The bifunctional PheA protein, having chorismate mutase and prephenate dehydratase (CMPD) activities, is one of the key regulatory enzymes in the aromatic amino acid biosynthesis in Escherichia coli, and is negatively regulated by an end-product, phenyalanine. Therefore, PheA protein has been thought as useful for protein engineering to utilize mass production of essential amino acid phenylalanine. To obtain feedback resistant PheA protein against phenylalanine, we mutated by using random mutagenesis, extensively screened, and obtained $pheA^{FBR}$ gene encoding a feedback resistant PheA protein. The mutant PheA protein contains substitution of Leu to Phe at the position of 118, displaying that higher affinity (about $290{\mu}M$) for prephenate in comparison with that (about $850{\mu}M$) of wild type PheA protein. Kinetic analysis showed that the saturation curve of $PheA^{FBR}$ against phenyalanine is hyperbolic rather than that of $PheA^{WT}$, which is sigmoidal, indicating that the L118F mutant enzyme has no cooperative effects in prephenate binding in the presence of phenylalanine. In vitro enzymatic assay showed that the mutant protein exhibited increased activity by above 3.5 folds compared to the wild type enzyme. Moreover, L118F mutant protein appeared insensitive to feedback inhibition with keeping 40% of enzymatic activity even in the presence of 10 mM phenylalanine at which the activity of wild type $PheA^{WT}$ was not observed. The substitution of Leu to Phe in CMPD may induce significant conformational change for this enzyme to acquire feedback resistance to end-product of the pathway by modulating kinetic properties.
The study analyzed the relationship between harmful cyanobacteria and physicochemical factors in Lake Juam from 2005 to 2018. The research locations were designated St. 1 (Juam-Dam) and St. 2 (Sinpyong). Harmful cyanobacteria was found in four genera (Microcystis sp., Anabaena sp., Aphanizomenon sp., Oscillatoria sp.). The average standing crops of harmful cyanobacteria in both locations were 2,575 cells mL-1 and 2,557 cells mL-1 from 2005 to 2011. Since 2012, there has been a significant decrease that the measurements were 42 cells mL-1 and 82 cells mL-1 from 2012 to 2018. To analyze the reason for the decrease in harmful cyanobacteria, Pearson's correlation and t-tests were performed on data collected during the summer period (June-September). Pearson's correlation showed a significantly positive correlation with total nitrogen(TN), outflow, and storage and a negative correlation with electrical conductivity. T-tests were conducted in two different periods and showed decreases in total nitrogen, electrical conductivity, and residence time. The average rainfall was decreased from 263.3 mm (2005-2011) to 219.9 mm (2012-2018) and total nitrogen was decreased from 0.912 mg L-1 (2005-2011) to 0.811 mg L-1 (2012-2018) and the same variability was seen in TP (total phosphorus). Therefore, it seems that the low-rainfall decreased the nutrients (TN) and variability in the TP, resulting in a decrease in harmful cyanobacteria in Lake Juam.
Present study have been performed to develop Bulnesia sarmienti as a functional food. Methanol, n-hexane, chloroform, ethyl acetate and butanol extracts of Bulnesia sarmienti contained total phenol by 5.81 to 7.47%. It is high content than fruits which were known as high contests of total phenol. The electron donating ability of the extract of Bulnesia sarmienti were increased along with increasing concentrations of extracts. At $500{\mu}g/mL\;and\;1000{\mu}g/mL$, the all extracts showde more than 80% of scavenging abilities, which means the equal effect of the antioxidant, BHT. Nitrite scavenging abilities were measured as follows: methanol, butanol, 5.53, 5.77% at $100{\mu}g/mL$, respectively. The ethyl acetate extract was 73.29% at $1000{\mu}g/mL$ which showed the highest activity and methanol, butanol, n-hexane, chloroform and water extract were 65.65, 65.02, 47.49, 52.51, 45.54% which also showed relatively high activities. The growth inhibitory effects of each solvent extract on tumor cell were as follows: test against SUN-1, the gastric carcinoma cell, exhibited the highest inhibitory effects at $100{\mu}g/mL$ where the n-hexane extract was 61.6%. The ethyl acetate and water extracts did not revealed any inhibitory effects. Hela, the uterine carcinoma cell, exhibited the highest inhibitory effects at $100{\mu}g/mL$ where the n-hexane extract was 75.1%. The water extracts did not revealed any inhibitory effects. HT-29, the colon carcinoma cell, also exhibited the highest inhibitory effects at $100{\mu}g/mL$ where n-hexane extract was 57.4%. In conclusion, Bulnesia sarmienti have been shown the antioxidant and antitumor effects, and that it is expected to be developed as functional foods.
Park, Kyung-Seo;Hong, Young-Pyo;Moon, Woon-Ki;Choi, Shin-Suk;An, Kwang-Guk
Korean Journal of Ecology and Environment
/
v.38
no.1
s.110
/
pp.73-82
/
2005
This study was conducted, based on the field survey and laboratory observations, to elucidate egg developmental processes and their characteristics of the Korean slender gudgeon, Squalidus gracilis majimae. For the experiments, the mature adults were collected at the Woongcheon-Cheon Stream and Boreung Reservoir located in Boreung City, Chungnam Province and eggs were obtained from the natural spawning area. Morphological characteristics of the egg and embryonic development were summarized as follows: The shape of the fertilized egg was spherical, adhesive and transparent. The fertilized egg was 2.9${\pm}$0.3 mm (n = 30) in mean diameter under water temperature of $26{\pm}1.5^{\circ}C$, light white in color and had no oil droplets. After 20 minutes from the time of fertilization, a blastodisc was formed and divided into two cells at 48 minutes after fertilization. The blastular stage occurred at 5 hours 40 minutes after fertilization and the gastrular stage was detected at 8 hours 41 minutes after fertilization. The beginning of embryo formation was observed at 12 hours 58 minutes after fertilization and optic vesicles and 9 somites were discovered at 17 hours 05 minutes after fertilization. Differentiation of brains and embryo wiggling were observed at 37 hours 27 minutes after fertilization. Heart beating and the formation of melanophores in optic vesicles were detected at 44 hours 46 minutes after fertilization. The formation of pectoral fins and melanophores in the body were discovered at 50 hours 36 minutes after fertilization. Hatching occurred at 57 hours 49 minutes after fertilization. The newly hatched larvae were 3.3${\pm}$0.2 mm (n = 120) in total length. We believe that these results may contribute the species and population conservations under the situation of accelerated water pollution and the decreases of its diversity.
Park, Myung-Hwan;Chung, Ill-Min;Kim, Baik-Ho;Hwang, Soon-Jin
Korean Journal of Ecology and Environment
/
v.41
no.spc
/
pp.99-106
/
2008
We examined the effects of crude and eight pure material (${\beta}$-sitosterol, ${\beta}$-sitosterol-${\beta}$-D-glucoside, 1-tetratriacontanol, hentriacontane, orizaterpenoid, stigmas-5-en-$3{\alpha}$ 26-diacetate, stearic acid, myristic acid), extracted from rice hull, on growth inhibition of toxic cyanobacterium, Microcystis aeruginosa NIER 10010. Strains of M. aeruginosa and Daphnia magna, obtained from the NIER (Korea) and BBE (Germany), were cultured in the CB medium with hard water. For all four treatment concentrations 0, 10, 100 and $1,000{\mu}g\;L^{-1}$) of the crude extract, the cell number of M. aeruginosa was reduced by $59{\sim}73%$ during the 7-day test period. Among eight kinds of pure extracts, ${\beta}$-sitosterol-${\beta}$-D-glucoside, hentriacontane and orizaterpenoid $(1,000{\mu}g\;L^{-1})$ exhibited relatively higher growth inhibition compared with other pure extracts. The mixture of three pure extracts (${\beta}$-sitosterol-${\beta}$-D-glucoside, hentriacontane and orizaterpenoid) showed the highest growth inhibition at $1,000{\mu}g\;L^{-1}$. Therefore, the synergistic effect was significantly highlighted by a mixture of the three pure extracts (p<0.05). Under the condition of $1,000{\mu}g\;L^{-1}$ in the crude extracts, D. magna exhibited survival rate by >85% for 96 hours. In conclusion, the growth inhibition of M. aeruginosa was probably attributed to the synergistic effect of various compounds extracted from the rice hull.
Journal of The Korean Society of Grassland and Forage Science
/
v.6
no.3
/
pp.157-163
/
1986
Synthesis and accumulation pattern of Weender components in orchardgrass (Dactylis glomerata L.) cv. Potomac and Baraula, perennial ryegrass (Lolium perenne L.) cv. Reveille and Semperweide and meadow fescue (Festuca pratensis Huds.) cv. Cosmos 11 and N.F.G. were studied under different growth environments and cutting managements. The field experiments were conducted as a split plot design with three cutting regimes of 6-7 cuts at grazing stage, 4-5 cuts at silage and 3 cuts at hay stage from 1975 to 1979 in Korea and West Germany. The results obtained are summarized as follows: 1. Air temperature, rainfalls and solar radiation were found to be an important meteorological factors influenced to synthesis and accumulation of Weender components. Under high temperature and strong solar radiation during summer season in Korea, accumulation of crude fiber and cell-wall constituents (NDF) in the plants, as average of all grass species and cutting regimes, were increased to about 30.1% and 48.7% from 27,9% and 42.9% in spring, respectively, while total nonstructural carbohydrates (TNC) were decreased to 1.52% in summer from 4.01% in spring. In West Germany, the concentration of Weeder components showed little seasonal variation. 2. Crude fiber and neutral detergent fiber (NDF) were shown higher concentration in orchardgrass than those of perennial ryegrass and meadow fescue, but N-free extractions and TNC as well as net energy value were less accumulated in orchardgrass. Orchardgrass contained lower net energy contents with 534 StE. 431 StE and 575 StE/kg for Suweon, Cheju and Freising, respectively, as compared with 624 StE (Suweon), 491 StE (Cheju) and 657 StE/kg (Freising) in meadow fescue.
This study was conducted to investigate the effects of taro powder on body weight and lipid metabolism in high fat and cholesterol fed mice. C57BL6 mice were divided into four groups; normal (normal) diet group, a high fat and cholesterol (HF) diet group, and high fat and cholesterol diet groups with 20% taro powder (HF-taro 20%) and 30% taro powder (HF-taro 30%). After 8 weeks, body weight in the HF group increased to 175% of the normal group, while those of HF-taro 20% and HF-taro 30% group decreased by 12.5 and 14% compared to the HF group. The HF-taro 20% and HF-taro 30% groups had significantly reduced epididymal adipose tissue weight. Plasma triglyceride, total cholesterol, and LDL-cholesterol concentrations were significantly higher in the HF group than those in the normal group, while their amounts were significantly diminished in the HF- taro 20% and HF- taro 30% groups. Furthermore, the leptin and insulin concentrations in blood plasma decreased significantly in the HF-taro 20% and HF-taro 30% groups. Accumulation of triglycerides and cholesterol in the liver was significantly inhibited in the HF- taro 20% and HF-taro 30% groups through liver weights, a blood analysis, and histological findings. These results suggest that taro powder may be an effective material for anti-obesity by reducing plasma triglyceride and cholesterol levels.
Park, Chang-Kyu;Tu, Qi;Cho, Ju-Hyun;Yu, Kwang-Won;Jeong, Heon-Sang;Lee, Hyeon-Yong;Jeong, Jae-Hyun
Korean Journal of Food Science and Technology
/
v.42
no.1
/
pp.82-89
/
2010
To obtain functional materials from a submerged culture of Hericium erinaceum, a suitable basal medium for flask culture was screened and the optimal culture conditions in a jar fermenter were investigated with the addition of ginseng extracts (GE) to the basal liquid medium. Of all tested basal liquid media, the mushroom complete medium (MCM) supplemented with 0.5% of GE produced the highest mycelial dry weight (MDW) of 5.91 g/L in the flask, which reached a plateau at $25^{\circ}C$, pH 5.5 after 10 days. The submerged culture conditions for the mass production of mycelia in a 50 L jar fermenter were also optimal at $25^{\circ}C$, pH 5.5, 120 rpm agitation speed and 0.4 vvm aeration rate. Under these conditions, the maximum MDW was produced, which reached a value of 4.28 g/L within 5 days. When we investigated the effects of the amount of GE in the MCM on the production of MDW in the jar fermenter, the addition of 5% GE (HE-GE-5) under the optimal culture conditions produced the maximum MDW (4.93 g/L). In addition, the crude polysaccharide of HE-GE-5 contained mainly neutral sugars (63.2%) with considerable amounts of uronic acid (19.3%) and a small amount of proteins (8.8%) and it had potent immunostimulation properties.
Predetermination of sex in livestock of offpring is in great demand and is of critical importance to providing for the most efficient production of the animal ariculture. Such a sexing techlology would also enhance the economy of conventional artificial insemination(AI) and aid the porcine industry. The purpose of this study was to evaluate the efficiency of enriching X-bearing porcine sperm using discontinuous percoll gradients and PCR mefhod. Semen was collected from mature boars of proven fertility center (AI center KimHae). Sperm was leaded on the isotonic discontinuous percoll gradient and then it was centrifuged at 120 ${\times}$ g for 20 minutes. After centrifugation, sperm included in each fraction were recovered (7${\times}$10$^6$ sperms/ml) and then sperm genomic DNA was extractedfor the PCR. SRY gene was used to evaluate the ratio between X and Y sperm in the separated fractions. Ju viro ffrtilization wascarried out by adding the unseparated sperm (control) or separated (experimental poop) to the matured oocytes in TCM-199. Embryos for sex determination were obtained at 2 cell stage and then was used for SRY gene amplification. After centrifugation of discontinuous percoll gradient, the most motile sperm was obtained at 95% fiaction (94.4% ${\pm}$ 5.1%, p < 0.01). The PCR analysis evaluated that 30%, 50% and 65% fractions were Y sperm rich, whereas 80% and 95% fractions were X sperm rich. PCR analysis with each porcineembryo showed that 33.3% of control and 66.7% of experimental group were determined as female embryos. In conclusion, in vitro matured oocytes inseminated with sperms (95% fraction) prepared by percoll gradient centrifugation showed high fertilization rates and female embryos than control sperms.
Lipopolysaccharide(LPS) was posttreated after the 14 day-pretreatment of Ulva lactuca fractions(ULF), and their correlativity to enzymatic activity alteration was investigated in the liver of rats. ULF was intraperitoneally administered into rats at dose of $1m{\ell}/kg$ of 100 mg/kg concentration for 14 days. On the day 15, $1m{\ell}/kg$ of LPS was injected. The corelativity was examined by measuring the changed values of superoxide dismutase(SOD) in mitochondrial fraction and catalase(CAT), glutathione peroxidase(GPx) in liver homogenate. The results showed that LPS treatment decreased the high values of SOD, CAT, GPx to the low values, but ULF pretreatment increased the low values of SOD, CAT, GPx to the high values. It was suggested that ULF, LPS and antioxidative enzymes like SOD, CAT, GPx had the corelativity of the high-low-high pattern and that the ULF pretreatment played the proper preventive role in the protection against the LPS treatment-induced enzymatic inactivity in the water fraction.
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