• Journal of Microbiology and Biotechnology
• /
• v.9 no.3
• /
• pp.292-296
• /
• 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
• /
• v.5 no.2
• /
• pp.65-69
• /
• 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.

• Korean Journal of Agricultural Science
• /
• v.14 no.2
• /
• pp.272-285
• /
• 1987

In order to obtain the data for the effective separation and purification of lysozyme from egg white, optimal conditions for the homogenization of egg white and lysozyme activities of some fresh hen eggs were examined. The recovery and purity of lysozyme isolated by the direct crystallization method, the adsorption with Bentonite and the batch method with Duolite were also investigated. The results obtained were summarized as follows; 1. On the homogenization of egg white, optimal stirring time and stirring rate for the measurement of the lysozyme activities were found to be 10 min. and 2,000 rpm respectively. 2. On the activities of lysozyme in the fresh hen eggs, the activities of W. Leghorn was greater than that of R. Island or Ogol fowl, and the activities of the thick white was a little greater than that of the thin white. 3. The residual lysozyme activities of the hen eggs stored at $4^{\circ}C$ was greater than that of the ones stored at $25^{\circ}C$ or $-20^{\circ}C$ over the storage periods. 4. Hen egg lysozyme recrystallized three times by the method of direct crystallization showed the recovery of 64.3% and the increase of 29 fold in specific activities. 5. By the adsorption method with bentonite, lysozyme in the egg white was adsorbed to 95.7%, and the elution rate of the ones adsorbed was 89.1%, and the increase in specific activities was 13 fold. 6. In the experiment exploying duolite as an adsorbent, Jysozyme in the egg white was obtained with the bound rate of 97%, the recovery rate of 84.8%, and was purified by 30fold.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.6 no.6
• /
• pp.410-418
• /
• 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.

• Proceedings of the Korea Crystallographic Association Conference
• /
• 2002.11a
• /
• pp.3-3
• /
• 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.

• Korean Journal of Food Science and Technology
• /
• v.30 no.2
• /
• pp.397-406
• /
• 1998

Hen egg white lysozyme (HEWL) is very valuable as a natural preservative in food processing due to its selective bactericidal activity. HEWL which traditionally isolated by crystallization or freeze drying was simply separated from 13 different hen egg white (HEW) proteins by a single-step ultrafiltration. Freeze dried HEW (0.25%, w/v) dissolved in a citrate-phosphate buffer (pH 4.6) was ultrafiltered with a PM30 membrane under various operating conditions, by changing concentration, temperature, transmembrane pressure $({\triangle}P_T)$, and stirring speed. Optimum separation conditions were decided when maximal flux was obtained. Under the optimum separation conditions, the effect of membrane material and fouling on flux as time passed as well as lysozyme concentration, protein concentration, specific activity (SA) in the permeate were measured. Best separation conditions of HEWL with PM30 membrane were sample concentration 0.25%, temperature $35^{\circ}C$, ${\Delta}P_T\;30\;psi$, and stirring speed 300 rpm. During the first 12 min, the flux of YM30 was higher, but at the steady-state it was lower than that of PM30. The SA of the PM30 permeate was over 2 times higher in spite of the lysozyme and protein concentration being lower than that of YM30 permeate. The flux of 5 times used PM30 decreased 30% compared to a new PM30, but both had the same tendency in flux decrease when time passed. Both of them reached a steady-state after 35 min and remained at 70% of the initial flux. In the PM30 permeate, the lysozyme concentration and SA were 110 units/mL and 2,821 units/mg protein, respectively. Therefore, PM30 membrane separation was very effective for separation of antimicrobial lysozyme.

• Food Science and Biotechnology
• /
• v.18 no.3
• /
• pp.706-711
• /
• 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
• /
• v.33 no.1
• /
• pp.1-8
• /
• 2008

It is well known that many antimicrobial proteins in saliva interact with each other. The purpose of the present study was to investigate the interactions of lysozyme with peroxidase in the aspects of enzymatic activity in vitro. The interactions of lysozyme with peroxidase were examined by incubating hen egg-white lysozyme(HEWL) with bovine lactoperoxidase(bLP). The influence of peroxidase system on lysozyme was examined by subsequent addition of potassium thiocyanate and hydrogen peroxide. Lysozyme activity was determined by turbidity measurement of a Micrococcus lysodeikticus substrate suspension. Peroxidase activity was determined with an NbsSCN assay. The Wilcoxon signed rank test was used to analyze the changes of enzymatic activities compared with their controls. bLP at physiological concentrations enhanced the enzymatic activity of HEWL(P < 0.05) and its effect was dependent on the concentration of peroxidase. However, HEWL did not affect the enzymatic activity of bLP. Thiocyanate did not affect the enzymatic activity of HEWL, either. The addition of potassium thiocyanate and hydrogen peroxide did not lead to additional enhancement of the enzymatic activity of HEWL. The changes of hydrogen peroxide concentration in the peroxidase system did not affect the enzymatic activity of HEWL. Collectively, despite an in vitro nature of our study, the results of the present study provide valuable information on the interactions of lysozyme and peroxidase in the aspects of enzymatic activity in oral health care products and possibly in the oral cavity.

• KSBB Journal
• /
• v.14 no.3
• /
• pp.279-283
• /
• 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.

• Journal of Oral Medicine and Pain
• /
• v.39 no.3
• /
• pp.90-95
• /
• 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.