• 제목/요약/키워드: I-gel

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김 가수분해물로부터 Angiotensin-I Converting Enzyme저해 Peptide의 분리$\cdot$정제 (Separation and Purification of Angiotensin-I Converting Enzyme Inhibitory Peptides from Layer Hydrolysate)

  • 이헌옥;김동수;도정룡;권대영
    • 한국수산과학회지
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    • 제34권2호
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    • pp.164-172
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    • 2001
  • 본 연구는 김의 Maxazyme NNP 가수분해물로부터 여러 단계의 column chromatography 및 HPLC를 통해 ACE 저해 peptide를 분리, 정제하여 이의 분자량과 amino acid sequence를 분석하였다. ACE 저해 효과가 큰 저분자 peptide의 함량이 많은 가수분해물을 얻기 위한 최적의 단백질 가수분해 효소를 선정하기 위하여 시험한 결과, Maxazyme NNP에 의한 가수분해물의 ACE 저해효과가 $37.2\%$로 가장 높게 나타났다 김의 효소 가수분해물로부터ACE 저해 peptide만을 효율적으로 분리하기 위한 추출 조건 시험에서, 색소 제거를 위해서는 diethylether 처리가, 다당류 및 고분자 단백질 제거를 위해서는 $70\%$ ethanol 처리가 각각 선정되었다. 김 가수분해물로부터 ACE 저해 peptide를 분리, 정제하기 위한 첫 단계로 ultrafiltration을 한 결과, 분자량 3,000 이하 분획물의 ACE 저해 효과가 $69.4\%$로 가장 높았으며, 이 분획물을 gel filtration chromatography (Sephadex G-25) , reverse-phase HPLC (ODS & Vydac C-18) 및 gel permeation chromatography (Superdex Peptide HR)와 같은 단계별 column chromatography를 행하여 최종적으로 단일 peptide peak들을 분리하였다. 이들 단일 peptide peak들의 분자량을 Electrospray-Mass Spectrometer로 측정한 결과, 각각 413.48 (S1O2V2V1P),346.86 (S1O2V2V2P) 그리고 320.32 (S2O6V3V1P) dalton이었으며, 이들 peptide의 amino acid sequence는 N-말단으로부터 아미노산 잔기를 분석한 결과, 각각 Val-Gln-Gly-Asn, Thr-Glu-Thr 및 Phe-Arg으로 확인되었다.

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수산발효식품 중의 Angiotensin-I 전환효소 저해제의 특성 -1. 멸치젓갈 중의 Angiotensin-I 전환효소 저해제의 특성- (Characteristics of Angiotensin-I Converting Enzyme Inhibitors Derived from Fermented Fish Product -1. Characteristics of Angiotensin-I Converting Enzyme Inhibitors Derived from Salted and Fermented Anchovy-)

  • 김선봉;이태기;박영범;염동민;김외경;변한석;박영호
    • 한국수산과학회지
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    • 제26권4호
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    • pp.321-329
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    • 1993
  • 수산자원의 기능특성 해명을 위한 연구의 일환으로 전통수산발효식품으로 널리 이용되고 있는 멸치젓갈에 함유되어 있는 angiotensin-I 전환효소저해물질을 추출하여 gel 여과에 의하여 분리된 획분들의 작용 및 특성에 대해서 살펴보았다. 그 결과를 요약하면 다음과 같다. 1. ACE 저해물질의 추출 조건을 검토하기 위하여 가장 높은 ACE 저해효과를 나타내는 숙성 60일차의 멸치젓갈을 ethanol 및 acetone의 농도를 각각 10, 25, 50 및 $80\%$로 하여 추출하였을 때 ethanol 농도가 $50\%$인 획분이 가장 우수한 저해효과를 나타내었다. 2. 멸치젓갈 숙성중 시료액의 $50\%$ ethanol 가용성 peptide-nitrogen 함량 및 ACE 저해효과는 아미노 질소 함량이 최고치에 달하는 숙성 60일차에 최대값을 나타내다가 그 후 다소 감소하는 경향을 나타내었다. 3. Gel 여과에 의한 멸치젓갈의 획분별 ACE 저해효과를 검토한 결과, 획분 C 및 D가 가장 높은 ACE 저해작용을 나타내어 이들을 rechromatography하여 분리한 단일 획분인 C'와 D'획분의 $IC_{50}$은 각각 97, $65{\mu}g$로 나타났다. 4. 분리한 단일 획분인 C'와 D'획분의 아미노산 조성은 다소 차이가 있었으며, C'획분은 threonine, glutamic acid 및 lysine의 함량이 많은 것으로 나타났으며, D'획분은 serine과 proline의 함량이 많은 것으로 나타났다.

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이차원전기영동법을 이용한 white muscle과 red muscle간의 단백질 발현양상의 비교분석 (Comparative Analysis of Muscle Proteome from Porcine White and Red Muscles by Two-dimensional Electrophoresis)

  • 김남국;조중호;추교선;박혜란;박범영;김언현;이창수
    • Journal of Animal Science and Technology
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    • 제45권5호
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    • pp.731-738
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    • 2003
  • 근육의 생화학적 특성을 단백질 수준에서 분석하기 위하여 3원교잡종 돼지 3개체를 선발하고, white muscle은 longissimus dorsi muscle을 red muscle은 soleus muslce을 분리하여 분석에 이용하였다. 각 근육조직은 수용성, 불수용성 단백질 및 총단백질로 분리하여 추출하였고, 이차원전기영동 분석을 위하여 17cm 길이의 immobilized pH gradient strip (Bio-Rad, 3-10NL)과 12% acrylamide gel을 이용하여 전개하였다. 각각의 gel은 coomassie stain과 silver stain을 통하여 가시화 하였고, PDQuest software을 통하여 단백질 발현양상을 분석하였다. 하나의 gel에서 평균 600개 이상의 단백질 spot을 관찰하였으며, 반복실험을 통하여 white muscle과 red muscle간에 발현의 차이를 보이는 5개의 단백질 spot을 확인할 수 있었다. 5개의 spot 중 4개의 단백질은 측정된 분자량과 pI값이 troponin I, T 및 myoglobin의 수치값과 유사한 것으로 확인되었다. 그러나, 1개의 spot은 오차범위 내에서 유사한 단백질을 확인할 수 없었다. 5개의 단백질 spot중 1개(spot 1)는 white muscle에서, 4개의 spot(spot 2~5)은 red muscle에서 높게 발현됨을 확인하였으며, 특히 spot 4의 경우 white muscle 보다 red muscle에서 평균 14.6배 높게 발현됨을 확인하였다. 본 연구는 근육의 생화학적 특성을 이해하는데 중요한 기초 자료로 활용될 수 있으며, 앞으로 white muscle과 red muscle의 단백질 발현 분석을 통하여 단백질 수준에서의 생화학적 특성에 관한 연구가 충분히 진행되어야 할 것이다.

The Concentration-Dependent Distribution of Tris(4,7'-diphenyl-1,10'-phenanthroline) Ruthenium (II) within Sol-Gel-Derived Thin Films

  • Lee, Joo-Woon;Cho, Eun-Jeong
    • Bulletin of the Korean Chemical Society
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    • 제32권8호
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    • pp.2765-2770
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    • 2011
  • Organic dye-doped glasses, viz., ruthenium (II) tris(4,7'-diphenyl-1,10'-phenanthroline) $[Ru(dpp)_3]^{2+}$ incorporated into thin silica xerogel films produced by the sol-gel method, were prepared and their $O_2$ quenching properties investigated as a function of the $[Ru(dpp)_3]^{2+}$ concentration (3-400 ${\mu}M$) within the xerogel. The ratio of the luminescence from the $[Ru(dpp)_3]^{2+}$-doped films in the presence of $N_2$ and $O_2$ ($I_{N2}/I_{O2}$) was used to describe the film sensitivity to $O_2$ quenching. ($I_{N2}/I_{O2}$ changed three-fold over the $[Ru(dpp)_3]^{2+}$ concentration range. Time-resolved intensity decay studies showed that there are two discrete $[Ru(dpp)_3]^{2+}$ populations within the xerogels (${\tau}_1$ ~ 300 ns; ${\tau}_2$ ~ 3000 ns) whose relative fraction changes as the $[Ru(dpp)_3]^{2+}$ concentration changes. The increased $O_2$ sensitivity that is observed at the higher $[Ru(dpp)_3]^{2+}$ concentrations is a manifestation of a greater fraction of the 3000 ns $[Ru(dpp)_3]^{2+}$ species (more susceptible to $O_2$ quenching). A model is presented to describe the observed response characteristics resulting from $[Ru(dpp)_3]^{2+}$ distribution within the xerogel.

Protein Profile of Embryonic Stem Cells Following Differentiation

  • D. I. Jin;Kang, H. S;Kim, H. J.;Lee, S. H.;Park, C. S.;K. S. Im;Lee, H. M.
    • 한국가축번식학회지
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    • 제26권4호
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    • pp.377-384
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    • 2002
  • To investigate the expression patterns of proteins and growth factor signals in differentiated rabbit embryonic stem (ES) cells, ES cells with confluent stage grown of feeder layer and differentiated cells into embryoid bodies (EB) without feeder cell were applied to protein gel and Western blotting analysis. There were 66kDa and 28kDa specifically expressed in differentiated ES cell but not in undifferentiated ES cell while 25kDa protein band showed up in only undifferentiated ES cells. Also there were some difference of protein bands in several area of gel between differentiated and undifferentiated ES cells such as about 100 kDa, 50kDa and 27kDa areas, but there was no difference in band pattern of one-dimensional gel analysis between mouse ES cells and rabbit ES cells. IGF-I receptor and EGF receptor were expressed in differentiated cells and undifferentiated cells. And ICF-I and EGF were not expressed in both differentiated and undifferentiated cells. These results indicated that ES cells express their own proteins to inhibit differentiation while EB cells synthesize different proteins to differentiate, and 16F-I receptor and EGF receptor were expressed in both ES and EB cells probably for the different functions.

닭 췌장 Protein Methylase II의 분리정제 및 성질 (Protein Methylase II from Chicken Pancreas: Purification and Properties)

  • 유태무;남궁석민;홍성렬;이향우
    • 약학회지
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    • 제35권6호
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    • pp.473-482
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    • 1991
  • Protein methylase II (S-adenosyl-L-methionine:protein carboxyl-O-methyltransferase; EC 2.1.1.24., PM II) was purified from chicken pancreas by subcellular fractionation, DEAE-cellulose chromatography, QAE-Sephadex A-50 chromatography, Sephadex G-75 chromatography, and Sephadex G-75 rechromatography. The purified PM II gave a single band upon polyarcrylamide gel electrophoresis both in the presence of SDS and in Tris glycine buffer without SDS. The pI value of purified PM II was identified as 5.7 on isoelectric focusing gel. Properties and activities of PM II were studied and the following results were obtained. 1) PM II from chicken pancreas was purified approximately 221-fold with a yield of 1.3%. 2) The purified PM II appear constituted of a single polypeptide chain of a molecular weight 46,800 daltons. 3) Hemoglobin exhibited the highest of methyl-accepting activity among the substrates tested. 4) The purified PM II has a $K_m$ of $4.67{\times}10^{-6}M$ and a $V_{max}$ of 37.5 pmoles of $methyl-^{14}C/min./mg$ enzyme for $SAM^{-14}CH_3$ as methyl donor in the presence of histone type II-As. 5) It is found that S-adenosyl-L-homocysteine is a competitive inhibitor for PM II with $K_i$ value of $3.23{\times}10^{-5}M$.

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Genomic Heterogeneity in Clinical Strains of Mycobacterium tuberculosis, M. terrae Complex, M. gordonae, M. avium-intracellulae Complex and M. fortuitum by Pulsed-Field Gel Electrophoresis

  • Kim, Jeong-Ran;Kang, Bong-Seok;Ko, Jeong-Heon;Park, Jin-Suk;Kim, Sang-Jae;Bai, Gil-Hwan;Chung, Tae-Ho;Nam, Kyung-Soo;Choi, Yong-Kyung;Choe, In-Sung;Chung, Tae-Wha;Lee, Young-Choon;Kim, Cheorl-Ho
    • BMB Reports
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    • 제29권6호
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    • pp.569-573
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    • 1996
  • Clinical strains of Mycobacterium tuberculosis, M. terrae complex, M. gordonae, M. avium-intracellulae complex, and M. fortuitum from Korean patients were isolated and analyzed by comparing large restriction fragment (LRF) patterns produced by digestion of genomic DNA with infrequent-cutting endonucleases like AsnI and XbaI. and pulsed-field gel electrophoresis (PFGE). Three M. tuberculosis, two M. terrae complex, two M. gordonae, two M. avium-intracellulae complex, and two M. fortuitum strains were compared by using AsnI and XbaI. and this allowed easy visual separation of all epidemiologically unrelated strains. PFGE exhibits different DNA restriction patterns which are easy to compare. Genome size of the strains roughly ranged from 3020 to 3335 kb. The LRF patterns are useful for epidemiologic studies of tuberculosis with regard to drug resistance.

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Sol-Gel 공정을 이용한 ZnO 쇼트키 다이오드의 제작 및 특성평가 (Fabrication and Characterization of ZnO Schottky Diode Using Sol-Gel Process)

  • 이득희;김경원;박기호;김상식;이상렬
    • 한국전기전자재료학회:학술대회논문집
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    • 한국전기전자재료학회 2010년도 하계학술대회 논문집
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    • pp.390-390
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    • 2010
  • We fabricate Schottky diodes with the contact between a sol-gel derived ZnO layer and Au that guarantees the expected Schottky contact due to the high work function. The formed single metal Schottky barrier shows characteristics comparable to the barrier formed by alloys. Au is deposited by thermal evaporation on a ZnO thin film that is optimally formed under sol-gel process conditions of a 1-mol zinc acetate concentration and a 3000-rpm coating speed. Possible defects. which can provide deleterious current paths. are minimized by patterning the deposited Au. The I-V curve verifies the formation of a Schottky contact. Measurements showed that the Schottky barrier height and leakage current at -5 V were 0.6 eV and $1{\times}10^{-12}A$. respectively.

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Microwave Sol-Gel Derived Ho3+/Yb3+ Co-Doped NaCaGd(MoO4)3 Phosphors and their Upconversion Photoluminescence

  • Lim, Chang Sung
    • 한국세라믹학회지
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    • 제53권4호
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    • pp.456-462
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    • 2016
  • $NaCaGd(MoO_4)_3:Ho^{3+}/Yb^{3+}$ ternary molybdates were successfully synthesized by microwave sol-gel method for the first time. Well-crystallized particles formed after heat-treatment at $900^{\circ}C$ for 16 h showed a fine and homogeneous morphology with particle sizes of $3-5{\mu}m$. Under excitation at 980 nm, the UC intensities of the doped samples exhibited strong yellow emissions based on the combination of strong emission bands at the 520-nm and 630-nm emission bands in the green and red spectral regions, respectively. The strong 520-nm emission band in the green region corresponds to the $^5S_2/^5F_4{\rightarrow}^5I_8$ transition of $Ho^{3+}$ ions, while the strong 630-nm emission band in the red region appears to be due to the $^5F_5{\rightarrow}^5I_8$ transition of the $Ho^{3+}$ ions. The optimal $Yb^{3+}:Ho^{3+}$ ratio was found at 9:1, as indicated by the composition-dependent quenching effect of $Ho^{3+}$ ions. The pump power dependence of the upconversion emission intensity and the Commission Internationale de L'Eclairage chromaticity coordinates of the phosphors were evaluated in detail.