• Journal of Microbiology and Biotechnology
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• v.22 no.11
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• pp.1557-1567
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• 2012

Glutathione reductase (GR, E.C. 1.6.4.2) is an important enzyme that reduces glutathione disulfide (GSSG) to a sulfydryl form (GSH) in the presence of an NADPH-dependent system. This is a critical antioxidant mechanism. Owing to the significance of GR, this enzyme has been examined in a number of animals, plants, and microbes. We performed a study to evaluate the molecular properties of GR (OsGR) from rice (Oryza sativa). To determine whether heterologous expression of OsGR can reduce the deleterious effects of unfavorable abiotic conditions, we constructed a transgenic Saccharomyces cerevisiae strain expressing the GR gene cloned into the yeast expression vector p426GPD. OsGR expression was confirmed by a semiquantitative reverse transcriptase polymerase chain reaction (semiquantitative RT-PCR) assay, Western-blotting, and a test for enzyme activity. OsGR expression increased the ability of the yeast cells to adapt and recover from $H_2O_2$-induced oxidative stress and various stimuli including heat shock and exposure to menadione, heavy metals (iron, zinc, copper, and cadmium), sodium dodecyl sulfate (SDS), ethanol, and sulfuric acid. However, augmented OsGR expression did not affect the yeast fermentation capacity owing to reduction of OsGR by multiple factors produced during the fermentation process. These results suggest that ectopic OsGR expression conferred acquired tolerance by improving cellular homeostasis and resistance against different stresses in the genetically modified yeast strain, but did not affect fermentation ability.

• Journal of The Korean Society of Grassland and Forage Science
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• v.36 no.3
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• pp.243-247
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• 2016

Arsenic (As) is a toxic element that easily taken up by plants root. Several toxic forms of As disrupt plant metabolism by a series of cellular alterations. In this study, we applied annealing control primer (ACP)-based reverse transcriptase PCR (polymerase chain reaction) technique to identify differentially expressed genes (DEGs) in alfalfa roots in response to As stress. Two-week-old alfalfa seedlings were exposed to As treatment for 6 hours. DEGs were screened from As treated samples using the ACP-based technique. A total of six DEGs including heat shock protein, HSP 23, plastocyanin-like domain protein162, thioredoxin H-type 1 protein, protein MKS1, and NAD(P)H dehydrogenase B2 were identified in alfalfa roots under As stress. These genes have putative functions in abiotic stress homeostasis, antioxidant activity, and plant defense. These identified genes would be useful to increase As tolerance in alfalfa plants.

• Microbiology and Biotechnology Letters
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• v.45 no.3
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• pp.271-275
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• 2017

The expression of Deinococcus radiodurans dr1558 and pprM genes was examined for enhanced lysine production in recombinant Corynebacterium glutamicum. These genes are known to confer high tolerance to pH and osmotic shock in Escherichia coli. D. radiodurans dr1558 and pprM genes were expressed in C. glutamicum by using 6 synthetic promoters of different strengths, to evaluate the effect of expression efficiency on lysine production. Recombinant C. glutamicum expressing DR1558 under the L26 and I64 promoters showed higher lysine production than that expressing DR1558 under other promoters. Similarly, recombinant C. glutamicum expressing PprM under same promoters (L26 and I64) showed a higher increase in lysine production compared to that expressing PprM under other promoters. In the absence of $CaCO_3$ in the medium, the expression of DR1558 or PprM also increased lysine concentration in C. glutamicum depending on the promoter used. Together, these results suggest that genes involved in radiation tolerance in D. radiodurans can be used to enhance production of amino acids and their derivatives.

• Proceedings of the Ginseng society Conference
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• 2002.10a
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• pp.55-65
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• 2002

Ginseng has been used as a key constituent in traditional medicine prescriptions for centuries. Other than its well-known anti-stress and adaptogenic properties, ginseng has also been shown to be very effective in treating age-related deterioration in metabolic and memory functions. Although it is generally believed that the saponin (GS) fraction of the ginseng root accounts for the bioactivity of ginseng, a direct demonstration on which ginsenoside does what is still generally lacking. In the past decade, our laboratory has endeavored to identify the active GS components involved in energy metabolism, memory, and anti-neurotoxicity. To examine the ergogenic effects of GS in enhancing aerobic capacity, rats were subjected to either severe cold ($40^{\circ}C$ under helium-oxygen, two hours) or exercise workload $(70\%\;VO_{2}max,$ to exhaustion). Acute systemic injection (i.p.) of ginseng GS (5-20 mg/kg) significantly elevated both the total and maximum heat production in rats and improved their cold tolerance. However, pretreating the animal with the optimal dose (10 mg/kg) of GS devoid of $Rg_1\;and\;Rb_1$ failed to elicit any beneficial effects in improving cold tolerance. This indicates that either $Rb_1\;and/or\;Rg_1$ may be essential in exemplifying the thermogenic effect of GS. Further studies showed that only pretreating the animals with $Rb_1(2.5-5\;mg/kg),\;but\;not\;Rg_l,$ resulted in an increase in thermogenesis and cold tolerance. In contrast to the acute effect of GS on cold tolerance, enhancement of exercise performance in rats was only observed after chronic treatment (4 days). Further, we were able to demonstrate that both $Rb_1\;and\;Rg_1$ are effective in enhancing aerobic endurance by exercise. To illustrate the beneficial effects of GS in learning and memory, a passive avoidance paradigm (shock prod) was used. Our results indicated that the scopolamineinduced amnesia can be significantly reversed by chronically treating (4 days) the rats with either $Rb_1\;or\;Rg_1$ (1.25 - 2.5 mg/kg). To further examine its underlying mechanisms, the effects of various GS on ${\beta}-amyloid-modulated$ acetylcholine (ACh) release from the hippocampal slices were examined. It was found that inclusion of $Rb_1$ (0.1 ${\mu}M$), but not $Rg_1$, can attenuate ${\beta}-amyloid-suppressed$ ACh release from the hippocampal slices. Our results demonstrated that $Rb_1\;and\;Rg_1$ are the key components involved in various beneficial effects of GS but they may elicit their effects through different mechanisms.