• 제목/요약/키워드: protein separation

검색결과 343건 처리시간 0.024초

역미셀계를 이용한 대두 단백질로부터 트립신 저해제의 분리에 관한 연구 (Separation of Soybean Trypsin Inhibitor Using Reverse Micellar System)

  • 조상우;최춘순;이준식
    • KSBB Journal
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    • 제4권2호
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    • pp.123-127
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    • 1989
  • 역미셀계를 이용하여 대두 트립신 저해제의 분리에 대하여 고찰한 바, 여러 완충 수용액 중에서 1.0M $C_aCl_2용액(pH3.0)과 1.0M NaCl용액(pH 11.5)이 단백질의 용해와 회수에 가장 적당하였다. 이 조건을 두 모델계와 하나의 실제시료에 적용해 본 결과, 7S 단백질과 트립신저해제로 구성된 모델계에서는 비활성이 두배이상 증가되었고, 선수용성 단백질과 트립신 저해제로 구성된 모델계에서는 1.6배의 비활성 증가를 볼 수 있었다. 광교대두로부터 추출한 전유출물의 경우, 1.56배의 비활성 증가가 있었다. 이들을 SDS-PAGE로 확인한 결과 모두 매우 순수한 단백질임을 알 수 있었다.

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형광과 여기광을 공간적으로 분리하는 바이오칩용 소형 형광측정시스템 (Miniature Biochip Fluorescence Detection System with Spatial Separation of Fluorescence from Excitation Light)

  • 김호성;김용권;박주한;이국녕;최재호
    • 대한전기학회논문지:전기물성ㆍ응용부문C
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    • 제54권8호
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    • pp.378-383
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    • 2005
  • We report the development of miniature fluorescence detection systems that employ miniature prism, mirrors and low coat CCD camera to detect the fluorescence emitted from 40 fluorescently-labeled protein patterns without scanner. This kind of miniature fluorescence detection system can be used in point of care. We introduce two systems, one uses prism+mirror block and the other uses prism and two mirrors. A large NA microscope eyepiece and low cost CCD camera are used. We fabricated protein chip containing multi-pattern BSA labeled with Cy5, using MEMS technology and modified the surface chemically to clean and to immobilize proteins. The measurements show that the combination of prism and mirrors can homogenize elliptical excitation light over the sample with higher optical efficiency, and increase the separation between excitation and fluorescence light at the CCD to give higher signal intensity and higher signal to noise ratio. The measurements also show that protein concentrations ranging from 10 ng/ml to 1000 ng/ml can be assayed with very small error. We believe that the proposed fluorescence detection system can be refined to build a commercially valuable hand-held or miniature detection device.

Fractionation of Chinese Cabbage Juice

  • Kim, Sang-Hun;Proydak, Nikolai
    • Agricultural and Biosystems Engineering
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    • 제6권2호
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    • pp.77-82
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    • 2005
  • The fractionation of green juice could be one of the ways to treat the green juice for saving the bio re-sources by using the basic processes of protein coagulation and separating juice coagulation into protein paste and brown juice and storing the final products. The fractionation of Chinese cabbage juice can be accomplished by applying the combine method of the formic acid with rate of 0.3% and the propionic acid with rate of 0.1 % added 4 hours later in the juice with maximum recovery of protein coagulation. The separation of coagulation into the protein paste and the brown juice completed in 6.5 hours by set up method in a special designed storage. The protein paste could be stored safely for 30days in anaerobic condition.

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친화성 막모듈에 의한 단백질 크로마토그래픽 특성 (Characteristics of Protein Chromatography by Affinity Membrane Mudule)

  • 이광진;염경호
    • KSBB Journal
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    • 제13권2호
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    • pp.125-132
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    • 1998
  • Protein affinity membrane was prepared via the coating of chitosan gel on the porous flat polysulfone membrane surface, followed by the immobilization f the reactive dye (Cibacron Blue 3GA) to the chitonsan gel. The maximum protein binding capacity of affinity membrane was about 70${\mu}g/cm^2$ determined by the batch adsorption experiments of human serum albumin (HSA). Using module of this membrane, the characteristics of protein chromatography were investigated through the experiments of elution and frontal chromatography of HSA. This membrane module promises as a chromatography column, since it represented a lower pressure drop and a greater reproducibility. The protein separation ratio was significantly influenced by the flow rate of mobile phase and the injection quantity of HSA. The dynamic protein binding capacity of module decreased from the equilibrium binding capacity with increasing flow rate and approached the value of 15 - 20 ${\mu}g/cm^2$ for flow rates above 6 mL/min.

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실크 정련 세리신 단백질의 분리특성과 응용(1) (Separation Performance and Application of Sericin Protein in Silk Degumming Solution(1))

  • 차진우;배기서;박인우;김용덕;홍영기
    • 한국염색가공학회지
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    • 제21권6호
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    • pp.56-63
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    • 2009
  • In this study we have evaluated the separation characteristics and concentration of sericin using tubular type ultrafiltration membrane in silk degumming solution that extracted from electrolytic reduction water process. Ultrafiltration membranes have used in sericin separation performance and the separation characteristics of membrane satisfied typical Hagen-Poiseuille equation. It had the increase of flux according to the increase of feed pressure and temperature in occasion of pure water flux. And also the flux and solute rejection had about $25{\sim}60{\ell}/m^2{\cdot}h$ and more than 95% in sericin feed solution with concentration 1.00~1.89% at feed pressure force of $3{\sim}8kgf/cm^2$ respectively. In addition, the separation performance of tubular type ultrafiltration membrane for silk degumming solution was very steady-state with long experiment time.

Separation and flux characteristics in cross-flow ultrafiltration of bovine serum albumin and bovine hemoglobin solutions

  • Hsiao, Ruey-Chang;Hung, Chia-Lin;Lin, Su-Hsia;Juang, Ruey-Shin
    • Membrane and Water Treatment
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    • 제2권2호
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    • pp.91-103
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    • 2011
  • The flux behavior in the separation of equimolar bovine serum albumin (BSA) and bovine hemoglobin (HB) in aqueous solutions by cross-flow ultrafiltration (UF) was investigated, in which polyacylonitrile membrane with a molecular weight cut-off (MWCO) of 100 kDa was used. BSA and HB have comparable molar mass (67,000 vs. 68,000) but different isoelectric points (4.7 vs. 7.1). The effects of process variables including solution pH (6.5, 7.1, and 7.5), total protein concentration (1.48 and 7.40 ${\mu}M$), transmembrane pressure (69, 207, and 345 kPa), and solution ionic strength (with or without 0.01 M NaCl) on the separation were examined. It was shown that the ionic strength had a negligible effect on separation performance under the conditions studied. Although BSA and HB are not rigid bodies, the flux decline in the present cross-flow UF did not result from the mechanism of cake filtration with compression. In this regard, the specific cake resistance when pseudo steady-state was reached was evaluated and discussed.

Pseudoalteromonas sp. 배양액으로부터 아가레이즈 분리를 위한 음이온교환 크로마토그래피 최적화 (Optimization of Anion-exchange Chromatography for the Separation of Agarase from Culture Broth of Pseudoalteromonas sp.)

  • 김유나;이재란;김무찬;김성배;장용근;홍순광;김창준
    • Korean Chemical Engineering Research
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    • 제49권6호
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    • pp.840-845
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    • 2011
  • 아가로오스 분해산물은 생리활성이 우수하여 의약품 및 기능성 화장품 원료로 사용될 뿐만 아니라 바이오에탄올 생산을 위한 효모 발효용 기질로 검토되고 있어 상당한 주목을 받고 있다. 이에 따라 고성능 아가레이즈 탐색에 대한 연구가 활발히 진행되고 있다. 최근 본 연구팀에서는 남해안에서 신규 아가레이즈를 생산하는 미생물인 Pseudoalteromonas sp.를 분리하였다. 본 연구에서는 음이온교환수지를 이용하여 미생물 배양액으로부터 아가레이즈를 분리 정제하기 위한 크로마토그래피 조건을 최적화 하였다. 황산암모늄을 첨가하여 배양 상등액으로부터 침전된 단백질을 회수하고 이를 투석하여 조효소액을 얻었다. 음이온 교환수지인 DEAE-Sepharose 레진이 충진된 칼럼에 조효소액을 로딩하여 아가레이즈를 분리하였다. 410 ${\mu}g$의 단백질 로딩 시 흡착에 적합한 평형 pH는 7.5~8.0 이었고, 적정한 resin의 부피는 3 mL였다. 등용매 용출에서 용출액 중의 NaCl 농도가 증가함에 따라 용출되는 단백질 양이 증가하여 400 mM의 NaCl에서 최대에 달하였다. 최종적으로, NaCl 농도를 1,000 mM까지 선형적으로 증가시키며 아가레이즈를 분리하였다. Lugol 용액을 이용한 염색법으로 용리액 중의 아가레이즈의 존재를 확인하였다.

Heterologous Expression and Optimized One-Step Separation of Levansucrase via Elastin-like Polypeptides Tagging System

  • Kang, Hye-Jin;Kim, Jin-Hee;Chang, Woo-Jin;Kim, Eung-Soo;Koo, Yoon-Mo
    • Journal of Microbiology and Biotechnology
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    • 제17권11호
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    • pp.1751-1757
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    • 2007
  • Elastin-like polypeptides (ELPs) undergo a reversible inverse phase transition upon a change in temperature. This thermally triggered phase transition allows for a simple and rapid means of purifying a fusion protein. Recovery of ELPs-tagged fusion protein was easily achieved by aggregation, triggered either by raising temperature or by adding salt. In this study, levansucrase has been used as a model enzyme in the development of a simple one-step purification method using ELPs. The levansucrase gene cloned from Pseudomonas aurantiaca S-4380 was tagged with various sizes of ELPs to functionally express and optimize the purification of levansucrase. One of two ELPs, ELP[V-20] or ELP[V-40], was fused at the C-terminus of the levansucrase gene. A levansucrase-ELP fusion protein was expressed in Escherichia coli $DH5{\alpha}$ at $37^{\circ}C$ for 18 h. The molecular masses of levansucrase-ELP[V-20] and levansucrase-ELP[V-40] were determined as 56 kDa and 65 kDa, respectively. The phase transition of levansucrase-ELP[V-20] occurred at $20^{\circ}C$ in 50 mM Tris-Cl (pH 8) buffer with 3 M NaCl added, whereas the phase transition temperature ($T_t$) of levansucrase-ELP[V-40] was $17^{\circ}C$ with 2 M NaCl. Levansucrase was successfully purified using the phase transition characteristics of ELPs, with a recovery yield of higher than 80%, as verified by SDS-PAGE. The specific activity was measured spectrophotometrically to be 173 U/mg and 171 U/mg for levansucrase-ELP[V-20] and levansucrase-ELP[V-40], respectively, implying that the ELP-tagging system provides an efficient one-step separation method for protein purification.