• 제목/요약/키워드: enzyme inhibitory

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Production of Dye-decolorizing Enzyme using Molasses-containing Medium

  • Lee, Tae-Ho;Shoda, Makoto
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2000년도 춘계학술발표대회
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    • pp.57-58
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    • 2000
  • Production of dye-decolorizing enzyme was investigated with the cultivation of Geotrichum candidum Dec1 (abbreviated to Dec1) using molasses as a cheap raw materials. Molasses was found to stimulate the enzyme production as well as a role of carbon source, because the production increased with molasses content up to 50 g/L. However, the severe inhibition of dye-decolorizing enzyme activity was observed even at low concentration of molasses 10 g/L when purified decolorizing peroxidase was used. Its inhibitory effect was reduced through the cultivation of Dec1. The fractions of molasses separated by a gel chromatography showed the different degrees of inhibition. As a way to reduce the inhibitory effect, the dilution of culture broth was examined, and the total decolorizing activity for Reactive blue 5 increased 7 times as much as that of original culture broth by 30 times dilution. On the basis of result, we proposed a process scheme which can fully utilize both positive and negative effect of molasses in dye-decolorizing process using molasses.

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Binding Subsites In the Active Site of $Zn^{2+}$-Glycerophosphocholine Cholinephosphodiesterase

  • Sok, Dai-Eun;Kim, Mee-Ree
    • BMB Reports
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    • 제28권2호
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    • pp.94-99
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    • 1995
  • The properties of binding sites in the active site of $Zn^{2+}$-glycerophosphocholine cholinephosphodiesterase were examined using substrates and inhibitors of the enzyme. Phosphodiesterase hydrolyzed p-nitrophenylphosphocholine, p-aminophenylphosphocholine, and glycerophosphocholine, but did not hydrolyze either acylated glycerophosphocholine or bis (p-nitrophenyl)phosphate, suggesting a size limitation for interaction with a glyceryl moiety-binding subsite. The hydrolysis of p-nitrophenylphosphocholine was competitively inhibited by glycerophosphocholine and p-aminophenylphosphocholine, while glycerophosphoethanolamine was a weak inhibitor. The enzyme was also inhibited by choline, but not by ethanolamine. Thiocholine, a much more potent inhibitor than choline, was more inhibitory than cysteamine, suggesting a strict specificity of an anionic subsite adjacent to a $Zn^{2+}$ subsite. Of all oxyanions tested, the tellurite ion was found to strongly inhibit the enzyme by binding to a $Zn^{2+}$ subsite. The inhibitory role of tellurite was synergistically enhanced by tetraalkylammonium salts, but not by glycerol. Deactivation of the enzyme by diethylpyrocarbonate was partially protected by choline, but not by glycerophosphate. It is suggested that the active site of phosphodiesterase contains three binding subsites.

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$\gamma$-Aminobutyrate Transaminase에 대한 Mycotoxin의 저해작용 (Inhibitory Actions of Mycotoxins on Brain $\gamma$-Aminobutyrate Transaminase)

  • Lee, Su-Jin;Lee, Kil-Soo;Choi, Soo-Young
    • 미생물학회지
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    • 제31권3호
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    • pp.224-229
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    • 1993
  • GABA transminase (4-aminobutyrate aminotransferase), which catalyzes the breakdown of the major inhibitory neurotransmitter, GABA, in mammalian brain, was inactivated by preincubation with the mycotoxin patulin. The time course of the reaction was significantly affected by the substrate .alpha.-ketoglutarate, which aforded complete protection against the loss of catalytic activity. The recovery from the inhibition of patulin by the addition of dithiothreitol (DTT) supports that patulin reacts with the sulfhydryl residue in the catalytic domain of the enzyme. The reconstitution of the reduced enzyme and apoenzyme with pyridoxal-5-P(PLP) was inhibited by another mycotoxin, penicilic acid. This mycotoxin may interact with lysyl residue of the enzyme. Therefore, it is postulated that the critical sulfhydryl and lysyl residues in the catalytic domain of the enzyme react with mycotoxin patulin and penicillic acid, respectively.

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Physiological Functionality and Enzyme Activity of Biomass from Pichia anomala Grown on Ginseng-Steaming Effluent

  • Kim, Na-Mi;So, Seung-Ho;Lee, Sung-Gae;Song, Jung-Eun;Seo, Dong-Soo;Lee, Jong-Soo
    • Mycobiology
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    • 제36권3호
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    • pp.148-151
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    • 2008
  • A novel biomass was prepared from Pichia anomala KCCM 11473, which grew well in ginseng-steaming effluent (GSE), and its physiological functionalities and enzyme activities were determined. When the strain was cultured in the GSE (pH 6.0) at 30$^{\circ}C$ for 48 h, 1.6 mg of biomass per ml-cultures was produced. The cell-free extract of the biomass showed high antihypertensive angiotensin I-converting enzyme inhibitory activity of 72.0% and anticholesteromia HMG-CoA reductase inhibitory activity of 46.5%. The cell-free extract also showed 13.0 U per ml and 8.5 U per ml of neutral protease activity and alkaline protease, respectively.

Effects of Polyacetylenes from Panax ginseng on Some Microsomal and Mitochondrial Enzymes

  • Kim, Young-Sook;Kim, Shin-Il;Hahn, Dug-Ryong
    • 생약학회지
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    • 제20권3호
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    • pp.154-161
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    • 1989
  • Effects of panaxydol, panaxynol and panaxytriol isolated from Panax ginseng C.A. Meyer on some enzyme activities were determined. Activities of ATPase, membrane-bound enzyme from Sarcoma 180 and rat liver were slightly inhibited by panaxydol. Activities of 5'-nucleotidase, membrane-bound enzyme and succinate cytochrome c reductase in mitochonidria from sarcoma 180 and rat livers were significantly inhibited in a dose-dependent manner by panaxynol. The inhibitory effects of panaxydol and panaxynol on succinate cytochrome c reductase activities were more potent than those on 5'-nucleotidase activities and panaxynol was found to be a very potent inhibitor of succinate cytochrome c reductase. Activities of glucose-6-phosphatase in endoplasmic reticulum from Sarcoma 180 and rat livers were not affected by all three polyacetylenes. These results suggested that the inhibitory effects of panaxydol and panaxynol on enzyme activities might contribute to their biological activities.

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백지근(白芷根) 성분(成分)이 간(肝)의 약물대사효소활성(藥物代謝酵素活性)에 미치는 효과(效果) (Effect of the Constituents of Angelicae dahuricae Radix on Hepatic Drug Metabolizing Enzyme Activity)

  • 신국현;김옥남;우원식
    • 생약학회지
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    • 제19권1호
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    • pp.19-27
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    • 1988
  • The hexane and ether extracts from the roots of Angelica dahurica caused a significant inhibition of hepatic drug-metabolizing enzyme (DME) activity. Through systematic fractionation by $SiO_2\;column$ and vacuum liquid chromatography monitoring by bioassays, three furanocoumarins, phellopterin, byakangelicin and tert-O-methylbyakangelicin were isolated as active principles. These components have biphasic responses, both inhibitory and inducing effects on DME system. Tert-O-methyl byakangelicin was found to have the strongest enzyme inhibitory potency.

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김 단백질 가수분해물의 Angiotensin Ⅰ 전환효소 저해 활성 (Angiotensin Ⅰ Converting Enzyme(ACE) Inhibitory Activities of Laver(Porphyra tenera) Protein Hydrolysates)

  • 김영명;도정룡;인재평;박종혁
    • 한국식품영양학회지
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    • 제18권1호
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    • pp.11-18
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    • 2005
  • Angiotensin Ⅰ converting enzyme(ACE) inhibitory activities of laver(Porphyra tenera) protein hydrolysates were investigated by enzymes used for hydrolysis, molecular fractions and drying methods. For the enzymatic hydrolysis, crude laver protein, separated by filtration of water extract of dried laver extracted with 20 times(w/v) water for 3 hours at boiling temperature, were hydrolyzed with three commercial protease, Pepsin, alcalase and maxazyme NNP at optimal conditions. The yield of hydrolysis and ACE inhibitory activities of which were high in order of pepsin, alcalase and maxazyme NNP. ACE inhibitory activities of laver hydrolysates by molecular levels were high in order of 3 kDa > 10 kDa > 3∼10 kDa, and the IC/sub 50/ ACE inhibitory activities by molecular lebels were 4 mg/mL(3 kDa), 5 mg/mL(total hydrolysate), and 20 mg/mL(10 kDa), respectively. The storage stability of dried laver hydrolysates at 20℃ were strongly affected by drying methods, hot air dried of which were much stabler than freeze-dried one.

재조합 대장균으로부터 항고혈압 Angiotensin I-Converting Enzyme 저해제의 특성연구 (Characterization of Antihypertensive Angiotensin I-Converting Enzyme Inhibitor from Recombinant E. coli)

  • 김재호;정승찬;이대형;이종수
    • 자연과학논문집
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    • 제16권1호
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    • pp.1-13
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    • 2005
  • 안지오텐신(ACE) 저해제는 항고혈 효과를 갖고 있으므로 오랫동안 고혈압의 예방이나 치료에 이용되어 왔다. 본 연구는 재조합 대장균으로부터 새로운 ACE 저해제를 생산하고 정제하며 나아가 이들이 구조-기능 관P를 규명하기 위해 수행되었다. Saccharomyces cerevisiae의 ACE 저해 펩타이드 유전자를 함유하고 있는 재조합 pGEX-4T-3을 대장균 BL21(DE3)로 형질전환 시켰다. 재조합 pGEX-4T-3을 갖고 있는 대장균 BL21(DE3)로부터 생산된 Glutathione-s 전이효소(GST) 융합 단백질을 얻어서 그중 ACE저해 펩타이드를 Sephadex G-25 컬럼 크로마토그래피로 정제하였다. 정제된 ACE 저해 펩타이드는 타이로신-아스파틱엑시드-그리신-글리신-발린-페닐알라린-아르기닌-발린-타이로신-트레오닌의 서열을 가진 새로운 decapeptide이었고 ACE에 대하여 경쟁적으로 저해하였다.

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Inhibition of Monoamine Oxidase B by Cigarette Smoke Constituents

  • Lim, Heung-Bin;Sohn, Hyug-Ok;Lee, Young-Gu;Moon, Ja-Young;Kang, Young-Kook;Kim, Yong-Ha;Lee, Un-Chul;Lee, Dong-Wook
    • 한국연초학회지
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    • 제19권2호
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    • pp.136-144
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    • 1997
  • Cigarette smoking is known to suppress both 1-methy14-phenyl-155,Ltetrahydropy-ridine (MPTP)-induced parkinsonism and idiopathic Parkinson's disease (PD). However, the precise mechanism underlying its protective action against PD is not clearly elucidated yet. In order to find possible clue on the mechanism of protective action of smoking, we investigated the inhibitory effect of cigarette smoke components on rat brain mitochondria1 monoamine oxidase B (MAO-B), responsible enzyme for the activation of MPTP to its toxic metabolitesr and identified the components having an inhibitory potency on this enzyme from cigarette smoke. Total 31 eligible constituents including nicotine were selected from cigarette smoke condensates via solvents partitioning and silica gel chromatographic separation, and inhibitory potencies of 19 components on MAO-B were determined. Hydroquinone and methylcatechol, the phenolic components, showed the strongest inhibitory potencies on MAO-B activity in the components tested. 3,4-Dihydroxybenzylamino, myosmine and indole in basic fracton, eugenol in phenolic fraction, and farnesol in neutral fraction also inhibited the enzyme activity dose-dependently. Among tobacco alkaloids tested only myosmine was effective for the inhibition of this enzyme. These results suggest that the decrease in MAO-B activity by such components derived from cigarette smoke seems to be related to the suppression of MPTP-induced neurotoxicity and to the less incidence of Parkinson's disease in smokers than in nonsmokers.

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전통된장으로부터 Angiotensin Converting Enzyme 저해물질의 분리 (Isolation of Angiotensin Converting Enzyme Inhibitor from Doenjang)

  • 김승호;이윤진;권대영
    • 한국식품과학회지
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    • 제31권3호
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    • pp.848-854
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    • 1999
  • 전통된장으로부터 안지오텐신전환효소(angiotensin converting enzyme; ACE)를 저해하는 물질을 추출하여 그 구조를 밝혀냈다. ACE 저해물질을 열수추출한 다음 gel permeation chromatography (GPC)를 통하여 ACE 저해작용이 큰 두 개의 큰 획분을 받았다. 앞획분은 90%와 70%의 ACE 저해효과를 나타내었으나 단일물질로 분리되지 않아 계속하여 역상 HPLC를 통하여 순수 분리를 하였다. 그러나 앞획분은 순수분리되지 않아 결국 2차원 전기영동/TLC를 통하여 분리한 결과 분자량이 759.63인 아미노기를 갖고 있는 비펩타이드 물질임이 밝혀졌다. 뒷획분은 다른 조건의 HPLC(reverse column과 $NH_2$, column)를 이용하여 순수분리에 성공하였다. 이중 ACE 저해효과가 큰 물질은 분자량 271.33인 dipeptide인 arginine-proline임을 밝혀냈다. 이물질의 ACE $IC_{50}$$92\;{\mu}M$이었다. 본 연구 결과는 대부분 ACE 저해물질이 3개 내지 7개 등의 긴 펩타이드임을 감안할 때, 짧은 dipeptide로 ACE 저해펩타이드가 한국의 전통된장에서 생산할 수 있음을 보여주고 있다.

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