• Title/Summary/Keyword: Lysozyme C

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Inhibitory effects of lysozyme on endothelial protein C 1receptor shedding in vitro and in vivo

  • Ku, Sae-Kwang;Yoon, Eun-Kyung;Lee, Hyun Gyu;Han, Min-Su;Lee, Taeho;Bae, Jong-Sup
    • BMB Reports
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    • v.48 no.11
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    • pp.624-629
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    • 2015
  • Lysozyme protects us from the ever-present danger of bacterial infection and binds to bacterial lipopolysaccharide (LPS) with high affinity. Beyond its role in the activation of protein C, the endothelial cell protein C receptor (EPCR) plays an important role in the cytoprotective pathway. EPCR can be shed from the cell surface, which is mediated by tumor necrosis factor-α converting enzyme (TACE). However, little is known about the effects of lysozyme on EPCR shedding. We investigated this issue by monitoring the effects of lysozyme on phorbol-12-myristate 13-acetate (PMA)-, tumor necrosis factor (TNF)-α-, interleukin (IL)-1βand cecal ligation and puncture (CLP)-mediated EPCR shedding and underlying mechanism. Data demonstrate that lysozyme induced potent inhibition of PMA-, TNF-α-, IL-1β-, and CLP-induced EPCR shedding. Lysozyme also inhibited the expression and activity of PMA-induced TACE in endothelial cells. These results demonstrate the potential of lysozyme as an anti-EPCR shedding reagent against PMA-mediated and CLP-mediated EPCR shedding.

Modification of Functionality for Lysozyme (Lysozyme의 기능성 개선)

  • Kim, Hyun-Ku
    • Applied Biological Chemistry
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    • v.37 no.6
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    • pp.456-462
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    • 1994
  • Lysozyme-dextran hybrids were prepared by incubated storage at $60^{\circ}C$ and 80% relative humidity for 16 days. The emulsifying properties of the hybrids were about 14 times higher than those of native lysozyme and were about 3 times higher than those of commercial emulsifiers. Lytic activity of the hybrids remained about 83% that of native lysozyme when mesured against Micrococcus lysodeikticus as a substrate. The excellent emulsifying properties of the hybrids were maintained even at pH 3 and were further improved at pH 10. The emulsifying properties of the hybrids were greatly improved by preheating the hybrids at $100^{\circ}C$. In addition the lysozyme-dextran hybrids showed an antimicrobial effect on Gram-negative bacteria.

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Molecular Cloning and Characterization of a New C-type Lysozyme Gene from Yak Mammary Tissue

  • Jiang, Ming Feng;Hu, Ming Jun;Ren, Hong Hui;Wang, Li
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.12
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    • pp.1774-1783
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    • 2015
  • Milk lysozyme is the ubiquitous enzyme in milk of mammals. In this study, the cDNA sequence of a new chicken-type (c-type) milk lysozyme gene (YML), was cloned from yak mammary gland tissue. A 444 bp open reading frames, which encodes 148 amino acids (16.54 kDa) with a signal peptide of 18 amino acids, was sequenced. Further analysis indicated that the nucleic acid and amino acid sequences identities between yak and cow milk lysozyme were 89.04% and 80.41%, respectively. Recombinant yak milk lysozyme (rYML) was produced by Escherichia coli BL21 and Pichia pastoris X33. The highest lysozyme activity was detected for heterologous protein rYML5 (M = 1,864.24 U/mg, SD = 25.75) which was expressed in P. pastoris with expression vector $pPICZ{\alpha}A$ and it clearly inhibited growth of Staphylococcus aureus. Result of the YML gene expression using quantitative polymerase chain reaction showed that the YML gene was up-regulated to maximum at 30 day postpartum, that is, comparatively high YML can be found in initial milk production. The phylogenetic tree indicated that the amino acid sequence was similar to cow kidney lysozyme, which implied that the YML may have diverged from a different ancestor gene such as cow mammary glands. In our study, we suggest that YML be a new c-type lysozyme expressed in yak mammary glands that plays a role as host immunity.

Lytic Action of Egg White Lysozyme Isolated from Ogol Fowl on Staphylococcus aureus Phage Type 29 (Staphylococcus aureus Phage Type 29에 대한 오골계 난백 Lysozyme의 용균성)

  • Oh, Hong Rock;Lee, Jong Soo;Kim, Chan Jo
    • Korean Journal of Agricultural Science
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    • v.14 no.2
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    • pp.286-294
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    • 1987
  • This experiment was carried out to investigate the bacteriolytic action of the egg white lysozyme isolated from Korean native Ogol fowl and to obtain the data for utilization of the enzyme as a food preservative. Staphyococcus aureus phage type 29 and Bacillus subtilis ATCC 6633 among the microorganisms tested were lyzed by the treatment with 0.05% lysozyme, but Staphylococcus aureus phage type 57 in addition to E. coli etc. was found to be a lysozyme- insensitive species. The lysis of S. aureus phage type 29 was maximized when incubated in nutrient broth (pH 7.0) at $37^{\circ}C$ for 24 hours and suspended it to absorbance 0.6 at 540nm in 0.05M sodium acetate but fer (pH 4.5) and then treated it with the 0.05% lysozyme for 30 min. at $30^{\circ}C$. It was found that the effect of 0.05% lysozyme in combination with 1% glycine on the growth inhibition of S. aurecus phage type 29 increased more 50% than that in the absence of glycine, but not effect with other any additeves and metal ions tested.

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Molecular Cloning and Characterization of Lysozyme II from Artogeia rapae and its Expression in Baculovirus-infected Insect Cells

  • Bang, In-Seok;Kang, Chang-Soo
    • Animal cells and systems
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    • v.11 no.2
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    • pp.175-182
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    • 2007
  • The lysozyme II gene of cabbage butterfly Artogeia rapae was cloned from fat body of the larvae injected with E. coli and its nucleotide sequence was determined by the RACE-PCR. It has an open reading frame of 414 bp nucleotides corresponding to 138 amino acids including a signal sequence of 18 amino acids. The estimated molecular weight and the isoelectric point of the lysozyme II without the signal peptide were 13,649.38 Da and 9.11, respectively. The A. rapae lysozyme II (ARL II) showed the highest identity (81%) in the amino acid sequence to Manduca sexta lysozyme among other lepidopteran species. The two catalytic residues ($Glu^{32}$ and $Asp^{50}$) and the eight Cys residue motifs, which are highly conserved among other c-type lysozymes in invertebrates and vertebrates, are also completely conserved. A phylogenetic analysis based on amino acid sequences indicated that the ARL II was more closely related to M. sexta, Hyphantria cunea, Heliothis virescens, and Trichoplusia ni lysozymes. The ARL II gene was expressed in Spodoptera frugiperda 21 insect cells and the recombinant ARL II (rARL II) was purified from cell-conditioned media by cation exchange column chromatography and reverse phase FPLC. The purified rARL II was able to form a clear zone in lysoplate assay against Micrococcus luteus. The lytic activity was estimated to be 511.41 U/mg, 1.53 times higher than that of the chicken lysozyme. The optimum temperature for the lytic activity of the rARL II was $50^{\circ}C$, the temperature dependency of the absolute lytic activity of rARL II was higher than that of the chicken lysozyme at low temperatures under $65^{\circ}C$.

The Effect of pH and Temperature on Lysozyme Separation in Ion-exchange Chromatography (이온교환크로마토그래피에서 라이소자임 분리에 미치는 pH와 온도 영향)

  • Ko, Kwan-Young;Kim, In-Ho
    • Korean Chemical Engineering Research
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    • v.52 no.1
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    • pp.98-105
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    • 2014
  • Lysozyme amounts to 0.3% in egg white and functions as an agent of cell lysis and activator of tissue reconstruction. Ion exchange chromatography is the most useful method of separation among affinity chromatography, ion exchange chromatography, and ultra-filtration. The aim of present study is to find the optimum pH and temperature for the separation of lysozyme in egg white within cation exchange gel filled glass column. And we compared results of experiments with those of simulations. Phosphate buffer was used, and pH and temperature were varied as 5~7 and $25{\sim}40^{\circ}C$ respectively. RP-HPLC was the tool for the retention time identification and quantitative analysis of lysozyme. OriginPro 8 measured the peak area of lysozyme chromatogram and quantified the eluted lysozyme. Largest amount of lysozyme was separated under the conditions of pH 5 and T $25^{\circ}C$.

The Preservative Effect of Egg White Lysozyme Added Surumi Products (난백 lysozyme에 의한 연제품의 방부 효과)

  • KIM Young-Man;LEE Byung-Ho;LEE Sang-Hoon;SHIN Il-Shik;LEE Tae-Shik
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.21 no.4
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    • pp.269-275
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    • 1988
  • Inhibitory effects on bacterial growth by using lysozyme and mixtures of it with other antibacterial substances (sodium hexametaphosphate and sodium pyrophosphate) were investigated against the 7 kinds of hacterial strains isolated from putrefied surumi products. The growth inhibitory concentrations of lysozyme and lysozyme + sodium hexametaphosphate + sodium pyrophosphate against the bacteria were added to kamaboko, imitation crab meat and fried surumi, then viable cell count, pH and VBN were examined during the storage at $30^{\circ}C$ for 7 days. Lysozyme showed growth inhibitions against 6 of 7 isolates and the inhibition effect of mixture of antibacterial substances was multiplied against all the isolates compare with those of its individual use. Growth inhibitory effect of the substances on the bacteria was high in order of lysozyme + sodium pyrophosphate + sodium hexametaphosphate, lysozyme + sodium hexametaphosphate, lysozyme + sodium pyrophosphate and lysozyme. The most effective inhibitory concentration of mixture of the antibacterial substances in kamaboko and imitation crab meat was $0.05\%$ of lysozyme, $0.5\%$ of sodium pyrophosphate and $0.1\%$ sodium hexametaphosphate. But the bacterial growth was slightly inhibited in fried surumi even if the same concentration of the dipped mixture and the effect of the mixture was less than that of $0.2\%$ sorbic acid.

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Protoplast Formation and Regeneration of Thermophilic Clostridium thermocellum and Clostridium thermohydrosulfuricum (고온성 Clostridium thermocellum과 Clostridium thermohydrosulfuricum의 원형질체 형성 및 재생)

  • 김욱한;정기택;이용현
    • Korean Journal of Microbiology
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    • v.28 no.4
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    • pp.304-310
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    • 1990
  • The conditions for protoplasts formation and regeneration of thermophilic anaerobic C. thermocellum and C. thermohydrosulfuricum were determined under the anaerobic growth conditions. The cells of C. thermocellum in initial exponential growth phase were identified to be the most suited for protoplast formation. The optimal conditions for protoplast formation were found to be at $37^{\circ}C$ for 2 hours with 0.5 mg/ml of lysozyme in TMG buffer (pH7.5). On the other hand, C. thermohydro-sulfuricum grown in the same medium but excluding glycine was optimally protoplasted at the same conditions but with 0.2 mg/ml of lysozyme. The protoplasts of both strains only subjected to lysozyme treatment of the short time were satisfactorily regenerated after 7-10 days incubation at $60^{\circ}C$ in regeneration medium containing 0.3-0.4 M sorbitol, 0.5% casamino acid, and high concentration of $CaCl_{2}$ and $MgCl_{2}$. The regeneration frequencies of the protoplasts of C. thermocellum and C. thermohydrosulfuricum were found to be very low level of $4.85{\times}10^{-3}$ and $4.23{\times}10^{-2}$, respectively. The nonregenerated L-form cells were also observed inregeneration medium together with regenerated cells.

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Antibacterial Activity of Lysozyme-Galactomannan Conjugate against Escherichia coli

  • Hwang, Jae-Kwan;Kim, Hyun-Jin;Park, Moon-Jung;Shin, Hae-Hun;Pyun, Yu-Ryang
    • Preventive Nutrition and Food Science
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    • v.3 no.4
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    • pp.320-323
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    • 1998
  • Lysozyme was covalentyl conjugated with galactomannan through a amino-carbonyl reaction between the lysine $\varepsilon$-amino groups of lysozyme and the reducing ends of galactomannan at a relative humidity of 79% and 6$0^{\circ}C$. The resulting lysozyme-galactomannan conjugate (LGC) was investigated for its antibacterial activity against Escherichia coli. Lysozyme alone did not exhibit antibacterial activity against E. coli. in contrast , significant bactericidal effect was observed for LGC, depending on the reaction temperature. The degree of conjugation between lysozyme and galactomannan was dependent on the incubation time, which affected the antibacterial efficiency against E. coli. This study demonstrated that the amino-carbonyl reaction between lysozyme and galactomannan could be a potential tool to modify lysozyme toward broadening its antibacterial spectrum to Gram-negative bacteria.

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Expression of c-Type Lysozyme from the Fleshy Shrimp Fenneropenaeus chinensis Is Upregulated Following Vibrio anguillarum and Lipopolysaccharide Injection

  • Qiao, Guo;Kim, Su-Kyoung;Cho, Yeong-Rok;Kim, Sukyoung;Jang, In-Kwon
    • Fisheries and Aquatic Sciences
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    • v.16 no.4
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    • pp.267-272
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    • 2013
  • Chicken-type lysozyme (c-lysozyme) is present in shrimp and is active against some bacteria. To further understand the regulation of c-lysozyme in the fleshy shrimp Fenneropenaeus chinensis, we determined the tissue-specific gene expression of c-lysozyme and the time-course of mRNA expression in response to Vibrio anguillarum and lipopolysaccharide (LPS) injection by quantitative reverse real-time polymerase chain reaction. The results showed that c-lysozyme was expressed in all tissues tested, including gill, eyestalk, eye, hemocytes, hepatopancreas, intestine, heart, and pleopod. It was most highly expressed in the intestine followed by the eyestalk, gill, hemocytes and hepatopancreas. The mRNA expression level began to decline in a short time after V. anguillarum challenge and was then upregulated by two fold or more at 24 h post injection (hpi) compared to that at 0 h. Expression was suppressed shortly after LPS injection and began to increase with higher levels of 5.8-, 5.2- and 8.4-fold at 24, 48, and 72 hpi, respectively. Higher expression was sustained and showed a gradual increasing trend until the end of the experiment (72 hpi). These results increase our understanding of the regulation of defense mechanisms and facilitate an evaluation of the effects of probiotics or immunostimulants in shrimp culture.