• 제목/요약/키워드: Partial purification

검색결과 190건 처리시간 0.028초

Drosophila sp.(robusta species group)의 난황 단백질의 분리 및 부분적 화학적 특성 (Isolation and Partial Chemical Characterization of the Yolk Proteins from Drosophila sp. (robusta species group))

  • Kim, Se-Jae;Gi
    • 한국동물학회지
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    • 제35권1호
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    • pp.17-22
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    • 1992
  • The three yolk polypeptides have been isolated and partially characterized. Their molecular weights of YPI, YP2, and YP3 were 48, 000, 47, 000, and 46, 000, respectivelv, as judged by SDS-polyacrvlamide gel electrophoresis. They have different digestion products upon in situ peptide mapping by limited proteolysis. Two-dimensional gel electrophoresis showed that their isoelectric points were heterogeneous from 5.92 to 6.54. And thew showed three different antigen-antibody reactions when each polvpeptides is reacted with antisera made to a mixture of all of three. These data reported here indicate that the yolk proteins are consisted of distinctive polypeptides in Drosophlla sp. (robusta species group).

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미국흰불나방 지방체에서 저장단백질-1의 수용체의 특성과 부분정제 (Characterization and Partial Purification of Storage Protein-i Receptor in the Fat Body of Hyphantria cunea)

  • 채권석;여성문;김학렬
    • 한국동물학회지
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    • 제38권4호
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    • pp.490-497
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    • 1995
  • 미국흰불나방 의지방체 조직을 [35S]-메타이오닌이 포하된 배지에서 조직배양한 결과, 저장단백질-1(SP-1)의 전용기부터 용 1일 사이에 지방체로 흡수됨을 알았다. CHAPS, Triton X-100 등의 계면활성제를 농도별로 처리하여 막단백질의 용해도를 스크리닝한 뒤, anti-SP-1 polyclonal 항체를 쓴 Western blotting과 ligand blotting, 그리고 in vitro reductive methylation으로 14C을 표지한 저장단백질-1을 사용한 fluorography 등으로 1개의 수용체 밴드를 확인하였다. 1% Triton X-100으로 용해시킨 부분정제하였고, SDS-PAGE에 의해서 분자량을 측정한 결과 약 80 kDa로 나타났고 isoelectric focusing 시행 결과 등전점은 약 6.1로 계산되었다. 수용체 분자는 환원조건과 비환원조건의 차이와 전기영동 중의 온도에 따라서 SDS-PAGE상의 뚜렷한 밴드 양상의 차이를 나타내었다.

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Bacillus subtilis가 생산하는 비특이적 $\beta$-fructofuransoidase의 부분정제 및 특성 (Partial Purification and Properties of Non-specific $\beta$ -fructofuranosidase Produced by Bacillus subtilis)

  • 송근섭;엄태붕
    • 한국미생물·생명공학회지
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    • 제18권5호
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    • pp.484-489
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    • 1990
  • Bacillus subtilis의 세포내 이눌라아제가 부분정제되고 그의 작용 모드와 일반적 특성이 조사되었다. 이 효소는 gel filtration에 의하여 분자량을 추정하였을 때 49,000이었고, 등진점은 5.2 이었다. 기질에 대한 친화성의 지표인 Km값은 설탕에 대해서는 10mM, 라피노오스에 대해서는 18mM 이었다. 이 효소는 산성쪽에서는 불안정한 단백질로서 pH6.6에서 최대 활성을 보였으며 최적온도는 10분간 반응시켰을 때 50'C였다. 이 효소의 작용모든는 이눌린같은 구조를 가지는 과당 중합체를 과당 끝부분으로부터 하나씩 잘라가는 exo-cleavage 형이었다.

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수삼으로부터 당단백질 인자의 부분정제와 특성연구 (Partial Purification and Characterization of a Glycoprotein Factor from Fresh Ginseng)

  • Kong, Yun-Cheung;Fong, Wing-Ping;Song, Myung-Eun;Ng, Kam-Hung;Ho, Dan-Dan;Ng, Ping-Chung
    • Journal of Ginseng Research
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    • 제14권2호
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    • pp.221-227
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    • 1990
  • The aqueous extract of fresh ginseng (Panax ginseng C.A. Meyer) contains a macromolecular fraction that showed mitogenic and co-mitogenic activities in human peripheral blood lymphocytes. Purification of the crude extract by size (ultrafiltration, Sephadex G-200) and charge (DEAE-cellulose, DEAE-Sepharose) yielded a semi.purified fraction (DS-3). This fraction contains at least three subgroups of anionic macromolecules with apparent molecular weight greater than 600 kilodaltons. It is a glycoprotein with a large amount of glucuronic acid. It acts as a mitogen in both T and B cells of human peripheral blood lymphocytes. It could also potentiate the mitogenic action of Concanavalin A in lymphocyte T cells. Such potentiation is not due to increased binding of Concanavalin A to the cell surface. Its mitogenic and co-mitogenic effects do depend on the presence of extracellular Ca2+.

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Purification and Characterization of an Antilisterial Bacteriocin Produced by Leuconostoc sp. W65

  • Oh, Se-Jong;Kim, Myung-Hee;Churey, John-J.;Worobo, Randy-W.
    • Journal of Microbiology and Biotechnology
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    • 제13권5호
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    • pp.680-686
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    • 2003
  • This study was carried out to characterize the antilisterial substances produced by Leuconostoc sp. W65 and to evaluate the effects of pH, temperature, and time on inhibitory activity using response surface methodology. Leucocin W65, an antilisterial substance produced by Leuconostoc sp. W65, markedly inhibited the growth of Listeria monocytogenes, L. innocua, and L. ivanovii, whereas other pathogens including Gram-negative bacteria were not susceptible. The pH was the most effective factor with regard to bacteriocin activity, while temperature and time of heat treatment had no significant effect. Fifty percent of inhibitory activity remained after 22.8 min at pH 4.2 and $121^{\circ}C$. Leucocin W65 was purified by ammonium sulfate precipitation, hydrophobic interaction chromatography, and tricine-SDS-PAGE. Compositional analysis originally estimated the peptide to be 56 amino acids in length without asparagine, glutamine, and tryptophane. The sequence of partial N-terminal amino acid residues of purified bacteriocin was identified as follows: $NH_{2}-XGXAGVXPXGGQQPXVPLXYP$.

마늘(Allium sativum L.)로부터 추출한 Inulinase의 부분정제 및 성질 (Partial Purification and Properties of Inulinase from Garlic(Allium sativum L.))

  • 이종수;권수진;이성훈;이김나미;유진영
    • 한국식품영양학회지
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    • 제10권3호
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    • pp.325-329
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    • 1997
  • 서산 6쪽 마늘 중의 inulinase를 추출하여 황산암모늄 침전가 Sephadex G-150여과 등을 통하여 9.1%의 수율로 부분정제하였다. 부분 정제된 inulinase는 4$0^{\circ}C$와 pH 6.0에서 inulin을 가장 잘 분해시켰고 7$0^{\circ}C$ 이하와 pH 5.0~8.0에서 안정하였다. 또한 이 효소는 Al3+, Mn2+, Hg2+, Cd2+ 및 EDTA에 의하여 심하게 실활되었고 inulin에 대한 Km값은 0.22%이었다.

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Purification and Characterization of Catalase-2 from Deinococcus radiophilus

  • Oh, Kyung-A;Lee, Young-Nam
    • BMB Reports
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    • 제31권2호
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    • pp.144-148
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    • 1998
  • A bifunctional catalase-peroxidase, designated catalase-2, of a UV resistant Deinococcus radiophilus was purified to electrophoretic homogeneity by both chromatographic and electrophoretic methods. Its molecular weight was 310 kDa and composed of a tetramer of 80 kDa subunits. The catalase-2 exerted its optimal activity at $30^{\circ}C$ and around pH 9. Its $K_m$ value for $H_{2}0_{2} $ was about 10 mM. It showed the typical ferric heme spectrum with maximum absorption at 403 nm which shifted to 419 nm in the presence of cyanide. The ratio of A40i' A2S0 was 0.48. Fifty percent inhibition of the enzyme activity was observed at $4.6{\times}10^{-6}$, $7.7{\times}10^{-6}$, and $3.0{\times}10^{-6}$ M of NaCN, $NaN_3$, and $NH_{2}OH$, respectively. The enzyme was thermostable and not sensitive to 3-amino-1,2,4-triazole. Treatment of the enzyme with ethanol-chloroform caused a partial loss (30%) of its activity. The catalase-2 was distinct from the Deinococcal bifunctional catalase-3 in a number of properties, particularly in its molecular structure and substrate affinity.

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Lactococcus sp. HY449가 생산한 Bacterisocin의 정제 (Purification and Partial Amino Acid Sequence of a Bacteriocin Produced by Lactococcus, sp. HY449)

  • 오세종;이상준;김경태;김상교;박연희;백영진
    • 한국미생물·생명공학회지
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    • 제29권3호
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    • pp.155-161
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    • 2001
  • Lactrococcus sp. HY449균줄르 M17-glucose broth에 배양하여 배양 상등액으로부터 propanol-actone 침전 ion-exchange chromatography gel-filtration chromatography 및 reverse-phase chroamtography 등을 통하여 비활성 25,600,000 BU/mg 인 순수한 bacteriocin 을 정제하였다. 정제 과정 주에서 ion-exchange chromatography 단계에 서는 35.3%의회수율이 7.3%로 감소하였다. Reverse-Phase chromatography에선 3.3%의 회수율을 보였고 활성도는 413.5배로 증가하였다. Tricine-SDS 전기영도 결과 bacteriocin 은 단일 밴드로 나타났으며, N-말단 아미노산 서열 분석을 수행한 결과 $NH_2$-IIe-Leu-Pro-GIn로 확인되었다. 아미노산조정 분석결과를 바탕으로 분자량을 예측한 결과 본 bacteriocin은 32개의 아미노산으로 이루어져 있으며 분자량은 3.6kDa인 것으로 추정되었다.

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메주에서 분리된 Enterococcus faecium MJ-14가 생산하는 박테리오신의 부분정제 (Partial Purification of Bacteriocin Produced by Enterococcus faecium MJ-14 Isolated from Meju)

  • 이종갑;이군자;임성미
    • 한국식품위생안전성학회지
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    • 제20권4호
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    • pp.211-216
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    • 2005
  • E. faecium MJ-14 균주의 배양 상등액에 $50\%$ 황산암모늄을 처리한 결과, 박테리오신 활성은 3,840 BU/mL으로 배양 상등액에 비해 6배 증가되었고 회수율은 약 $60\%$에 이르렀다. 이온교환크로마토그래피에 의해 용출된 획분 중 $48\∼76$번에서 항균 활성이 나타났으며, 용출물의 비활성은 35.7배 증가하였고, 회수율은 약 $41\%$였다. 그리고 겔 크로마토그래피에 의해 비활성 127,293 BU/mg인 박테리오신 단백질을 정제하였으며, 이 때 정제도는 114배, 수율은 $36\%$였다. 정제된 박테리오신 단백질을 SDS-FACE한 결과. 항균 활성을 나타내는 4.3 kDa과 5.8 kDa의 분자량을 확인하였다.

도시하천의 환경특성과 친자연적 계획전략 - 춘천시 공지천을 대상으로 - (Environmental Characteristics and Nature-friendly Planning Strategies for an Urban Stream - The Case of Chuncheon's Gongji Stream -)

  • 조현길;안태원
    • 한국조경학회지
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    • 제34권3호
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    • pp.1-11
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    • 2006
  • This study analyzed characteristics of natural and human environments in Chuncheon's Gongji stream, and suggested nature-friendly planning strategies for self-purification of water quality, biodiversity improvement and conservative waterfront recreation. The environmental analysis included streambed structures, floodplain soils, water quality, vegetation, wildlife, and human facilities. Natural colonization of vegetation for the middle section of the study stream was obstructed by a straightened concrete revetment of baseflow channel, and vehicle movement and concrete parking lots across the floodplain. These human disturbances also deteriorated the naturalness of the stream landscape and limited habitation of bird species. However, natural sedimented wetlands in half of the channel width for the lower section of the stream contributed to a desirable vegetational landscape and greater bird occurrence. Based on BOD measurements, water quality of the stream fell under class $II{\sim}III$ of the stream water-quality standard, but it was worse around sewage outlets due to incomplete sewage collection especially during the dry season. Dominant fish species included typical inhabitants of good water-quality streams that are tolerant of adverse habitat changes. Nature-friendly planning strategies were established based on analysis of the environmental characteristics. They focused on not merely spatial zoning and layout divided into four zones - preservation, partial preservation, conservation and use -, but close-to-nature channel revetment techniques, natural water-purification facilities, biotope diversification, and water-friendly recreation and circulation. Strategies pursued both renewal of stream naturalness and hydraulic stability of streamflow by minimizing transformation of natural channel micro-topography and biotope, and by reflecting natural traces of streambed structures such as revetment scour and sedimentation.