• Korean Chemical Engineering Research
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• v.55 no.2
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• pp.237-241
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• 2017

Acetoin is variously applicable platform chemical in chemical and food industry. In this study, Klebsiella pneumoniae was engineered for acetoin production using metabolic engineering. From the recombinant Klebsiella pneumoniae (KMK-05) producing 2,3-butanediol, budC and dhaD genes encoding two 2,3-butanediol dehydrogenases were deleted to reduce 2,3-butanediol production. Furthermore, a transcriptional regulator, AcoK, was deleted to reduce the expression levels of acetoin degrading enzyme. Lastly, NADH oxidase was overexpressed for adjusting intracellular redox balance. The resulting strain (KJW-03-nox) produced considerable amount of acetoin, with concentration reaching 51 g/L with 2.6 g/L/h maximum productivity in 36 h fed-batch fermentation.

• The Korean Journal of Pharmacology
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• v.20 no.1 s.34
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• pp.1-11
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• 1984

The present study was performed to determine oxygen metabolites involved in cell-free MPO/$H_2O_2/Cl^-$ system by observing the effects of their scavengers on NADH oxidation and ethylene production from methional by the action of MPO prepared from human leukocytes. It was clearly demonstrated that NADH was oxidized by the cell-free MPO/$H_2O_2/Cl^-$ system as evidenced by complete inhibition of the oxidation of the substrate in the presence of eiher azide or catalase, or by omitting $Cl^-$. The MPO-mediated oxidation of NADH was completely abolished by a $^1O_2$ quencher, DABCO but not by $OH{\cdot}$ scavengers, mannitol, benzoate, formate and methanol. In ethylene assay, no ethylene was detected from methional in the MPO/$H_2O_2/Cl^-$ system with evident production of the gas by xanthine-oxidase and $Cu^{++}-H_2O_2$ systems which are suggested to generate $OH{\cdot}$. From the results obtained, it is concluded that $^1O_2$ is a major mediator with exclusion of $OH{\cdot}$ involvement in the cell-free MPO-mediated oxidation.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.36 no.6
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• pp.573-577
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• 2003

Effects of 4-nonylphenol (4-NP) on mixed function oxygenase (MFO) in the kidneies of rockfish (Sebastes schlegeli) were investigated. The cytochrome P45O (CYP) contents and NADPH cytochrome P450 reductase, NADH cytochrome b5 reductase and 7-ethoxyredorufin-O-deethylase (EROD) activities of microsome, glutathione S-transferase (GST) activity of cytosol in rockfish exposed to 4-NP for 7 days using an intraperitoneal injection (25 mg/kg) were quantitatively determined. The GST activity of rockfish exposed to 4-NP were higher, up to 5.2 times higher, than those in the control fish. The activities of NADPH-and NADH-dependent reductases were inhibited. On the other hand, CYP contents and EROD activity of the 4-NP exposed fish demonstrated neither an increasing or decreasing trend.

• Journal of Embryo Transfer
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• v.25 no.1
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• pp.35-42
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• 2010

Many studies propose that dysfunction of mitochondrial proton-translocating NADH-ubiquinone oxidoreductase (complex I) is associated with neurodegenerative disorders, such as Parkinson's disease and Huntington's disease. Mammalian mitochondrial proton-translocating NADH-quinone oxidoreductase (complex I) consists of at least 46 different subunits. In contrast, the NDI1 gene of Saccharomyces cerevisiae is a single subunit rotenone-insensitive NADH-quinone oxidoreductase that is located on the matrix side of the inner mitochondrial membrane. With a recombinant adeno-associated virus vector carrying the NDI1 gene (rAAV-NDI1) as the gene delivery method, we were able to attain high transduction efficiencies even in the human epithelial cervical cancer cells that are difficult to transfect by lipofection or calcium phosphate precipitation methods. Using a rAAV-NDI1, we demonstrated that the Ndi1 enzyme is successfully expressed in HeLa cells. The expressed Ndi1 enzyme was recognized to be localized in mitochondria by confocal immunofluorescence microscopic analyses and immunoblotting. Using digitonin-permeabilized cells, it was shown that the NADH oxidase activity of the NDI1-transduced HeLa cells were not affected by rotenone which is inhibitor of complex I, but was inhibited by flavone and antimycin A. The NDI1-transduced cells were able to grow in media containing rotenone. In contrast, control cells that did not receive the NDI1 gene failed to survive. In particular, in the NDI1-transduced cells, the yeast enzyme becomes integrated into the human respiratory chain. It is concluded that the NDI1 gene provides a potentially useful tool for gene therapy of mitochondrial diseases caused by complex I deficiency.

• Reproductive and Developmental Biology
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• v.35 no.4
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• pp.427-434
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• 2011

Mitochondria diseases have been reported to involve structural and functional defects of complex I-V. Especially, many of these diseases are known to be related to dysfunction of mitochondrial proton-translocating NADH-ubiquinone oxidoreductase (complex I). The dysfunction of mitochondria complex I is associated with neurodegenerative disorders, such as Parkinson's disease, Huntington's disease, and Leber's hereditary optic neuropathy (LHON). Mammalian mitochondrial proton-translocating NADH-quinone oxidoreductase (complex I) is largest and consists of at least 46 different subunits. In contrast, the NDI1 gene of Saccharomyces cerevisiae is a single subunit rotenone-insensitive NADH-quinone oxidoreductase that is located on the matrix side of the inner mitochondrial membrane. The Saccharomyces cerevisiae NDI1 gene using a recombinant adeno-associated virus vector (rAAV-NDI1) was successfully expressed in AML12 mouse liver hepatocytes and the NDI1-transduced cells were able to grow in media containing rotenone. In contrast, control cells that did not receive the NDI1 gene failed to survive. The expressed Ndi1 enzyme was recognized to be localized in mitochondria by confocal immunofluorescence microscopic analyses and immunoblotting. Using digitonin-permeabilized cells, it was shown that the NADH oxidase activity of the NDI1-transduced cells was not affected by rotenone which is inhibitor of complex I, but was inhibited by antimycin A. Furthermore, these results indicate that Ndi1 can be functionally expressed in the AML12 mouse liver hepatocytes. It is conceivable that the NDI1 gene is powerful tool for gene therapy of mitochondrial diseases caused by complex I deficiency. In the future, we will attempt to functionally express the NDI1 gene in mouse embryonic stem (mES) cell.

• Environmental Analysis Health and Toxicology
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• v.13 no.3_4
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• pp.105-115
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• 1998

Reactive oxygen species (ROS) are highly reactive molecules due to their unpaired electron. They have been suspected as one of the major tissue damage inducers in biological metabolic systems. Antioxidant enzymes, such as catalase and superoxide dismutase, could not repair all the oxidative damages resulting from those excessive toxic ROS. It is, therefore, urgent to develop effective antioxidants to relieve from the oxidatire damages. In this study antioxidative effects were investigated by using two flavonoids such as quercetin and naringenin and a flavonoid-rich extract, Ginkgo biloba extract in combination with paraquat that is known as a strong generator of oxygen radicals. The results are summeringed as follows: 1. To assess radical scavenging ability reduction concentrations (IC$_{50}$) of 1,1-diphenyl-2-picrylhydrazine (DPPH) within 15 minutes were measured. The values of the IC$_{50}$ of quercetin and Ginkgo biloba extract were 15.4 $\mu$M and 13.2$\mu$g/ml, respectively. Their radical removing activities showed concentration-dependent manners. 2. In the hydrogen peroxide assay by using PMS-NADH system, quercetin, naringenin and Ginkgo biloba extract led to removing hydrogen peroxide in concentrationdependent manner whose removing abilities at 100$\mu$M or 100 $\mu$g/ml were 75.6, 25.8 and 26.0%, respectively. 3. In the hydrogen peroxide-induced rat blood hemolysis assay all three compounds led to similar effects whose hemolysis inhibition ratios at 100$\mu$M or 100$\mu$g/ml were 68.0, 5.14 and 55.8%, respectively. 4. In the xanthinee oxidase assay by measuring degree of NADH oxidation in the presence of hypoxanthine and xanthinee oxidase, both quercetin and Ginkgo biloba extract showed excellent activities showing 42.8 and 24.2% inhibiting xanthine oxidase activity at 100$\mu$M or 100$\mu$g/ml concentrations, respectively.

• Parasites, Hosts and Diseases
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• v.53 no.6
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• pp.731-735
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• 2015

A 2-year-old female donkey (Equus asinus) was euthanized in the Pathology Department of Firat University, Elazig, Turkey. Necropsy disclosed the presence of 7 hydatid cysts distributed throughout the lung parenchyma. One of those cysts represented the parasite material of the present study and was molecularly identified through sequencing of a fragment of cytochrome c oxidase subunit 1 (CO1) and nicotinamide adenine dinucleotide dehydrogenase subunit 1 (NADH1) gene, as Echinococcus equinus. The generated CO1 sequence supports the presence of the dominant haplotype as has been described in Europe and Africa. The NADH1 sequence was found similar to sequences reported in equids in Egypt and the United Kingdom. The molecular identification of E. equinus in a donkey is being reported for the first time in Turkey.

• Microbiology and Biotechnology Letters
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• v.26 no.1
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• pp.7-14
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• 1998

To study the oxygen tolerance mechanism of bifidobacteria, we have studied the growth of cells, the activities of the enzymes which were related with oxygen, such as catalase, superoxide dismutase(SOD), NADH oxidase, and NADH peroxidase, and cellular fatty acid compositions of Bifidobacterium adolescentis and B. longum under anaerobic and aerated (microaerobic and aerobic) conditions. B. longum grew relatively well under the microaerobic conditions, whereas the growth of B. adolescentis was inhibited under the same aerated conditions. B. adolescentis had extremely low level of NADH oxidative enzymes while B. longum had the relatively high level of NADH oxidative enzymes, whose activities were dramatically increased from 3.7 to 11.4 times by microaerobic condition but not in B. adolescentis. The activity of SOD was unexpectedly high in B. adolescentis compared with in B. longum under anaerobic and aerated conditions. The activities of catalase were not detected in all samples tested in this study. We also found that normal $C_{l6:0}$ and $C_{18:1}$ were the major fatty acids in B. adolescentis and B. longum under anaerobic and aerated conditions. 2.2-14.1% $C_{l9:0}$ cyclo fatty acid was detected only in B. longum and the fatty acid was increased by the addition of the aeration. The $C_{l9:0}$ cyclic fatty acid was identified as a cis 9, 10-methylene octadecanoic acid, which was different from lactobacillic acid in the cyclized site. 6.6%-24.6% of dimethyl acetals (DMA) which came from plasmalogen were observed in the B. adolescentis and B. longum grown under anaerobic condition, and the components were notably decreased in the cells grown under the aerated conditions. It is believed that NADH oxidative enzymes play an important role to detoxify oxygen metabolites of Bifidobacteriurn spp. under anaerobic and microaerobic conditions. Independently from oxidative enzymes, it seems that oxygen stress may induce the change of the level of cellular fatty acids showing an increase of $C_{l9:0}$ cyclo in B. longum and a decrease of $C_{l8:1}$ of plasmalogen in B. longum and B. adolescentis to adapt in environment.

• The Korean Journal of Pharmacology
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• v.21 no.1
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• pp.1-11
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• 1985

The effects of feeding indole, indole-3-carbinol and benzofuran (all at 5 mmole/kg body wt./day) on various hepatic microsomal and cytosolic enzyme activities involved in xenobiotic metabolism have been compared. Benzofuran was found to elevate the activities of many enzymes both in microsomes (e.g., aniline hydroxylase, 7-ethoxycoumarin O-deethylase, p-nitrophenol UDPGA-transferase and epoxide hydrolase) and in cytosol (e.g., glutathione reductase, glutathione S-transferase, NADH:quinone reductase and UDP-glucose dehydrogenase). The structures of indole and indole-3-carbinol are similar to benzofuran except for the substitution of nitrogen with oxygen atom within the furan ring. Results showed that the activities of UDPGA-transferase and NADH:quinone reductase were not elevated by these indole compounds. While the chemical structure of these two indole compounds are identical except for the presence of the carbinol (methanol) group in indole-3-carbinol, there were marked differences in the types and activities of microsomal enzymes that were enhanced. Among the microsomal enzyme activities determined, indole elevated only the NADPH:cytochrome c reductase, while indole-3-carbinol increased several mixed function oxidase and particularly the epoxide hydrolase activities. Based on the chemical structures of tested compounds and the observed results, possible explanations for the mechanisms involved in elevating epoxide hydrolase activity by benzofuran and indole-3-carbinol are discussed.

• Korean Journal of Environmental Biology
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• v.22 no.1
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• pp.177-183
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• 2004

Cytochrome P45O (CYP) contents and 7-ethoxyresorufin-O-deethylase (EROD) activity were determined in hepatic microsome of olive flounder (Paralichthys olivaceus) exposed to tributyltin chloride (TBTC), tributyltin oxide (TBTO), and triphenyltin chloride (TPTC). In addition, effects of in vivo (intraperitoneal injection of 7.5 mg $kg^{-1}$ BW) exposure of flounder to TPTC on CYP, NADPH cytochrome c reductase, NADH cytochrome b5 yeductase and EROD levels were measured. In in vitro exposure of hepatic microsome to organotins, TBTC, TBTO and TPTC reduced CYP contents and inhibited EROD activity. The TPTC was the strongest inhibitor, which is followed by TBTO and TBTC. The degree of inhibition, especially EROD acitivity, depended on the exposure duration. In addition, all the target enzymes in flounder were inhibited by TPTC with the in vivo exposure to TPTC. As EROD activity was the most sensitive to the inhibitions and demonstrated good reproducibility of the results, it could be used as a helpful tool toy monitor effects of organotin compounds on mixed funciton oxygenase system in marine fish.