• Journal of Microbiology and Biotechnology
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• v.1 no.3
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• pp.151-156
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• 1991

In order to construct a yeast strain having high ethanol tolerance together with good lactose fermentation ability, the protoplast fusion using Saccharomyces cerevisiae STV 89 and Kluyveromyces fragilis CBS 397 was carried out. Auxotrophic mutants of K. fragilis were obtained as a selection marker by treatment of ethylmethane sulfonate. The best mutant for protoplast fusion was selected based on the capabilities of ${\beta}-galactosidase$ production and lactose fermentation. The protoplast fusion using polyethylene glycol and calcium chloride solution led to the fusion frequence of $3{\times}10^{-6}$ and a number of fusants were obtained. Among these fusants, a fusant F-3-19 showed the best results in terms of ethanol tolerance, ${\beta}-galactosidase$ activity and lactose fermentation. The performance of lactose fermentation and ethanol tolerance by this fusant were better than those of K. fragilis. Study on the ethanol tolerance having relation to fatty acid composition and intracellular ethanol concentration revealed that the fusant F-3-19 had a higher unsaturated fatty acids content and accumulated less amount of intracellular ethanol compared with a parent of K. fragilis.

• YAKHAK HOEJI
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• v.51 no.5
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• pp.343-349
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• 2007

To investigate the effect of ethanol extract of Saxifraga stolonifera MEERBURGH on skin care, we measured anti-oxidant and anti-aging activity. S. stolonifera ethanol extract itself had anti-oxidant activity in a dose-dependent manner in DPPH radical scavenging. Silica dose-dependently increased the intracellular ROS generation in RAW 264.7 cells. S. stolonifera ethanol extract inhibited silica-induced intracellular superoxide anion generation, $H_2O_2$ and hydroperoxide generation in RAW 264.7 cells. S. stolonifera ethanol extract significantly inhibited both hyaluronidase and elastase activity, also significantly inhibited MMP-1(collagenase) activity as well. In NIH 3T3 fibroblast cells, S. stolonifera ethanol extract significantly increased collagen-like polymer synthesis, which suggesting the S. stolonifera ethanol extract might be used as hydration and anti-wrinkle agents. From the above results, it is suggested that the main ingredients of S. stolonifera ethanol extract play an important role in anti-oxidant and anti-aging activity.

• Korean Journal of Pharmacognosy
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• v.48 no.3
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• pp.219-225
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• 2017

We investigated the anti-oxidative effect of the blueberry duke (Vaccinium corymbosum L., Ericaceae) ethanol extract in Caenorhabditis elegans model. The ethanol extract of blueberry duke showed relatively significant DPPH radical scavenging and superoxide quenching activities. To prove antioxidant activity of the extract, we checked the activities of superoxide dismutase (SOD), catalase, intracellular ROS, and oxidative stress tolerance in C. elegans. In addition, to verify if the increased stress tolerance of C. elegans by treating with the extract was due to regulation of stress-response genes, we checked SOD-3 expression using a transgenic strain. As a consequence, the blueberry duke ethanol extract increased SOD and catalase activities of C. elegans, and reduced intracellular ROS accumulation in a dose-dependent manner. Besides, blueberry duke ethanol extract-treated CF1553 worms showed higher SOD-3::GFP intensity.

• Journal of Microbiology and Biotechnology
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• v.27 no.9
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• pp.1649-1656
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• 2017

In simultaneous saccharification and fermentation (SSF) for production of cellulosic biofuels, engineered Saccharomyces cerevisiae capable of fermenting cellobiose has provided several benefits, such as lower enzyme costs and faster fermentation rate compared with wild-type S. cerevisiae fermenting glucose. In this study, the effects of an alternative intracellular cellobiose utilization pathway-a phosphorolytic pathway based on a mutant cellodextrin transporter (CDT-1 (F213L)) and cellobiose phosphorylase (SdCBP)-was investigated by comparing with a hydrolytic pathway based on the same transporter and an intracellular ${\beta}$-glucosidase (GH1-1) for their SSF performances under various conditions. Whereas the phosphorolytic and hydrolytic cellobiose-fermenting S. cerevisiae strains performed similarly under the anoxic SSF conditions, the hydrolytic S. cerevisiae performed slightly better than the phosphorolytic S. cerevisiae under the microaerobic SSF conditions. Nonetheless, the phosphorolytic S. cerevisiae expressing the mutant CDT-1 showed better ethanol production than the glucose-fermenting S. cerevisiae with an extracellular ${\beta}$-glucosidase, regardless of SSF conditions. These results clearly prove that introduction of the intracellular cellobiose metabolic pathway into yeast can be effective on cellulosic ethanol production in SSF. They also demonstrate that enhancement of cellobiose transport activity in engineered yeast is the most important factor affecting the efficiency of SSF of cellulose.

• Journal of Microbiology and Biotechnology
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• v.1 no.1
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• pp.6-11
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• 1991

This study deals with investigation of the ethanol tolerance of yeast strains with respect to fatty acid composition and intracelluar ethanol concentration during alcohol fermentation. The cell viabilities and fermentation abilities of Saccharomyces cerevisiae and Kluyveromyces fragilis were improved by aeration and addition of unsaturated fatty acids into growth medium. Aeration decreases the accumulation of ethanol, while increases unsaturated fatty acid contents inside yeast cells. Thus it was found that oxygen and unsaturated fatty acids play decisive roles in the increase of ethanol tolerance of yeasts.

• Toxicological Research
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• v.16 no.1
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• pp.63-66
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• 2000

The effects of acute ethanol on the high K+ induced $Ca^{2+}}$ signals were examined from primary cultures of cerebellar granule neurons. $Ca^{2+}}$ signals were measured with Calcium Green-1 based microscopic video imaging. Because $Ca^{2+}}$ signal was low in most of granule neurons without stimuli, high KCI was used for depolarization. In most case, acute exposure to ethanol reduced the peak amplitude of the $Ca^{2+}}$ signals, induced by high K+, even though low concentration of ethanol(2~10mM) was used and the effects lasted more than 30min. In was also possible to see differences of ethanol inhibition, i.e. the temporal pattern of $Ca^{2+}}$ signal reductions and the strength of inhibition of $Ca^{2+}}$ signals in cerebellar granule neurons. These results indicate that low concentration of ethanol has diverse actions on the $Ca^{2+}}$ signals in cerebellar granule neurons.

• KSBB Journal
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• v.7 no.1
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• pp.33-37
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• 1992

This study is to investigate the ethanol tolerance of yeast strains related to fatty acid composition and intracellular ethanol concentration for various fermentation temperatures. The maximum accumulation of ethanol in the cells was decreased by lowering the fermentation temperature, while unsaturated fatty acid content was increased by decreasing the fermentation temperature. Thus, we found that the increase of ethanol accumulation in cells resulted in the decrease of unsaturated fatty acid content. Therefore, it was suggested that the composition of unsaturated fatty acids in the cell membrane be strongly related to the diffusion of ethanol from cell to medium.

• Fisheries and Aquatic Sciences
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• v.15 no.1
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• pp.21-27
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• 2012

Ground seawater in the east area of the volcanic Jeju Island contains abundant minerals. We investigated the metabolic activity of electrodialyzed, desalted ground seawater (EDSW) from Jeju in both cultured cells and animals. The addition of EDSW to the culture medium (up to 20%, v/v) reduced the leakage of lactate dehydrogenase and increased MTT activity in CHO-IR cells. EDSW (10%) promoted insulin-induced glucose consumption in L6 muscle cells as well as the activities of the liver ethanol-metabolizing enzymes, alcohol dehydrogenase and aldehyde dehydrogenase. Moreover, EDSW suppressed palmitate-induced intracellular fat accumulation in human hepatoma $HepG_2$ cells. Activities of AMP-stimulated protein kinase and acetyl CoA carboxylase, enzymes that modulate fat metabolism, were altered by EDSW in $HepG_2$ cells toward the suppression of intracellular lipid accumulation. EDSW also suppressed hepatic fat accumulation induced by a high-fat diet in mice. Taken together, EDSW showed beneficial metabolic effects, including the enhancement of ethanol metabolism and insulin-induced glucose consumption, and the suppression of intrahepatic fat accumulation.

• Development and Reproduction
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• v.15 no.1
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• pp.31-39
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• 2011

Change in intracellular $Ca^{2+}$-concentration ($[Ca^{2+}]_i$) is an essential event for egg activation and further development. $Ca^{2+}$ ion is originated from intracellular $Ca^{2+}$-store via inositol 1,4,5-triphosphate receptor and/or $Ca^{2+}$ influx via $Ca^{2+}$ channel. This study was performed to investigate whether changes in $Ca^{2+}$/calmodulin dependent protein kinase II (CaM KII) activity affect $Ca^{2+}$ influx during artificial egg activation with ethanol using $Ca^{2+}$ monitoring system and whole-cell patch clamp technique. Under $Ca^{2+}$ ion-omitted condition, $Ca^{2+}$-oscillation was stopped within 30 min post microinjection of porcine sperm factor, and ethanol-induced $Ca^{2+}$ increase was reduced. To investigate the role of CaM KII known as an integrator of $Ca^{2+}$- oscillation during mammalian egg fertilization, CaM KII activity was tested with a specific inhibitor KN-93. In the eggs treated with KN-93, ethanol failed to induce egg activation. In addition, KN-93 inhibited inward $Ca^{2+}$ current ($I_{Ca}$) in a time-dependent manner in whole-cell configuration. Immunostaining data showed that the voltage-dependent $Ca^{2+}$ channels were distributed along the plasma membrane of mouse egg and 2-cell embryo. From these results, we suggest that $Ca^{2+}$ influx during fertilization might be controlled by CaM KII activity.

• Journal of Microbiology and Biotechnology
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• v.28 no.6
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• pp.917-930
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• 2018

Intracellular synthesis of silver/silver chloride nanoparticles (Ag/AgCl-NPs) using Meyerozyma guilliermondii KX008616 is reported under aerobic and anaerobic conditions for the first time. The biogenic synthesis of Ag-NP types has been proposed as an easy and cost-effective alternative for various biomedical applications. The interaction of nanoparticles with ethanol production was mentioned. The purified biogenic Ag/AgCl-nanoparticles were characterized by different spectroscopic and microscopic approaches. The purified nanoparticles exhibited a surface plasmon resonance band at 419 and 415 nm, confirming the formation of Ag/AgCl-NPs under aerobic and anaerobic conditions, respectively. The planes of the cubic crystalline phase of the Ag/AgCl-NPs were confirmed by X-ray diffraction. Fourier-transform infrared spectra showed the interactions between the yeast cell constituents and silver ions to form the biogenic Ag/AgCl-NPs. The intracellular Ag/AgCl-NPs synthesized under aerobic condition were homogenous and spherical in shape, with an approximate particle size of 2.5-30nm as denoted by the transmission electron microscopy (TEM). The reaction mixture was optimized by varying reaction parameters, including temperature and pH. Analysis of ultrathin sections of yeast cells by TEM indicated that the biogenic nanoparticles were formed as clusters, known as nanoaggregates, in the cytoplasm or in the inner and outer regions of the cell wall. The study recommends using the biomass of yeast that is used in industrial or fermentation purposes to produce Ag/AgCl-NPs as associated by-products to maximize benefit and to reduce the production cost.