• Journal of Plant Biotechnology
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• v.2 no.2
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• pp.61-71
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• 2000

By screening a cDNA library of auxin-treated mung bean (Vigna radiata L.) hypocotyls, we have isolated two full-length cDNA clones, pVR-ACS6 and pVR-ACS7, for 1-aminocyclopropane-1-carboxylate (ACC) synthase, the rate-limiting enzyme in the ethylene biosynthetic pathway. While PVR-ACS6 corresponds to the previously identified PCR fragment pMBA1, pVR-ACS7 is a new cDNA clone. A comparison of deduced amino acid sequences among auxin-induced ACC synthases reveal that these enzymes share a high degree of homology (65-75%) to VR-ACS6 and VR-ACS7 polypeptides, but only about 50% to VR-ACS1 polypeptide. ACS6 and ACS7 are specifically induced by auxin, while ACS1 is induced by cycloheximide, and to lesser extent by excision and auxin treatment. Results from nuclear run-on transcription assay and RNA gel blot studies revealed that all three genes were transcriptionally active displaying unique patterns of induction by IAA and various hormones in etiolated hypocotyls. Particularly, 24-epibrassinolide (BR), an active brassinosteroid, specifically enhanced the expression of VR-ACS7 by distinct temporal induction mechanism compared to that of IAA. In addition, BR synergistically increased the IAA-induced VR-ACS6 and VR-ACS7 transcript levels, while it effectively abolished both the IAA- and kinetin-induced accumulation of VR-ACS1 mRNA. In light-grown plants, VR-ACS1 was induced by IAA in roots, whereas W-ACS6 in epicotyls. IAA- and BR-treatments were not able to increase the VR-ACS7 transcript in the light-grown tissues. These results indicate that the expression of ACC synthase multigene family is regulated by complex hormonal and developmental networks in a gene- and tissue-specific manner in mung bean plants. The VR-ACS7 gene was isolated, and chimeric fusion between the 2.4 kb 5'-upstream region and the $\beta$-glucuronidase (GUS) reporter gene was constructed and introduced into Nicotiana tobacum. Analysis of transgenic tobacco plants revealed the VR-ACS7 promoter-driven GUS activity at a highly localized region of the hypocotyl-root junction of control seedlings, while a marked induction of GUS activity was detected only in the hypocotyl region of the IAA-treated transgenic seedlings where rapid cell elongation occurs. Although there was a modest synergistic effect of BR on the IAA-induced GUS activity, BR alone failed to increase the GUS activity, suggesting that induction of VR-ACS7 occurs via separate signaling pathways in response to IAA and BR.

• Microbiology and Biotechnology Letters
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• v.45 no.3
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• pp.226-235
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• 2017

Carotenoids such as phytoene, lycopene, and ${\beta}-carotene$ are used as food colorants, animal feed supplements, and for human nutrition and cosmetic purposes. Previously, we reported the isolation of a novel marine bacterium, Kocuria gwangalliensis, which produces a pink-orange pigment. Phytoene desaturase (CrtI), encoded by the gene crtI, catalyzes lycopene formation from phytoene and is an essential enzyme in the early steps of carotenoid biosynthesis. CrtI is one of the key enzymes regulating carotenoid biosynthesis and has been implicated as a rate-limiting enzyme of the pathway in various carotenoid synthesizing organisms. Here, we report the cloning of the crtI gene responsible for lycopene biosynthesis from K. gwangalliensis. The gene consisted of 1,584 bases encoding 527 amino acid residues. The nucleotide sequence of the crtI gene was compared with that of other species, including Kocuria rhizophila and Myxococcus xanthus, and was found to be well conserved during evolution. An expression plasmid containing the crtI gene was constructed (pCcrt1), and Escherichia coli cells were transformed with this plasmid to produce a recombinant protein of approximately 57 kDa, corresponding to the molecular weight of phytoene desaturase. Lycopene biosynthesis was confirmed when the plasmid pCcrtI was co-transformed into E. coli containing the plasmid pRScrtEB carrying the crtE and crtB genes required for lycopene biosynthesis. The results from this study will provide valuable information on the primary structure of K. gwangalliensis CrtI at the molecular level.

• Journal of Animal Science and Technology
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• v.48 no.5
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• pp.719-726
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• 2006

Bovine whey protein are rich in cysteine, which is the rate limiting amino acid for synthesis of antioxidant glutathione(GSH). Some strains of Lactobacillus caseihas been reported to contain high level of GSH in cell extracts. The objective ofthis study was to determine whether enzymatically hydrolyzed whey protein isolate(WPI) and cell extract of Lb. casei HY2782 could increase intracellular GSH concentrations and protect against oxidant induced cell death in human prostate epithelial cell line (designated as RWPE1, and PC3MMM2 cells). Treatment of RWPE1 cellsandPC3MMM2 cells with hydrolyzed WPI (500g/ml) significantly increased GSH by28.2% and38.4% respectively. Compared with control cells receiving no hydrolyzed WPI(P<0.05). hydrolyzed WPI and Lb casei HY2782 cell extracts significantly protected RWPE1 and PC3MMM2 cellsfrom oxidant induced cell death compared with controls receiving no WPI. DNA damage associated with oxidant treatment was demonstrated by single cell gel (SCG) electrophoresis.

• Horticultural Science & Technology
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• v.30 no.4
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• pp.417-425
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• 2012

Salinity stress is one of the most serious factors limiting the productivity of agricultural crops. The aim of this study was to assess inter-cultivar (intraspecific) variation for salinity tolerance in six Korean rapeseed (Brassica napus L.) cultivars at the seedling stage. The effect of three different salinity stress levels (EC 4, 8, and 16 $dS{\cdot}m^{-1}$) on seedlings of six cultivars was investigated through leaf size, leaf dry weight, and leaf chlorosis. At the highest salinity level (16 $dS{\cdot}m^{-1}$), the mean decrease of leaf dry weight in 'Sunmang', 'Tammi', 'Tamla', 'Naehan', 'Youngsan', and 'Halla' was about 56.2, 56.9, 78.4, 79.3, 77.4, and 80.9%, respectively. 'Tammi' and 'Sunmang' showed much less reduction in leaf dry weight than all the other cultivars. In addition, diluted seawater treatments increased the occurrence of leaf chlorosis in six cultivars. At EC 8 and 16 $dS{\cdot}m^{-1}$, 'Naehan', 'Youngsan', and 'Halla' showed a higher level of leaf chlorosis than 'Tammi' 'Sunmang', and 'Tamla'. On the basis of these results, six cultivars were placed into salinity-tolerant and sensitive groups. 'Tammi' and 'Sunmang' were the salinity-tolerant cultivars, while 'Naehan', 'Halla', 'Youngsan', and 'Tamla' were the salinity-sensitive cultivars. 'Tammi' and 'Naehan' rated as the most tolerant and most sensitive cultivar, respectively. To further analyze protein expression profiles in 'Tammi' and 'Naehan', 2-D proteomic analysis was performed using the plants grown under diluted seawater treatments. We identified eight differentially displayed proteins that participate in photosynthesis, carbon assimilation, starch and sucrose metabolism, amino acid metabolism, cold and oxidative stress, and calcium signaling. The differential protein expressions in 'Tammi' and 'Naehan' are likely to correlate with the differential growth responses of both cultivars to salinity stress. These data suggest that 'Tammi' is better adapted to salinity stressed environments than 'Naehan'.

• Molecules and Cells
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• v.38 no.4
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• pp.373-379
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• 2015

Pyruvate kinase M2 isoform (PKM2), a rate-limiting enzyme in the final step of glycolysis, is known to be associated with the metabolic rewiring of cancer cells, and considered an important cancer therapeutic target. Herein, we report a novel PKM2 activator, PA-12, which was identified via the molecular docking-based virtual screening. We demonstrate that PA-12 stimulates the pyruvate kinase activity of recombinant PKM2 in vitro, with a half-maximal activity concentration of $4.92{\mu}M$, and effectively suppresses both anchorage-dependent and -independent growth of lung cancer cells in non-essential amino acid-depleted medium. In addition, PA-12 blocked the nuclear translocalization of PKM2 in lung cancer cells, resulting in the inhibition of hypoxia response element (HRE)-mediated reporter activity as well as hypoxia-inducible factor 1 (HIF-1) target gene expression, eventually leading to the suppression of cell viability under hypoxia. We also verified that the effects of PA-12 were dependent on PKM2 expression in cancer cells, demonstrating the specificity of PA-12 for PKM2 protein. Taken together, our data suggest that PA-12 is a novel and potent PKM2 activator that has therapeutic implications for lung cancer.

• Journal of Nutrition and Health
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• v.8 no.1
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• pp.15-37
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• 1975

I. Subject of the study A study on the nutritive value and utilization of powdered seaweeds. II. Purpose and Importance of the study A. In Korea the shortage of food will be inevitable by the rapidly growing population. It will be very important study to develop a new food from the seaweeds which were not used hitherto for human consumption. B. The several kinds of seaweeds have been used by man in Korea mainly as side-dishes. However, a properly powdered seaweed will enable itself to be a good supplement or mixture to certain cereal flours. C. By adding the powdered seaweed to any cereals which have long been staple foods in this country the two fold benefits; saving of cereals and change of dietary pattern, will be secured. III. Objects and scope of the study A. Objects of the study The objects will come under four items. 1. To develop a powdered seaweed as a new food from the seaweeds which have been not used for human consumption. 2. To evaluate the nutritional quality of the products the analysis for chemical composition and animal feeding experiment will be conducted. 3. Experimental cocking and accepability test will be conducted for the powdered products to evaluate the value as food stuff. 4. Sanitary test and also economical analysis will be conducted for the powdered products. B. Scope of the study 1. Production of seaweed powders Sargassum fulvellum growing in eastern coast and Sargassum patens C.A. in southern coast were used as the material for the powders. These algae, which have been not used for human consumption, were pulverized through the processes of washing, drying, pulverization, etc. 2. Nutritional experiments a. Chemical composition Proximate components (water, protein, fat, cellulose, sugar, ash, salt), minerals (calcium, phosphorus, iron, iodine), vitamins (A, $B_1,\;B_2$ niacin, C) and amino acids were analyzed for the seaweed powders. b. Animal feeding experiment Weaning 160 rats (80 male and 80 female rats) were used as experimental animals, dividing them into 16 groups, 10 rats each group. Each group was fed for 12 weeks on cereal diet (Wheat flour, rice powder, barley powder, potato powder, corn flour) with the supplementary levels of 5%, 10%, 15%, 20% and 30% of the seaweed powder. After the feeding the growth, feed efficiency ratio, protain efficiency ratio and ,organs weights were checked and urine analysis, feces analysis and serum analysis were also conducted. 3. Experimental cooking and acceptability test a. Several basic studies were conducted to find the characteristics of the seaweed powder. b. 17 kinds of Korean dishes and 9 kinds of foreign dishes were prepared with cereal flours (wheat, rice, barley, potato, corn) with the supplementary levels of 5%, 10%, 15%, 20% and 30% of the seaweed powder. c. Acceptability test for the dishes was conducted according to plank's Form. 4. Sanitary test The heavy metals (Cd, Pb, As, Hg) in the seaweed powders were determined. 5. Economical analysis The retail price of the seaweed powder was compared with those of other cereals in the market. And also economical analysis was made from the nutritional point of view, calculating the body weight gained in grams per unit price of each feeding diet. IV. Results of the study and the suggestion for application A. Chemical composition 1. There is no any big difference in proximate components between powders of Sargassum fulvellum in eastern coast and Sargassum patens C.A. in southern coast. Seasonal difference is also not significant. Higher levels of protein, cellulose, ash and salt were found in the powders compared with common cereal foods. 2. The levels of calcium (Ca) and iron (Fe) in the powders were significantly higher than common cereal foods and also rich in iodine (I). Existence of vitamin A and vitamin C in the Powders is different point from cereal foods. Vitamin $B_1\;and\;B_2$ are also relatively rich in the powders.'Vitamin A in ·Sargassum fulvellum is high and the levels of some minerals and vitamins are seemed4 to be some influenced by seasons. 3. In the amino acid composition methionine, isoleucine, Iysine and valine are limiting amino acids. The protein qualities of Sargassum fulvellum and Sargassum patens C.A. are seemed to be .almost same and generally ·good. Seasonal difference in amino acid composition was found. B. Animal feeding experiment 1. The best growth was found at.10% supplemental level of the seaweed Powder and lower growth rate was shown at 30% level. 2. It was shown that 15% supplemental level of the Seaweed powder seems to fulfil, to some extent the mineral requirement of the animals. 3. No any changes were found in organs development except that, in kidney, there found decreasing in weight by increasing the supplemental level of the seaweed powder. 4. There is no any significant changes in nitrogen retention, serum cholesterol, serum calcium and urinary calcium in each supplemental level of the seaweed powder. 5. In animal feeding experiment it was concluded that $5%{\sim}15%$ levels supplementation of the seaweed powder are possible. C. Experimental cooking and acceptability test 1. The seaweed powder showed to be utilized more excellently in foreign cookings than in Korean cookings. Higher supplemental level of seaweed was passible in foreign cookings. 2. Hae-Jo-Kang and Jeon-Byung were more excellent than Song-Pyun, wheat cake, Soo-Je-Bee and wheat noodle. Hae-Je-Kang was excellent in its quality even as high as 5% supplemental level. 3. The higher levels of supplementation were used the more sticky cooking products were obtained. Song-Pyun and wheat cake were palatable and lustrous in 2% supplementation level. 4. In drop cookie the higher levels of supplementation, the more crisp product was obtained, compared with other cookies. 5. Corn cake, thin rice gruel, rice gruel and potato Jeon-Byung were more excellent in their quality than potato Man-Doo and potato noodle. Corn cake, thin rice gruel and rice gruel were excellent even as high as 5% supplementation level. 6. In several cooking Porducts some seaweed-oder was perceived in case of 3% or more levels of supplementation. This may be much diminished by the use of proper condiments. D. Sanitary test It seems that there is no any heavy metals (Cd, Pb, As, Hg) problem in these seaweed Powders in case these Powders are used as supplements to any cereal flours E. Economical analysis The price of the seaweed powder is lower than those of other cereals and that may be more lowered when mass production of the seaweed powder is made in future. The supplement of the seaweed powder to any cereals is also economical with the criterion of animal growth rate. F. It is recommended that these seaweed powders should be developed and used as supplement to any cereal flours or used as other food material. By doing so, both saving of cereals and improvement of individual's nutrition will greatly be achieved. It is also recommended that the feeding experiment for men would be conducted in future.