• Title/Summary/Keyword: cofactor

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Ferric Reductase Activity of the ArsH Protein from Acidithiobacillus ferrooxidans

  • Mo, Hongyu;Chen, Qian;Du, Juan;Tang, Lin;Qin, Fang;Miao, Bo;Wu, Xueling;Zeng, Jia
    • Journal of Microbiology and Biotechnology
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    • v.21 no.5
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    • pp.464-469
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    • 2011
  • The arsH gene is one of the arsenic resistance system in bacteria and eukaryotes. The ArsH protein was annotated as a NADPH-dependent flavin mononucleotide (FMN) reductase with unknown biological function. Here we report for the first time that the ArsH protein showed high ferric reductase activity. Glu104 was an essential residue for maintaining the stability of the FMN cofactor. The ArsH protein may perform an important role for cytosolic ferric iron assimilation in vivo.

Screening of New Antibiotics Inhibiting Bacterial Enoyl-Acyl Carrier Protein Reductase (Fabl) (세균의 지방산 생합성 효소 (Enoyl-Acyl Carrier Protein Reductase, FabI)를 저해하는 새로운 항균물질의 스크리닝)

  • 곽진환
    • YAKHAK HOEJI
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    • v.46 no.1
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    • pp.24-29
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    • 2002
  • Enoyl-Acyl Carrier Protein Reductase (Fabl) of bacteria is hem as an important target for new antibacterial drugs and plays a determinant role in completing cycles of elongation in type-H fatty acid synthase system. In this study, a fabI gene from Staphylococcus aureus 6538p cloned in pET-l4b vector and FabI protein was over-produced in Escherichaia coli BL2l (DE3). $NH_2$-terminal His-tagged FabI protein was purified by nickel-nitrilotriacetic acid (Ni-NTA) metalaffinity chromatography Purified 6xHis-tagged FabI showed a catalytic activity on tram - 2 - octenoyl - N -acethlcysteamine by utilizing NADPH as a cofactor. For the discovery of new FabI inhibitors from chemical libraries, a target-oriented screening system using a 96-well plate was developed. About 10,000 chemical libraries from Korea Chemical Bank wore tested in this screening system, and 26 chemicals (0.25%) among them showed an inhibitory activity against FabI enzyme. This result showed that a new screening system can be used for the discovery of new FabI inhibitors.

DA 11004, a synthetic IDPc inhibitor, inhibits the high fat high sucrose diet-induced obesity in C57BL/6 mice.

  • Jeong, Mi-Young;Shin, Chang-Yell;Sohn, Jin-Bup;Lee, In-Ki;Son, Mi-Won;Bae, Cheol-Jun;Byun, Jong-Soo;Kim, Dong-Sung;Kim, Soon-Hae
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.129.2-130
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    • 2003
  • Recently. it has been known that NADPH-dependent isocitrate dehydrogenase (IDPc) involves in the obesity through production of NADPH, an important cofactor. DA-11004 is a synthetic potent IDPc inhibitor that $IC_{50}$ for IDPc is 1.49$\mu\textrm{M}$ (0.9$\mu$g/ml). The purpose of this study was to evaluate the effects of DA-11004 on the high fat high sucrose (HF)-induced obesity in C57BL/6 mice. (omitted)

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Site-Directed Mutagenesis of Ile91 of Restriction Endonuclease EcoRV: Dramatic Consequences on the Activity and the Properties of the Enzyme

  • Moon, Byung-Jo;Vipond, I. Barry;Halford, Stephen E.
    • BMB Reports
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    • v.29 no.1
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    • pp.17-21
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    • 1996
  • Ile91 of restriction endonuclease EcoRV, which has not been known to take part directly in catalytic activity, was substituted with Leu by site-directed mutagenesis. The Ile91Leu mutant shows over 1000-fold less activity than the wild type EcoRV under standard reaction condition. The metal ion dependency of the reaction was altered. In contrast to the wild type EcoRV, the mutant prefers $Mn^{2+}$ to $Mn^{2+}$ as the cofactor. In $Mn^{2+}$ buffer the mutant is as active as the wild type enzyme in $Mn^{2+}$ buffer. Like the wild type enzyme, the mutant shows an unspecific binding of DNA in gel shift experiments. In contrast to the wild type enzyme, the mutant did not cleave at noncognate sites of DNA under star condition.

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Toluene Monooxygenase의 Peroxide shuntting에 의한 TCE와 PCE 분해 특성

  • 류두현;김형수;최용욱;김용미;이경애;유재수;조현
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2004.09a
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    • pp.277-280
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    • 2004
  • TCE and PCE, suspected carcinogens, are the most common groundwater pollutant from extensive use as a solvent and degreaser. Escherichia coli TGI pBSKAN TOM Green and E. coli TGI pBSKAN ToMO, which were used DNA shuffling technique, produce Toluene-o-monooxygenase(TOM) and toluene-o-xylene- monooxygenase(ToMO). These cells and enzymes are degrading TCE and PCE, TOM and ToMO are needed to cofactor, such as NADH, NADPH and other cofactors. Used TCE and PCE degrading microorganisms experiment the contaminated material removal efficiency. A shunting test used NAD and Hydrogen peroxide.

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Activities of Sulfhydryl-Related and Phenylpropanoid-Synthesizing Enzymes during Leaf Development of Arabidopsis thaliana

  • Sa, Jae-Hoon;Park, Eun-Hee;Lim, Chang-Jin
    • BMB Reports
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    • v.31 no.6
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    • pp.554-559
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    • 1998
  • Activities of glutathione- and thioredoxin-related enzymes and phenylpropanoid-synthesizing enzymes were measured and compared in the developing leaves of Arabidopsis thaliana. Phenylalanine ammonia-lyase activity is maximal in the leaves of 2-wk-grown Arabidopsis. Tyrosine ammonia-lyase activity is maximal in the leaves of 3-wk-grown and 4-wk-grown Arabidopsis. Activity of thioitransferase, an enzyme involved in the reduction of various disulfide compounds, is higher in younger leaves than in older ones. A similar pattern was obtained in the activity of thioredoxin, a small protein known as a cofactor of ribonucleotide reductase and a regulator of photosynthesis. Activity of glutathione reductase is also higher in the younger leaves. Malate debydrogenase activity remains relatively constant during the development of Arabidopsis leaves. The results offer preliminary information for further approach to elucidate the mechanism of growth-dependent variations of these enzymes.

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Maintenance of cellular tetrahydrobiopterin homeostasis

  • Kim, Hye-Lim;Park, Young-Shik
    • BMB Reports
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    • v.43 no.9
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    • pp.584-592
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    • 2010
  • Tetrahydrobiopterin (BH4) is a multifunctional cofactor of aromatic amino acid hydroxylases and nitric oxide synthase (NOS) as well as an intracellular antioxidant in animals. Through regulation of NOS activity BH4 plays a pivotal role not only in a variety of normal cellular functions but also in the pathogenesis of cardiovascular and neurodegenerative diseases, which develop under oxidative stress conditions. It appears that a balanced interplay between BH4 and NOS is crucial for cellular fate. If cellular BH4 homeostasis maintained by BH4 synthesis and regeneration fails to cope with increased oxidative stress, NOS is uncoupled to generate superoxide rather than NO and, in turn, exacerbates impaired BH4 homeostasis, thereby leading to cell death. The fundamental biochemical events involved in the BH4-NOS interplay are essentially the same, as revealed in mammalian endothelial, cardiac, and neuronal cells. This review summarizes information on the cellular BH4 homeostasis in mammals, focusing on its regulation under normal and oxidative stress conditions.

Alterations in dopamine and glutamate neurotransmission in tetrahydrobiopterin deficient spr-/- mice: relevance to schizophrenia

  • Choi, Yong-Kee;Tarazi, Frank I.
    • BMB Reports
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    • v.43 no.9
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    • pp.593-598
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    • 2010
  • Tetrahydrobiopterin ($BH_4$) is a pivotal cofactor for enzymes responsible for the synthesis and release of monoamine neurotransmitters including dopamine and serotonin as well as the release of glutamate. Deficiencies in $BH_4$ levels and reduced activities of $BH_4$-associated enzymes have been recently reported in patients with schizophrenia. Accordingly, it is possible that abnormalities in the biochemical cascades regulated by $BH_4$ may alter DA, 5-HT and Glu neurotransmission, and consequently contribute to the pathophysiology of different neuropsychiatric diseases including schizophrenia. The development of a novel strain of mutant mice that is deficient in $BH_4$ by knocking out the expression of a functional sepiapterin reductase gene (spr -/-) has added new insights into the potential role of $BH_4$ in the pathophysiology and improved treatment of schizophrenia.

Simultaneous Utilization of Two Different Pathways in Degradation of 2,4,6-Trinitrotoluene by White Rot Fungus Irpex lacteus

  • 김현영;송홍규
    • Korean Journal of Microbiology
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    • v.38 no.4
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    • pp.250-250
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    • 2002
  • This study confirmed that white rot fungus Irpex lacteus was able to metabolize 2,4,6-trinitrotoluene (TNT) with two different initial transformations. In one metabolic pathway of TNT a nitro group was removed from the aromatic ring of TNT. Hydride-Meisenheimer complexes of TNT (H/sup -/-TNT), colored dark redo were confirmed as the intermediate in this transformation by comparison with the synthetic compounds. 2,4-Dinitrotoluene as a following metabolic product was detected, and nitrite produced by denitration of $H^-$-TNT supported this transformation. In the other TNT pathway, nitro groups in TNT were successively reduced to amino groups via hydroxylamines. Hydroxylamino-dinitrotoluenes and amino-dinitrotoluenes were identified as the intermediates. The activity of a membrane-associated aromatic nitroreductase was detected in the cell-free extract of I. lacteus. This enzyme catalyzed the nitro group reduction of TNT with NADPH as a cofactor, Enzyme activity was not observed in the presence of molecular oxygen.

Changes in Serum Insulin-Like Growth Factor Binding Protein-5 of Osteopenia (골감소증 환자의 혈청중 Insulin-Like Growth Factor Binding Protein-5의 변화)

  • Kim, Young;Nam, Taek-Jeong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.3
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    • pp.493-499
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    • 2000
  • 골감소증 환자의 혈청중에 존재하는 IGFBP-5의 존재와 변화를 검토한 결과, 골감소즈 환자에서는 정상 대조군에 비해 IGFBP-5가 감소하였는데 이 변화는 IGFBP-5의 분해효소가 작용하기 때문인 것으로 나타났다. IGFBP-5에 작용하는 단백질분해효소 저해제중 metallo계인 EDTA 및 1,10-phenanthroline과 seriner계인 aprotinin, heparin, heparin cofactor 2(HC2), heparine+HC2가 IGFBP-5에 대해 저해효과가 크므로 metalloprotease이면서 serine protease의 성질을 가지는 효소들이 IGFBP-5에 작용하였다. IGF-I과 IGF-II 그리고 insulin은 효소 활성에 아무런 영향이 없었다. IGFBP-5의 zymography에서 정상인과 골감소증 환제어서 180 kDa 크기의 band가 나타났고, gelatin zymography에서 정상 대조군의 경우 66 kDa과 97 kDa 정도의 band가 확인되었고 골감소증 환자의 경우는 69 kDa의 band가 확인되었다.

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