• Korean Journal of Food Science and Technology
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• v.37 no.6
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• pp.951-956
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• 2005

Selection of oxygen-tolerant strains and elucidation of their oxygen tolerance mechanism were crucial for effective use of bifidobacteria. Oxygen-tolerant bifidobacteria were able to significantly remove environmental oxygen (oxygen removal activity) as compared to oxygen-sensitive strains. Most oxygen removal activity was inhibited by heat treatment and exposure to extreme pH (2.0) of bifidobacterial cell. NADH oxidase was major enzyme related to oxygen removal activity. Oxygen-tolerant bifidobacteria possessed high NADH peroxidase activity level to detoxify $H_2O_2$ formed from reaction of NADH oxidase. Addition of oxygen to anaerobic culture broth significantly increased activities of HADH oxidase and NADH peroxidase within 1hr and rapid increment of oxygen concentration was prevented. Results showed NADH oxidase and NADH peroxidase of oxygen-tolerant bifidobacteria played important roles in elimination of oxygen and oxygen metabolite $(H_2O_2)$.

• Journal of Microbiology and Biotechnology
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• v.7 no.5
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• pp.356-359
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• 1997

To study the relationship between oxygen tolerance and enzyme activity in the oxygen metabolism of bifidobacteria, the activities of catalase, superoxide dismutase (SOD), NADH oxidase and NADH peroxidase from six typical bifidobacteria and other bacteria were assayed by spectrophotometry. Catalase activity was hardly detected in any of the bifidobacteria tested. SOD activity was detected in every species including the Clostridium species. In particular SOD activity was notably high in the aerosensitive Bifidobacterium adolescentis. This fact indicates that SOD activity is not a critical factor to ensure aerotolerance. Aerosensitive B. adolescentis showed very low NADH oxidative enzyme activity whereas other aerotolerant bifidobacteria exhibited considerable activity for the enzymes. It seems that detoxification of $H_2O_2$ by NADH oxidative enzymes might be an important factor in improving for aerotolerant bifidobacteria survival rates in an oxygen environment.

• Journal of Microbiology and Biotechnology
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• v.14 no.5
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• pp.919-923
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• 2004

The sk10 isolated from kimchi was identified as W. kimchii on the basis of l6s-rDNA sequencing. Studies were made to analyze the metabolic flux shift of the sk10 on glucose under aerobic growth conditions. The sk10 produced 38.2 mM acetate, 16.3 mM ethanol, and 33.2 mM lactate under aerobic conditions, but 2.4 mM acetate, 48.0 mM ethanol, and 44.1 mM lactate under anaerobic conditions. The NADH peroxidase (NADH-dependent hydrogen peroxidase) activity of sk10 grown under aerobic conditions was 11 times higher than that under anaerobic conditions. Under the low ratio of $NADH/NAD^+$, the metabolic flux toward lactate and ethanol was shifted to the flux through acetate kinase without NADH oxidation. The kinds of enzymes and metabolites of sk10 were close to those in the pathway of Leuconostoc sp., but the metabolites produced under aerobic growth conditions were different from those of Leuconostoc sp. The stoichiometric balance calculated using the concentrations of metabolites and substrate was about 97%, coincident with the theoretical values under both aerobic and anaerobic conditions. From these results, it was concluded that the metabolic flux of W. kimchii sk10 was partially shifted from lactate and ethanol to acetate under aerobic conditions only.

• Journal of Microbiology and Biotechnology
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• v.14 no.5
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• pp.914-918
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• 2004

This study was carried out to analyze the metabolic flux of W. kimchii sk10 on pyruvate and ethanol as a carbon source. The sk10 grown on ethanol produced acetate under aerobic conditions rather than under anaerobic conditions. The lactate and acetate were produced on ethanol plus pyruvate by the sk10 grown under aerobic and anaerobic conditions, respectively. The resting cell of sk10 produced 99.1 mM acetate and 17.3 mM lactate under aerobic conditions and 51.1 mM acetate and 62.4 mM lactate under anaerobic conditions from ethanol plus pyruvate, respectively. This result is thought to be due to the difference in the $NADH/NAD^+$ ratio depending on the growth conditions. The 11-fold overproduction of NADH peroxidase results in a low $NADH/NAD^+$ratio under aerobic growth conditions. At the low $NADH/NAD^+$ ratio, the metabolic flux of pyruvate toward lactate has to be shifted to a flux toward acetate without NADH oxidation to $NAD^+$, and ethanol oxidation to acetate coupled to $NAD^+$ reduction to NADH has to be activated.

• Korean Journal of Veterinary Research
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• v.31 no.1
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• pp.41-47
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• 1991

The protective of influences of sodium selenitc ($Na_2SeO_3$) against the methemoglobinemia with sodium nitrite were investigated on hemoglobin, methemoglobin, glutathione peroxidase and NADH-methemoglobin reductase activity in rabbits which were given 0,1,3 and 9ppm sodium selenite of drinking water for a week. Dietary selenium did not alter total hemoglobin in the blood of rabbits. Selenium was found to decrease nitrite-induced methemoglobin in a dose-dependent manner. The glutathione peroxidase activity was also increased by selenium in all the experimental groups. However, the NADH-methemoglobin reductase activity by selenite did not show significant differences as concerns the methemoglobinemia. These results showed that selenium could inhibit nitrite-induced methemoglobinemia. Its influence of inhibition is suggested that the effect of the reduction of methemoglobin was greatly stimulated by glutathione peroxidase activity.

• Korean Journal of Plant Tissue Culture
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• v.25 no.4
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• pp.277-282
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• 1998

Whereas cationic extracellular peroxidases (PODs) were observed in the suspension cultures of rose (Rosa sp. L. cv Pual's scarlet) grown under normal conditions, new anionic isozymes were induced within 24 hr by the treatment of low host-specific elicitor (10 mg glucan/L media) prepared from yeast cell wall. Prominent anionic (pI 6.1) and cationic POD (pI 8.4) were purified and characterized to understand the physiological role of the enzymes. Both enzymes were purified (ca.200 fold) by the ammonium sulfate precipitation, ion exchange chromate-graphy and gel filtration chromatography. The Km values of the purified anionic POD for ferulic acid and $\textrm{H}_2\textrm{O}_2$ were 4.64 mM and 0.72 mM, whereas those of the cationic POD were 1.38 mM and 0.48 mM, respetively. The activity of the anionic POD as NADH oxidase was twice higher than that of cationic POD. The NADH oxidation in the anionic POD fraction was inhibited by 60% on the addition of 0.1 mM coniferyl alcohol, while that in the cationic fraction was inhibited by 15%.

• Korean Journal of Veterinary Service
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• v.30 no.3
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• pp.291-304
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• 2007

Mutants of an obligate aerobic bacterium, Vitreoscilla, that have deficiency in heat-labile catalase-peroxidase hydroperoxidase I (HPI) were created by EMS treatment. The catalase-peroxidase HPI-deficient mutant showed substantially lower peroxidase activity in exponential and mid-stationary phase compared with the wild type strain. In late stationary phase, the mutant exhibited no peroxidase activity. Peroxidase deficiency in the mutant was revealed by polyacrylamide gels stained for peroxidase activity. Characteristically, catalase levels in the mutant increased about 14- and 8-fold during growth in the exponential and stationary phases, respectively, compared to those in the wild type, suggesting a compensatory effect for protection from $H_2O_2$ toxicity. The mutant showed differences in physiology from the wild type: retardation in growth rate and decrease in oxygen consumption. Both the wild type and the catalase-peroxidase HPI-deficient mutant of Vitreoscilla had lower growth rates in media containing increasing $H_2O_2$ concentrations. However, the mutant exhibited an additionally decreased growth rate after 6 to 8 h of growth compared to the wild type. The wild type was resistent up to 20 mM $H_2O_2$, whereas the mutant was very sensitive to high concentrations of exogenous $H_2O_2$. Although elevated catalase levels would provide protection of the bacteria from the deleterious effect of $H_2O_2$, it did not appear to be complete. Cell-free extracts of the mutant showed decreased NADH oxidation rates and higher accumulation of $H_2O_2$ during this oxidation. These results may account for the impaired growth and earlier onset of death phase by the catalase-peroxidase HPI-deficient mutant of Vitreoscilla.

• 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.

• Toxicological Research
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• v.9 no.1
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• pp.23-33
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• 1993

In the present study, the effects of allopurinol on paraquat toxicity were investigated in paraquat-treated rats. The surivals of paraquat-treated rats were increased by allopurinol treatment. The contents of glutathione in liver and kidney were significantly decreased by paraquat, but restored by allopurinol. The activity of xanthine oxidase was significantly reduced but NADH dehydrogenase was not changed by allopurinol teatment. The activities of catalase, SOD and glutathione peroxidase in liver were significantly decreased by paraquat but catalase was restored by allopurinol treatment.

• Journal of Microbiology and Biotechnology
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• v.19 no.7
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• pp.666-674
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• 2009

An electrochemical bioreactor (ECB) composed of a cathode compartment and an air anode was used in this study to characterize the ethanol fermentation of Zymomonas mobilis. The cathode and air anode were constructed of modified graphite felt with neutral red (NR) and a modified porous carbon plate with cellulose acetate and porous ceramic membrane, respectively. The air anode operates as a catalyst to generate protons and electrons from water. The growth and ethanol production of Z. mobilis were 50% higher in the ECB than were observed under anoxic nitrogen conditions. Ethanol production by growing cells and the crude enzyme of Z. mobilis were significantly lower under aerobic conditions than under other conditions. The growing cells and crude enzyme of Z. mobilis did not catalyze ethanol production from pyruvate and acetaldehyde. The membrane fraction of crude enzyme catalyzed ethanol production from glucose, but the soluble fraction did not. NADH was oxidized to $NAD^+$in association with $H_2O_2$reduction, via the catalysis of crude enzyme. Our results suggested that NADH/$NAD^+$balance may be a critical factor for ethanol production from glucose in the metabolism of Z. mobilis, and that the metabolic activity of both growing cells and crude enzyme for ethanol fermentation may be induced in the presence of glucose.