$ (ACHT), CD68(KP-1) and FcyRIII(CD16). The mesothelial cells were isolated from patients with liver cirrhosis and pleural effusion, and short-term cultured in RPMI 1640 media containing 10% heat inactivated fetal calf serum and 1% identical supernatant fluid of the patients' transudates. The results obtained are as follows 1. The cultured-reactive mesothelial cells were positive for the protein of cytoskeleton such as cytokeratin and vimentin, but negative for desmin and actin. The resting mesothelial cells showed positive reactions for cylokeratin, but negative for vimentin, desmin and actin. 2. The primary antibodies to the cytokeratin were strongly reactive for CK1, CK8 and CK18 but negative for CK7 and CK19 in both reactive and resting mesothelial cells. 3. Resting mesothelial cells showed negative reactions for CEA, but strong positive reactions in cultured-reactive mesothelial cells. 4. The markers for the monocytes/histiocytes(CD11b, CD14, CD16, CD68, Iysozyme and $\alpha1-antitrypsin$ and $\alpha1-antichymotrypsin$) were nonreactive in resting mesothelial cells, but lysozyme and $\alpha1-antitrypsin$ were weakly reactive in reactive and proliferative mesothelial cells. 5. MHC Class II molecule(HLA-DR antigen) was negative in both resting and reactive mesothelial cells. These results suggest that the short-term cultured, reactive mesothelial cells show a newly aberrant expression of the vimentin and calcine-embryonic antigen. The reason of the aberrant expression of the intermediate filament and oncofetal antigen in reactive and proliferative mesothelial cells should be further evaluated.

• Korean Journal of Food Science and Technology
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• v.22 no.6
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• pp.711-715
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• 1990

Continuous column chromatographic separation of lysozyme from egg white was investigated. A weak acid type cation exchange resin, Duolite C-464, was used because of high lysozyme recovery and ease of column operation in this experiment. The resin was equilibrated at $pH\;7.9{\pm}0.1$ in Na+form. Continuous lysozyme separation was processed by repeating cycles(one cycle : resin equilibration, flow egg white, rinse, lysozyme elution) in automated preparative Liquid Chromatography(LC) system(column size ; i.d. 50 mm, resin bed volumn ; 1020 ml). At comparison of UV levels in rinse end point and elution end point of every cycle, the UV levels of rinse end point are maintained below 30% for 19 cycles and that of elution end point are also maintained below 30% for 17 cycles, stably, but was increased above 50% after 18 cycle. That indicated the eluting ability of lysozyme was reduced conspicuously after 18 cycle in continuous cycling process. The recovery of lysozyme was maintained above 90% from one to 17 cycle, but was decreased to 72% and 65% in 18 cycle and 19 cycle, respectively.

• Korean Journal of Food Science and Technology
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• v.28 no.3
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• pp.399-404
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• 1996

Encapsulation of lysozyme using lecithin vesicles and its stimilated release properties were studied. Lecithin vesicles were prepared by the dehydration-rehydration (DR)method. The highest encapsulation efficiency (EE) value of 80.1% was obtained by sonicating the multilamellar vesicles (MLVs) at 100 KHz for 120 min in bath sonicator. The value of entrapment progressively increased with the concentration of lysozyme, while the EE value decreased with the increase of enzyme concentration up to 50mg per 100mg per 100mg of lecithin, and then became nearly constant. At the pH of 5.9, only a small amount of lysozyme was released from DR vesicles during incubation at $37^{\circ}C$ As the pH decreased to 3.0, lysozyme was released more rapidly. Lysozyme release was accelerated for 24h and reached a plateau after 72h incubation with 1% Tween 80, $Ca^{2+}$ gave a pulse-like-release in the first hour, which was followed by a slow release.

• Korean Journal of Poultry Science
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• v.16 no.3
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• pp.157-167
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• 1989

Pretreatment of egg white and ion exchange resins was attempted to separate lysozyme from egg white efficiently. Apparent viscosity of egg white could be decreased to 3cp by homogenization for 30 minutes at 2, 000rpm and ultrasonication for 45 minutes. The result of testing adsorption capacity of lysozyme was as follows； CM-Sephadex C-25 ＞Duolite C464＞Amberlite C-50＞Dowex MSC-1＞Amberlite IRC-50＞Amberlite IRC-84. Although CM-Sephadex C-25 showed highest adsorption capacity of lysozyme, egg white could not eluted easily. Duolite Cf64 was selected based on relatively high lysozyme adsorption and good egg white eluting property for separation of egg white lysozyme. Na$^{＋}$ form of Duolite C-464 was most effective on adsorption of Iysozyme. To separate lysozyme from egg white efficiently rinse buffer and eluting solution were selected 0.1M sodium phosphate buffer at pH 6.5 and 10% ammonium sulfate respectively. After separating lysozyme from egg white, foaming power of egg white was decreased to 85.3%. Color of egg white gel was not changed while hardness of egg white gel was decreased by 30% after separating lysozyme. However, elasticity of egg white gel was increased by 13% in lysozyme-separated egg white.