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.
Kim, C.H.;Park, B.K.;Park, J.G.;Kim, H.S.;Sung, K.I.;Shin, J.S.;Ohh, S.J.
Journal of Animal Science and Technology
/
v.47
no.5
/
pp.759-768
/
2005
The study was designed to estimate the in vitro rumen by-pass rate of both chromium methionine chelate as an organic supplement and $ClCl_3$ as an inorganic supplement. Rumen by-pass rates of the supplements were evaluted by comparing ruminal metabolites in rumen fluid and Cr and methionine contents in the body of ruminal microorganism. For in vitro digestion examination, basic nutrients for ruminal microbes were supplied with 7g(DM) of feed, 2g of rice straw, and 2g of corn silage per each incubation jar. Three treatments including Control(no supplementation of Cr), T1(1000ppb supplementation of $ClCl_3$) and T2(chromium methionine chelate supplementation equivalent to 1000ppb of Cr content) were prepared with five replications per each treatment. pH of T2 was lower than that of Control and T1 regardless of incubation time. Ammonia content was higher in T2 than in Control and T1 during first 6 hours of incubation. However, the ammonia content in Control was remained low after 6 hours. Total volatile fatty acids(VFA) content in control was increased constantly as incubation time was extended. Therefore, VFA content in T1 and T2 were significantly lower (P<0.05) than those of Control. Dry matter recovery rate by ruminal microorganism was the lowest in T1, however ruminal microbial population was increased most efficiently in T2 during 12 hours of in vitro incubation. Cr concentrations in the body of ruminal microbes were not different(P>0.05) between Control and T2, but it was significantly high in T1(P<0.05). Contents of methionine and cystine in ruminal microbes also were not different between Control and T2(P>0.05), but it was relatively low in T1. Based on the above results, the chromium methionine chelate was believed to by-pass rumen and could remain intact until it reaches small intestine compared to inorganic chromium. This results implies that chromium methionine chelate could be more effective to function in the small intestine of ruminant animals.
About the phenomenon of being imaged of everything, the scholars of the humanities who had studied on the simple reason structure in a text have been in a big agony how accept it. Especially, semiologists have studied about this for a long time and the points at issues of Saussure, Peirce as well as Umbeto Eco are more outstanding. Being based upon his philosophic interesting from medieval esthetics to modern semiotics, Eco was very concerned about the field of general esthetics and poputar arts like television and cartoons. He connected the mutual open-relations between 'signifier' and 'signified' debated in Semiotics with the open and vague modern arts and regarding it as a deviation from the custom, intensively studied the film-media. Saussure is a representative figure of semiotics and explained Sign and the character of semiotics as the division into two parts such as signifier/ signified, form/ substance, langue/ parole, synchrony/ diachrony. The triadic semiotics (the theory that Sign is composed of the triadic structure like sign, referent and interpretant) of Peirce put the new item- 'interpretant' in sign and referent to connect them and open the possibility to introduce time in to the Sign. In this paper, I try to analyze a cartoon film in the semiotic structure with the systemic, reasonable and logical approach and analysis as as possible. While the images shown through a film were depended on the romantic and impressional judge in the past, due to semiotics, it' s quite possible to correlate the procedure of symbolization to social coherence so that we analyze the incredible power of images to suck audiences with the systemetic Sign. I accept all ot film-images including a cartoon film as not the simple esthetic arts but a social custom and system, want to serve as a aid to properly understand world and humanbeings and prevent the film-image from being mystic. A cartoon and a cartoon film which were begun with the link of a text and an illustration give shape to all of images such as materials, places and even thoughts with a cartoon icon existed in only a cartoon. A cartoon and a cartoon film simply and exquistely conceptualize the complex and vague attribute of an organic creature and extend them infinetly beyond language. However, it can be exploited as a mysticism to temptate the general public and a faking material. In addition to that, it can distort our world-knowledge engaging a political power and the massive power of mass media. In this paper, being based on semiotics to approach a cartoon film in a scientific and organic system, I conclude that a non-linguistic cartoon expression is entangled with the manifold signs and implies the supplementary meanings just like a regular linguistic expression. It remarks that the iconic images of a cartoon film are composed of the social codes and can be analyzed on grounds of a linguistic system.
Acid mine drainage (AMD) from abandoned mine sites typically has low pH and contains high level of various heavy metals, aggravating ground- and surface water qualities and neighboring environments. This study investigated removal of heavy metals in a biological treatment system, mainly focusing on the removal by adsorption on a substrate material. Bench-scale batch experiments were performed with a mushroom compost to evaluate the adsorption characteristics of heavy metals leached out from a mine tailing sample and the role of SRB in the overall removal process. In addition, adsorption experiments were perform using an artificial AMD sample containing $Cd^{2+}$, $Cu^{2+}$, $Pb^{2+}$ and $Zn^{2+}$ to assess adsorption capacity of the mushroom compost. The results indicated Mn leached out from mine tailing was not subject to microbial stabilization or adsorption onto mushroom compost while microbially mediated stabilization played an important role in the removal of Zn. Fe leaching significantly increased in the presence of microbes as compared to autoclaved samples, and this was attributed to dissolution of Fe minerals in the mine tailing in a response to the depletion of $Fe^{3+}$ by iron reduction bacteria. Measurement of oxidation reduction potential (ORP) and pH indicated the reactive mixture maintained reducing condition and moderate pH during the reaction. The results of the adsorption experiments involving artificial AMD sample indicated adsorption removal efficiency was greater than 90% at pH 6 condition, but it decreased at pH 3 condition.
Lee, Jae Won;Kim, Ji Yoon;Kim, Hee gon;Hur, Kyung Jin;Kwon, Chan Hyeok;Hur, Jang Hyun
Korean Journal of Environmental Agriculture
/
v.36
no.1
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pp.57-62
/
2017
BACKGROUND: Pesticide residue analysis is essentially required for safety evaluation of agricultural products. Bistrifluron and chlorantraniliprole have been currently considered as potentials to deeply evaluate their residues in agricultural products because they are frequently found in strawberry. This work was performed to investigate the residual patterns of bistrifluron and chlorantraniliprole in strawberry after harvest. METHODS AND RESULTS: Strawberry was treated with bistrifluron and chlorantraniliprole 0, 1, 2, 3, 5, 7 and 10 days before harvest under greenhouse conditions. The strawberry samples were subjected to solvent and solid phase extractions followed by LC-MS/MS analysis. There covery percentages of bistrifluron and chlorantraniliprole for tified in the control samples ranged from approximately 82 to 103% with the method limit of 0.005 mg/kg. The concentrations of bistrifluron and chlorantraniliprole in strawberry samples decreased significantly in 10 days after treatment, giving the safety levels of 0.04 to 0.06 mg/kg at 10 days after application, as considered maximum residue limit. The half-lives of bistrifluron and chlorantraniliprole based on first order kinetics were determined to 6.3 days and 6.4 days, respectively. CONCLUSION: Bistrifluron and chlorantraniliprole are suggested to use in strawberry 10 days before harvest to reach residual safety levels.
Scuticociliatosis has badly settled one of most damaging diseases during the seedling production process of olive flounder. Paralichthys olivaceus in Korea. We isolated a new type of Scuticociliate from flounder. The parasite metamorphoses to ciliate and cyst phases with each other by environmental changes and survive for a relatively long span. The ciliate was measured average 41.8 ${\mu}m$ in length and 21.0 ${\mu}m$ in width, and cyst was 17.0 ${\mu}m$ and 13.5 ${\mu}m$, respectively. Nutritional condition was determined as a major parameter of metamorphosing between ciliate and cyst stages. The ciliate transforms to a cyst stage because of food shortage, and the cyst returns to a ciliate stage with a favorite environmental condition and shows active growth and reproduction. The ciliate multiplied at the maximal density of $2.9 {\times} 10^5 {m\ell}^{-1}$cells in vitro cultivation at $15 ^{\circ}C$temperature using MS BHI medium and bacterial food sources. The ciliate could be proliferated at a 2.5 to $30 ^ \circ}C$ temperature range, pH 6 to 9, and 1 to 55 ppt salinity. Particularly, it survived over one week at $0 ^{\circ}C$temperature showing a high resistance against unfavorable environmental conditions. And the cyst survived for 320 days in the condition of $5 ^{\circ}C$with no feeding, but its survival period was markedly shortened in higher temperature conditions. The chemotherapeutants (formalin and hydrogen peroxide) were clarified as effective chemicals against the ciliate during in vitro trials, but the effect of therapeutants differed in proportion, depending upon the density and the bathing time of chemical compounds.
Overwintering and succession of phytoplankton community with physicochemical and biological characteristics were investigated in pilot culture system. Water and phytoplankton samples were collected twice a week from February 23 to June 28, 2016. A total of 17 overwintering taxa including cyanophyceae, chlorophyceae, bacillariophyceae were identified in the experimental group in winter (February), and these overwintering species showed a marked succession pattern along with environment changes. In the process of phytoplankton succession, a total of 56 species in 28 genera were identified in two (experimental, control) pilot culture system. In the experimental group, 52 phytoplankton species in 24 genera were identified, and the number of taxa was highest in chlorophyceae (35 species), followed by Bacillariophyceae (9 species), Cyanophyceae (5 species) and others (3 species). In the control group, 25 phytoplankton species in 14 genera were classified and these taxa consisted of 17 chlorophyceae, 3 cyanophyceae, 2 Bacillariophyceae and 3 others. The standing crops ranged from 40 to $325,450cells\;mL^{-1}$ in the experimental group, and from 900 to $37,100cells\;mL^{-1}$ in the control group, respectively. The dominant species were represented by Monoraphidium minutum, Microcystis aeruginosa, Rhodomonas lacustris, Ankyra judai and Chlorella vulgaris in the experimental group; and M. minutum and Coenochloris cf. pyrenoidosa in the control group. In conclusion, overwintering and succession of predominant phytoplankton species developed due to interactions of internal environmental factors(physicochemical and biological factors) in the pilot culture system.
Conditions for artificial culture of Lemna Paucicostata and its nutritional values were examined in this study. Lemna P. was cultured using artificial wastewater and a bioreactor (total volume $2,630\;cm^3$, working volume $2,240\;cm^3$) was operated at conditions of 6,250 lux and $28^{\circ}C$. Water flow affected the growth of Lemna P.: growth rate was very high (more than $1.1\;d^{-1}$) at a condition of no-water movement, but it was very low (less than $0.15\;d^{-1}$) when water moved slowly. The growth of Lemna P. was higher in $16h\;d^{-1}$ light cycle than in Sand $24h\;d^{-1}$, and it was also severely affected by the initial $NH_4$-N levels of wastewater. The growth rate of Lemna P. was high in lower $NH_4$-N level, indicating that the growth rate is in inverse proportion to $NH_4$-N concentration in wastewater. However, the contents of crude protein (CP) of Lemna P. were proportional to the initial $NH_4$-N concentration. The CP contents of Lemna P. cultured at 2, 10, 50 and 100 $NH_4$-N mg $L^{-1}$ was 18, 24, 37, 43%, respectively, showing the Lemna P. cultured at 50 and $100\;mg\;L^{-1}$ had similar protein contents to linseed (CP 35%), cottonseed (CP 38%) and soybean (CP 45%). Fat, protein, fiber, NDF and ADF contents of Lemna P. harvested at conditions of $16h\;d^{-1}$ light cycle and less than $2\;mg\;L^{-1}$ of $NH_4$-N level was 2.8, 18, 27, 20, 41 and 65.7%, respectively. Since the growth rate of Lemna P. was very high (more than $1.1\;d^{-1}$) at those conditions, it was convinced that mass production of valuable protein and fiber sources are feasible. In particular, since the Lemna P. has unsaturated fatty acids found mainly in animal fat as well as beneficial fatty acids to health such as C18:ln9c, C18:2n6c, C20:5n3 and C22:2, the Lemna P. biomass would be a highly valuable alternative feed source to grains.
Kim, Eun-Sic;An, Jun-Geon;Kim, Gi-Beum;Shim, Won-Joon;Joo, Chang-Kyu;Kim, Moon-Koo
Journal of the Korean Society for Marine Environment & Energy
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v.15
no.3
/
pp.247-256
/
2012
Stable carbon isotope ratio of oil components are known to be unaffected by weathering processes and thus has been widely used to determine the origin of spilled oil. In this study, molecular index and composition of stable carbon isotope in 15 crude oils and petroleum product were analyzed and used as oil fingerprints to determine the discriminating power of each fingerprinting method among target crude oils. Through the fingerprints of alkane distribution only Bintulu and B-C(1%) were distinguishable from other crude oils. The pristane/phytane ratio can classify the crude oils into three groups but differentiation of crude oils within a group was impossible using the ratio. The crude oils of A.L., A.S.L., Foroozan and B-C(1%) were differentiated from the other oils using PAH source recognition indexes of C2D/C2P and C3D/C3P. The usage of 4-mD/1-mD and 2/3-mD/1-mD ratio was able to distinguish A.S.L., Bintulu and Oman from the other crude oils. However the PAH source recognition ratios in the other crude oils were similar and thus they were impractical to be used for source identification among the target crude oils. Stable carbon isotope ratios of alkanes were able to uniquely specify each crude oil in the plot of ${\delta}^{13}C_{C21}$ and ${\delta}^{13}C_{C25}$ except A.L., A.M., Qatar-Marine, B-C(1%). The oil fingerprinting method using stable carbon isotope ratios of individual alkane compounds showed more discriminating power among the target crude oils than the conventional source recognition indexes of PAHs or alkanes.
Recently many studies have reported that total and bioavailable androgens reduced in male and female athletes and that physical exercise reduces the body weight and increases the reproductive abnormalities such as oligomenorrhea, anovulation, inadequate luteal phase, and delayed puberty in women by the inhibition of the hypothalamic-pituitary-gonadal (HPG) axis . In addition, high mileage endurance 겨nning, psychological stress, and military endurance training in men also reduce the secretion of reproductive hormones. To investigate the efffcts of physical endurance exercise on the secretion of reproductive hormones in men, androgenic hormones from adrenal glands and testis were measured in serum by the conventional radioimmunoassays after long-term (more than3 months), short-term (1 week), and acute (1${sim}$2 hours) physical exercises. Androgenic hormones from adrenal glands and testis such as total testosterone (TT), free testosterone (fT), dehydroepiandrosterone (DHEA), and androstenedione (A) decreased after thesestrenuous endurance trainings, whereas ACTH, cortisol, and dehydroepiandrosterone sulfetes (DHEAS) increased. Conadotropins (LH and FSH) were not idluenced by the physical exercises. Based upon the present results, we assume that the decrease in adrenal and testicular androgens by physical endurance exercises might be associated with the reproductive abnormalities in athletes by unknown factor(s) in addition to the HPG axis disturbance.
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