• 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.

• Preventive Nutrition and Food Science
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• 제5권2호
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• pp.65-69
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• 2000

Hen egg-white lysozyme was conjugated with 7~9 mers xyloglucan hydrolysates(MW-1,400) at 6$0^{\circ}C$ and 79% relative humidity for 3 days. SDS-PAGE showed that the conjugation between lysozyme and the oligosaccharide began from 1-day incubation, and three molecules of carbohydrate chains were attached to a protein molecule after 30day incubation. The enzymatic activity of lysozyme was totally conserved in the neoglycoprotein, when measured by using glycol chitin as substrate. Besides, the emulsifying properties of lysozyme were vastly improved by the conjugation with the oligosaccharide, in which emulsifying activity of the neoglycoprotein was five times higher than that of native one.

• 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.

• 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.

• 한국결정학회:학술대회논문집
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• 한국결정학회 2002년도 정기총회 및 추계학술연구발표회
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• pp.3-3
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• 2002

Many studies on crystal growth mechanisms of the hen egg-white lysozyme protein crystals have mainly performed by optical microscopy and atomic force microscopy (AFM). As results, two types of growth mechanisms, which are a two-dimensional nucleation mechanism and a spiral growth mechanism, were identified. However, there was no direct evidence of grown-in screw dislocations at the spiral sites. We first observed the screw dislocations in tetragonal lysozyme crystals using synchrotron X-ray topography. In addition, to confirm the characteristics of dislocations, we have observed some elastic constants in lysozyme crystals in terms of the sound velocity measurement by pulse echo methods. Tetragonal hen egg-white lysozyme crystals were grown by the concentration gradient method. The crystals were grown in test tubes, with an inner diameter of 8 ㎜ and 80 ㎜ in length, held vertically. The test tubes were kept at 23C for 2 weeks. The maximum size of crystals were 3×3×4 ㎟. The high quality crystals were examined by Laue topography with a water filter using synchrotron radiation. Figure is a X-ray topograph. Several straight screw dislocations were observed. We also determined Burgers vector to be a [110] direction. The measurement of sound velocity was performed by the digital signal processing method. the crystals were placed in stainless steel vessel, which was filled with lysozyme solution used for crystal growth. We observed the longitudinal sound velocity along the [110] direction in the tetragonal is obtained to be 1817 ㎧. Therefore, Young modulus and shear modulus were evaluated to be 2.70 Gpa and 1.02 Gpa, respectively, if we assumed Poisson ratio is 0.33. These results will be discussed at the meeting.

• 한국식품과학회지
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• 제30권2호
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• pp.397-406
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• 1998

난백 lysozyme은 박테리아 세포벽을 선택적으로 분해하므로 식품 가공 공정에 있어서 천연 식품보존제로서의 이용 가치가 높다. 기존의 결정화와 냉동건조 방법 대신 PM30 막을 사용하여 한외여과함으로써 13종의 난백 단백질로부터 single-step에 의해 lysozyme을 분리하고자 하였다. 냉동 건조한 lysozyme을 pH 4.6 citrate-phosphate buffer에 녹여 PM30 막으로 한외여과시 시료 농도, 온도, 막 횡단 압력, 교반속도 등의 운용 조건을 변화시켜주면서 flux를 최대화하는 최적 막분리 조건을 구하였다. 최적 막 분리 조건하에서 시간이 경과함에 따라 막 재질과 막 침착이 flux에 미치는 효과를 측정하였고, 막 분리 여액내 단백질 농도와 lysozyme 농도, 비활성도를 측정하였다. PM30 막을 이용한 난백 lysozyme의 막 분리 최적 조건은 시료농도 0.25%, 온도 $35^{\circ}C$, 막 횡단 압력 30 psi, 교반속도 300 rpm 이었다. 막 분리 초기 12분까지는 YM30 막의 flux가 더 높았으나, 정상상태에서의 flux는 PM30 막이 더 높았다. PM30 막의 분리 여액내 단백질 농도와 lysozyme 농도는 YM30 막에 비해 낮았으나, 비활성도는 2배 이상 높았다. 5회 막 분리한 PM30 막은 새 PM30 막에 비하여 flux가 약 30% 감소하였으나 시간 경과에 따른 flux 감소 경향은 거의 동일하였으며, 막 분리 35분 경과 후 정상상태에 이르러 초기 flux 약 70%를 유지하였다. 막 분리 여액내 lysozyme 농도와 비활성도는 각각 110 units/mL, 2,821 units/mg protein이었으므로, PM30 막을 이용한 한외여과 공정은 천연 항균 효소제인 lysozyme을 분리하는데 매우 효과적인 방법이었다.

• Food Science and Biotechnology
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• 제18권3호
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• pp.706-711
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• 2009

Lysozyme was treated with digestive enzymes and the production of interleukin 8 (IL-8) was measured in Caco-2 cell with the peptides from lysozyme upon stimulating with lipopolysaccharide (LPS) to investigate the overall anti-inflammatory activity of lysozyme when it is in digestive tracts. Lysozyme reduced IL-8 production, and the peptides from pepsin hydrolysis of lysozyme had the similar effect. The products of trypsin digestion of lysozyme had no effect on the reduction of IL-8 production while those of pepsin-trypsin hydrolysis did. The effectiveness of lowering IL-8 production was not different by time of the peptide addition. When Caco-2 cells were pre-incubated with peptides for 24 hr, the reduction effects were observed from the peptides from pepsin hydrolysis, indicating that some of the peptides are still remaining in the cells. Therefore, it can be concluded that the IL-8 reduction effect of lysozyme against LPS still remained even after the pepsin and trypsin hydrolysis.

• Journal of Oral Medicine and Pain
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• 제33권1호
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• pp.1-8
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• 2008

타액에 존재하는 여러 항균물질은 상호작용을 통하여 부가적이거나 상승작용을 나타내고 때로는 저해작용을 나타내는 것으로 알려져 있다. 본 연구는 in vitro 상에서 lysozyme과 peroxidase 사이의 상호작용을 효소활성 측면에서 조사하고자 시행되었다. Lysozyme과 peroxidase의 상호작용은 hen egg-white lysozyme(HEWL)과 bovine lactoperoxidase(bLP)를 혼합하는 방법으로 조사되었고, peroxidase system이 lysozyme에 미치는 영향은 potassium thiocyanate와 hydrogen peroxide를 추가적으로 첨가하는 방법으로 조사되었다. Lysozyme 활성은 Micrococcus lysodeikticus 기질용액의 혼탁도 변화를 측정하는 방법으로 측정되었고, peroxidase 활성은 NbsSCN 법으로 측정되었다. Wilcoxon signed rank 법을 이용하여 lysozyme과 peroxidase 효소활성 변화를 대조군과 비교하였다. 생리적 농도범위에서 bLP는 HEWL의 효소활성을 증가시켰으며(P < 0.05), 그 효과는 bLP의 농도증가에 따라 영향을 받았다. 하지만 HEWL는 bLP의 효소활성에 영향을 주지 못하였다. Thiocyanate는 HEWL의 효소활성에 영향을 주지 못하였고, potassium thiocyanate와 hydrogen peroxide를 추가한 peroxidase system도 HEWL 효소활성의 추가적인 상승을 유도하지는 못하였으며, hydrogen peroxide의 농도변화도 영향을 미치지 못하였다. 본 연구의 결과는 타액 항균 물질을 포함하고 있는 구강건강용품이나 구강에서 lysozyme과 peroxidase의 상호작용을 이해하는데 필요한 중요한 정보를 제공해준다.

• KSBB Journal
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• 제14권3호
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• pp.279-283
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• 1999

본 연구에서는 난백에 포함된 라이소자임의 컬럼 크로마토그래피에서의 breakthrough behavior 실험을 통해 최적 크로마토그래피 분리조건을 결정하였다. 다양한 양이온 교환수지의 라이소자임 breakthrough behavior를 비교한 결과 Cellufine C-200(Amicon, USA)이 라이소자임 흡착에 가장 우수한 성능을 보였다. Cellufine C-200 수지에서 이온강도, pH, 선속도 등이 난백 라이소자임의 breakthrough behavior에 미치는 영향을 비교한 결과 conductivity 2.75 mS/cm, pH 7, 선속도 0.635cm/min의 최적 조건을 얻었다. 본 최적조건에서 prep 규모 컬럼 크로마토그래퍼를 이용 난백으로부터 라이소자임을 분리하였다. 정제된 라이소자임과 Sigma 라이소자임의 specific activity는 비슷하였으며 SDS-PAGE 분석 결과 본 연구에서 정제된 라이소자임이 Sigma 라이소자임보다 impurity 단백질들을 적게 포함하고 있음을 확인하였다.

• Journal of Oral Medicine and Pain
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• 제39권3호
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• pp.90-95
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• 2014

Purpose: To investigate viscosity and wettability of hyaluronic acid (HA) solutions according to supplementation of lysozyme and/or peroxidase, and different ionic strength and pH conditions. Methods: Solutions containing HA were prepared using distilled deionized water (DDW) and simulated salivary buffer (SSB) in different conditions. Different concentrations of hen egg-white lysozyme and bovine lactoperoxidase was added into HA solutions. HA solutions with antimicrobials in different ionic strength and pH conditions were prepared. Viscosity was measured using cone-and-plate digital viscometer at six different shear rates and wettability on acrylic resin and Co-Cr alloy was determined by contact angle. Results: The viscosity values of HA dissolved in DDW were decreased in order of HA, HA containing lysozyme, HA containing peroxidase, and HA containing lysozyme and peroxidase. The viscosity values for HA in DDW were decreased as the concentration of lysozyme and/or peroxidase increased. However, the viscosity values for HA in SSB showed no significant changes according to the concentration of lysozyme and/or peroxidase. The viscosity values of HA solutions were inversely proportional to ionic strength and pH. The contact angle of HA solutions showed no significant differences according to tested surface materials, addition of lysozyme and/or peroxidase, and different ionic strength and pH conditions. Contact angles on acrylic resin by HA solutions in all tested conditions were much higher than those by human saliva. Conclusions: The rheological properties of HA supplemented with lysozyme and/or peroxidase in different ionic strength and pH conditions were objectively confirmed, indicating the possibility of HA with lysozyme and/or peroxidase as main components in the development of effective saliva substitutes.