• 자연과학논문집
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• 제14권2호
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• pp.83-91
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• 2004

박테리오 파이지 K11 lysozyme은 최근에 우리 실험실에서 클로닝 되었으며, 숙주균주의 세포벽을 분해하는 고유의 lysozyme활성과 박테리오 파아지 K11 RNA 중합효소의 전사반응을 억제하는 활성을 가지고 있는 것으로 확인되었다. 이미 잘 연구된 박테리오 파아지 T7 lysozyme의 경우 클로닝되고 분리 정제된 T7 lysozyme 단백질의 3차 구조 및 T7 RNA 중합효소와의 결합양상에 대하여 밝혀졌다. 따라서 우리 실험실에서는 K11 lysozyme과 K11 RNA 중합효소와의 결합 정도 및 그 특성을 파악할 목적으로 yeast two hybrid 시스템을 통하여 K11 RNA 중합효소와 K11 lysozyme의 단백질-단백질 상호작용을 알아보고자 하였다. LexA 시스템을 이용하여 LexA DNA 결합 부위를 갖고 있는 pLexA에 K11 lysozyme 유전자를 삽입하여 prey로 하였따. 활성 부위로는 B42 융합 단백질을 만드는 pJG4-5에 K11 RNA 중합효소의 결합은 생체 밖에서 reporter 유전자인 lacZ와 leu2의 발현으로 확인되었으며 이들의 결합정도와 결합부위에 대한 연구들은 진행중에 있다.

• 자연과학논문집
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• 제17권1호
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• pp.55-64
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• 2006

이 연구에서는 6개의 히스티딘이 첨부된 파지 K11 라이소자임의 제조 분리 및 특성을 알아보고자 하였다. 첨부된 히스티딘에 의한 이 효소의 활성도의 변화는 없었으며, 효소 활성의 최적 pH는 7.2-7.4 이었다. 여러 다른 종류의 양이온 존재하의 활성도를 측정한 결과 칼슘과 마그네슘 이온에 의해 효소 활성이 완전히 억제되었으나, 아연이나 나트륨 이온은 효소의 활성도를 이 이온들이 없을 때와 같은 정도로 유지하였다. 단지 100 mM 이상의 아연 이온 농도에서 K11 라이소자임 효소 활성도를 완전히 억제하였다.

• BMB Reports
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• 제48권11호
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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.

• BMB Reports
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• 제40권4호
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• pp.539-546
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• 2007

The lysozymes encoded by bacteriophage T7 and K11 are both bifunctional enzymes sharing an extensive sequence homology (75%). The constructions of chimeric lysozymes were carried out by swapping the N-terminal and C-terminal domains between phage T7 and K11 lysozymes. This technique generated two chimeras, T7K11-lysozyme (N-terminal T7 domain and C-terminal K11 domain) and K11T7-lysozyme (N-terminal K11 domain and C-terminal T7 domain), which are both enzymatically active. The amidase activity of T7K11-lysozyme is comparable with the parental enzymes while K11T7-lysozyme exhibits an activity that is approximately 45% greater than the wild-type lysozymes. Moreover, these chimeric constructs have optimum pH of 7.2-7.4 similar to the parental lysozymes but exhibit greater thermal stabilities. On the other hand, the chimeras inhibit transcription comparable with the parental lysozymes depending on the source of their N-terminals. Taken together, our results indicated that domain swapping technique localizes the N-terminal region as the domain responsible for the transcription inhibition specificity of the wild type T7 and K11 lysozymes. Furthermore, we were able to develop a simple and rapid purification scheme in purifying both the wild-type and chimeric lysozymes.

• 한국식품과학회지
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• 제24권1호
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• pp.11-13
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• 1992

MRS 배지에 Lactobacillus plantarum을 접종하고 200ppm lysozyme에 0.1% glycine 또는 0.1%lysine을 단독 및 혼합 첨가하여 균의 성장 억제효과를 조사하였다. 각 첨가구 모두 대조구에 비하여 접종균 성장 억제효과가 높았으며, 특히 lysozyme과 glycine 혼합첨가구는 그 효과가 현저하므로 각각을 농도별로 조합첨가하였을 때 상승 또는 병용효과가 있었다. Lysozyme과 glycine 각각의 농도가 증가할수록 성장 억제효과는 증가하였으며, lysozyme 200ppm 이상과 glycine 0.5% 이상의 조합에서는 100% 성장 억제효과를 나타내었다. 또한 배지에 120ppm lysozyme과 0.1, 1, 10mM의 EDTA를 단독 및 혼합첨가하고 균의 성장 억제효과를 조사하였으며, 모든 첨가구에서 대조구보다 높은 억제효과를 나타내었고, 120ppm 이상의 lysozyme과 0.8mM 이상의 EDTA 혼합첨가구에서는 97% 억제효과가 인정되었다.

• Asian-Australasian Journal of Animal Sciences
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• 제29권11호
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• pp.1601-1607
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• 2016

This study was conducted to determine the effect of lysozyme addition on in vitro rumen fermentation and to identify the lysozyme inclusion rate for abating methane ($CH_4$) production. An in vitro ruminal fermentation technique was done using a commercial concentrate to rice straw ratio of 8:2 as substrate. The following treatments were applied wherein lysozyme was added into 1 mg dry matter substrate at different levels of inclusion: Without lysozyme, 2,000, 4,000, and 8,000 U lysozyme. Results revealed that, lysozyme addition had a significant effect on pH after 24 h of incubation, with the highest pH (p<0.01) observed in 8,000 U lysozyme, followed by the 4,000 U, 2,000 U, and without lysozyme. The highest amounts of acetic acid, propionic acid (p<0.01) and total volatile fatty acid (TVFA) (p<0.05) were found in 8,000 U after 24 h of incubation. The $CH_4$ concentration was the lowest in the 8,000 U and the highest in the without lysozyme addition after 24 h of incubation. There was no significant differences in general bacteria, methanogen, or protozoan DNA copy number. So far, addition of lysozyme increased the acetate, propionate, TVFA, and decreased $CH_4$ concentration. These results suggest that lysozyme supplementation may improve in vitro rumen fermentation and reduce $CH_4$ emission.

• 자연과학논문집
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• 제11권1호
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• pp.23-26
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• 1999

Lysozyme에 X선 조사는 단백질-물 혼합물의 상분리 온도를 증가시키며, 온도에 따른 광산란량은 증가하였다. 사포닌 첨가는 상분리 온도를 감소시키며, 온도에 따른 광산란량은 현저하게 감소됨을 관측하였다.

• KSBB Journal
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• 제14권6호
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• pp.661-664
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• 1999

Lysozyme 수용액으로부터 역미셀을 이용한 Lysozyme 추출공정에서 주요변수인 pH, 이온강도, AOT 농도, 추출시간 등을 실험을 통하여 연구하였으며, 미셀내의 수분함량과 lysozyme 농도는 비례관계임을 KCl 농도변화에 따른 결과로서 알 수 있었다. 본 연구에 사용된 lysozyme의 등전점이 약 11.2로 음이온 계면활성제를 사용할 경우 추출공정에 유리함을 알 수 있었고, 추출공정(forward extraction)에서의 최적조건은 pH 6-9일 때, 그리고 0.1 M 염농도와 50 mM이하의 AOT 농도에서 가장 높은 lysozyme 추출 효율을 얻을 수 있었다. 그리고 역추출(back extraction)에서는 pH 12이상, 1M 염농도에서 Lysozyme의 가장 높은 추출 효율을 나타내었다.

• Journal of Microbiology and Biotechnology
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• 제9권3호
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• pp.292-296
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• 1999

The high density mixed mode adsorbent known by the trade name of Mimo-AD was used to purify lysozyme directly from the hen egg white (HEW). The homogenized hen egg white was treated with the adsorbent in a stirred vessel for lysozyme adsorption, and then the adsorbent, easily separated from the HEW by sedimentation, was packed into a column. The remaining HEW and contaminant proteins were removed by washing with pH 11 distilled water in an expanded-bed state, and subsequently the elution was performed with pH 12 distilled water in a packed-bed state. By this simple and rapid adsorption, washing, and elution procedure, lysozyme was purified to＞95％ with an overall recovery yield of 66％. This process offers a great potential for industrial application by allowing the extraction of lysozyme while retaining the commercial value of HEW.

• Animal cells and systems
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• 제11권2호
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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 $65^{\circ}C$.