• Molecules and Cells
• /
• v.40 no.3
• /
• pp.230-242
• /
• 2017

In the Arabidopsis genome, approximately 80 MAP3Ks (mitogen-activated protein kinase kinase kinases) have been identified. However, only a few of them have been characterized, and the functions of most MAP3Ks are largely unknown. In this paper, we report the function of MAP3K16 and several other MAP3Ks, MAP3K14/15/17/18, whose expression is salt-inducible. We prepared MAP3K16 overexpression (OX) lines and analyzed their phenotypes. The result showed that the transgenic plants were ABA-insensitive during seed germination and cotyledon greening stage but their root growth was ABA-hypersensitive. The OX lines were more susceptible to water-deficit condition at later growth stage in soil. A MAP3K16 knockout (KO) line, on the other hand, exhibited opposite phenotypes. In similar transgenic analyses, we found that MAP3K14/15/17/18 OX and KO lines displayed similar phenotypes to those of MA3K16, suggesting the functional redundancy among them. MAP3K16 possesses in vitro kinase activity, and we carried out two-hybrid analyses to identify MAP3K16 substrates. Our results indicate that MAP3K16 interacts with MKK3 and the negative regulator of ABA response, ABR1, in yeast. Furthermore, MAP3K16 recombinant protein could phosphorylate MKK3 and ABR1, suggesting that they might be MAP3K16 substrates. Collectively, our results demonstrate that MAP3K16 and MAP3K14/15/17/18 are involved in ABA response, playing negative or positive roles depending on developmental stage and that MAP3K16 may function via MKK3 and ABR1.

• Korean Journal of Oriental Medicine
• /
• v.15 no.3
• /
• pp.83-91
• /
• 2009

$\beta$-glucan is a fiber-type complex sugar (polysaccharide) derived from the cell wall of baker's yeast, oat and barley fiber, and many medicinal mushrooms, such as maitake. The primary uses of $\beta$-glucan are to enhance the immune system, to lower blood cholesterol levels and to treat tumor. $\beta$-glucan has no systemic toxicity in mice, therefore it needed clinical trail to prove efficacy and safety for human. The subjects total 56 healty volunteers were divided into two groups including taken $\beta$-glucan tablet group and placebo group. Subjects were taken two tablets per oral for 4 weeks. They had agreed to take part in this experiment, and didn't take any other clinical trail products. After 4 weeks blood of subjects were checked. The check list are TNF-$\alpha$, INF-$\gamma$, IL-2, IL-4, total WBC, differential WBC, RBC, hemoglobin, platelet, MCV, MCH, MCHC, HCT, Na, K, Ca, Cl, AST, ALT, ALP, $\gamma$-GTP, total protein, triglyceride, total cholesterol, total bilirubin, albumin, uric acid, creatinine, BUN, pH, protein, glucose, ketone body, blood, bilirubin. We evaluated efficacy by cytokines that compare before and after taking. Collected data were analyzed as two sample t-test, chi-square test and ANOVA using SAS V.9.1.This study results are that in TNF-$\alpha$ of $1^{st}$ efficacy measurement item, all of two groups figure were increased significantly compare to before figure. In IL4 of $2^{nd}$ efficacy measurement item, experimental group figure were decreased significantly but placebo group figure were increased. The conclusions show that based on the above results, $\beta$-glucan has favorable effect to enhance immune system, especially IL4 results showed that it has effect to improve the allergic immune system.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.38 no.1
• /
• pp.76-82
• /
• 2009

Wheat is the most widely cultivated cereal and an important source of dietary protein worldwide. Wheat allergy, defined as an adverse immunologic reaction to wheat, encompasses a broad spectrum of disorders with different pathomechanisms and clinical manifestation. The Nuruk, a traditional Korean Koji for brewing, was made with wheat flour and fermenting microbes such as bacteria, yeast and mold. The strains grown on Nuruk secrete various enzymes as amylase and protease. By the activation of such enzymes, starch and proteins in Nuruk are hydrolyzed to sugar and amino acid. Therefore, it is supposed to reduce allergic proteins in wheat. To study quality properties and degradation degree of allergenicity in Nuruk by fermentation, we investigated the changes of general ingredients and allergenicity in Nuruk during fermentation. Moisture contents was decreased from 24.2% to 13.6% during fermentation. Crude lipid and protein contents were gradually increased during fermentation. After 15 days of fermentation, reducing sugar and total sugar contents were reached its maximum level, and they were 27.45% and 39.00%, respectively. Acid and neutral protease activity were significantly increased during fermentation, but alkaline protease activity was not detected. ${\alpha}$-amylase activity was gradually increased and showed maximum level about 2,833.00 U/g after 15 days of fermentation. Glucoamylase activity was the highest level about 497.9 U/g after 10 days of fermentation. The increase of these proteolytic and saccharogenic enzyme activities will provide efficient condition for production of rice wine. Also, protein fractions were isolated from Nuruk, and degradation of these proteins during fermentation were confirmed by SDS-PAGE. IgE immunoblotting using patient's sera with wheat allergy was performed to confirm allergenic protein in Nuruk. These results as fermentation of Nuruk will provide a useful tool for developing safer wheat products to prevent wheat allergy.

• Journal of Life Science
• /
• v.29 no.3
• /
• pp.369-375
• /
• 2019

Microtubules form through the polymerization of ${\alpha}-$ and ${\beta}-tubulin$, and tubulin transport plays an important role in defining the rate of microtubule growth inside cellular appendages, such as the cilia and flagella. Heterotrimeric kinesin 2 is a molecular motor member of the kinesin superfamily (KIF) that moves along the microtubules to transport multiple cargoes. It consists of two motor subunits (KIF3A and KIF3B) and a kinesin-associated protein 3 (KAP3), forming a heterotrimeric complex. Heterotrimeric kinesin 2 interacts with many different binding proteins through the cargo-binding domains of the KIF3s, but these binding proteins have not yet been specified. To identify these proteins for KIF3A, we performed yeast two-hybrid (Y2H) screening and found a specific interaction with ${\beta}2-tubulin$ (Tubb2), a microtubule component. Tubb2 was found to bind to the cargo-binding domain of KIF3A but did not interact with KIF3B, KIF5B, or kinesin light chain 1 in the Y2H assay. The carboxyl-terminal region of Tubb2 is essential for interaction with KIF3A. Other Tubb isoforms, including Tubb1, Tubb3, Tubb4, and Tubb5, also interacted with KIF3A in the Y2H screening. However, ${\alpha}1-tubulin$ (Tuba1) did not interact with KIF3A. In addition, an antibody to KIF3A specifically co-immunoprecipitated the KIF3B and KAP3 associated with Tubb2 from mouse brain extracts. In combination, these results suggest that a heterotrimeric kinesin 2 motor protein is capable of binding to tubulin and may transport it in cells.

• The Korean Journal of Food And Nutrition
• /
• v.24 no.3
• /
• pp.367-375
• /
• 2011

For this study, Korean-type Koumiss was made by the fermentation of mixed cultures, in which yeast, Kuyveromyces, and microflora, Streptococcus thermophiles and Lactobacillus bulgaricus, were inoculated into 10% skimmed milk with added whey powder(control: A, 2%: B, 4%: C, 6%: D, and 8%: E). Fat, protein, lactose, titratable acidity, pH, the number of lactic acid bacteria, the number of yeast, alcohol content, volatile fatty acids, volatile free amino acids and minerals were measured in the products. The results were as follows: As the dosage of whey powder increased, fat increased from 0.74% in the control to 2.30% in sample E, protein increased from 2.95% in the control to 4.39% in sample E and lactose increased from 3.10% in the control to 7.43% in sample E. Titratable acidity and pH increased gradually. The number of lactic acid bacteria increased from $10^9\;cfu/m{\ell}$ in the control to $3.8{\times}10^9\;cfu/m{\ell}$ in sample E, and the number of yeast increased from $6.1{\times}10^7\;cfu/m{\ell}$ in the control to $1.65{\times}10^8\;cfu/m{\ell}$ in sample E, according to the increase of whey powder content. For alcohol content, the average values were 0.863%, 0.967%, 0.890%, 1.290%, and 1.313% for the control and samples B, C, D, and E, respectively. As the dosage of whey powder increased, alcohol content showed a tendency to gradually increase. The average alcohol content of E was 1.313 and this was higher than the alcohol content of Kazahstana-type Koumiss with 1.08%. Sixteen types of free amino acids were detected. Glycine was the lowest in the control at $0.38mg/m{\ell}$ and sample E contained $0.64mg/m{\ell}$. Histidine was also low in the control at $0.42mg/m{\ell}$ and sample E contained $0.65mg/m{\ell}$. On the other hand, glutamic acid was highest at $4.13mg/m{\ell}$ in the control whereas sample E had $6.96mg/m{\ell}$. Proline was also high in the control at $1.71mg/m{\ell}$ in control, but E contained $2.80mg/m{\ell}$. Aspartic acid and leucine were greater in sample E than in the control. For volatile free fatty acids, content generally had a tendency to increase in the control, and samples B, C, D, and E. Content of acetic acid gradually increased from $12,661{\mu}g/100m{\ell}$ in the control to $37,140{\mu}g/m{\ell}$ in sample E. Butyric acid was not detected in the control and was measured as $1,950{\mu}g/100m{\ell}$ in sample E. Caproic acid content was $177{\mu}g/100m{\ell}$ in the control and $812{\mu}g/100m{\ell}$ in sample E, and it increased according to the increase of whey powder content. Valeric acid was measured in a small amount in the control as $22{\mu}g/100m{\ell}$, but it was not detected in any other case. Mineral contents of Ca, P, and Mg increased from 1,042.38 ppm, 863.61 ppm, and 101.28 ppm in the control to 1,535.12 ppm, 1,336.71 ppm, and 162.44 ppm in sample E, respectively. Na content was increased from 447.19 ppm in the control to 1,001.57 ppm in sample E. The content of K was increased from 1,266.39 ppm in the control to 2,613.93 ppm in E. Mineral content also increased with whey powder content. In sensory evaluations, the scores increased as whey powder content increased. Flavor was lowest in the control with 6.3 points and highest in E with 8.2 points. Body and texture were highest at 4.2 points in the control, which did not have added whey powder. In the case of appearance, there were no great differences among the samples.

• Journal of Aquaculture
• /
• v.10 no.3
• /
• pp.327-334
• /
• 1997

This study was conducted with Israeli strain of common carp to determine apparent protein, lipid and gross energy digestibility coefficients and apparent calcium and phosphorus availabilities of the following feedstuffs : steam-dried white fish meal (WFM-S), flame-dried white fish meal (WFM-F), brown fish meal (BFM), soybean meal (SBM), corn gluten meal (CGM), sesame meal (SSM), wheat flour (WFL), brewer's yeast (BY) and monocalcium phosphate (MCP). Test diets consisting of a 70:30 mixture of refoernce diet to test ingredient were utilized with chromic oxide as an external indicator. Fecal collection was made during 10 days before the end of feeding trial lasted for 27 days. The best weight gain and feed utilization were found in fish fed MCP containing test diet. Protein digestibility was high for most ingredients, ranging from 81.2% for BY to 91.9% for SBM, with the exception of SSM which was 77.6%. Lipid digestibility coefficients ranged 74% for CGM to 85.8% for two kinds of white fish meal. Digestible energy coefficients were generally high for fish meals, ranging from 77.4% for WFM-S to 81.3% for WFM-F, whereas those for plant feedstuffs were significantly (P<0.05) lower, from 64.5% for CGM to 70.6% for BY. Phosphorus availability from fish meals was variable with a low of 6.6% for WFM-F and a high of 13.6% for WFM-S. Phosphorus availability from SBM and BY was 7.3% and 57.2%, respectively. Calcium and phosphorus availabilities from MCP were 84.6% and 80.6%, respectively. These results provide more precise information concerning nutrients and energy utilization of Israeli strain of common carp and will allow ingredient substitutions in practical diet formulations based on levels of availabile protein and phosphorus.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.7 no.3
• /
• pp.105-114
• /
• 1974

Identification and change of microflora during the fermentation of anchovy Engraulis japonica, under the halophilic circumstance were investigated. The change of salinity and pH in meat and juice which decide the environment for microorganism and decomposition of nitrogenous compound which functions as a nutrient source were also discussed by measuring the content of total-N, amino-N, nonprotein-N, TMA and VBN, The fresh anchovy was mixed with rock salt (20 percent w/w) and stocked for six months. Through the fermentation lag phase of viable cells extended for 20 days that was obviously larger compared with other circumstances, hereafter increased to reach the maximum value of $5\times10^4$ total count per gram at 35 day stock. The stationary phase proceeded for 25 days. 540 strains were isolated and among them 11 genus of bacteria, 3 genus of yeasts, were identified and other 2 yeast strains of unidentified. At the initial stage of fermentation, Pseudomonas, and Helobacterium prevalently grew, at the middle stage, they disappeared rapidly and Pediococcus and yeasts completely dominated, where they are assumed to get directly involved with fermentation of fish, The PH value tended to decrease in the progress of fermentation and at 100 day stock it showed the minimum value of 5.5 to 5.6 in both meat and juice. The highest salinity of meat decreased to 18 percent, while in juice it decreased to 28 percent since 50 days stock. The content of total-N in meat gradually decreased to 2.8 percent, while in juice it increased to 2.3 percent at 100 day stock, However nonprotein-N was 1.8 percent and amino-N was 1.1 Percent. Since 100 days stock, the increasing rate of amino-M is too low it could be judged to entered the final stage of fermentation, In the first 20 days stock, the increase of VBN and TMA can be explained by the growth of putrefactive bacteria such as pseudomonas on the meat before salts penetrate into the fish meat, while reincrement after 100 days stock, is explained by decomposition of free amino acid due to the reactions of bacteria and enzymes.

• Journal of Life Science
• /
• v.18 no.8
• /
• pp.1053-1058
• /
• 2008

Glutathione is a well known chemotherapeutic agent for liver disease and is a popular nutritional supplement in the United States. Previous our studies reported the suppressive effects of glutathione-enriched Saccharomyces cerevisiae FF-8 strain (FF-8GY) on carbon tetrachloride- and alcohol-induced hepatotoxicity. The primary objective of this study was to investigate the comparative effects of FF-8GY and commercially available glutathione-enriched yeast extract (GYE) against the oxidative stress in alcohol-induced fatty liver of rats. The lipid peroxidative index (thiobarbituric acid-reactive substances, TBARS) and antioxidant status (reduced glutathione level) were used to monitor those protective roles of FF-8GY or GYE treatment. When the rat was treated alcohol, the TBARS levels in the whole liver and the subfractions of microsomal and mitochondria were significantly increased but these were significantly decreased by FF-8GY treatment and tended to be lowered by GYE treatment. The concentration of hepatic glutathione is known to be closely associated with antioxidant system and this was slightly deplete in the alcohol-induced rats, but this was recovered by treating with FF-8GY. However, the glutathione concentration was more significantly decreased in the GYE supplementation in alcohol feeding rats. Alcohol treatment also negatively affected the serum total protein and albumin, but these were significantly increased near normal levels in FF-8GY coadministered rats. These results suggest that glutathione-enriched Saccharomyces cerevisiae FF-8 strain may have positively mediate the alcohol-induced oxidative stress, and this effect was more pronounced in FF-8GY compared to GYE.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.32 no.6
• /
• pp.791-797
• /
• 1999

This study was conducted in order to evaluate nutritive values of yeasts (Kluyveromyces fragilis and Candida utilis) according to growth stages (early log phase, log phase, stationary phase and death phase) and chemical treatment of their cell wall, Proximate, amino acids, fatty acids and nucleotides composition of the yeast samples was determined. Crude protein content was high in K. fragilis ($48\~59\%$) compared to C. utilis ($26\~43\%$). Crude lipid and fiber contents of the yeasts were below than $1.6\%$ and $3.3\%$, respectively. Conposition of aspartic acid, glycine, proline, leucine, Iysine and valine of K. fragilis were higher than those of C. utilis, and glutamic acid and arginine of C. utilis were higher than those of K. fragilis. Proximate and amino acids composition was not siginificantly influenced by growth stage of the yeasts. Major fatty acids of the yeasts in all growth stages were $C_{10-18}$. $C_{16-18}$ contents were relatively high in the early log or log phase and $C_{10-12}$ contents were relatively high in the stationary or death phase. However, n-3 highly unasturated fatty acids (C$\ge$20) in the all growth stages were not observed. This result indicated that these yeast strains could not be adequate as a dietary lipid source for marine fish. Composition of nucleotides and their related compounds (ATP ADP AMP, IMP and inosine) in the early log phase yeasts were lower than those in the log, stationary and death phase yeasts.

• Proceedings of the Korean Society for Applied Microbiology Conference
• /
• 2005.06a
• /
• pp.154-164
• /
• 2005

Saccharomyces cerevisiae KNU5377 is a thermotolerant strain, which can ferment ethanol from wasted papers and starch at 40$^{\circ}C$ with the almost same rate as at 30$^{\circ}C$. This strain showed alcohol fermentation ability to convert wasted papers 200 g (w/v) to ethanol 8.4% (v/v) at 40$^{\circ}C$, meaning that 8.4% ethanol is acceptable enough to ferment in the industrial economy. As well, all kinds of starch that are using in the industry were converted into ethanol at 40$^{\circ}C$ with the almost same rate as at 30$^{\circ}C$. Hyperthermic cell killing kinetics and differential scanning calorimetry (DSC) revealed that exponentially growing cells of this yeast strain KNU5377 were more thermotolerant than those of S. cerevisiae ATCC24858 used as a control. This intrinsic thermotolernace did not result from the stability of entire cellular components but possibly from that of a particular target. Heat shock induced similar results in whole cell DSC profiles of both strains and the accumulation of trehalose in the cells of both strains, but the trehalose contents in the strain KNU5377 were 2.6 fold higher than that in the control strain. On the contrary to the trehalose level, the neutral trehalase activity in the KNU5377 cells was not changed after the heat shock. This result made a conclusion that though the trehalose may stabilize cellular components, the surplus of trehalose in KNU5377 strain was not essential for stabilization of whole cellular components. A constitutively thermotolerant yeast, S. cerevisiae KNU5377, was compared with a relatively thermosensitive control, S. cerevisiae ATCC24858, by assaying the fluidity and proton ATPase on the plasma membrane. Anisotropic values (r) of both strains were slightly increased by elevating the incubation temperatures from 25$^{\circ}C$ to 37$^{\circ}C$ when they were aerobically cultured for 12 hours in the YPD media, implying the membrane fluidity was decreased. While the temperature was elevated up to 40$^{\circ}C$, the fluidity was not changed in the KNU5377 cell, but rather increased in the control. This result implies that the plasma membrane of the KNU5377 cell can be characterized into the more stabilized state than control. Besides, heat shock decreased the fluidity in the control strain, but not in the KNU5377 strain. This means also there's a stabilization of the plasma membrane in the KNU5377 cell. Furthermore, the proton ATPase assay indicated the KNU5377 cell kept a relatively more stabilized glucose metabolism at high temperature than the control cell. Therefore, the results were concluded that the stabilization of plasma membrane and growth at high temperature for the KNU5377 cell. Genome wide transcription analysis showed that the heat shock responses were very complex and combinatory in the KNU5377 cell. Induced by the heat shock, a number of genes were related with the ubiquitin mediated proteolysis, metallothionein (prevent ROS production from copper), hsp27 (88-fold induced remarkably, preventing the protein aggregation and denaturation), oxidative stress response (to remove the hydrogen peroxide), and etc.