• Title/Summary/Keyword: isozymes

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Biodegradation of 2,4,5-Trichlorophenol Using Cell-Free Culture Broths of Phanerochaete chrysosporium

  • Choi, Sueh-Yung;Moon, Seung-Hyeon;Lee, Jae-Suk;Gu, Man-Bock
    • Journal of Microbiology and Biotechnology
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    • v.10 no.6
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    • pp.759-763
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    • 2000
  • Cell-free culture broth of Phanerochaete chrysosporium has been adopted to biologically degrade 2,4,5-trichlorophenol. Two different medium compositions of nitrogen-sufficient and nitrogen-limited were compared for their distribution of isozymes, activity of lignin peroxidase, and production of oxalate. The two different culture broths were tested for their ability to degrade 2,4,5-trichlorophenol, and the biodegradation efficiency was estimated in terms of the disappearance of 2,4,5-trichlorophenol. The degradation efficiency for the nitrogen-limited culture broth was higher than that of the nitrogen-sufficient culture broth, since the nitrogen-limited culture broth induced lignin peroxidases (LiPs) and manganese peroxidases (MnPs), and contained sufficient oxalate for producing necessary radicals. Finally, the possible mechanism of 2,4,5-CP degradation using the nitrogen-limited culture broth was proposed.

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Genetic Diversity and Spatial Structure of Symplocarpus renifolius on Mt. Cheonma, Korea

  • Jeong, Ji-Hee;Park, Yu-Jin;Kim, Zin-Suh
    • Korean Journal of Plant Resources
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    • v.20 no.6
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    • pp.530-539
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    • 2007
  • Genetic variation and structure of 9 subpopulations of Symplocarpus renifolius Schott ex Tzvelev on Mt. Cheonma, in Korea, were determined via starch-gel electrophoresis. The genetic diversity at 10 loci for 8 isozymes ($P_{99}=66%,\;A=2.26,\;H_o=0.212,\;H_e=0.230$) was found to be considerably higher than that seen in other long-lived perennial plants. On the whole, the genotype frequencies were in accordance with Hardy-Weinberg expectations. Approximately 5%($\theta=0.049$) of the total variability was among subpopulations. The high levels of observed genetic diversity in S. renifolius were attributed to a universal outcrossing system and other specific factors like differences in age classes and widely scattered individuals around the main distribution. Heterozygosity was highest at a mid-range of elevation($450m{\sim}600m$). The lowest heterozygosity at lower elevation was attributed to the possible origin of seeds transported by water from upstream regions during the monsoon season. Spatial structure in a subpopulation evidenced a strong autocorrelation between closer individuals within $3{\sim}4m$ of distance. This was assumed to be attributable to the restricted seed dispersal characteristics of S. renifolius. In accordance with the findings generated in this study, some implications regarding the conservation of S. renifolius at the Mt. Cheonma were also presented.

The Beneficial Effect of Trolox on Sepsis-Induced Hepatic Drug Metabolizing Dysfunction

  • Park, Sang-Won;Lee, Sun-Mee
    • Archives of Pharmacal Research
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    • v.27 no.2
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    • pp.232-238
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    • 2004
  • Trolox is a hydrophilic analogue of vitamin E. The aim of this study was to investigate its effects on hepatic injury, especially alteration in cytochrome P450 (CYP)-dependent drug metabolism during polymicrobial sepsis. Rats were subjected to polymicrobial sepsis by cecal ligation and puncture (CLP). The rats were treated intravenously with Trolox (2.5 mg/kg) or vehicle, immediately after CLP. Serum aminotransferases and lipid peroxidation levels were markedly increased 24 h after CLP. This increase was attenuated by Trolox. Total CYP content and NADPH-P450 reductase activity decreased significantly 24 h after CLP. This decrease in CYP content was attenuated by Trolox. At 24 h after CLP, there was a significant decrease in the activity of these CYP isozymes: CYP1A1, 1A2, 2B1, and 2E1. However, Trolox differentially inhibited the decrease in CYP isozyme activity. Trolox had little effect on the decrease in CYP1A1 activity but Trolox significantly attenuated decreases in CYP1A2 and 2E1 activities. In fact, Trolox restored CYP2B1 activity to the level of activity found in control rats. Our findings suggest that Trolox reduces hepatocellular damage as indicated by abnormalities in hepatic drug-metabolizing function during sepsis. Our data also indicates that this protection is, in part, caused by decreased lipid peroxidation.

Substrate and Inhibitor Specificities of Esterase in Lucilia illustris Meigen (연두금파리 Esterase의 기질 및 억제제 특이성에 관한 연구)

  • 유종명;박천배이형철
    • The Korean Journal of Zoology
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    • v.39 no.2
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    • pp.190-197
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    • 1996
  • The optimal conditions and substrate specificity of whole body esterase (EST) activity, effects of inhibitors (Eserine, Paraoxon, p-HMB, DDVP, DFP) on the enzyme, and ontogenv of the isozymes were determined in Lucilio ilfustris Meisen. The optimal temperature was $45^{\circ}C$ regardless of kind of reacted substrate, $\alpha-naphthyl$ acetate $(\alpha-Nal,$ a.naphthvl butylate $(\alpha-N),$ and Pnaphthyl acetate $(\beta-Na),$ but the optimal pH showed some regioselectivitv to naphthvl group of the esters; PH 7.0 for Iform, pH 7.5 for a-form. The maximum reaction rate was recorded at about 2.5 $\times$ 10's M of PNa and etNa, but 1.0 $\times$ 10'S M of $\alpha-Nb.$ Among the five EST inhibitors tested, DDVP was the most powerful. However, distinction of the relative specificity of inhibitors between three body parts, head, thorax, and abdomen, was shouts, representing differences in the distribution and activity of isozvmes. Of 12 carboxyl-esterases (CE), 8 cholinesterases (ChE) and 2 arvlesterases (ArE) identified based on their inhibitor specificity throughout the development, two larval and prepupal stage specific ChEs, no pupal specific, and 2 CEs.2ChEs. and one ArE adult specific isozvmes were confirmed.

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In vitro Metabolism of Pentoxifylline Metabolite M-l in Human Liver Microsomes (인체 간 microsome에서 pentoxifylline 대사체 M-1의 시험관내 대사)

  • 신혜순
    • YAKHAK HOEJI
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    • v.43 no.6
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    • pp.834-842
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    • 1999
  • The metabolism and pharmacokinetics of M-l, which is metabolite of pentoxifylline, have been studied in human liver microsomes. Biphasic kinetics was observed from the Eadie-Hofstee plot for the formation of both metabolites of M-l. For the kinetics of pentoxifylline, mean values of $V_{max1}{\;}and{\;}V_{max2}$ were 1,648 and 5,622 nmol/min/mg protein, and the estimated values of $K_{ml}{\;}and{\;}K_{m2}$ were 0.180 and 4.829 mM, respectively. For M-3, mean values of $V_{max1}{\;}and{\;}V_{max2}$ were 0.062 and 0.491 nmol/min/mg protein, and estimated values of $K_{ml}{\;}and{\;}K_{m2}$ were 0.025 and 1.216 mM. The formations of pentoxifylline and M-3 from M-1 were indentified by using several selective inhibitors of cytochrome P450 isoformes at 0.05-5 mM concentration of M-1 in human liver microsomes. For the analysis of low (0.05 mM) concentration of M-1, where the affinity was expected as low, indicated that CYPlA2 and CYP3A4 were major P450 isoforms responsible for pentoxifylline and M-3 formation. CYP3A4 and CYP2A6 appeared to be P450 isoforms responsible for M-3 formation at high (5 mM) concentration of M-1.

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Rabbit Liver and Lung Microsomal Metabolism of $\beta$-Nicotyrine:Isozyme Specificities toward the Oxidation of $\beta$-Nicotyrine

  • ;Mark K. Shigenaga
    • Environmental Mutagens and Carcinogens
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    • v.9 no.2
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    • pp.87-96
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    • 1989
  • Studies on the biodisposition of beta-nicotyrine by lung and liver microsomes was examined in order to provide a better understanding of its fate in this tissue. beta-nicotyrine (100$\mu$M) was incubated with microsomes (1 mg/ml) prepared from New Zealand White rabbits. The rate of oxidation observed in lung microsomal incubations was 1.7 nmoles $\beta$-nicotyrine oxidized mg$^{-1}$ min$^{-1}$ compared with 2.7 nmoles $\beta$-nicotyrine oxidized mg$^{-1}$ min$^{-1}$ by the liver microsomal preparation. However, when these rates were expressed as a function of cytochrome P-450 content, the specific activity of the metabolic oxidation catalyzed by lung (8.3 nmoles $\beta$-nicotyrine oxidized nmole cytochrome P-450$^{-1}$ min$^{-1}$) was approxiamtely 4 times greater than liver microsomes (2.3 nmoles $\beta$-nicotyrine oxidized nmole cytochrome P-450$^{-1}$ min$^{-1}$). Isozyme studies on the oxidation of $\beta$-nicotyrine employed several methods of altering activities of specific isozymes present in pulmonary microsomes, including the use of the isozyme 2 and 6 specific inhibitor $\alpa$-methyl ABT, metabolic inhibitor(MI) complex formation. The results of this inhibition study would appear to indicate the $\beta$-nicotyrine is metabolized predominantly by pulmonary isozyme 5.

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Relationship of Seed Germination and Lipoxygenase Activity in Soybean

  • Lee, Suk-Ha;Son, Beom-Young;Lee, Yeong-Ho;Lee, Hong-Suk
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.47 no.2
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    • pp.123-126
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    • 2002
  • Lipoxygenase might be associated with seed deterioration by catalyzing the incorporation of molecular oxygen into fatty acids and generating free radicals. This study was performed to determine whether seed lipoxygenase activity would alter soybean seed longevity. In this study, germination percentage of lipoxygenase-lacking cultivar Jinpumkong2 (lx1lx1lx2lx2lx3lx3) was lower than that of Taekwangkong (Lx1Lx1Lx2Lx2Lx3Lx3). Segregation ratio for the three lipoxygenase isozymes of the F2-derived from the cross between Taekwangkong and Jinpumkong2 was fitted to 9 (Lx1Lx2Lx3) : 3 (Lx1Lx2lx3) : 3 (lxllx2Lx3) : 1 (lx1lx2lx3), suggesting the tight linkage between the Lx1 and Lx2 loci. Germination percentages varied widely but not differed among lipoxygenase isozyme types of F$_3$ seeds before and after accelerated aging. Seed coat of Jinpumkong2 was damaged severely following accelerated aging, whereas that of Taekwangkong was not. Thus, seed of lipoxygenase-lacking soybean cultivar, Jinpumkong2 showed greater deterioration compared with that of the normal Taekwangkong. However, the presence or absence of lipoxygenase activity had no effect on soybean germination.

Mechanism of P Solubilization in Vermicompost Treated Red Lateritic Soils

  • Pramanik, Prabhat;Chakraborty, Hritesh;Kim, Pil-Joo
    • 한국환경농학회:학술대회논문집
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    • 2011.07a
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    • pp.188-195
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    • 2011
  • Red lateritic soils are typically low in total organic carbon (TOC) and available phosphorus (AP) content and continuous fertilization is required to obtain desired crop yield. In this experiment, cattle manure in three forms (air-dried, composted and vermicomposted) were applied to red lateritic soil to study their effect on TOC and AP content of soil and probable mechanism of P-solubilization as affected by these treatments were also studied. Vermicompost was the most effective to solubilize insoluble P in red lateritic soil (Alfisols) as compared to other organic amendments (air-dried cattle manure and compost). The highest SPA in vermicompost-treated soil attributed to the comparatively higher concentration of all the three SPA isozymes in these soils. The maximum P-solubilization in these soils might be attributed to the highest SPA and presence of several organic acids like citric, lactic and oxalic acids in vermicompost-treated soils. Since, vermicompost application also increased TOC, mineralizable N and exchangeable K content of soil, vermicompost could be considered as the most rational organic amendment to improve chemical properties of red lateritic soils.

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Substrate Specificity of Cellulase from Aspergillus niger (Aspergillus niger가 생산하는 섬유소 분해효소의 기질에 대한 특이성)

  • Oh, Tae-Kwang;Park, Kwan-Hwa;Shin, Hyun-Kyung;Kim, Ze-Uook
    • Applied Biological Chemistry
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    • v.28 no.3
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    • pp.162-166
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    • 1985
  • Three isozymes of Carboxymethyl Cellulase $(FI^*,\;FII^*,\;FIII)$ and two fractious of ${\beta}-1,4-D-Cellobiohydrolase$(CI, CIl) from Aspergillus niger were purified by Sephadex G-150, DEAE-Sephadex and Sephadex G-75 column chromatography. From the results of enzymatic hydrolysis and X-ray diffraction, ${\beta}-1,4-D-Cellobiohyarolase$ has a high activity toward highly crystalline cellulose such as filter paper and acts synergistically with Cx enzyme.

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Function of NADPH Oxidases in Diabetic Nephropathy and Development of Nox Inhibitors

  • Lee, Sae Rom;An, Eun Jung;Kim, Jaesang;Bae, Yun Soo
    • Biomolecules & Therapeutics
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    • v.28 no.1
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    • pp.25-33
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    • 2020
  • Several recent studies have reported that reactive oxygen species (ROS), superoxide anion and hydrogen peroxide (H2O2), play important roles in various cellular signaling networks. NADPH oxidase (Nox) isozymes have been shown to mediate receptor-mediated ROS generation for physiological signaling processes involved in cell growth, differentiation, apoptosis, and fibrosis. Detectable intracellular levels of ROS can be induced by the electron leakage from mitochondrial respiratory chain as well as by activation of cytochrome p450, glucose oxidase and xanthine oxidase, leading to oxidative stress. The up-regulation and the hyper-activation of NADPH oxidases (Nox) also likely contribute to oxidative stress in pathophysiologic stages. Elevation of the renal ROS level through hyperglycemia-mediated Nox activation results in the oxidative stress which induces a damage to kidney tissues, causing to diabetic nephropathy (DN). Nox inhibitors are currently being developed as the therapeutics of DN. In this review, we summarize Nox-mediated ROS generation and development of Nox inhibitors for therapeutics of DN treatment.