• Title/Summary/Keyword: Lysozyme

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Further Studies on the Specificity of the N- and C-terminal Antigenic Determinant of Hen Egg-white Lysozyme (계난백(鷄卵白) Lysozyme의 N-말단(末端)과 C-말단(末端) 항원결정기(抗原決定基)에 대한 연구(硏究))

  • Ha, Youn-Mun
    • The Journal of the Korean Society for Microbiology
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    • v.12 no.1
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    • pp.19-32
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    • 1977
  • The specificity of the N- and C-terminal antigenic determinant($P_{17}$: sequence $Lys^1-{cys-}^6-Asn^{27},\;{Trp^{12}}_2-Cys^{127}-Leu^{129}$) of hen egg-white lysozyme(HL) was studied in more detail. In a Scatchard plot of the binding of $^{14}C$-acetyl HL with guinea pig purified anti-$P_{17}$ antibody experimental values bent sharply aear r=1. This suggests of two antibody populations with different affinities for HL or possible steric hindrance in the binding of a second HL molecule to the second binding site of the antibody molecule. The antigenic activities of various peptides were tested by measuring their inhibition of the binding of $^{14}C-acetyl-P_{17}$ with the antibody, Only $P_{17}$ and $P_{17}t$(sequence $Lys^1-cys^6-Homoser^{12},\;Trp^{123}-Cys^{127}-Leu^{128})$) were inhibitory, with $K_1$ values of $2.0{\times}10^4$ and $8.1{\times}10^3$, respectively. These results indicate that the direct binding site of $P_{17}$ to anti-$P_{17}$ antibody may be located in the terminal portion of $P_{17}$ (sequence $Lys^1-Cys^6-Homoser^{12},\;Trp^{123}-Cys^{127}-Leu^{129})$) while the rest of $P_{17}$ may be important in maintaining the conformation of this determinant. The single disulphide bond involved in this determinant is essential for manifestation of immunological activity.

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Adsorption Properties of the Lysozyme and Albumin with Physicochemical Properties of the Contact Lens (콘택트렌즈의 물리화학적 특성에 따른 라이소자임과 알부민의 흡착 특성)

  • Sung, Yu-Jin;Ryu, Geun-Chang;Jun, Jin
    • Journal of Korean Ophthalmic Optics Society
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    • v.18 no.3
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    • pp.261-270
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    • 2013
  • Purpose: Adsorption properties of lysozyme and albumin according to physiochemical properties of commercial contact lens classified with the FDA categories and a contact lens fabricated in the laboratory were investigated. Methods: The contact lens were prepared using HEMA(2-hydroxyethyl methacrylate) and TRIM(3-(trimethoxysilyl) propyl methacrylate) in a cast mold. Artificial tears containing lysozyme and albumin were prepared. We measured the amounts of protein adsorbed on the each lenses with varying adsorbed time (48 hour) and the pH range (6, 6.8, 7.4, 8.2, 9) of artificial tear. Amount of the proteins absorbed on the contact lenses were measured by using HPLC. Results: Time to reach the equilibrium of protein adsorption for silicone hydrogel lens was taken longer than hydrogel lens. The amount of adsorbed both lysozyme and albumin at equilibrium were greater for the hydrogel lens than the silicone hydrogel lens, and larger for the ionic lens than the non-ionic lens. Lysozyme was more adsorbed on the higher water content of contact lens, whereas albumin was more adsorbed on the lower water content of contact lens. Only lysozyme was adsorbed on the Group IV hydrogel lens of ionic higher water content. The adsorption of protein on contact lens increased with pH of artificial tears as close to the isoelectric point of each protein. Conclusions: The adsorption amount of lysozyme is more affected by the ionic strength of the contact lens surface than the water content of contact lens. Albumin adsorption is more affected by water content than the ionic strength of the contact lens surface. For the adsorption of proteins on the silicone hydrogel lens, the pore size, determined both by the number of Si atoms and the chemical structure of the silicone-containing monomers, as well as the polarity of contact lens should be also considered.

Statistical Optimization of the Lysis Agents for Gram-negative Bacterial Cells in a Microfluidic Device

  • Kim, Young-Bum;Park, Ji-Ho;Chang, Woo-Jin;Koo, Yoon-Mo;Kim, Eun-Ki;Kim, Jin-Hwan
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.4
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    • pp.288-292
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    • 2006
  • Through statistically designed experiments, lysis agents were optimized to effectively disrupt bacterial cells in a microfluidic device. Most surfactants caused the efficient lysis of Gram-positive microbes, but not of Gram-negative bacteria. A Plackett-Burman design was used to select the components that increase the efficiency of the lysis of the Gram-negative bacteria Escherichia coli. Using this experimental design, both lysozyme and benzalkonium chloride were shown to significantly increase the cell lysis efficiency, and ATP was extracted in proportion to the lysis efficiency. Benzalkonium chloride affected the cell membrane physically, while lysozyme destroyed the cell wall, and the amount of ATP extracted increased through the synergistic interaction of these two components. The two-factor response-surface design method was used to determine the optimum concentrations of lysozyme and benzalkonium chloride, which were found to be 202 and 99 ppm, respectively. The lysis effect was further verified by microscopic observations in the microchannels. These results indicate that Gram-negative cells can be lysed efficiently in a microfluidic device, thereby allowing the rapid detection of bacterial cells using a bioluminescence-based assay of the released ATP.

Optimization of Chromatographic Separation of Lysozyme from Homogenate of Hen Egg White by Comparison of Breakthrough Behavior (파과분석(Breakthrough behavior) 비교에 의한 난백으로부터 라이소자임 크로마토그래피 분리 최적화)

  • 김원경;정봉현
    • KSBB Journal
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    • v.14 no.3
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    • pp.279-283
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    • 1999
  • We have compared the breakthrough behavior of lysozyme contained in fresh han egg white on various cation exchagers, and the adsorbent, known by the trade name Cellufine C-200 (Amicon), has shown the best performance. The effects of ion strength, pH, and linear flow rate on the breakthrough behavior were examined using the Cellufine C-200 adsorbent. The optimal conductivity, pH and linear flow rate were determined from the breakthrough behavior and found to be 2.75 mS/cm, 7.0, and 0.635 cm/min, respectively.

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Effect of Quercetin on the Activity and mRNA Expression of Antioxidant Enzymes and Physiological Responses in Olive Flounder (Paralichthys olivaceus) Exposed to Cadmium

  • Shin, H.S.;Yoo, J.H.;Min, T.S.;Lee, J.;Choi, C.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.6
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    • pp.742-749
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    • 2010
  • We investigated the antioxidant efficacy of quercetin (0% Diet 1, 0.25% Diet 2, and 0.5% Diet 3) pretreatment for 30 and 60 days in response to cadmium (Cd) toxicity in the olive flounder, and measured the plasma lysozyme activity to understand the immune effects of quercetin. The lysozyme activity with Diets 2 and 3 was higher than with Diet 1. Based on this result, to examine the immune ability and antioxidant role of quercetin, we exposed olive flounder fed quercetin to Cd and then measured the expression and activity of antioxidant enzymes (superoxide dismutase (SOD) and catalase (CAT)) and lipid peroxidation (LPO). With Diets 2 and 3, the expression and activity of antioxidant enzymes and the $H_2O_2$ concentration were lower than with Diet 1. In addition, the LPO levels were lower than with Diet 1, which protected the cell membrane. Therefore, quercetin removed the reactive oxygen species (ROS) produced by Cd, indicating that quercetin has antioxidant ability. In addition to its antioxidant ability, quercetin has immune effects.

The Effects of Acute Osmotic Stress on Innate Immunity of Nile Tilapia (Oreochromis niloticus)

  • Choi, Sang-Hoon;Park, Kwan-Ha
    • Fisheries and Aquatic Sciences
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    • v.13 no.4
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    • pp.343-349
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    • 2010
  • The effects of osmotic stress on the non-specific immune response of Nile tilapia, Oreochromis niloticus, were investigated. Osmoregulatory mechanism of tilapia has been studied, but less information is available about innate immune response of O. niloticus faced with hyperosmolality. Acute osmotic stress was elicited by transferring tilapia from freshwater (FW) to 24 psu seawater (SW). Non-specific immune parameters including lysozyme activities of plasma and head kidney (HK), alternative complement pathway (ACP) activity in plasma, phagocytic capacities of spleen and HK immune cells, and respiratory burst activity of immune cells in both HK and spleen were analyzed. Lysozyme activities were increased at 1 h and 30 h after transfer to SW, but decreased at 10 h after SW transfer. Conversely, ACP activity increased 10 h after SW transfer. Phagocytic capacity increased slightly at 1 h and 5 h after SW transfer, and respiratory burst activity showed an increase in superoxide release at 10 h after SW transfer. Taken together, these results indicate that the exposure of tilapia to hyperosmotic conditions has immunostimulatory effects on cellular and humoral immune reactions.

Effect of Korean Mistletoe (Viscum album Coloratum) on the Non-Specific Immune Responses in Japanese Eel (Anguilla japonica)

  • Cho, Young-Hye;Choi, Dae-Han;Choi, Sang-Hoon
    • Journal of fish pathology
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    • v.18 no.1
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    • pp.59-66
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    • 2005
  • In the present paper, the immunostimulatory effects of Korean mistletoe (Viscum album Coloratum) on the non-specific immune responses of Japanese eel (Anguilla japonica) were examined. Eel were innoculated with mistletoe, Freund's complete adjuvant (FCA), or phosphate-buffered saline (PBS) as a control into their peritoneal cavities. The number of nitrobule tetrazolium (NBT)-positive cells in the head kidney of fish was significantly increased by the second day post-injection of mistletoe. ROI products were more enhanced in mistletoe-injected fish kidney leucocytes than in FCA-injected ones. The level of lysozyme activity detected in the serum of fish 2 days after injection with mistletoe was also significantly higher than that found in the serum of the control fish. The appropriate concentration of mistletoe to induce the highest level of serum lysozyme activity was revealed to 1000${\mu}g$/200 g of fish. In phagocytic activity assay, mistletoe-sensitized eel kidney phagocytes captured more zymosan than did the control fish. Korean mistletoe appeared to be a good activator of the non-specific immune responses of Japanese eel.

The Effecs of Preservatives on the Shelf-life of Boiled Scallop[Patinopecten yessoensis(Jay)] (보존제 첨가에 의한 자숙가리비의 보존 효과)

  • 김상무;조순영
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.27 no.1
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    • pp.69-74
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    • 1998
  • Scallop, Patinopecten yessoensis (Jay), cultivated in the cold east coast of Kangwon region in Korea, is expected to be produced to about 50,000 tons in 2000 year. First of all, scallop should be exuviated to process for new goods. In this study, preservatives such s sorbate, lactate, benzoic acid, and lysozyme were to extend the shelf-life of exuviated scallop boiled with steam for 10 min. The addition of preservatives retarded the decrease in pH, and inhibited the productions of VBN, TMA, and TBA of exuviated scallop stored at $5^{\circ}C$ in the latter period of storage. They also inhibited the microbial growth, especially in the first period of storage. The estimated values of shelf-life of boiled and exuviated scallops for control, sorbate, lactate, benzoic acid, and lysozyme were about 22, 26, 29, 25, and 27 days, respectively.

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Induction of Bactericidal Substance from the Immunized Larval Haemolymph of L. illustris (면역유충 체액으로 부터 항균물질의 유도)

  • 육순학;장정순
    • Korean Journal of Microbiology
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    • v.25 no.4
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    • pp.309-317
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    • 1987
  • It was known that normal-haemolymph from the 3rd instar larvae of Lucillia illustris contain a lysozyme (or lysozyme-like substance) with bactericidal activity to fram positive bacteria, and the bactericidal activity of injured-haemolymph was increased significantly after injuring the body wall. To elucidate the defence mechanism of insect against the nonpathogenic bacteria, the immune-haemolymph against Escherichia coli K-12 was prepared after immunization. The bactericidal activity between injured and immune-haemolymph was compared, and it was revealed that the immune-haemolymph showed higher titer of bactericidal activity to fram positive bacteria as well as to Escherichia coli. The bactericidal substance from the immune-haemolymph was purified through a successive chromatographies on Sephacryl S-300 and CM-Sepharose CL-6B, and it was characterized as a basic protein in nature with heat stable property at acidic conditions.

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Physiological Responses of the Chicken Grunt Parapristipoma trilineatum to High Water Temperature Stress (사육수의 고수온 스트레스가 벤자리(Parapristipoma trilineatum)에 미치는 생리학적 영향)

  • Kim, Ki-Hyuk;Hong, Sung-Won;Moon, Hye-Na;Yeo, In-Kyu
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.51 no.6
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    • pp.714-719
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    • 2018
  • We investigated the effects of water temperature on physiological parameters in the chicken grunt Parapristipoma trilineatum. At high temperature, the aspartate aminotransferase (AST) and the alanine aminotransferase (ALT) levels were increased, suggesting that high temperature induced hepatic damage. In addition, total protein (TP) was high at high water temperatures, which were considered stressful in the breeding environment. At high water temperatures, triglycerides (TG) were low due to increased metabolic activity, which decreased the blood TG levels as TG were used as an energy source. There was no significant difference in the plasma osmolality or the blood ion concentrations with water temperature. In generally, lysozyme, a factor in innate immunity, increased with water temperature. However, lysozyme activity tended to decrease with increasing water temperature, but the difference was not significant. These results suggested that the decrease of biophylaxis at high temperature was affect the growth or survival of the population.