• Title/Summary/Keyword: Lysozyme adsorption

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Effects of Egg White and Ion Exchange Resin Pretreatment on Separation of Egg White Lysozyme (난백 및 이온교환수지의 전처리 조건이 난백 Lysozyme의 추출에 미치는 영향)

  • 유익종;이성기;김경환;민병용
    • Korean Journal of Poultry Science
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    • v.16 no.3
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    • pp.157-167
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    • 1989
  • Pretreatment of egg white and ion exchange resins was attempted to separate lysozyme from egg white efficiently. Apparent viscosity of egg white could be decreased to 3cp by homogenization for 30 minutes at 2, 000rpm and ultrasonication for 45 minutes. The result of testing adsorption capacity of lysozyme was as follows; CM-Sephadex C-25 >Duolite C464>Amberlite C-50>Dowex MSC-1>Amberlite IRC-50>Amberlite IRC-84. Although CM-Sephadex C-25 showed highest adsorption capacity of lysozyme, egg white could not eluted easily. Duolite Cf64 was selected based on relatively high lysozyme adsorption and good egg white eluting property for separation of egg white lysozyme. Na$^{+}$ form of Duolite C-464 was most effective on adsorption of Iysozyme. To separate lysozyme from egg white efficiently rinse buffer and eluting solution were selected 0.1M sodium phosphate buffer at pH 6.5 and 10% ammonium sulfate respectively. After separating lysozyme from egg white, foaming power of egg white was decreased to 85.3%. Color of egg white gel was not changed while hardness of egg white gel was decreased by 30% after separating lysozyme. However, elasticity of egg white gel was increased by 13% in lysozyme-separated egg white.

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Direct Purification of Lysozyme from Hen Egg White Using High Density Mixed Mode Adsorbent

  • KIM, WON KYUNG;BONG HYUN CHUNG
    • Journal of Microbiology and Biotechnology
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    • v.9 no.3
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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.

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Development of Hyaluronic Acid-Functionalized Hydrogel Lens and Characterization of Physical Properties and Lysozyme Adsorption (Hyaluronic acid의 첨가방법에 따른 하이드로겔 콘택트렌즈의 물리적 특성과 lysozyme 흡착량 비교)

  • Lim, Hwa-lim;Kim, Ho-joong;Jun, Jin
    • Journal of Korean Ophthalmic Optics Society
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    • v.20 no.3
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    • pp.285-291
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    • 2015
  • Purpose: The hydrogel lenses have been functionalized with HA(Hyaluronic Acid) using two different methods: construction of an IPN(Interpenetrating Polymer Networks) and formation of CCB(Chemical Covalent Bonding). The lysozyme adsorption and physical properties such as optical transmittance and water content of the hydrogel lenses have been investigated in order to determine whether method is suitable for the application potentials in contact lens industry. Methods: HA have been added to the hydrogel lenses prepared in the Lab using the two different method, e.g. IPN and CCB. The optical transmittance was measured in the wavelength range of 300~800 nm. The water content was measured by the gravimetric method using 0.9% NaCl saline solution. The amounts of adsorbed lysozyme on the contact lenses was analyzed by HPLC after incubation for 12h in artificial tears. Results: The water content of the HA added hydrogel contact lenses was increased, and the lens made by IPN method showed higher water content than the lens made by CCB method. The optical transmittance was over 90% both before and after addition of HA. Comparing the lysozyme adsorption reduction ratio, contact lens manufactured by IPN method was 60.0%, and the lens made by CCB method was 40.4%. Conclusions: CCB method is appropriate to distribute the functional material evenly throughout the lens, whereas IPN method is effective for the case of giving the functionality on the lens surface without phase separation.

Adsorption Properties of the Lysozyme and Albumin with Physicochemical Properties of the Contact Lens (콘택트렌즈의 물리화학적 특성에 따른 라이소자임과 알부민의 흡착 특성)

  • Sung, Yu-Jin;Ryu, Geun-Chang;Jun, Jin
    • Journal of Korean Ophthalmic Optics Society
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    • v.18 no.3
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    • pp.261-270
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    • 2013
  • Purpose: Adsorption properties of lysozyme and albumin according to physiochemical properties of commercial contact lens classified with the FDA categories and a contact lens fabricated in the laboratory were investigated. Methods: The contact lens were prepared using HEMA(2-hydroxyethyl methacrylate) and TRIM(3-(trimethoxysilyl) propyl methacrylate) in a cast mold. Artificial tears containing lysozyme and albumin were prepared. We measured the amounts of protein adsorbed on the each lenses with varying adsorbed time (48 hour) and the pH range (6, 6.8, 7.4, 8.2, 9) of artificial tear. Amount of the proteins absorbed on the contact lenses were measured by using HPLC. Results: Time to reach the equilibrium of protein adsorption for silicone hydrogel lens was taken longer than hydrogel lens. The amount of adsorbed both lysozyme and albumin at equilibrium were greater for the hydrogel lens than the silicone hydrogel lens, and larger for the ionic lens than the non-ionic lens. Lysozyme was more adsorbed on the higher water content of contact lens, whereas albumin was more adsorbed on the lower water content of contact lens. Only lysozyme was adsorbed on the Group IV hydrogel lens of ionic higher water content. The adsorption of protein on contact lens increased with pH of artificial tears as close to the isoelectric point of each protein. Conclusions: The adsorption amount of lysozyme is more affected by the ionic strength of the contact lens surface than the water content of contact lens. Albumin adsorption is more affected by water content than the ionic strength of the contact lens surface. For the adsorption of proteins on the silicone hydrogel lens, the pore size, determined both by the number of Si atoms and the chemical structure of the silicone-containing monomers, as well as the polarity of contact lens should be also considered.

Preparation and Characteristics of P(AN-co-MA) Membrane Imprinted with Lysozyme Molecules (라이소자임 분자각인 P(AN-co-MA) 막의 제조와 특성)

  • Min, Kyoung Won;Yoo, Anna;Youm, Kyung Ho
    • Membrane Journal
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    • v.31 no.3
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    • pp.219-227
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    • 2021
  • Molecularly imprinted membrane (MIM) is a porous polymer membrane incorporating with the molecular recognizing sites. In this study, the supporting P(AN-co-MA) asymmetric membrane was prepared by nonsolvent induced phase separation (NIPS) method. And then, MIM with lysozyme template sites was prepared using the surface imprinting method on the P(AN-co-MA) asymmetric membrane introducing a photoactive iniferter and then photo-grafting. The P(AN-co-MA) asymmetric membrane was modified with 3-chloropropyltrimethoxysilane and dithiocarbamate as a photoactive iniferter. To prepare a lysozyme imprinted membrane, the modified P(AN-co-MA) membrane was copolymerized with acrylamide as a functional momomer, N,N'-methylene bisacrylamide as a crosslinker and lysozyme as a template in the UV irradiation environment. The lysozyme imprinted MIM was analyzed by using SEM, FT-IR and EDS measurements. Its results confirm that all the P(AN-co-MA) membranes have an asymmetric structure and the iniferter group is successfully introduced on the membrane surface. The process parameters were adjusted to obtain MIM having the excellent lysozyme adsorption. The maximum lysozyme adsorption capacity reaches at 2.7 mg/g, which is 13 times higher than that of the non imprinted membrane (NIM). The permselective membrane filtration experiments of ovalbumin to lysozyme show that the P(AN-co-MA) MIM preferentially bounds a greater amount of lysozyme.

Studies on the Isolation of Lysozyme from the Egg White and the Determination of It's Bacteriolytic Activities (난백(卵白) Lysozyme의 분리법(分離法) 및 용균활성(溶菌活性)의 측정(測定)에 관(關)한 연구(硏究))

  • Her, Yong;Oh, Hong Rock
    • Korean Journal of Agricultural Science
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    • v.14 no.2
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    • pp.272-285
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    • 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.

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Fabrication and Characterisation of a Novel Pellicular Adsorbent Customised for the Effectvie Fluidised Bed Adsorption of Protein Products

  • Sun, Yam;Pacek, Andrzej W.;Nienow, Alvin W.;Lyddiatt, Andrew
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.6
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    • pp.419-425
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    • 2001
  • A dense pellicular solid matrix has been fabricated by coating 4% agarose gel on to dense zironia-silica(ZS) spheres by watr-in-oil emulsification . The agarose evenly laminated the ZS bead to a depth of 30㎛, and the resultin gpellicular assembly was characterised by densities up to 2.39g/mL and a mean particle dimeter of 136 ㎛. In comparative fluidisation tests, the pellicular solid phase exhibited a two-fold greater flow velocity than commercial benchmark ad-sorbents necessary to achieve common values of bed expansion. Furthermore, the perlicular parti-cles were characterised by improved qualities of chromatographic behaviour, particularly with re-spect to a three-fold increase in the apparent effective diffusivity of lysozyme within a pellicular assembly modified with Cibacron Blue 3GA. The properties of rapid protein adsorption/desorp-tion were attributed to the physical design and pellicular deployment of the reactive surface in the solid phase. When combined with enhanced feedstock throughput, such practical advantages recommend the pellicular assembly as a base matrix for the selective recovery of protein products from complex, particulate feedstocks(whole fermentation broths, cell disruptates and biological extracts).

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Preparation and Characterization of Silicone Hydrogel Lens Containing Poly(ethylene glycol) (PEG를 포함한 실리콘 수화젤 렌즈의 제조 및 특성)

  • Jang, Ha-Na;Chung, Youn-Bok;Kim, Sung-Soo
    • Polymer(Korea)
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    • v.33 no.2
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    • pp.169-174
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    • 2009
  • Silicone hydrogels incorporated with poly(ethylene glycol)(PEG) were prepared and characterized to evaluate the effects of PEG on contact lenses. The silicone hydrogels were copolymerized with methacryloxypropyl tris(trimethylsiloxy) silane (TRIS), methyl methacrylate (MMA), N,N-dimethyl acrylamide (DMA) and PEG-containing monomers such as poly(ethylene glycol) methyl ether methacrylate (PEG- MEM). The silicone hydrogels were characterized using Fourier transform infrared spectroscopy (FT-IR), electron spectroscopy of chemical analysis (ESCA), and scanning electron microscopy (SEM). Water absorbance, water contact angle and light transmittance of the silicone hydrogels were evaluated. The experiments of protein adsorption were also carried out to evaluate the protein adsorption in tears. The peak intensity of C-O bond was increased by the incorporation of PEG-containing monomers and thus PEG incorporation into silicone hydrogels could be confirmed. Phase separation was not shown by the SEM observation of the cross-section of silicone hydrogels. Water absorbancy was increased, while water contact angle and light transmittance were decreased with increasing incorporation of the PEG-containing monomers. The absorption of proteins in tears, albumin, lysozyme and $\gamma$-globulin, on the surface of silicone hydrogels was decreased with increasing incorporation of the PEG-containing monomers.

Affinity Filtration Chromatography of Proteins by Chitosan and Chitin Membranes: 1. Preparation and Characterization of Porous Affinity Membranes (키토산 및 키틴 막에 의한 단백질의 친화 여과 크로마토그래피: 1. 다공성 친화 막의 제조와 특성 평가)

  • Youm Kyung-Ho;Yuk Yeong-Jae
    • Membrane Journal
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    • v.16 no.1
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    • pp.39-50
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    • 2006
  • Porous chitosan and chitin membranes were prepared by using silica particles as porogen. Membrane preparation was achieved via the following three steps: (1) chitosan film formation by casting an chitosan solution containing silica particles, (2) preparation of porous chitosan membrane by dissolving the silica particles by immersing the film into an alkaline solution and (3) preparation of porous chitin membrane by acetylation of chitosan membrane with acetic anhydride. The optimum preparation conditions which could provide a chitosan and chitin membranes with good mechanical strength and adequate pure water flux were determined. To allow protein affinity, a reactive dye (Cibacron Blue 3GA) was immobilized on porous chitosan membrane. Binding capacities of affinity chitosan and chitin membranes for protein and enzyme were determined by the batch adsorption experiments of BSA protein and lysozyme enzyme. The maximum binding capacity of affinity chitosan membrane for BSA protein is about 22 mg/mL, and that of affinity chitin membrane for lysozyme enzyme is about 26 mg/mL. Those binding capacities are about $several{\sim}several$ tens times larger than those of chitosan and chitin-based hydrogel beads. Those results suggest that the porous chitosan and chitin membranes are suitable in affinity filtration chromatography for large scale separation of proteins.

Antimicrobial Hydrogel Contact Lens Containing Alginate

  • Lee, Hyun-Mee;Kim, Jong-Ki;Cho, Tae-Sub
    • Bulletin of the Korean Chemical Society
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    • v.32 no.12
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    • pp.4239-4243
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    • 2011
  • Biocompatible hydrogels from 2-hydroxyethyl methacrylate (HEMA) monomer containing various amount of alginate in the presence and absence of hydrophilic methacrylic acid (MAA) were synthesized in order for biomedical application. The antimicrobial effect and interaction with proteins for hydrogels were investigated in this study. MAA was introduced because it was expected to increase the amount of water content in the polymer which is an important factor for biocompatibility, and alginate was expected to enhance the antimicrobial activity. The antimicrobial effect against S. aureus and E. coli increased for all hydrogels as the amount of alginate and MAA contained. Presence of MAA further enhances the antimicrobial effect. Amount of adsorption of bovine serum albumin (BSA) increased with increasing concentration of alginate whether MAA was present or not. Contrarily, the amount of lysozyme was not affected with increasing alginate concentration in the absence of MAA, while it decreased in the presence of MAA.