• Applied Biological Chemistry
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• v.13 no.3
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• pp.237-242
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• 1970

Fifty strains of mold which isolated from the various sources were screened for the production of Naringinase which hydrolyse naringin, the 7-rhamnoe-glucoside of 4'.5.7. - trihydroxyflavanonin, the main bitter principle of citrus fruits and grape fruits. Of the 4 strains yielded naringinase with significant activity, S-1 strain was selected on the criterion of industrial application, and some properties of crude naringinase of this S-1 was investigated. The results obtained were as follows. 1. Naringenase obtained from S-1 strain has optimum pH range from 3.0 to 5.0 for its activity. 2. Production of naringinase was increased on the addition of naringin to the medium. 3. Hydrolysis of naringin with approporiate concentration of naringinase was carried out linerly up to 80% on the 0.1% substrate solution. 4. The optimum temperature for its activity was $50^{\circ}C$, and this enzyme was inactivated 80% of its total activity at $70^{\circ}C$ for 10 minutes, 40% at $60^{\circ}C$ for 30 minutes. But signifiant decrease of activity were not occurred by heat treatment at $50^{\circ}C$ for 2 hours. 5. Crude enzyme of the naringinase obtained from S-1 strain was competitively inhibited by addition of glucose on the substrate, and inhibitor constant of the glucose on the this enzyme was 1.5 Mol, and inhibition rate were linearly increased according to the increase of sucrose concentration and 56% of its total activity was inhibited at 1 Mol sucrose solution.

• Journal of Korean Society of Environmental Engineers
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• v.30 no.7
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• pp.700-704
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• 2008

Nitrite and free ammonia have been known as substrate inhibitors in anaerobic ammonium oxidation. To reduce inhibitory effect of these substrates, the NH$_3$-N/NO$_2$-N ratio in the influent could be properly controlled in anaerobic ammonium oxidation process. Five kinds of NH$_3$-N/NO$_2$-N ratio were assayed in batch to find optimum NH$_3$-N/NO$_2$-N ratio, curtailing substrate inhibition. As the results of batch test, the highest T-N removal efficiency of 88% was obtained at 1.00 : 1.30 of NH$_3$-N/NO$_2$-N ratio. In addition, rate constants for ammonium and nitrite in zero-order kinetics were found to be the highest values as 7.66 mg/L$\cdot$hr and 11.89 mg/L$\cdot$hr at 1.00 : 1.30 ratio, respectively. However, as for the specific anammox activity, the ratio of NH$_3$-N/NO$_2$-N ratio was recommended as 1 : 1.15 which can maintain the highest SAA during continuous operation and preclude the accumulation of nitrite in the reactor.

• Journal of Korean Society of Forest Science
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• v.39 no.1
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• pp.57-63
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• 1978

Enzymatic hydrolysis of the substrate from Alnus hirsuta (Spach) Rupr (8-14years) was investigated using cellulase preparations of Trichoderma viride Pers. ex. Fr. SANK 16374 and conduced on the optimum reaction conditions of the cellulase on saccharification. The crude cellulase was produced by the submerged culture process and produced in the culture fluid was salted out quantitatively by the use of ammonium sulfate. The method of delignification from wood(Saw dust) was treated by the peracetic acid (PA) method. Reducing sugar was determined by the dinitrosalicylic acid (DNS) method. The results were summerized as follows; 1. The optimum pH of cellulase was 5.0 and the range of stability with respect to pH was generally from 4.0 to 6.0 2. The optimum temperature of cellulase was generally $40^{\circ}C$, but reducing sugar formation did not show significent differences at 5% levels in the reaction temperature from $40^{\circ}C$ to $50^{\circ}C$. 3. The redusing sugar were increased with increase of cellulase concentration. 4. The reducing sugar were decreased with increase of substrate concentration. 5. Fructose was a very good inhibitor of the enzyme from Trichoderma viride, but glucose inhibition was generally weak.

• The Korean Journal of Pesticide Science
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• v.7 no.2
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• pp.100-107
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• 2003

To improve the growth inhibitory activity against the shoot and root of rice plant (Oryza sativa L) and barnyard grass (Echinochloa crus-galli), a series of N-substituted phenyl-3,4-dimethylmaleimdes derivatives as substrates were synthesized and then their the inhibitory activities of protoporphyrinogen oxidase (1.3.3.4), protox were measured. The quantitative structure-activity relationships (QSAR) between structures and the inhibitory activities were studied quantitatively using the 2D-QSAR method. And also, molecular sharp similarity between the substrate derivatives and protogen, substrare of protox enzyme were studied. The activities of the two plants indicated that barnyard grass had a higher activity than the rice plant and their correlation relationships have shown in proportion for each. Accordingly, the results of SARs suggest that the electron donating groups as $R_2=Sub.X$ group will bind to phenyl ring because the bigger surface area of negative charged atoms in the substrate molecule derivatives may increase to the higher the activity against barnyard grass. Based on the molecular shape similarity, when the derivatives and protogen, subsbrate of protox enzyme were superimposed by atom fitting, the similarity indices (S) were above 0.8 level but the correlation coefficients (r) between S values and the activities showed not good.

• Korean journal of applied entomology
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• v.37 no.2
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• pp.179-185
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• 1998

The effect of physiological factors of beet armyworm, Spodoptera exigua (Hubner), on esterase variation was analyzed by comparing electrophoretic esterase isozymes. Each esterase isozyme was also characterized by substrate and inhibitor specificities. A total of 28 esterase isozymes were separated on 10% nondenaturing polycarylamide gel electrophoresis (PAGE). These isozymes were denoted from El to E28 according to cathodal migration distances. There was a variation in esterase isozymes among developmental stages. Larvae and pupae had more isozymes than did adults. Eggs had only eight isozymes. The isozymes of El and E2 were specific only in the first instar larvae. Esterases also showed variation according to different tissues. More kinds of esterase isozymes were found in epidermis and gut tissues than in hemolymph and fat body. Some isozymes were specific in epidermis (from El to E6), gut (E10, El 1, E25, E26, and E27), and hemolymph (E18). Among 10 naphthyl esters, a-naphthyl propionate was the most reactive substrate to the esterase isozymes. The isozymes were classified into cholinesterases (El0 and E24), arylesterases (E4, E9, E17, E19, E21, and E23), and carboxylesterases (the others) on the basis of inhibition by the esterase inhibitors-eserine, dichlorovos, moncrotophos, and paraoxon.

• The Journal of Internal Korean Medicine
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• v.32 no.4
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• pp.565-583
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• 2011

Objectives : This study investigated the biochemical mechanisms of anti-proliferative and pro-apoptotic effects of the water extract of Dan-Seon-Tang (DST) in human leukemia U937 cells. Methods : U937 cells were exposed to DST and growth inhibition was measured by MTT assay. Results : Exposure of U937 cells to DST resulted in the growth inhibition in a concentration-dependent manner. This inhibitory effect was associated with morphological changes and apoptotic cell death such as formation of apoptotic bodies, increased populations of apoptotic-sub G1 phase and induction of DNA fragmentation. The induction of apoptotic cell death in U937 cells by DST was associated with up-regulation of death receptor 4 (DR4) and down-regulation of Bid, surviving and cellular inhibition of apoptosis protein-2 (cIAP-2) expression. DST treatment also induced the proteolytic activation of caspase-3, caspase-8 and caspase-9, and a concomitant degradation of caspase-3 substrate proteins such as poly (ADP-ribose) polymerase (PARP), phospholipase (PLC)-${\gamma}1$, ${\beta}$-catenin and DNA fragmentation factor 45/inhibotor of caspase activated DNAse (DFF45/ICAD). Furthermore, apoptotic cell death by DST was significantly inhibited by caspase-3 specific inhibitor z-DEVD-fmk, demonstrating the important role of caspase-3. Conclusions : These findings suggest that herb prescription DST may be a potential chemotherapeutic agent for the control of human leukemia U937 cells; further study is needed to identify the active compounds.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.19
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• pp.8107-8113
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• 2014

The aim of this study was to investigate the effects of olanzapine on growth inhibition as well as autophagy in glioma cells in vitro and in vivo. The proliferation of both LN229 and T98 glioma cells, measured by MTT assay, was suppressed in a concentration-dependent and time-dependent manner. Moreover, apoptosis of both cells was significantly increased with the treatment of olanzapine as evidenced by increased Bcl-2 expression, Hoechst 33258 staining and annexinV-FITC/PI staining. Olanzapine treatment also enhanced activation of autophagy with increased expression of LC3-II, expression of protein p62, a substrate of autophagy, being decreased. The growth inhibition by olanzapine in both glioma cell lines could be blocked by co-treatment with 3-MA, an autophagy inhibitor. Furthermore, olanzapine effectively blocked the growth of subcutaneous xenografts of LN229 glioma cells in vivo. The increased level of protein LC3-II and decreased level of p62 followed by a decreased level of Bcl-2, suggesting that autophagy may contribute to apoptosis. In addition, reduced proliferation of glioma cells was shown by a decrease of Ki-67 staining and increased caspase-3 staining indicative of apoptosis in mouse xenografts. These results indicated that olanzapine inhibited the growth of glioma cells accompanied by induction of autophagy and apoptosis both in vitro and in vivo. Olanzapine-induced autophagy plays a tumor-suppressing role in glioma cells.

• Journal of Applied Biological Chemistry
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• v.44 no.3
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• pp.135-139
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• 2001

To observe the changes in isotopic composition (${\delta}^{15}N$) of $NO_3{^-}$ during denitrification, an incubation experiment using soil treated with nitrification inhibitor (2-chloro-6-trichloromethyl-pyridine) under water-saturated condition was conducted for 153 h. The $NO_3-N$ concentration decreased from 73.3 to $20.6mg\;kg^{-1}$ during the incubation period, with denitrification rate constant of $0.00905h^{-1}$, and ${\delta}^{15}N$ values of $NO_3-N$ increased from +0.9 to +25.5‰ with decreasing the $NO_3-N$ concentration. The increase in the ${\delta}^{15}N$ values of $NO_3-N$ is due to kinetic isotope fractionation, which always results in $^{15}N$ enrichment of the substrate. The isotopic fractionation factor calculated in this study was 1.0196, an indication that 1.96% more $^{14}NO_3{^-}$ reacted at a given time interval than a comparable number of $^{15}NO_3{^-}$. The ${\delta}^{15}N$ values measured through the incubation study showed a good agreement with the results calculated from the Fochts isotope fractionation model. Our results suggest that when the ${\delta}^{15}N$ of $NO_3{^-}$ is used for tracing the fate of N, the kinetic isotope fractionation associated with denitrification must be taken into consideration.

• Korean Journal of Animal Reproduction
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• v.19 no.1
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• pp.55-63
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• 1995

The putative totipotency germ cells has a relative abundance of alkaline phosphatases. Thus, histological staining of AP activity offers a new route to isolate totipotent cells and also provides insights into culture systems of these cells. Furthermore, the AP staining technique is simple and fast, requires only the napthol AS/MS substrate in combination with trapping diazonium salts such as fast red or fast blue. However, our unexpected finding was that AP staining of mouse ES cells were detected in the undifferentiaed epiblast-derived cells as well as several types of differentiating cells. This findings are different from results of Talbot et al. (1993) reported usefulness of the AP staining and implies that histological staining of AP may not by useful to determine undifferentiaed state or totipotency of ES cells. Thus, we have investigated the patterns of AP expression by RT-PCR in order to identify a marker of undifferentiated ES/primordial germ (PG) cells. In RT-PCR analysis, embryonic (E)-AP was detected only in undifferentiated ES cells, but intestinal(I)-AP was not detected in all of the examined ES and PG cells. In addition, nonspecific (NS)-AP wasdetected in undifferentiated PG cell from day 7, 5 to 13 of gestation. Histological activity of AP in ES cells was completely suppressed by addition of L-phenylalanine (Phe), L-homoarginine (Har), and L-phenylalanylglycylglycine (PheGlyGly) as an inhibitor, but RT-PCR showed the same results as in the absence of an inhibitors. Our findings suggested that expression of E-AP and NS-AP may use as a marker to determine the undifferentiated status in ES and PG cells.

• Korean Journal of Food Science and Technology
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• v.23 no.4
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• pp.414-419
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• 1991

Polyphenol oxidase from Jerusalem artichoke(Helianthus tuberosus L.) tubers was partially purified by precipitation with ammonium sulfate, followed by gel filtration on Sephadex G-100. The enzyme showed maximal activity at pH 6.5 and $4^{\circ}C$. Kinetic studies indicated $K_{m}$ value of 3 mM for catechol and activation energy of 72.6 kcal/mole. As for substrate specificity of polyphenol oxidase the enzyme showed high affinity towards diphenol compounds, but not towards monophenols. The enzamatic browning was completely inhibited at 1 mM concentration of L-ascorbic acid, sodium hydrosulfite and L-cystein(HCl). The activity of polyphenol oxidase in 0.1 M potassium phosphate buffer(pH 6.5) was fairly stable for a week at $4^{\circ}C$, while it decreased remarkably at $25^{\circ}C$.