• 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.

• Journal of Pharmaceutical Investigation
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• 제35권2호
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• pp.89-94
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• 2005

The micron or nano-sized lysozyme as a model protein drug was prepared using solution enhanced dispersion by supercritical fluid (SEDS) process at various conditions (e.g., solvent, temperature and pressure) to investigate the feasibility of pulmonary protein drug delivery. The lysozyme particles prepared were characterized by laser diffraction particle size analyzer, scanning electron microscopy (SEM) and powder X-ray diffractometry (PXRD). The biological activity of lysozyme particles after/before SEDS process was also examined. Lysozyme was precipitated as spherical particles. The precipitated particles consisted of 100 - 200 nm particles. Particle size showed the precipitates to be agglomerates with primary particles of size $1\;-\;5 \;{\mu}m$. The biological activity varied between 38 and 98% depending on the experimental conditions. There was no significant difference between untreated lysozyme and lysozyme after SEDS process in PXRD analysis. Therefore, the SEDS process could be a novel method to prepare micron or nano-sized lysozyme particles, with minimal loss of biological activity, for the pulmonary delivery of protein drug.

• Asian-Australasian Journal of Animal Sciences
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• 제28권12호
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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.

• Journal of Microbiology and Biotechnology
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• 제9권5호
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• pp.576-581
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• 1999

Effects of signal sequences, protein sizes and dissolved oxygen on the secretion of human lysozyme from a recombinant yeast were experimentally characterized. The systems consisted of Saccharomyces cerevisiae host SEY2102 that was transformed with two different plasmids. These plasmids were identical with an exception to the plasmid pMC614, which contained the native yeast MFα1 sequence and the plasmid pMC632 with the non-native rat α-amylase signal sequence. The expression of human lysozyme was controlled by the ADHI promoter. The native yeast MFαl signal sequence was more efficient than the non-native rat α-amylase signal sequence in directing the secretion of human lysozyme. Lysozyme secreted with the α-amylase signal was retained inside the cells and released to the medium very slowly, thereby causing a lower cell growth rate and a decreased product secretion rate. Lysozyme was secreted more efficiently than invertase, which is an order of magnitude bigger in molecular size compared to lysozyme, which was under the direction of the MFαl signal sequence, suggesting that protein sizes may affect the secretion efficiency. When expressed in anaerobic conditions in the medium where the ADHI promoter was derepressed, the amount of lysozyme secreted was about twice higher than that of the aerobic culture. However, the secretion rates were identical. This result showed that the dissolved oxygen level may affect the efficiency of protein secretion only, and not the secretion rate of the product protein.

• Applied Biological Chemistry
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• 제35권3호
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• pp.191-195
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• 1992

강낭콩 잎에서 에틸랜에 의하여 유도되는 분자량 30KD인 염기성 키틴분해효소를 정제하고 그 항균활성을 연구하였다. 이 단백질은 chitinase 활성과 lysozyme 활성을 가졌으며, Aspergillus fumigatus, Botrytis cinerea, Fusarium oxysporum, Rhizoctonia solani의 균사 생장을 억제하였다. 그러나 함께 실험한 2종류의 미생물 chitinase, 달걀 lysozyme, 파파야 protease는 이들에 대한 항균작용을 갖지 않았다. 이상의 결과는 lysozyme 활성을 가진 식물 chitinase가 병원균의 균사생장을 억제하여 자신을 방어할 수 있음을 시사한다.

• Applied Biological Chemistry
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• 제37권6호
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• pp.456-462
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• 1994

라이소자임-덱스트란 hybrids는 상대 습도 80%와 $60^{\circ}C$에서 16일간 유지시켜 만들었다. 라이소자임-덱스트란 hybrids의 유화성은 라이소자임보다 약 14배 정도, 시판유화제보다는 약3배 정도 높았다. 그 hybrids의 효소 활성도는 기질로 Micrococcus lysodeiticus 세포벽을 사용한 결과 라이소자임의 약 83% 정도였다. 그 hybrids의 뛰어난 유화성은 pH 3에서도 유지되었고 pH 10에서는 더욱 개선되었다. $100^{\circ}C$로 가열 처리함으로써 유화성은 크게 향상되었다. 또한 라이소자임-덱스트란 hybrids 이 그람음성세균에 대하여 항균효과를 나타냈다. 이 결과들은 라이소자임-덱스트란 hybrids가 식품에서 보존료와 유화제로 사용될 수 있다고 사료된다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제6권6호
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• pp.410-418
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• 2001

The effects of several variables on the refolding of hen egg white lysozyme have been studied, Lysozyme was denatured in both urea, and guanidine hydrochloride(GuHCl), and batch refolded by dilution (100 to 1000 fold) into 0.1 M Tris-HCI, pH 8.2 mM EDTA 3 mM reduced glutathione and 0.3 mM oxidised glutathions. Refolding was found to be sensitive to temperature, with the highest refolding yield obtained at 50$\^{C}$. The apparent activation energy for lysozyme re-folding wasf ound to be 56kJ/mol, Refolding by dilution results in low concentrations of both de-naturant and reducing agent species. It was found that the residual concentrations obtained dur-ing dilution(100-fold dilution:[GuHCI]=0.06 mM, [DTT]=0.15 mM) were significant and could inhibit lysozyme refolding. This study has also shown that the initial protein concentration (1-10mg/mL) that is refolded is an important parameter. In the presence of residual GuHCl and DTT higher refolding yields were obtained when starting from higher initial lysozyme concentra-tions. This trend was reversed when residual denaturant components were removed from the re-folding buffer.

• 농업과학연구
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• 제14권2호
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• pp.272-285
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• 1987

난백내(卵白內) lysozyme을 분리(分離)하기 전(前)에 난백(卵白)의 균질(均質) 최적조건(最適條件)과 원료난(原料卵)의 lysozyme활성(活性)을 검토(檢討)하였으며, 직접(直接) 결정법(結晶法)과 Bentonite 및 Duolite의 흡착법(吸着法)을 실시(實施)하여 균질(均質)된 난백(卵白)으로부터 lysozyme의 회수율(回收率)과 순도(純度)를 조사(調査)한 결과(結果)는 다음과 같다. 1. 균질(均質)된 난백(卵白)의 효소활성(酵素活性)을 측정(測定)함에 있어서 난백(卵白)의 균질(均質) 최적조건(最適條件)은 교반시간(攪拌時間)이 10분(分), 교반속도(攪拌速度)는 2,000rpm이었다. 2. 원료난(原料卵)의 lysozyme활성(活性)에서 품종중(品種中) W. Leghorn 종(種)이 높은 활성(活性)을 보였고, 난백(卵白) 부위중(部位中) 농후난백(濃厚卵白)이 수양난백(水樣卵白)보다 약간 높은 활성(活性)을 보였으며, $4^{\circ}C$에 보존(保存)된 계란(鷄卵)의 활성잔존율(活性殘存率)은 $25^{\circ}C$나 $-20^{\circ}C$에 비하여 더 높았다. 3. 직접(直接) 결정법(結晶法)에 의하여 난백(卵白)으로부터 분리(分離)된 3차(次) 결정화(結晶化)된 lysozyme의 비활성(比活性)은 29배까지 증가되었으며, 이때의 회수율(回收率)은 64.3%이었다. 4. Bentonite의 흡착법(吸着法)에 의하여 lysozyme의 흡착율(吸着率)은 95.7%, 용출율(溶出率)은 89.1%이었고, 난백(卵白)으로부터 분리(分離)된 lysozyme의 회수율(回收率)은 85.3%이었으나, 이때의 비활성(比活性)이 13배 밖에 증가되지 않았다. 5. Duolite의 batch법(法)에 의하여 난백(卵白) $100m{\ell}$에 Duolite 20g로 90분간(分間) 흡착(吸着)시키고 3시간(時間) 용출(溶出)시켰을 때, 그 흡착율(吸着率)은 97%, 회수율(回收率)이 84.8%이었으며, 이때의 비활성(比活性)은 30배 증가되었다.

• 한국식품과학회지
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• 제22권6호
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• pp.711-715
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• 1990

식품 및 의약품 원료로 이용되고 있는 Iysozyme을 난백으로부터 이온교환 크로마토그래프를 이용하여 연속추출하였다. 사용된 수지는 지금까지 수율, 순도, 작동용이성에서 가장 효과적이라고 밝혀져 있는 Duolite C-464 양이온교환수지를 선택하여 $Na^+$형으로 $pH\;7.9{\pm}0.1$로 평형화하여 사용하였다. 조제용 자동 Liquid Chromatography(LC) system(column size ; i.d. 50 mm, bed volumn ; 1020 ml)에서 수지 평형화, 난백접촉, rinse, lysozyme용출 순으로 cycle을 연속적으로 반복하면서 시행하고 그 재현성을 관찰하기 위해 각 cycle별 rinse끝점과 용출끝점의 UV level을 비교하였다. 그 결과 rinse끝점은 19 cycle까지 30% 이하를 유지하였다. 용출 끝점은 17 cycle까지는 30% 이하에서 비교적 안정하였으나 18 cycle 이후 부터는 50% 이상으로 용출능력이 현저하게 감소하였다. 또한, 회수율 비교에서 17cycle까지는 90% 이상을 유지하다가 18cycle에서는 72%, 19 cycle에서는 65%로 격감하였다. 본 추출 정제 lysozyme은 전기영동상 단일 band로 나타났고, densitometer로 측정한 순도는 99% 이상이었다.

• 대한의생명과학회지
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• 제24권2호
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• pp.116-124
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• 2018

Lysozyme is known as a key substance of the innate immunity and have antibacterial effect in the mucosal tissues, especially middle ear. Aquaporin (AQP) functions as water movement in the tissue and has been expected to be participated in the inflammatory responses. In the present study, we investigated to reveal association of lysozymes and AQPs in otitis media. The gene expression of lysozyme genes, homo sapiens lysozyme (hLYZ), homo sapiens lysozyme M (hLYZ M), and homo sapiens lysozyme G like-2 (hLYGH), and AQP genes (AQP 0 - AQP 12) were measured from postauricular skin, mastoid mucosa, inflamed mastoid mucosa, and middle ear mucosa. The hLYZ, hLYZ M and hLYGH gene were expressed in mastoid mucosa, inflamed mastoid mucosa, middle ear mucosa. Of AQP genes, all AQP gene except AQP 3 gene were expressed in the tissue of middle ear. Among them, AQP 4, AQP 8, AQP 9, AQP 10, AQP 11 and AQP 12 were highly expressed in the inflamed mastoid mucosa and normal mastoid mucosa (P<0.001). Interestingly, expression levels of AQP 4, AQP 9, and AQP 12 gene were significantly higher in the inflamed mastoid mucosa compared to normal middle ear mucosa (P<0.05). These results suggest that lysozyme and AQPs could be associated with inflammatory response in the middle ear.