• Title/Summary/Keyword: monooxygenase

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Effects of Tributyltin in vitro on Hepatic Monooxygenase System in Marine Fishes (유기주석화합물이 해산 어류의 간장 MFO 효소계에 미치는 영향)

  • 전중균;이미희;이지선;심원준;이수형;허형택
    • Korean Journal of Environmental Biology
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    • v.21 no.1
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    • pp.18-25
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    • 2003
  • Effects of tributyltin chloride (TBTC) in vitro on mixed function oxygenase (MFO) system on liver microsome of eight marine fish species were investigated. To determine the effects on MFO system, cytochrome P45O (CYP) and cytochrome b5 con-tents, activities of two reductases (NADH-cytochrome b5 reductase and NADPH-cy-tochrome P450 reductase) and four dealkylation enzymes (EROD, PROD, MROD and ECOD) were measured in fish microsoms exposed to TBTC for 20 min. The WP content was reduced to 10% of the control group in 6 out of 8 species exposed to TBTC, whereas there was no significant change in the cytochrome bs content. the response of NAD(P)H dependant reductases depended on fish species. The dealkylation enzyme activities in microsome were also apparently inhibited by TBTC. The degree of inhibition was different among fish species and four enzymes. The EROD activities in eight species were decreased to the range of 1∼65% of control group.

Mechanisms of Parathion Resistance in a Ethyl Fenitrothion-Selected Yumenoshima III Strain of House Flies (페니트로치온 도태 Yumenoshima 저항성 집파리에 있어서의 파라치온 저항성 메카니즘)

  • ;;Toshio SHONO
    • Korean journal of applied entomology
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    • v.35 no.3
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    • pp.254-259
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    • 1996
  • The biochemical factors responsible for parathion resistance in a ethyl fenitrothion-selected Yumenoshima I11 (EF-30) strain of the housefly were examined. Great difference (167-fold) in the Iso was observed between the resistant EF-30 (R) and susceptible SRS (S) strains in vitro, suggesting that altered acetylcholinesterase (AChE) in the housefly strain was an important factor in the resistance. The in vitro degradative activity of parathion and paraoxon in both strains was associated with the microsomal and soluble fractions and required NADPH and reduced glutahione (GSH), respectively. The R strain possessed higher activity for GSH S-transferase than the S strain, and this enzyme appears to be important in the resistance mechanism. The R strain was highly resistant to parathion (101,487-fold), but substitution of the methoxy group for ethoxy group decreased the resistance level (25,914-fold) and parathion could be a substrate of GSH S-transferase. It is concluded that the combination of some factors (altered AChE, and enhanced activity of cytochrome P450 dependent monooxygenase and GSH S-transferase) could be sufficient to account for the extremely high level of resistance to parathion and parathion-methyl, although a possible involvement of other factor(s) can not be excluded.

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Characterization of ent-Kaurenoic Acid 13-Hydroxylase in Steviol Biosynthesis of Stevia rebaudiana Bertoni (Stevia rebaudiana Bertoni의 Steviol 생합성 효소 ent-Kaurenoic Acid 13-Hydroxylase의 특성)

  • Shibata, Hitoshi;Kim, Keun-Ki
    • Applied Biological Chemistry
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    • v.40 no.6
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    • pp.501-507
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    • 1997
  • Chloroplasts isolated from Stevia rebaudiana Bertoni leaves contained an enzyme activity which catalyzed hydroxylation of ent-kaurenoic acid (ent-kaur-16-en-19-oic acid; ent-KA) to steviol (ent-13-hydroxy kaur-16-en-19-oic acid), the diterpenoid carboxylic alcohol which is the aglycone of sweet stevioside-related glycosides. $[^(14)C]-methylated$ ent-KA was used to localize ent-KA hydroxylase. $[^(14)C]-methyl-KA$ was most actively was transformed into methyl-steviol in chloroplast. The enzymatic activity was found in stroma fraction but not in thylakoid membrane in Stevia rebaudiana Bertoni. However, ent-KA 13-hydroxylase activity was not detected in stroma fraction of either Spinacia oleracea and Solidago altissima. The reaction products using $[^(14)C]-methyl-KA$ were purified and identified on TLC autoradiogram. The hydroxylation of ent-KA from stromal protein to form steviol required NADPH and oxygen. FAD and riboflavin stimulated the enzyme activity 1.5-and 1.7-fold, respectively. It also turned out that the activity of this enzyme using methyl-KA as a substrate was 16.7% that of ent-KA. The purified ent-KA 13-hydroxylase did not act on t-cinnamic acid, 4-hydroxyphenyl acetic acid, choline and resorcinol, known as monooxygenase and hydroxylase substrates.

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Responses in Hepatic Xenobiotic Metabolizing Enzymes and Sex Hormones of Yellowfin Goby Acanthogobius flavimanus in Nakdong Estuary (낙동강 하구에서 채집한 문절망둑 Acanthogobius flavimanus의 간장 약물대사효소계와 성호르몬 농도)

  • Lee, Ji-Seon;Jeong, Jee-Hyun;Han, Chang-Hee;Shim, Won-Joon;Jeon, Joong-Kyun
    • Korean Journal of Environmental Biology
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    • v.26 no.2
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    • pp.87-93
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    • 2008
  • To assess effects of contaminants on fish in Nakdong river, feral yellowfin goby Acanthogobius flavimanus were caugt in two different sites and its hepatic monooxygenase enzyme, including cytochrome P450 (CYP), NADPH-cytochrome P450 reductase (P450R), NADH-cytochrome b5 reductase (b5R), ethoxyresorufin deethylase (EROD), glutathione S-transferase (GST) were quantitatively determined. Gonadosomatic index (GSI), hepatosomatic index (HSI) and three sex steroid hormone (17$\beta$-estradiol, E2; testosterone, T; 11-ketotestosterone, 11-KT) levels of the fish were also investigated. HSI of fish from polluted site (site 1) were significantly higher than that of unpolluted site (site 2), but GSI levels were significantly lower in polluted site. No significant differences in plasma 11-KT and T levels were observed in two sites surveyed. E2 level was, however, significantly (p<0.05) higher in female fish from site 1 than site 2. In addition, hepatic EROD activity and CYP level of site 1 fish were lower than those of site 2 fish, whereas relatively high levels of P450R, b5R and GST activities were found in site 1. The results imply that yellowfin goby, especially female fish in Nakdong river estuary are affected from contaminants disrupting sex steroid hormone system.

Characteristic study and isolation of Bacillus subtilis SRCM 101269 for application of cow manure (우분 적용을 위한 Bacillus subtilis SRCM 101269의 분리 및 특성 연구)

  • Jeon, SaeBom;Oh, HyeonHwa;Uhm, Tai-Boong;Cho, Jae-Young;Yang, Hee-Jong;Jeong, Do-Youn
    • Korean Journal of Microbiology
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    • v.52 no.1
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    • pp.74-83
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    • 2016
  • Bacillus subtilis SRCM 101269 having safety and amo gene isolated from Korean traditional fermented food and their investigated characterization to apply the cow manure such as cellulase and xylanase activities, 16S rRNA sequencing, and ability of removal of livestock manure odor. Cow manure application results for the removal of livestock manure odor, the ammonia gas was reduced more than two-folder compared to the control group after 6 days, and reduced to less than 10 ppm after 9 days. In the case of cow manure added fowl droppings and other wood-based mixture components, ammonia gas maintained constant after 3 days of fermentation. However, in the case of sample inoculated B. subtilis SRCM 101269, ammonia gas reduced in course of fermentation time, and concentration of hydrogen sulfide also reduced for 65 ppm. Changes of nitrite concentration according to fermentation time no showed different for cow manure, however nitrite concentration in mixed livestock manure increased when compared to control. And then sulfate concentration in cow manure decreased, and no showed different when compared to the initial fermentation. No apparent change of sulfate concentration in mixed livestock manure detected. Through the previously studies, B. subtilis SRCM 101269 has high potential in industrial application manufacturing the cow manure as removal of livestock manure odor.

The Roles of Arachidonic Acid and Calcium in the Angiotensin II-induced Inhibition of $Na^+$ Uptake in Renal Proximal Tubule Cells

  • Park, Soo-Hyun;Koh, Hyun-Joo;Lee, Yeun-Hee;Son, Chang-Ho;Park, Min-Kyoung;Lee, Young-Jae;Han, Ho-Jae
    • The Korean Journal of Physiology and Pharmacology
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    • v.3 no.1
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    • pp.83-91
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    • 1999
  • Angiotensin II (ANG II) has a biphasic effect on $Na^+$ transport in proximal tubule: low doses of ANG II increase the $Na^+$ transport, whereas high doses of ANG II inhibit it. However, the mechanisms of high dose ANG II-induced inhibition on $Na^+$ uptake are poorly understood. Thus the aim of the present study was to investigate signal transduction pathways involved in the ANG II-induced inhibition of $Na^+$ uptake in the primary cultured rabbit renal proximal tubule cells (PTCs) in hormonally defined serum-free medium. ANG II $(10^{-9}\;M)-induced$ inhibition of $Na^+$ uptake was blocked by losartan $(10^{-8}\;M,\;AT_1\;antagonist),$ but not by PD123319 $(10^{-8}\;M,\;AT_2\;antagonist)$ (P<0.05). ANG II-induced inhibition of $Na^+$ uptake was also completely abolished by neomycin $(10^{-4}\;M,$ PLC inhibitor), W-7 $(10^{-4}\;M,$ calmodulin antagonist), and $AACOCF_3\;(10^{-6}\;M,\;PLA_2\;inhibitor)$ (P<0.05). ANG II significantly increased $[^3H]arachidonic$ acid (AA) release compared to control. The ANG II-induced $[^3H]AA$ release was blocked by losartan, $AACOCF_3,$ neomycin, and W-7, but not by PD123319. ANG II-induced $[^3H]AA$ release in the presence of extracellular $Ca^{2+}$ was greater than in $Ca^{2+}-free$ medium, and it was partially blocked by TMB-8 $(10^{-4}\;M,$ intracelluar $Ca^{2+}$ mobilization blocker). However, in the absence of extracellular $Ca^{2+},$ it was completely blocked by TMB-8. In addition, econazole $(10^{-6}\;M,$ cytochrome P-450 monooxygenase inhibitor) and indomethacin $(10^{-6}\;M,$ cyclooxygenase inhibitor) blocked ANG II-induced inhibition of $Na^+$ uptake, but NGDA $(10^{-6}\;M,$ lipoxygenase inhibitor) did not affect it. In conclusion, $PLA_2-mediated$ AA release is involved in ANG II-induced inhibition of $Na^+$ uptake and is modulated by $[Ca^{2+}]_i$ in the PTCs.

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Modulation of Cytochrome P-450 Induction by Long-Term Supplementation with Korean Red Ginseng to Rats (홍삼의 장기 투여가 흰쥐 간 대사효소 유도에 미치는 영향)

  • Lim, Heung-Bin
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.6
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    • pp.437-443
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    • 2007
  • This study was carried out to investigate the effect of dietary supplementation with red ginseng water-extracts on the induction of microsomal cytochrome P-450 in rats. Phenobarbital (PB) and 3-methylcholanthrene (3-MC), P-450 inducers, were administered to 3- or 12-month old rats received red ginseng extracts (25 mg/kg) from 6 weeks to 12 months for 3 days. PB and 3-MC increased levels of P-450, P-450 reductase, ethoxycoumarin O-deethylase, benzphetamine N-demethylase and glutathione-S-transferase in the liver of rats. However, chronic administration of red ginseng significantly reduced these increase of enzyme levels induced by P-450 inducers. Chronic administration of red ginseng did not affect the induction of cytochrome $b_5$ and NADH cytochrome $b_5$ reductase by P-450 inducers. It is suggested that the induction of cytochrome P-450 system in the liver in relation to xenobiotics toxicity can be modulated by long-term supplementation with Korean red ginseng to rats.

Metagenomics analysis of methane metabolisms in manure fertilized paddy soil (메타게놈 분석을 이용한 돈분뇨 처리에 의한 논토양에서 메탄대사에 미치는 영향 조사)

  • Nguyen, Son G.;Ho, Cuong Tu;Lee, Ji-Hoon;Unno, Tatsuya
    • Korean Journal of Microbiology
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    • v.52 no.2
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    • pp.157-165
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    • 2016
  • Under flooded rice fields, methanogens produce methane which comes out through rice stalks, thus rice fields are known as one of the anthropogenic sources of atmospheric methane. Studies have shown that use of manure increases amount of methane emission from rice. To investigate mechanisms by which manure boosts methane emission, comparative soil metagenomics between inorganically (NPK) and pig manure fertilized paddy soils (PIG) were conducted. Results from taxonomy analysis showed that more abundant methanogens, methanotrophs, methylotrophs, and acetogens were found in PIG than in NPK. In addition, BLAST results indicated more abundant carbohydrate mabolisetm functional genes in PIG. Among the methane metabolism related genes, PIG sample showed higher abundance of methyl-coenzyme M reductase (mcrB/mcrD/mcrG) and trimethylamine-corrinoid protein Co-methyltransferase (mttB) genes. In contrast, genes that down regulate methane emission, such as trimethylamine monooxygenase (tmm) and phosphoserine/homoserine phosphotransferase (thrH), were observed more in NPK sample. In addition, more methanotrophic genes (pmoB/amoB/mxaJ), were found more abundant in PIG sample. Identifying key genes related to methane emission and methane oxidation may provide fundamental information regarding to mechanisms by which use of manure boosts methane emission from rice. The study presented here characterized molecular variation in rice paddy, introduced by the use of pig manure.

Molecular and Ecological Analyses of Microbial Community Structures in Biofilms of a Full-Scale Aerated Up-Flow Biobead Process

  • Ju, Dong-Hun;Choi, Min-Kyung;Ahn, Jae-Hyung;Kim, Mi-Hwa;Cho, Jae-Chang;Kim, Tae-Sung;Kim, Tae-San;Seong, Chi-Nam;Ka, Jong-Ok
    • Journal of Microbiology and Biotechnology
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    • v.17 no.2
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    • pp.253-261
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    • 2007
  • Molecular and cultivation techniques were used to characterize the bacterial communities of biobead reactor biofilms in a sewage treatment plant to which an Aerated Up-Flow Biobead process was applied. With this biobead process, the monthly average values of various chemical parameters in the effluent were generally kept under the regulation limits of the effluent quality of the sewage treatment plant during the operation period. Most probable number (MPN) analysis revealed that the population of denitrifying bacteria was abundant in the biobead #1 reactor, denitrifying and nitrifying bacteria coexisted in the biobead #2 reactor, and nitrifying bacteria prevailed over denitrifying bacteria in the biobead #3 reactor. The results of the MPN test suggested that the biobead #2 reactor was a transition zone leading to acclimated nitrifying biofilms in the biobead #3 reactor. Phylogenetic analysis of 16S rDNA sequences cloned from biofilms showed that the biobead #1 reactor, which received a high organic loading rate, had much diverse microorganisms, whereas the biobead #2 and #3 reactors were dominated by the members of Proteobacteria. DGGE analysis with the ammonia monooxygenase (amoA) gene supported the observation from the MPN test that the biofilms of September were fully developed and specialized for nitrification in the biobead reactor #3. All of the DNA sequences of the amoA DGGE bands were very similar to the sequence of the amoA gene of Nitrosomonas species, the presence of which is typical in the biological aerated filters. The results of this study showed that organic and inorganic nutrients were efficiently removed by both denitrifying microbial populations in the anaerobic tank and heterotrophic and nitrifying bacterial biofilms well-formed in the three functional biobead reactors in the Aerated Up-Flow Biobead process.

Biodegradation of Gasoline Oxygenate MTBE(Methyl tert-Butyl Ether) by Butane-Utilizing Bacteria (부탄분해미생물에 의한 가솔린첨가제 MTBE(Methyl tert-Butyl Ether) 분해)

  • 장순웅;백승식;이시진
    • Journal of Soil and Groundwater Environment
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    • v.6 no.3
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    • pp.31-41
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    • 2001
  • In this study, we have examined the potential degradation of MTBE(methyl tert-butyl ether) by pure culture ENV425 and mixed culture obtained from gasoline contaminated soil using n-butane as the sources of carbon and energy. The results described in this study suggest that MTBE is degraded cometabolically by ENV425 and mixed culture grown on n-butane. Butane and MTBE degradation was completely inhibited by acetylene, which indicated that both substrates were degraded by butane monooxygenase. These cultures grown on n-butane generated TBA (tert-butyl alcohol) as a metabolite of MTBE oxidation. TBA Production was accounted 54.7% and 58.6% for MTBE oxidation by ENV425 and mixed culture, respectively. In resting cell experiments, however, TBA and TBF were detected as the oxidation products of MTBE by ENV425 and mixed culture. The observed maximal MTBE degradation rates were 52.3 and 62.3 (nmol MTBE degraded/hr/mg TSS) by ENV425 and mixed culture, respectively, and the observed maximal transformation yields ($T_y$) were 44.7 and 34.0 (nmol MTBE degraded/$\mu$mol n-butane utilized), and the observed maximal transformation capacities ($T_c$) were 199 and 226 ($\mu$mol MTBE degraded/mg TSS used).

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