• Title/Summary/Keyword: enzyme inhibitors

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Synthesis and Characterization of Fructooligosaccharides Using Levansucrase with a High Concentration of Sucrose

  • Seo Eun-Seong;Lee Jin-Ha;Choi Jae-Young;Seo Mi-Young;Lee Hee-Sun;Chang Seuk-Sang;Lee Hyung-Jong;Choi Jeong-Sik;Kim Doman
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.5
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    • pp.339-344
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    • 2004
  • A method for synthesizing branched fructo-oligosaccharides (BFOS) with a high concentration of sucrose ($1{\~}3$ M) was developed using levansucrase prepared from Leuconortoc mesenteroides B-1355C. The degree of polymerization of oligosaccharides synthesized according to the present method ranged from 2 to over 15. The synthesized BFOS were stable at a pH ranges of 2 to 4 under $120^{\circ}C$. The percentage of BFOS in the reaction digest was $95.7\%$ (excluding monosaccharides; $4.3\%$ was levan). BFOS reduced the insoluble glucan formation by Streptococcus sobrinus on the surfaces of glass vials or stainless steel wires in the presence of sucrose. They also reduced the growth and acid productions of S, sobrinus. Oligosaccharides can be used as sweeteners for foods such as beverages requiring thermo- and acid-stable properties and 3s potential inhibitors of dental caries.

Characterization of Acid Phosphatase from Welsh Onion (파의 Acid Phosphatase의 특성)

  • Kim, Gi-Nahm;Kim, Suk-Ji;Kim, Seok-Hwan;Park, In-Shik
    • Korean Journal of Food Science and Technology
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    • v.28 no.4
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    • pp.663-667
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    • 1996
  • Acid phosphatase (EC 3.1.3.2) from Welsh onion was partially purified by Sephacryl S-200 gel filtration and CM-Sepharose CL-6B ion exchange chromatography. The optimum pH and temperature of acid phosphatase from green onion were pH 5.5 and $60^{\circ}C$, respectively. The enzyme was the most stable at pH 6.0 and unstable above pH 9.0. The activation energy of the enzyme was determined to be 4.86kcal/mole. The enzyme utilized p-nitrophenyl phosphate most as a best substrate among tested possible substrates, while 5'-GMP and 5'-IMP were poor substrates for the enzyme. $K_{m.app.}$ of the enzyme with p-nitrophenyl phosphate as a substrate was identified as 0.87mM. Among metal ions and inhibitors tested, $Cr^{+++},\;Zn^{++},\;Cu^{++}$, molybdate and metavanadate ions inhibited the enzyme reaction drastically.

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A Case of Angioedema Induced by Angiotensin-Converting Enzyme Inhibitor (Angiotensin-Converting Enzyme Inhibitor(ACE Inhibitor)에 의해 유발된 안면부 맥관부종(angioedema) 치험례)

  • Hsia, Yu-Chun;Jung, Ki-Yong;Baik, Jong-Woo;Kim, Dong-Woo;Park, Jong-Hyung;Jun, Chan-Yong;Choi, You-Kyung
    • The Journal of Internal Korean Medicine
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    • v.28 no.2
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    • pp.399-407
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    • 2007
  • Angioedema is a localized transient swelling of sudden onset that can occur in the face, lips, tongue, hand, feet, respiratory system and gastrointestinal system. Angioedema is classified as allergy, hereditary, idiopathic or induced by medication as like aspirin, nonsteroidal anti-inflammatory agents, opiates, antibiotics, and angiotensin-converting enzyme. Angiotensin-converting enzyme inhibitors are widely prescribed for hypertension and heart failure. This drug is commonly associated with angioedema which may be potentially life threatening. We experienced a case of angioedema induced by ACE inhibitor (angiotensin-converting enzyme inhibitor) in a 74-year-old patient who took ACE inhibitor to control hypertension during hospitalization. We thought the angioedema in the face had relation to myenzhong (面腫, mienjong) in oriental medicine. Weiling-tang (Wiryung-tang) was effective for angioedema in the face. As a result the symptoms disappeared rapidly. After 6 days, the patient's symptoms had notably improved.

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Acidophilic Tannase from Marine Aspergillus awamori BTMFW032

  • Beena, P.S.;Soorej, M.B.;Elyas, K.K.;Sarita, G. Bhat;Chandrasekaran, M.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.10
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    • pp.1403-1414
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    • 2010
  • Aspergillus awamori BTMFW032, isolated from sea water, produced tannase as an extracellular enzyme under submerged culture conditions. Enzymes with a specific activity of 2,761.89 IU/mg protein, a final yield of 0.51%, and a purification fold of 6.32 were obtained after purification through to homogeneity, by ultrafiltration and gel filtration. SDS-PAGE analyses, under nonreducing and reducing conditions, yielded a single band of 230 kDa and 37.8 kDa, respectively, indicating the presence of six identical monomers. A pI of 4.4 and a carbohydrate content of 8.02% were observed in the enzyme. The optimal temperature was found to be $30^{\circ}C$, although the enzyme was active in the range of $5-80^{\circ}C$. Two pH optima, pH 2 and pH 8, were recorded, although the enzyme was instable at a pH of 8, but stable at a pH of 2.0 for 24 h. Methylgallate recorded maximal affinity, and $K_m$ and $V_{max}$ were recorded at $1.9{\times}10^{-3}$M and 830 ${\mu}Mol$/min, respectively. The impacts of a number of metal salts, solvents, surfactants, and other typical enzyme inhibitors on tannase activity were determined in order to establish the novel characteristics of the enzyme. The gene encoding tannase, isolated from A. awamori, was found to be 1.232 kb, and nucleic acid sequence analysis revealed an open reading frame consisting of 1,122 bp (374 amino acids) of one stretch in the -1 strand. In silico analyses of gene sequences, and a comparison with reported sequences of other species of Aspergillus, indicate that the acidophilic tannase from marine A. awamori differs from that of other reported species.

Synthesis of Angiotensin Converting Enzyme Inhibitors (안지오텐신 변환효소(變換酵素) 억제(抑制) 작용(作用) 물질(物質)의 합성(合成))

  • Lee, Hee-Joo;Kim, Young-Sook;Chang, Young;Lee, Jong-Ran;YunChoi, Hye-Sook
    • YAKHAK HOEJI
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    • v.28 no.6
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    • pp.313-319
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    • 1984
  • Phenylurea, phenylthiourea, benzylcarbamate and toluenesulfonate of L-alanyl-L-proline(comp. $3{\sim}6$) were synthesized and their effects against angiotensin-converting enzyme (ACE) were tested. Comp. 3 showed only mild inhibitory activities against ACE while comp. $4{\sim}6$ were inert indicating that those functional groups were not suitable for interactions with ACE. Ortho-hydroxy- or ortho-carboxy-benzamide of proline (comp. 7) and phenylalanine (comp. 8 and 9) were also tested. Of the benzamides, ortho-hydroxy function was unsuitable to exert inhibitory activities against ACE. Ortho-carboxy group of 9 seemed to have mild interactions with active site of ACE possibly because of the shorter distance between the amide and ortho-carboxyl group of the compound than the corresponding two active sites of the enzyme.

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Production and Characterization of Antihypertensive Angiotensin I-Converting Enzyme Inhibitor from Pholiota adiposa

  • Koo Kyo-Chul;Lee Dae-Hyoung;Kim Jae-Ho;Yu Hyung-Eun;Park Jeong-Sik;Lee Jong-Soo
    • Journal of Microbiology and Biotechnology
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    • v.16 no.5
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    • pp.757-763
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    • 2006
  • Angiotensin I-converting enzyme (ACE) inhibitors have generally been very useful to remedy or prevent hypertension. This study describes the extraction and characterization of an ACE inhibitor from the fruiting body of Pholiota adiposa ASI 24012, which can be used as an antihypertensive drug. The maximal ACE inhibitory activity $(IC_{50};0.25mg)$ was obtained when the fruiting body of Pholiota adiposa ASI 24012 was extracted with distilled water at $30^{\circ}C$ for 12 h. After the purification of ACE inhibitor with ultrafiltration, Sephadex G-25 column chromatography, and reverse-phase HPLC, an active fraction with an $IC_{50}$ of 0.044 mg was obtained. The purified ACE inhibitory peptide was a novel pentapeptide, showing very little similarity to other ACE inhibitory peptide sequences. The molecular mass of the purified ACE inhibitor was estimated to be 414 daltons with a sequence of Gly-Glu-Gly-Gly-Pro, and showed a clear antihypertensive effect on spontaneously hypertensive rats (SHR) at a dosage of 1 mg/kg.

Inhibition of Various Proteases by MAPI and Inactivation fo MAPI by Trypsin

  • Lee, Hyun-Sook;Kho, Yung-Hee;Lee, Kye-Joon
    • Journal of Microbiology and Biotechnology
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    • v.10 no.2
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    • pp.181-186
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    • 2000
  • MAPI (microbial alkaline protease inhibitor) was isolated from cultrue broth of Streptomyces chromofuscus SMF28. The Ki values of MAPI for the representative serine proteases such as chymotrypsin and proteinase K were 0.28 and $0.63{\;}\mu\textrm{M}$, respectively, and for the cysteine proteases cathepsin B and papain were 0.66 and $0.28{\;}\mu\textrm{M}$, respectively. These data indicate that MAPI is not a potent selective inhibitor of serine or cysteine proteases. Progress curves for the inhibition of three proteases by MAPI exhibithe characteristic patterns; MAPI exhibited slow-binding inhibition of cathepsin B. It was rapidly associated with chymotrypsin before the addition of substrate and then reactivation of MAPI-inhibited enzyme was investigated in the presence of substrate. On the other hand, MAPI-proteinase K interaction was typical for those classical inhibitors. When MAPI was incubated with trypsin, there was an extensive reduction in the ingibitory activities of MAPI corresponding to 66.5% inactivation of MAPI, indicating that trypsin-like protease may play a role in the decrease of the inhibitory activity during cultivation.

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Molecular Docking Studies of Wolbachia Endosymbiont of Brugia Malayi's Carbonic Anhydrase Using Coumarin-chromene Derivatives Towards Designing Anti-filarial Agents

  • Malathy, P.;Jagadeesan, G.;Gunasekaran, K.;Aravindhan, S.
    • Journal of Integrative Natural Science
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    • v.9 no.4
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    • pp.268-274
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    • 2016
  • Filariasis causing nematode Brugia malayi is shown to harbor wolbachia bacteria as symbionts. The sequenced genome of the wolbachia endosymbiont from B.malayi (wBm) offers an unprecedented opportunity to identify new wolbachia drug targets. Hence the enzyme carbonic anhydrase from wolbachia endosymbiont of Brugia malayi (wBm) which is responsible for the reversible interconversion of carbon dioxide and water to bicarbonate and protons (or vice versa) is chosen as the drug target for filariasis. This enzyme is thought to play critical functions in bacteria by involving in various steps of their life cycle which are important for survival, The 3D structure of wBm carbonic anhydrase is predicted by selecting a suitable template using the similarity search tool, BLAST. The BLAST results shows a hexapeptide transferase family protein from Anaplasma phagocytophilum (PDB ID: 3IXC) having 77% similarity and 54% identity with wBm carbonic anhydrase. Hence the above enzyme is chosen as the template and the 3D structure of carbonic anhydrase is predicted by the tool Modeller9v7. Since the three dimensional structure of carbonic anhydrase from wolbachia endosymbiont of Brugia malayi has not yet solved, attempts were made to predict this protein. The predicted structure is validated and also molecular docking studies are carried out with the suitable inhibitors that have been solved experimentally.

Structure-Activity Relationships Study of Angiotensin Converting Enzyme Inhibitor Captopril Derivatives: Importance of Solution Moleculnr Dynamics Study (Angiotensin 변환 효소 억제제인 Captopril 유도체들의 구조와 활성관계 연구: 수용액상의 분자동력학적 연구의 중요성)

  • 지명환;윤창노;진창배;박종세
    • Biomolecules & Therapeutics
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    • v.2 no.1
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    • pp.34-38
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    • 1994
  • In order to investigate the structure-activity relationships of the stereoisomers of angiotensin converting enzyme inhibitors, captopril and its derivatives were selected as model compounds. In vitro enzymatic activities of them depend on the symmetry at the asymmetric carbons. Especially, the alanyl carbon should have the S configuration to be biologically active. But the demethylated captopril having the achiral carbon also shows the activity although it is less active than captopril. Seven stereoisomers of captopril and its derivatives were chosen and their acidic and ionic forms were used for molecular dynamics simulations. Four computer simulations were practiced for each model compound in order to obtain the good condition for simulation to explain the experimental structure-activity relationships. From the computer simulation results, relativistic movements of three well-known pharmacophoric sites, carboxylate carbon, carbonyl oxygen, and sulfur atoms, were analyzed. Good results were obtained from the aqueous solution molecular dynamics simulation with ionic forms of model compounds. Active model compounds have the pharmacophoric areas of 6.08 to 6.38 $\AA$$^2$and the similarity in the geometrical data. But inactive ones have the largely deviated values of 4.51 to 4.87 $\AA$$^2$from those of active ones.

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Purification and characterization of catalase-3 of deinococcus radiophilus

  • Lee, In-Jeong;Lee, young-Nam
    • Journal of Microbiology
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    • v.33 no.3
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    • pp.239-243
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    • 1995
  • Deinococcus radiophilus, an UV resistant bacterium seemed to contain three issoenzymes of catalase. Among them, the samllest and most abundant species in cell-free extract, catalase-3 which also exhibited peroxidase activity was purified to electrophoretic homogeneity (145-fold purification) by chromatographic procedures. Its molecular weight was 155 kDa composed of four 38 kDa subunits. The $K_{m}$ value of catalase-3 for H$\_$2/O$\_$2/ was approximately 0.5 mM. This enzyme showed a typical ferric heme spectrum with maximum absorption at 405 nm. Upon binding to cyanide, the 405 nm peak shifted to 420 nm. Catalase-3 was very sensitive to inhibitors of heme proteins, such as cyanide, azide and hydroxylamine. A ratio of A$\_$405/A$\_$28O/ was 0.5 Catalase-3 was active over a wide range of pH, between pH 7 and 10. The enzyme was rather heat-labile and partially sensitive to edthanol-chloroform treatment, but resistant to 3-amino-1, 2, 4-triazole. Catalase-3 of D. radiophilus, which is a bifunction catalatic peroxidatic enzyme seemed to share certain molecular properties with the typical catalase and the catalase-[roxidase along with its own unique features.

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