• 제목/요약/키워드: protein separation

검색결과 341건 처리시간 0.029초

Development of Extraction Process of Protein-bound Polysaccharides from Inonotus obliquus Mycelia (차가버섯 균사체로부터 단백다당체의 추출 공정 확립)

  • Park, Nam-Kyu;Chun, Gie-Taek;Jeong, Yong-Seob
    • KSBB Journal
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    • 제27권3호
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    • pp.177-185
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    • 2012
  • Inonotus obliquus mushroom, which is a fungus belonging to Hymenochaetaceae family, is known to grow on birth trees in colder northern climates and to be a fungal parasite that draws nutrients out of living trees rather than from the ground. For the separation of protein-bound polysaccharide (PBP) from the culture broth and mycelium of Inonotus obliquus, three well known extraction methods namely hot water, ultrasound and microwave were used. The best extraction conditions to separate the PBP (64.94 mg/g) from mycelium by microwave were found to be for 1 hour and $150^{\circ}C$. The possibility for concentration of extracted PBP solution by using membrane was also studied. The extracted PBP solution was concentrated effectively by using an ultrafiltration membrane and the molecular weight cut off (MWCO) is 30 KDa. It was observed that a concentration by the ultrafiltration membrane is essential not only for the development of clean separation technology but also for enhanced production of PBP. As a result, we have shown that PBP in the final concentrated solution showed approximately 10 times higher than that in the crude solution by application of the developed separation systems. The separation yield of PBP was about 89.79% by gel filtration of purification steps and the purified product was confirmed to be PBP by using FT-IR.

Separation of Calcium-binding Protein Derived from Enzymatic Hydrolysates of Cheese Whey Protein

  • Kim, S.B.;Shin, H.S.;Lim, J.W.
    • Asian-Australasian Journal of Animal Sciences
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    • 제17권5호
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    • pp.712-718
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    • 2004
  • This study was carried out to separate the calcium-binding protein derived from enzymatic hydrolysates of cheese whey protein. CWPs (cheese whey protein) heated for 10 min at $100^{\circ}C$ were hydrolyzed by trypsin, papain W-40, protease S, neutrase 1.5 and pepsin, and then properties of hydrolysates, separation of calcium-binding protein and analysis of calcium-binding ability were investigated. The DH (degree of hydrolysis) and NPN (non protein nitrogen) of heated-CWP hydrolysates by commercial enzymes were higher in trypsin than those of other commercial enzymes. In the result of SDS-PAGE (sodium dodecyl sulphate polyacrylamide gel electrophoresis), $\beta$-LG and $\alpha$-LA in trypsin hydrolysates were almost eliminated and the molecular weight of peptides derived from trypsin hydrolysates were smaller than 7 kDa. In the RP-HPLC (reverse phase HPLC) analysis, $\alpha$-LA was mostly eliminated, but $\beta$-LG was not affected by heat treatment and the RP-HPLC patterns of trypsin hydrolysates were similar to those of SDS-PAGE. In ion exchange chromatography, trypsin hydrolysates were shown to peak from 0.25 M NaCl and 0.5 M NaCl, and calcium-binding ability is associated with the large peak, which was eluted at a 0.25 M NaCl gradient concentration. Based on the results of this experiment, heated-CWP hydrolysates by trypsin were shown to have calcium-binding ability.

Salt-Induced Protein Precipitation in Aqueous Solution: Single and Binary Protein Systems

  • Kim, Sang-Gon;Bae, Young-Chan
    • Macromolecular Research
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    • 제11권1호
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    • pp.53-61
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    • 2003
  • A molecular-thermodynamic model is developed for the salt-induced protein precipitation. The protein molecules interact through four intermolecular potentials. An equation of state is derived based on the statistical mechanical perturbation theory with the modified Chiew's equation for the fluid phase, Young's equation for the solid phase as the reference system and a perturbation based on the protein-protein effective two body potential. The equation of state provides an expression for the chemical potential of the protein. In a single protein system, the phase separation is represented by fluid-fluid equilibria. The precipitation behaviors are simulated with the partition coefficient at various salt concentrations and degree of pre-aggregation effect for the protein particles. In a binary protein system, we regard the system as a fluid-solid phase equilibrium. At equilibrium, we compute the reduced osmotic pressure-composition diagram in the diverse protein size difference and salt concentrations.

Synthesis of Wood Adhesive Derived from the Milk Protein and the Blocked Isocyanate

  • Ha, Yuna;Lee, Sang-Min;Lee, Hyang-Yeol
    • Journal of the Korean Applied Science and Technology
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    • 제30권3호
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    • pp.551-559
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    • 2013
  • To investigate the adhesion effect of sodium bisulfite content in making blocked isocyanate, wood adhesive PB-1, PB-2, PB-3 and PB-4 containing sodium bisulfite content of 15%, 22.5%, 30% and 37.5% were synthesized respectively. As a result, when the amount of sodium bisulfite increased in adhesives, the tensile strength was found to be proportionally increased. The final adhesive mixtures were manufactured using a two-components system which are prepared by mixing two separate protein and BI solutions due to the precipitate in the adhesives. As PVA was added to adhesives to increase tenacity, the plywood dehiscence phenomenon after cold pressing process was declined. By addition of PVA, the tensile strength was improved up to $6.5{\sim}7kgf/cm^2$ with BI/protein ratio from 1:6 up to 1:8. Phase separation between milk fat and aqueous layer was disappeared after addition of emulsifier, Tween 20. Additon of Tween 20 showed tensile strength up to $5{\sim}6.5 kgf/cm^2$ at NCO/protein ratio of 1:12 ~ 1:14 without phase separation.

Enhancement of Protein Separation by Electric Field Applied to Ultrafiltration

  • Shin, Chun-Hwa;Son, Dong-Ho;Lee, Yun-Hee;Koo, Ja-Kyung;Jang, Dong-Il;Cho, Nam-Jun
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2005년도 생물공학의 동향(XVII)
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    • pp.554-557
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    • 2005
  • Ultrafiltration has been performed to separate proteins, which is almost unique method of protein separation in mass production scale. The problems of its low selectivity and decline in permeation flux resulted from gel formation on the membrane surface have been greatly improved by an applied electric field across the membrane. The applied electric field promoted or hindered the permeation of protein through membranes depending on the electric charge of protein molecules in aqueous solution. With the effects of electric field, the permeation flux and the selectivity of the ultrafiltration could be improved significantly.

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Removal of Waste Generated by Flounder (Paralichthys olivaceus) in Aquarium using a Foam Separator (활어수조에서 넙치 사육시 포말분리장치를 이용한 오염물 제거)

  • SHIN Jeong-Sik;LEE Chang-Kuen;JEONG Ho-Su;LEE Min-Su;LEE Jin-Kyung;SUH Keun-Hack
    • Korean Journal of Fisheries and Aquatic Sciences
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    • 제37권6호
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    • pp.498-504
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    • 2004
  • Removal of waste generated by Paralichthys olivaceus in the seawater aquarium using a foam separator was investigated. Protein concentration without a foam separator continuously increased until 3 days after stocking and reached at 25 mg/L after 5 days stocking, but protein concentration became lower than the initial protein concentration (2.5 mg/L) with a foam separator. The trends of other fish wastes such as ammonia, total suspended solids (TSS) and chemical oxygen demand (COD) were similar to protein. Dissolved oxygen (DO) in the aquarium decreased below 6.0 mg/L without a foam separator, but with a foam separator the average DO in the aquarium was 7.3 mg/L. Foam separation with the increase of superficial air velocity (SAV) was more effective than that with the fixed SAV. This study showed that wastewater. treatment of seawater aquarium using a foam separator is effective method for a fish waste removal and oxygen supply.

Separation and Purification of Lysozyme from Chicken Eggwhite Through Ultrafiltration (한외여과를 통한 난백 중 라이소자임의 분리정제)

  • Koo Ja-Kyung;Son Dongho;Jun Hoejin;Lee Yunhee;Cho Namjun;Jang Dong Il
    • Membrane Journal
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    • 제15권2호
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    • pp.121-131
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    • 2005
  • Separation and purification of lysozyme from chicken egg white was studied using ultrafiltration. We have obtained experimental data through the cellulose membranes with the molecular weight cut off (MWCO) of 10 kDa, 30 kDa and 100 kDa in a stirred ultrafiltration device. Certain amounts of egg white were dissolved into 20 mM phosphate buffers of pH 6, 7 and 8 to make protein solutions of $1\%,\;2\%,\;3\%\;and\;10\%$ concentration. Permeation flux increased with increasing MWCO of the membrane. Permeation flux increased with increasing transmembrane pressure (TMP) and decreasing the protein concentration. As the MWCO of membrane decreased, the selectivity increased. The selectivity increased with increasing TMP and protein concentration of the solution.

An Iterative Spot Matching for 2-Dimensional Protein Separation Images (반복 점진적 방법에 의한 2차원 단백질 분리 영상의 반점 정합)

  • Kim, Jung-Ja;Hoang, Minh T.;Kim, Dong-Wook;Kim, Nam-Gyun;Won, Yong-Gwan
    • Journal of Biomedical Engineering Research
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    • 제28권5호
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    • pp.601-608
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    • 2007
  • 2 Dimensional Gel Electrophoresis(2DGE) is an essentialmethodology for analysis on the expression of various proteins. For example, information for the location, mass, expression, size and shape of the proteins obtained by 2DGE can be used for diagnosis, prognosis and biological progress by comparison of patients with the normal persons. Protein spot matching for this purpose is comparative analysis of protein expression pattern for the 2DGE images generated under different conditions. However, visual analysis of protein spots which are more than several hundreds included in a 2DGE image requires long time and heavy effort. Furthermore, geometrical distortion makes the spot matching for the same protein harder. In this paper, an iterative algorithm is introduced for more efficient spot matching. Proposed method is first performing global matching step, which reduces the geometrical difference between the landmarks and the spot to be matched. Thus, movement for a spot is defined by a weighted sum of the movement of the landmark spots. Weight for the summation is defined by the inverse of the distance from the spots to the landmarks. This movement is iteratively performed until the total sum of the difference between the corresponding landmarks is larger than a pre-selected value. Due to local distortion generally occurred in 2DGE images, there are many regions in whichmany spot pairs are miss-matched. In the second stage, the same spot matching algorithm is applied to such local regions with the additional landmarks for those regions. In other words, the same method is applied with the expanded landmark set to which additional landmarks are added. Our proposed algorithm for spot matching empirically proved reliable analysis of protein separation image by producing higher accuracy.

Prefractionation and Enrichment for the Analysis of Low Aboundance Proteome (극미세 단백질 분석을 위한 프로테옴 분획 농축 기술)

  • 지재웅;변상요
    • KSBB Journal
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    • 제16권5호
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    • pp.435-441
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    • 2001
  • In spite of the powerfulness for the simultaneous study of proteome expression and post-translational modification, 2-D PAGE has inevitable limitation on detect low aboundant proteins. Since many of the low abundant proteins are likely to have very important regulatiory functions in cells, separation and analysis of low copy number proteins is an important issue in proteome studies and challenge for 2-D techniques. Among various methods developed to detect low abundant proteins, electrophoretic protein prefractionation, chromatographic protein prefractionation, and subcellular fractionation are explained in this paper. Their practical strengths and weaknesses are also explained with current research trends.

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