• 제목/요약/키워드: Hemoproteins

검색결과 11건 처리시간 0.025초

사람의 간에서 Ethanol에 의해 유발되는 hemoprotein들의 확인 및 부분정제 (Identification and Partial Purification of Ethanol-Induced Hemoproteins in Human Liver)

  • 박성우;서배석;진광호
    • 분석과학
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    • 제8권2호
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    • pp.117-124
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    • 1995
  • 지속적인 음주로 인하여 간경변증으로 사망한 human 간에서 ethanol에 의해서 유도되는 것으로 추측되는 hemoprotein들을 확인 및 부분정제하였다. 이 hemoprotein을 정제하기 위하여 Mohamed 등의 방법을 변형하여 단백질을 정제하였고, SDS-PAGE 및 spectrum 양상을 관찰하였다. Triton N-101을 처리한 crude extract를 준비하여 CO gas를 bubbling시킨 후 Octyl-Sepharose CL-4B column chromatography에서 0.06% Lubrol PX로 용출한 다음 0.25% Lubrol PX로 용출하였다(Fig. 2). 0.06% Lubrol PX로 용출한 active fraction을 Hydroxyapatite와 DEAE-Sephadex A-25 column으로 정제하였다(Fig. 3, 4). 정제한 단백질을 12.5% SDS-PAGE를 실시한 결과 분자량은 대조군으로 사용한 흰쥐 간에서 정제한 단백질의 분자량은 55 KDa와 52 KDa였고, 돌연사한 사람의 간에서 정제한 단백질의 분자량은 62 48KDa이며, 간경변증으로 사망한 사람의 간에서 정제한 단백질의 분자량은 54KDa였고(Fig. 5). Cytochrome P450 함량은 20.8nmol/mg protein이며 회수율은 약 4.1%이고, 이들의 최대흡수 파장은 446nm이었다(Fig. 6).

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경유의 Model solution에서 고정화효소를 이용한 Dibezothiophene의 산화

  • 허정찬;성현태;류근갑
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2001년도 추계학술발표대회
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    • pp.517-520
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    • 2001
  • Fossil fuels such as coal and crude oil contain various organic sulfur compounds. Combustion of these fuels emit sulfur oxides which are considered as msjor air pollutants causing acid rain problem. Among various organic sulfur compounds, aromatic sulfur compounds of thiophenes which constitute major sulfur fractions of heavy oils are not easily removed by hydrodesulfurization. Many peroxidase and hemoproteins are known to oxidize dibenzothiophene (DBT) to dibenzothiophene-sulfoxide(DBT - sulfoxide) then dibenzothiophene- sulfone (DBT-sulfone). The oxidation of DBT by the immobilized hemoproteins in n-octane was increased significantly when the hemoproteins were deposited on celites of the particle size between 0.75 - 1.0 mm and a conventional substrates. such as t-butyl hydroperoxide and cumene hydroperoxide. In anhydrous organic solvents with log P values larger than 4.0 DBT was completely oxidized by cumene hydroperoxide catalyzed by cytochrome c deposited on celites.

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Lightening up Light Therapy: Activation of Retrograde Signaling Pathway by Photobiomodulation

  • Kim, Hong Pyo
    • Biomolecules & Therapeutics
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    • 제22권6호
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    • pp.491-496
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    • 2014
  • Photobiomodulation utilizes monochromatic (or quasimonochromatic) light in the electromagnetic region of 600~1000 nm for the treatment of soft tissues in a nondestructive and nonthermal mode. It is conceivable that photobiomodulation is based upon the ability of the light to alter cell metabolism as it is absorbed by general hemoproteins and cytochrome c oxidase (COX) in particular. Recently it has been suggested radiation of visible and infrared (IR) activates retrograde signaling pathway from mitochondria to nucleus. In this review, the role of COX in the photobiomodulation will be discussed. Further a possible role of water as a photoreceptor will be suggested.

Heme 단백질의 Model로서의 Hemin 착물에 관한 $^1H$ NMR 연구 ($^1H$ NMR Study of mono-and di-cyanide ligated Hemin Complexes as Models of Hemoproteins)

  • 이강봉;김남준;권지혜;이재성;최영상
    • 분석과학
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    • 제7권4호
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    • pp.505-515
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    • 1994
  • DMSO(dimethylsuloxide-$d_6$) 용액 속에 존재하는 CN/CN 리간드의 hemin 착물과 CN/DMSO의 hemin 착물이 $^1H$ NMR로 기록되어지고 분석되어졌다. Hemin으로의 CN 착물화 과정은 온도에 따라 변화함을 NMR 스펙트럼이 보여 주며, 한 개의 CN 리간드에서 두 개의 CN 리간드착물로 바뀌는 과정의 열역학함수는 ${\Delta}H^{\circ}=736.6cal/mol$${\Delta}S^{\circ}=16.4eu$인 흡열과정을 나타낸다. CN/DMSO의 hemin 착물은 Curie behavior로부터의 벗어남은 high-spin 성격의 존재를 나태내고, 이는 Fe-DMSO 결합이 순간적으로 깨짐을 의미하며, 이러한 CN/DMSO hemin 착물이 한 개의 axial ligand가 약한 heme 단백질의 전자 및 분자구조의 model complex로 작용할 수 있음을 보여 준다.

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Overexpression of Shinorhizobium meliloti Hemoprotein in Streptomyces lividans to Enhance Secondary Metabolite Production

  • Kim, Yoon-Jung;Sa, Soon-Ok;Chang, Yong-Keun;Hong, Soon-Kwang;Hong, Young-Soo
    • Journal of Microbiology and Biotechnology
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    • 제17권12호
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    • pp.2066-2070
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    • 2007
  • It was found that Shinorhizobium meliloti hemoprotein (SM) was more effective than Vitreoscilla hemoglobin (Vhb) in promoting secondary metabolites production when overexpressed in Streptomyces lividans TK24. The transformant with sm (sm-transformant) produced 2.7-times and 3-times larger amounts of actinorhodin than the vhb-transformant in solid culture and flask culture, respectively. In both solid and flask cultures, a larger amount of undecylprodigiocin was produced by the sm-transformant. It is considered that the overexpression of SM especially has activated the pentose phosphate pathway through oxidative stress, as evidenced by an increased NADPH production observed, and that it has promoted secondary metabolites biosynthesis.

Oxidation of Dibenzothiophene Catalyzed by Surfactant-Hemoprotein Complexes in Anhydrous Nonpolar Organic Solvents

  • Ryu, Keun-Garp;Chae, Young-Rae;Kwon, O-Yul
    • Journal of Microbiology and Biotechnology
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    • 제13권5호
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    • pp.647-650
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    • 2003
  • In anhydrous organic solvents, the complexes formed between AOT (dodecylbenzene sulfuric acid sodium salt) and hemoproteins, such as hemoglobin, myoglobin, or cytochrome c, displayed remarkably higher activity than the hemoprotein powders to oxidize dibenzothiophene, a model compound of organic sulfurs contained in fossil fuels. In slightly hydrophobic organic solvents, such as ethyl acetate and butyl acetate, dibenzothiophene was completely oxidized catalytically by the cytochrome c-AOT complex with cumene hydroperoxide (${\alpha},{\alpha}-dimethylbenzyl$ hydroperoxide) as an oxidant. In highly hydrophobic organic solvents, such as decane and hexadecane, however, the activity of the cytochrome c-AOT complex decreased, presumably due to the aggregation of the hemoprotein-AOT complex in these solvents.

Bacillus subtilis HmoB is a heme oxygenase with a novel structure

  • Park, Seong-Hun;Choi, Sa-Rah;Choe, Jung-Woo
    • BMB Reports
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    • 제45권4호
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    • pp.239-241
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    • 2012
  • Iron availability is limited in the environment and most bacteria have developed a system to acquire iron from host hemoproteins. Heme oxygenase plays an important role by degrading heme group and releasing the essential nutrient iron. The structure of Bacillus subtilis HmoB was determined to 2.0 ${\AA}$ resolution. B. subtilis HmoB contains a typical antibiotic biosynthesis monooxygenase (ABM) domain that spans from 71 to 146 residues and belongs to the IsdG family heme oxygenases. Comparison of HmoB and IsdG family proteins showed that the C-terminal region of HmoB has similar sequence and structure to IsdG family proteins and contains conserved critical residues for heme degradation. However, HmoB is distinct from other IsdG family proteins in that HmoB is about 60 amino acids longer in the N-terminus and does not form a dimer whereas previously studied IsdG family heme oxygenases form functional homodimers. Interestingly, the structure of monomeric HmoB resembles the dimeric structure of IsdG family proteins. Hence, B. subtilis HmoB is a heme oxygenase with a novel structural feature.

중금속류가 취절편의 Amylase 분비에 미치는 영향 (Effect of Heavy Metals on the Secretion of Amylase in Rat Pancreatic Fragments)

  • 김혜영;김원준
    • 대한약리학회지
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    • 제17권2호
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    • pp.31-36
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    • 1981
  • Heavy metals which are present as trace elements in human body have been known to modify various enzymatic reaction. These metals can be essential or non-essential. Zinc, copper and calcium are essential in maintaining some biological processes, whereas non-essential metals such as cadmium, lead and mercury produce accumulatve toxic effect. Cadmium accumulated in pancreas can cause toxicity and damage of pancreatic cells, thereby influencing CHO metabolism. Lead compounds are known to produce toxic effects on the kidney, digestive system and brain fellowed by inhibition of activity of ${\rho}-aminolevulinic$ acid and biosynthesis of hemoproteins and cytochrome. Evidence has been accumulated that zinc not only acts as a cofactor in enzyme reaction but also prevents toxic effect induced by heavy metal such as copper and cadmium. To demonstrate the effect of heavy metals on pancreatic secretion, part of uncinate pancreas was taken and incubated in Krebs-Ringer bicarbonate buffer with heavy metals used. Additional treatment with CCK-OP was performed when needed. After incubation during different period of time, medium was analyzed for amylase activity using Bernfeld's method. The present study was attempted in order to elucidate the effect of several kinds of heavy metal on exocrine pancreatic secretion in vitro. The results obtained are as follows: 1) CCK-OP stimulated significantly amylase release from pancreatic fragments in vitro. 2) CCK-OP response of amylase release from pancreatic fragments was inhibited by treatmant with cadmium, especially high doses of cadmium. 3) CCK-OP response of amylase release from pancreatic fragments was inhibited when pretreated with $10^{-4}M$ copper chloride. 4) Lead chloride at the concentration of $10^{-3}M\;and\;10^{4}M$ stimulated the basal amylase release in vitro but CCK-OP response did not augment by lead chloride. 5) Zine chloride did not affect amylase release from pancreatic fragment in vitro. From the results mentioned above, it is suggested that CCK-OP response was inhibited it the amylase release from pancreatic fragments pretreated with cadmium and copper chloride.

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정어리표피중에 존재하는 지방산화촉진물질의 검색확인 및 그 특성구명 (Characterization of Lipid Prooxidants in Sardine Skin)

  • 조순영;모리사토시;엔도야스시;후지모토겐시로
    • 한국수산과학회지
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    • 제25권6호
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    • pp.501-510
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    • 1992
  • 정어리표피 중에 존재하는 지방산화촉진물질의 검색확인 및 그 특성구명을 시도하였다 정어리표피중의 산화촉진물질은 0.05M 인산염 완충액으로 거의 모두 추출되었고, 이 조추출액에 대해 Sephadex G-200, DEAE-Sephadex A-50 및 CM-Sephadex A-50 등으로 연속적인 분리를 행하였다. 정어리표피 중의 주 산화촉진물질은 열에 극히 불안정하고, 지방산화촉진의 최적 작용 pH가 7이며, 특징적인 Sohret흡수를 나타내는 점등으로부터 햄을 가진 효소임을 알 쑤 있었다. 특히 이들은 기질인 유리 및 에스테르화 리놀산에 모두 잘 작용하여 빠른 속도로 공역과산화물을 형성하였다. 이상의 결과로부터 정어리표피 중의 주 지방산화촉진물의 본태는 새로운 형태의 리폭시지네이스유사 효소인 것으로 추정되었다.

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Sebekia benihana에서 Streptomyces coelicolor SCO4967 유전자 도입을 통한 하이드록실 사이클로스포린 A의 생전환 (Improvement of Cyclosporin A Hydroxylation in Sebekia benihana by Conjugational Transfer of Streptomyces coelicolor SCO4967, a Secondary Metabolite Regulatory Gene)

  • 김현범;이미진;한규범;김응수
    • 한국미생물·생명공학회지
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    • 제38권4호
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    • pp.475-480
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    • 2010
  • Actinomycetes are Gram-positive soil bacteria and one of the most important industrial microorganisms due to superior biosynthetic capabilities of many valuable secondary metabolites as well as production of various valuable bioconversion enzymes. Among them are cytochrome P450 hydroxylase (CYP), which are hemoproteins encoded by a super family of genes, are universally distributed in most of the organisms from all biological kingdoms. Actinomycetes are a rich source of soluble CYP enzymes, which play critical roles in the bioactivation and detoxification of a wide variety of metabolite biosynthesis and xenobiotic transformation. Cyclosporin A (CyA), one of the most commonly-prescribed immunosuppressive drugs, was previously reported to be hydroxylated at the position of 4th N-methyl leucine by a rare actinomycetes called Sebekia benihana, leading to display different biological activity spectrum such as loss of immunosuppressive activities yet retaining hair growth-stimulating side effect. In order to improve this regio-selective CyA hydroxylation in S. benihana, previously-identified several secondary metabolite up-regulatory genes from Streptomyces coelicolor and S. avermitilis were heterologously overexpressed in S. benihana using an $ermE^*$ promoter-containing Streptomyces integrative expression vector. Among tested, SCO4967 encoding a conserved hypothetical protein significantly stimulated region-specific CyA hydroxylation in S. benihana, implying that some common regulatory systems functioning in both biosynthesis and bioconversion of secondary metabolite might be present in different actinomycetes species.