• 제목/요약/키워드: Purification and characterization

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마황으로부터 췌장 Cholesterol Esterase 저해물질 분리 및 규명 (Purification and Characterization of the Inhibitory Principle aganist Pancreatic Cholesterol Esterase from Ephedra herba)

  • 김희숙;조은정;류병호;송병권;이태훈;서판길;류성호
    • 한국식품영양과학회지
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    • 제28권4호
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    • pp.816-821
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    • 1999
  • Cholesterol esterase(pCEH, pancreas cholesterol ester hydrolase, E.C.3.1.1.13) which is secreted from pancreas has been known as an important lipase for cholesterol uptake. cholesteryl acyl esters from a diet must be hydrolyzed to free cholesterol and fatty acid by cholesterol esterase before the absorption in small intestine. For the development of inhibitory substances from natural source, we screened many extracts of oriental herbs for the inhibition of cholesterol esterase in vitro. The ethanol extract of Ephedra herba showed strong inhibitory activity. Solvent fractionation and silica gel column chromatography with the extract lead to the purification of the inhibitory principle in Ephedra herba. Crystallized inhibitor was identified as ( ) ephedrine by using UV, FT IR, 1H NMR, 13C NMR and GC/Mass. These results suggest that ( ) ephedrine can be used as a potential lead compound for the development of inhibitor for cholesterol uptake by cholesterol esterase inhibition.

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Purification and Characterization of Haemolymph Ferritin from the Cricket, Gryllus bimaculatus

  • Seo, Dong-Hwan;Nam, Kung-Pil;Kim, Iksoo;Ryu, Kang-Sun;Yun, Chi-Young
    • 한국잠사학회:학술대회논문집
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    • 한국잠사학회 2003년도 제46회 춘계 학술연구 발표회
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    • pp.80-80
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    • 2003
  • The isolation and purification of ferritin from the cricket, Gryllus bimaculatus haemolymph were accomplished by anion exchange column chromatography using HiTrap Q column (1.6 $\times$ 4 cm, Amersham Phamacia Co.), 7$0^{\circ}C$ heat and acid treatment, and gel filtration column chromatography using G4000SW column (0.75 $\times$ 60cm, TOSOH Co.) by FPLC system. (omitted)

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대구의 간 단백질의 효소적 가수분해물로부터 안지오텐신 I 전환효소 저해 펩타이드의 분리.정제 및 특성 (Purification and Characterization of Angiotensin I Converting Enzyme lnhibitory Peptides from Enzymatic Hydrolysate of Cod Liver Protein)

  • 최영일;박표잠;최정호;변희국;정인철;문성훈;김세권
    • 생명과학회지
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    • 제10권2호
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    • pp.140-149
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    • 2000
  • In order to utilize marine processing waste which would normally be discarded, cod liver protein was hydrolysed by ${\alpha}$-chymotrysin, and the hydrolysate was investigated for the new angiotensin I converting enzyme (ACE) inhibitor. Thy hydrolysate was separated into three major types, with molecular weight cut-off (MWCO) values less than 10 kDa, 5 kDa and 1 kDa of ultrafiltration membranes, respectively. ACE inhibitory peptides were isolated from the fractions passed through MWCO 1 kDa membrane, and purified by using ion-exchange chromatography on a SP-Sephadex C-25 column, gel filtration on a Sephadex G-15 column, and HPLC on an ODS column. The purity was identified with capillary electrophoresis. The amino acid sequences of two peptides were Met-Ile-Pro-Pro-Tyr-Tyr (IC50=10.9 ${\mu}$M) and Gly-Leu-Arg-Asn-Gly-Ile (IC50=35.0 ${\mu}$M)

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Purification and Characterization of Crystalins by Aqueous Two-Phase Extraction

  • Bermudez, Ondrea;Forciniti, Daniel
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제6권6호
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    • pp.395-401
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    • 2001
  • Crystallins are a family of water-soluble proteins that constitute up to 90% of the wa-ter-soluble proteins in mammalian eye lenses, We present in this paper an alternative purification method for these proteins using polyethylene glycol/dextran aqueous two-phase extraction. Un-der the appropriate conditions, we were able to recover the γ-crystallin fraction essentially free of the remaining proteins. High concentrations of salt at a neutral pH maximize the recovery of γ-crystallins in the top phase and minimize the contamination by the other proteins present in the lenses. The proposed protocol decreases the separation time by about 50%. The complex partition behavior observed for these proteins reflects a delicate balance between protein/phase-forming species(various polymers and salts) and protein interactions. This is evidenced, in part, by the role played by the largest proteins in this group as a "pseudo"phase-forming species.

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Characterization and diagnosis of YMV-K strains infecting Chinese yam(Dioscorea opposita Thunb. cv. Dung-Gun-Ma) in Korea. (oral)

  • D. K. Kang;H. Y. Shin;J. H. Sung;Park, J. H.;M. U. Chang
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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    • pp.127.2-127
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    • 2003
  • YMV-K strains were purified from D. oppostita Thunb. tv. Dung-Gun-Ma showing mosaic symptom on their leaves. YMV-K strains were filamentous particles of 780nm in length and induced cytoplasmic disorder such as inclusion body formation. Nucleotide and amino acid sequences of 5'-UTR, Pl and CP of YMV-K strains shared 80.8, 64.7 and 98.3% identity respectively to JYMV J1 in the mean value. Purification of YMV-K strains according to JYMV purification method(S. Fuji) was conducted to product antiserum. With antiserum against YMV-K strains, the Various diagnosis methods such as IC-RT-PCR, DIBA, RIPA and indirect-ELISA were used to detect YMV-K strains in Chinese yam plant.

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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
    • 고려인삼학회:학술대회논문집
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    • 고려인삼학회 1990년도 Proceedings of International Symposium on Korean Ginseng, 1990, Seoul, Korea
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    • pp.79-85
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    • 1990
  • The aqueous extract of fresh ginseng (Panax ginseng C.A. Meyer) contains a macromolecular fraction that showed mitogenic and comitogenic activities in human peripheral blood Iymphocytes. 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 Iymphocyte 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 the $\beta$-Adrenergic Receptor from Rat Cerebral Cortex

  • 신찬영;김희진;노민수;함경수;강현삼;고광호
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1994년도 춘계학술대회 and 제3회 신약개발 연구발표회
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    • pp.292-292
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    • 1994
  • Catecholamines acting through ${\beta}$-adrenergic receptors regulate a wide range of metabolic activities in mammalian tissue. Of the various receptors coupled to adenylate cyclase, the ${\beta}$-adrenergic receptors are the most extensively characterized and have been purified from both nonmammal ian and mammal inn sources. However, most studies of the molecular properties of ${\beta}$-adrenergic receptors have been confined to peripheral tissues. Less progress has been achieved in characterizing the brain ${\beta}$-adrenergic receptor The goal of the present study was, therefore, to purify and characterize the neurotransmitter receptor proteins. To achieve this goal, the following stepwise experiments were performed. At first, the membrane-bound ${\beta}$-adrenergic receptors were. solubi1ized from brain tissue. Secondly, conditions for affinity chromatography were determined to purify the solubilized receptors effectively. Finally, the large-scale purification was performed and the characteristics of the purified ${\beta}$-adrenergic receptor were examined.

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Purification and Characterization of Protein Carboxyl O-Methyltransferase from Porcine Spleen

  • Yoon, Sung-Pil;Son, Min-Sik;Han, Jeung-Whan;Lee, Hyang-Woo;Hong, Sung-Youl
    • BMB Reports
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    • 제30권6호
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    • pp.410-414
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    • 1997
  • We purified a protein carboxyl O-methyltransferase (protein methylase II) from porcine spleen to homogeneity. The molecular weight of the porcine spleen protein methylase II (ps-PM II) was estimated to be 27,500 daltons on SDS-PAGE. Amino acid sequence of N-terminal 28 residues for ps-PM II was identified. Amino-terminal three amino acid residues of ps-PM II were deleted when compared to those of other protein carboxyl methytransferase. S-Adenosyl-L-homocysteine competitively inhibits ps-PM II with a K, value of $1.63{\times}10^{-7}M$. Myelin basic protein exhibited the highest methyl-accepting capacity among the proteins tested.

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Purification and Characterization of a Laccase from Cerrena unicolor and Its Reactivity in Lignin Degradation

  • Kim, You-Sung;Cho, Nam-Seok;Eom, Tae-Jin;Shin, Woon-Sup
    • Bulletin of the Korean Chemical Society
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    • 제23권7호
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    • pp.985-989
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    • 2002
  • For efficient biopulping process, very active and stable lignase is essential. Laccase is one of the best enzyme in terms of environmentally benign processes, since the enzyme uses oxygen as an oxidant to degrade lignin and produces no hamful prod ucts. We could purify a laccase homogeneously from Cerrena unicolor in a very active state. It shows characteristic absorption feature with blue band at λmax = 604 ㎚. Molecular weight of the enzyme is 57,608 which could be accurately determined by MALDI/TOF MS. The enzyme has 2.8 copper ions per enzyme implying apoenzymes might exist together. The enzyme is active in lignin degradation and the activity increases 4 times in the presence of ABTS as a mediator.

Purification and Characterization of Chitinase from a Marine Bacterium, Vibrio sp. 98CJ11027

  • Park, Shin-Hye;Lee, Jung-Hyun;Lee, Hong-Kum
    • Journal of Microbiology
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    • 제38권4호
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    • pp.224-229
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    • 2000
  • Chitin-degrading marine bacterial strain 98CJ11027 was isolated from bryozoa from the coastal area of Cheju Island, Korea, and identified as a member of the genus Vibrio. The molecular mass of the main extracellular chitinase (chitinase I), purified from strain 98CJ11027, was estimated to be 98 kDa. The optimal condition for chitinase I activity is pH 6.0 and 45$^{\circ}C$. The activity was inhibited by Fe$\^$+2/ and Cu$\^$+2/. Chitinase I displayed the hydrolysis type of chitobiosidase and catalyzed reversed hydrolysis leading to the synthesis of tetraacetylchitotetraose.

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