• Title/Summary/Keyword: cytochromes

Search Result 24, Processing Time 0.029 seconds

Electron Transport System for Carbon Monoxide Oxidation in Acinetobacter sp.1 (Acinetobacter sp.1의 일산화탄소 산화를 위한 전자전달계)

  • 김영민;조진원
    • Korean Journal of Microbiology
    • /
    • v.24 no.1
    • /
    • pp.46-50
    • /
    • 1986
  • Experiments with particulate fractions of Acinetobacter sp. 1 revealed that coeuzyme $Q_{10}$ is not the physiolohical electron acceptor, and that cytochromes of a, b, c, and o types are found in cells grown with carbon monoxide (CO) as the sole source of carbon and energy. It was found that cytochromes of b and o types, but not the a and c types, are functional in CO oxidation. Nicotinamide adenine dinucleotide (phosphate) is not involved in CO oxidation.

  • PDF

Properties of Electron Carriers in the Process of Methanol Oxidation in a New Restricted Facultative Marine Methylotrophic Bacterium, Methylophaga sp. MP

  • Koh, Moon-Joo;Kim, Chun-Sung;Kim, Yun-A;Choi, Hack-Sun;Cho, Eun-Hee;Kim, Eung-Bin;Kim, Young-Min;Kim, Si-Wouk
    • Journal of Microbiology and Biotechnology
    • /
    • v.12 no.3
    • /
    • pp.476-482
    • /
    • 2002
  • Methanol dehydrogenase (MDH) and c-type cytochromes from marine methanol-oxidizing bacterium, Methylophaga sp. MP, were purified and characterized. The native MDH had a molecular mass of 148 kDa and its isoelectric point was 5.5. Two c-type cytochromes, $c_L\;and\;c_H$, were found, and their isoelectric points were 3.4 and 8.0, respectively. The purified MDH had higher thermal stability than that of the other soil methylotrophic bacteria. The electron flow rate from MDH to cytochrome $c_L$was higher than that from MDH to cytochrome $c_H$, indicating that the physiological primary electron acceptor for MDH is cytochrome $c_L$. The electron transfer from MDH to phenazine ethosulfate (PES, artificial electron acceptor) in the two dye (PES/DCPIP)-linked assay system was not inhibited by NaCl, whereas the electron flow from MDH to cytochrome $c_L$ in the cytochrome/DCPIP-linked assay system was suppressed significantly by NaCl. Metal chelating agents such as EDTA showed the same effects on the MDH activity.

Effect of Copper on the Growth and Methanol Dehydrogenase Activity of Methylobacillus sp. Strain SK1 DSM 8269

  • Kim, Si W.;Kim, Young M.
    • Journal of Microbiology
    • /
    • v.34 no.2
    • /
    • pp.172-178
    • /
    • 1996
  • Methylobacillus sp. strain SK1, which grows only on methanol, was found to grow in the absence of added copper. The doubling time (t$_{d}$ = 1.3 h) of the bacterium growing at the exponential growth phase at 30.deg.C in the absence of copper was the same as that of the cell growing in the presence of copper. The bacterium growing after the exponential phase in the absence of copper, however, grew faster than the cell growing in the presence of copper. Cells harvested after thee arly stationary phase in the presence of copper were found to exhibit no methanol dehydrogenase (MDH) activity, but the amount and subunit structure of the enzyme in the cells were almost the same as that in cells harboring active MDH. Pellets of the cells harvested after the early stationary phase in the presence of copper were pale green. Cell-free extracts prepared from cells harvested at the early stationary phase in the presence of copper were pink and exhibited MDH activity, but it turned dark-green rapidly from the surface under air. The green-colored portions of the extracts showed no MDH activity and contained c-type cytochromes that were oxidized completely. The inactive MDH activity and contained c-type cytochromes that were oxidized completely. The inactive MDH proteins in the green portions were found to have antigenic sites identical to those of the active one as the inactive MDHs in cells grown in the presence of copper. The bacterium was found to accumulate copper actively during the exponential growth phase. MDH prepared from cells grown in the presence or absence of copper was found to be more stable under nitrogen gas than under air. Methanol at 10 mM was found to enhance the stability of the MDH under air.r.

  • PDF

Temperature Effect on the Functional Expression of Human Cytochromes P450 2A6 and 2E1 in Escherichia coli

  • Yim Sung-Kun;Ahn Taeho;Jung Heung-Chae;Pan Jae-Gu;Yun Chul-Ho
    • Archives of Pharmacal Research
    • /
    • v.28 no.4
    • /
    • pp.433-437
    • /
    • 2005
  • Human cytochromes P450 (GYP) 2A6 and 2E1 are of great interest because of their important roles in the oxidation of numerous drugs and carcinogens. Bacterial expression systems, especially Escherichia coli cells, have been widely used for the production of various GYP enzymes in order to obtain high yield of proteins. The expression methods usually employ longer culture time (30-72 h) at lower temperature (usually under $30^{\circ}C$). Expression levels of GYPs 2A6 and 2E1 at $37^{\circ}C$ were compared to those at $28^{\circ}C$, which is a usual temperature used in most bacterial expression systems for human GYP expression. Within 18 h the expression levels of GYPs 2A6 and 2E1 reached up to 360 and 560 nmol per liter culture at $37^{\circ}C$, respectively, which are compatible with those of 36 h culture at $28^{\circ}C$. The activities of GYPs expressed at $37^{\circ}C$ were also comparable to those expressed at $28^{\circ}C$. The present over-expression system can be useful for rapid production of large amounts of active human GYPs 2A6 and 2E1 in E. coli.

Menadione-Modified Anodes for Power Enhancement in Single Chamber Microbial Fuel Cells

  • Ahmed, Jalal;Kim, Sunghyun
    • Bulletin of the Korean Chemical Society
    • /
    • v.34 no.12
    • /
    • pp.3649-3653
    • /
    • 2013
  • As anode fabrication with different materials has been proven to be a successful alternative for enhancing power generation in the microbial fuel cells, a new approach to improved performance of MFCs with the use of menadione/carbon powder composite-modified carbon cloth anode has been explored in this study. Menadione has formal potential to easily accept electrons from the outer membrane cytochromes of electroactive bacteria that can directly interact with the solid surface. Surface bound menadione was able to maintain an electrical wiring with the trans-membrane electron transfer pathways to facilitate extracellular electron transfer to the electrode. In a single chamber air cathode MFC inoculated with aerobic sludge, maximum power density of $1250{\pm}35mWm^{-2}$ was achieved, which was 25% higher than that of an unmodified anode. The observed high power density and improved coulomb efficiency of 61% were ascribed to the efficient electron shuttling via the immobilized menadione.

ROLES OF HUMAN LIVER CYTOCHROMES P450 3A4 AND 1A2 IN THE OXIDATION OF MYRISTICIN

  • Yun, Chul-Ho;Lee, Hye-Suk;Lee, Hee-Yong;Yim, Sung-Kun;Kim, Keon-Hee;Yea, Sung-Su
    • Proceedings of the PSK Conference
    • /
    • 2003.04a
    • /
    • pp.137.1-137.1
    • /
    • 2003
  • Myristicin, 1-allyl-3, 4-methylenedioxy-5-methoxybenzene, is a naturally occurring alkenylbenzene compound. It is found in nutmag, mace, parsley, carrot, black pepper, many natural oils and flavoring agents. The aim of this work was to identify the form(s) of human liver cytochrome P450 (P450) involved in the hepatic transformation of myristicin to its major metabolite, 5-allyl-1-methoxy-2, 3-dihydroxybenzene (M1). (omitted)

  • PDF

Sex-related Differences in Rat Hepatic Cytochromes P450 Expression Following Treatment with Phenobarbital or 3-Methylcholanthrene

  • Lee, Yoon-Sook;Park, Sang-Shin;Kim, Nak-Doo
    • Archives of Pharmacal Research
    • /
    • v.15 no.1
    • /
    • pp.78-86
    • /
    • 1992
  • The induction of hepatic cytochromes P450 and metabolic effects have been examined in male and female Sprague-Dawley rats following treatment with either phenobarbital or 3-methylcholanthrene. Hepatic cytochrome P450 levels were higher in males than in females by ~40%. Treatment of male and female rats with phenobarbital or 3-methylcholanthrene resulted in an ~1.6 and 2-fold increase, respectively, in heptic microsomal cytochrome P450 levels in both sexes, relative to untreated animals. Immunoblot analyses were performed to compare sex-related changes in P450 levels. Hepatic P45IIB1 levels in males were greater than those in females following phenobarbital treatment. 3-Methylcholanthrene-induced male hepatic microsomes exhibited greater levels of P450 females failed to exhibit a band. Mab PCN 2-13-1 against P-45-IIIA recognized an intense in uninduced microsomes from female rats. The levels of P450IIIA in males were increased 2 to 3-fold following treatment with phenobarbital, while the increase of IIIA levels in females by phenobarbital was minimal, as monitored by immunoblot analysis. Solid phase radiommunoassay using monoclonal antibodies supported the results of immunoblot analysis. Phenobarbital treatment caused a 6.5-fold increase in the monoclonal iantibody binding to IIBI in males, whereas treatment of females with phenobarbital resulted of IA levels by 3-methylcholanthrene was also greater in females than in males (10-vs. 8-fold) although the levels of induced IA were comparable inboth sexes, as assessed by radiommunoassay. Radioimmunoassay also showed that hepatic IIEI level was 1.5-fold higher in males than in females and that either phenobarbital or 3-methylcholanthrene treatment caused 80% to 40% decrease in IIEL levels, relative to control, in both sexes. Sex-related metabolic activities were examined in hepatic microsomes. Hexobarbital hydroxylase activity was 2-to 3-fold higher in uninduced microsomes from males than that from females. This hydroxylase activity was increased 2-and 3-fold in males and females, respectively, following phenobarbital treatment, as compared to controls. Addition females produced 64% and 84% inhibition of hexobarbital oxidation, respectively. Aryl hydrocarbon hydroxylase activity was increased -12 and 26-fold in males and females respectively. Following phenobarbital treatment, as compared to controls. Addition of anti-P450IIB1 antibody to phenobarbital-induced hepatic microsomes from males and females produced 64% and 84% inhibition of hexobarbital oxidation, respectively. Aryl hydrocarbon hydroxylase activity was increased -12 and 26 fold in males and females, respectively, following 3-methylcholanthrene treatment relative to controls. The anti-P-450IA antibody inhibitable rate of aryl hydrocarbon hydroxylase activity was comparable in both sexes following 3-methylcholanthrene treatment relative to controls. The anti-P450LA antibody inhibitable rate of aryl hydrocarbon hydroxylase activity was comparable in both sexes following 3-methylcholanthrene treatment (-70%). These results demonstrate that levels of hepatic P450IIB1 or P450IA are greater in male than in female for untreated, phenobarbital-or-3-methylcholanthrene treated rats. In addition, the relative for untreated phenobarbital-or 3-methylcholanthrene treated rats. In addition, the relative increase of phenobarbital-or 3-methylcholanthrene treated rats. In addition, the relative increase of phenobarbital-or 3-methylcholanthrene treated rats. In addition, the relative increase of P450IIB1 or IA phenobarbital or 3-methylcholanthrene is more significant in females.

  • PDF

Effect of Enzymatic Methylation of Proteins on Their Isoelectric Points

  • Park, Kwang-Sook;Frost, Blaise F.;Lee, Hyang-Woo;Kim, Sang-Duk;Paik, Woon-Ki
    • Archives of Pharmacal Research
    • /
    • v.12 no.2
    • /
    • pp.79-87
    • /
    • 1989
  • Enzymatic methylation of arginine and lysine residues of several cytochrome c and lysine residue of calmodulin always resulted in lowering of their respective isoelectric points (pI). Employing cytochromes c derived from various sources, we examined a possible relationship between the degree of amino acid sequence degeneracy and the magnitude of change in the pI values by enzymatic methylation, and found that there was no correlation between these two parameters. By constructing space-filling models of oligopeptide fragments adjacent to the potential methylation sites, we have noted that not all the methylatable residues are able to form hydrogen bonds prior to the methylation. Two preparations of yeast apocytochrome c, one chemically prepared by removing heme from holocytochrome c and the other by translating yeast iso-1-cytochrome c mRNA in vitro, exhibited slightly higher Stokes radii than the homologous holocytochrome c, indicating relatively 'relaxed or open' conformation of the protein. However, when the in vitro synthesized methylated apocytochrome c was compared with the unmethylated counter-part, the Stokes radius of the latter was found to be larger.

  • PDF

Effects of Pretreated Korean Red Ginseng on Carbon Tetrachloride and Galactosamine-induced Hepatotoxicity in Rats (홍삼의 전처리에 의한 사염화탄소 및 갈락토사민 유발 간독성에 대한 보호효과)

  • Lee, Chung-Kyu;Kim, Na-Young;Han, Yong-Nam;Choi, Jong-Won
    • Journal of Ginseng Research
    • /
    • v.27 no.1
    • /
    • pp.1-10
    • /
    • 2003
  • In present study, we examined whether or not the pretreatment of Korean Red Ginseng (KRG) could protect hepatotoxicity induced by carbon tetrachloride (CCl$_4$) and D-galatosamine (GalN). For this study, we not only tested activity of various plasma enzymes (AST, ALT, SDH, LDH), which are used as indicators of liver disease, but also checked the change of liver components such as lipid, glutathione and cytochromes content, and several liver enzyme activity. Pretreatment of KRG for two weeks significantly reduced the elevated plasma enzyme activities induced by CCl$_4$ and GalN. Pretreatment of KRG also restored the hepatic enzymes, malonedialdehyde formation, and depletion of reduced glutathione content induced by CCl$_4$ and GalN to near normal level. However, ${\gamma}$-glutamylcysteine synthetase activity was lot affected by KRG. These results suggest that KRG shows the hepatoprotective effect by reducing lipid peroxidation, by reducing the activity of free radical generating enzymes, and by preserving the hepatic glutathione.

In-silico and In-vitro based studies of Streptomyces peucetius CYP107N3 for oleic acid epoxidation

  • Bhattarai, Saurabh;Niraula, Narayan Prasad;Sohng, Jae Kyung;Oh, Tae-Jin
    • BMB Reports
    • /
    • v.45 no.12
    • /
    • pp.736-741
    • /
    • 2012
  • Certain members of the cytochromes P450 superfamily metabolize polyunsaturated long-chain fatty acids to several classes of oxygenated metabolites. An approach based on in silico analysis predicted that Streptomyces peucetius CYP107N3 might be a fatty acid-metabolizing enzyme, showing high homology with epoxidase enzymes. Homology modeling and docking studies of CYP107N3 showed that oleic acid can fit directly into the active site pocket of the double bond of oleic acid within optimum distance of $4.6{\AA}$ from the Fe. In order to confirm the epoxidation activity proposed by in silico analysis, a gene coding CYP107N3 was expressed in Escherichia coli. The purified CYP107N3 was shown to catalyze $C_9-C_{10}$ epoxidation of oleic acid in vitro to 9,10-epoxy stearic acid confirmed by ESI-MS, HPLC-MS and GC-MS spectral analysis.