• Applied Biological Chemistry
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• v.50 no.4
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• pp.321-326
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• 2007

An organophosphorus insecticide, chlorpyrifos, has been of a great concern due to persistence, toxicity and accumulation in soils and groundwaters. This study deals with degradation efficiency and dechlorination kinetics of chlorpyrifos by various types of zerovalent irons (ZVIs) for effective remediation of the soils contaminated with chlorinated pesticides. Chlorpyrifos degradation rate was increased with increasing ZVI treatment amount and reaction time. The degradation rate and dechlorination kinetics of chlorpyrifos increased in the order of mZVI > nZVI > cZVI in solutions and soils. Dechlorination number value of chlorpyrifos by cZVI, nZVI and mZVI treatment exhibited 1.08, 3.09 and 3.18, respectively. In soils, degradation efficiency and kinetics of chlorpyrifos significantly were affected by moisture content because of the limited contact between ZVIs and chlorpyrifos. These results suggest that nanosized and functionalized mZVI could be effectively applied to degradation of chlorinated pesticides in the soil and aqueous environments.

• Journal of Korean Society on Water Environment
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• v.18 no.2
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• pp.201-212
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• 2002

The objective of this study was to investigate the treatment efficiency, kinetics, and morphological characteristics of biofilm in upflow $Biobead^{(R)}$ process, a kind of biological aerated filter(BAF). The $Biobead^{(R)}$ system showed high removal rates of $COD_{Mn}$(76~83%), $BOD_5$(67~88%) and SS(71~91%) for food wastewater with high salt concentration ($>4,000mg/{\ell}$) under short reaction times(2~3hrs). Even at aerobic condition, the system had high treatment efficiency for both T-N (51~63%) and T-P(62~81%). The removal kinetics of $COD_{Mn}$, $BOD_5$, T-N, T-P, and $Cl^-$ in the $Biobead^{(R)}$ system showed a plug-flow pattern with reaction rate constants($hr^{-1}$) of 0.58, 0.63, 0,30, 0.48, and 0.38 respectively. A backwashing process to remove excess biomass and filtered solids was needed at least once during 22-hour operation at $0.5kg\;BOD\;m^{-3}{\cdot}d^{-1}$ loading. At the higher loading($1.0kg\;BOD\;m^{-3}{\cdot}d^{-1}$) the backwashing interval was shorten by 8 hours. The COD, BOD, T-N, and T-P were removed from 43 to 66% only by aerobic biodegradation. The SS was removed over 70% by the filtering of $Biobead^{(R)}$ media in the treatment system. The first one of three serial Biobead reactors showed the highest removal values for $COD_{\alpha}$(52.3%), $COD_{Mn}$(38.8%), BOD(62.5%), and T-N(40.0%). The SS and T-P had the highest removal values(47.5% and 29.2%) at the second one of the serial reactors. The biofilm had non-homogeneous spatial distribution and the colonies were embedded in the sunk area of the Biobead. The thickness of the biofilm was very thin ($5.0{\sim}29.4{\mu}m$) compared to the biofilm thickness($200{\sim}300{\mu}m$) used in other BAF systems.

• Animal cells and systems
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• v.3 no.2
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• pp.229-236
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• 1999

The recA gene plays a central role in genetic recombination and SOS DNA repair in Escherichia coli (E. coli). We have previously identified a 42 kDa RecA-like protein inducible by a variety of DNA damages from a fluorescent Pseudomonas strain sp. and characterized its inducible kinetics. In the present study, we cloned and characterized the gene encoding the RecA-like protein by immunological screening of Pseudomonas genomic expression library using polyclonal E. coli anti-RecA antibodies as a probe. From 10$^{5}$ plaques screened, five putative clones were finally isolated. Southern blot analysis indicated that four clones had the same DNA inserts and the recA-like gene was located within the 3.2 kb EcoRI fragment of Pseudomonas chromosomal DNA. In addition, the cloned recA-like gene was transcribed into an RNA transcript approximately 1.1 kb in size, as judged by Northern blot analysis. The cellular level of RNA transcript of the cloned recA-like gene was increased to an average of 5.15- fold upon treatment with DNA damaging agents such as ultraviolet (UV)- light, nalidixic acid (NA), methyl methanesulfonate (MMS), and mitomycin-C (MMC). These results suggest that the cloned gene is inducible by DNA damage similarly to the recA gene in E. coli. However, the cloned gene did not restore the DNA damage sensitivity of the E. coli recA-mutant.

• Journal of Applied Biological Chemistry
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• v.63 no.3
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• pp.267-274
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• 2020

The dissipation characteristics and kinetics of fungicide mandipropamid and insecticide thiamethoxam in lettuce under greenhouse conditions were investigated at three different lettuce-growing fields for estimating the pre-harvest residue limits (PHRLs). The analytical methods were fully validated for the quantitation of pesticide residues using High-Performance Liquid Chromatography-Photo Diode Array detector or Ultraviolet-Visible Detector and applied to real samples. The lettuces suitable for shipment were harvested during 10 days including pre-harvest interval after treatment at the recommended dose by safe-use guidelines. The initial mean residues in different fields were 6.68-17.87 and 4.96-8.31 mg/kg for mandipropamid and thiamethoxam, respectively, which decreased to 16-54 and 14-44% in 10 days. The clothianidin, a metabolite of thiamethoxam, was detected in <0.02 to 0.37 mg/kg. The dissipation of both pesticides followed first-order kinetics over a period of 10 days after application. Based on the residue data, the mean dissipation rate constant (λ) and biological half-lives (T_1/2) were estimated to be -0.1060 and 6.5 days of mandipropamid and -0.1236 and 5.6 days of thiamethoxam. The PHRLs for lettuce on the 10th and 5th day before harvesting were calculated to be 63.24 and 43.56 mg/kg for mandipropamid, and 44.66 and 25.88 mg/kg for thiamethoxam, with -0.0746 and -0.1091 of the upper 95% confidence intervals of dissipation rate constant, respectively. This work would be useful as guidance for adjusting the shipment date and contribute to stabilizing the income of farmers in Korea.

• Journal of Microbiology and Biotechnology
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• v.22 no.3
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• pp.343-351
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• 2012

In this paper, the kinetics of a cloned special glucosidase, named ginsenosidase type III hydrolyzing 3-O-glucoside of multi-protopanaxadiol (PPD)-type ginsenosides, were investigated. The gene (bgpA) encoding this enzyme was cloned from a Terrabacter ginsenosidimutans strain and then expressed in E. coli cells. Ginsenosidase type III was able to hydrolyze 3-O-glucoside of multi-PPD-type ginsenosides. For instance, it was able to hydrolyze the 3-O-${\beta}$-D-(1${\rightarrow}$2)-glucopyranosyl of Rb1 to gypenoside XVII, and then to further hydrolyze the 3-O-${\beta}$-D-glucopyranosyl of gypenoside XVII to gypenoside LXXV. Similarly, the enzyme could hydrolyze the glucopyranosyls linked to the 3-O-position of Rb2, Rc, Rd, Rb3, and Rg3. With a larger enzyme reaction $K_m$ value, there was a slower enzyme reaction speed; and the larger the enzyme reaction $V_{max}$ value, the faster the enzyme reaction speed was. The $K_m$ values from small to large were 3.85 mM for Rc, 4.08 mM for Rb1, 8.85 mM for Rb3, 9.09 mM for Rb2, 9.70 mM for Rg3(S), 11.4 mM for Rd and 12.9 mM for F2; and $V_{max}$ value from large to small was 23.2 mM/h for Rc, 16.6 mM/h for Rb1, 14.6 mM/h for Rb3, 14.3 mM/h for Rb2, 1.81mM/h for Rg3(S), 1.40 mM/h for Rd, and 0.41 mM/h for F2. According to the $V_{max}$ and $K_m$ values of the ginsenosidase type III, the hydrolysis speed of these substrates by the enzyme was Rc>Rb1>Rb3>Rb2>Rg3(S)>Rd>F2 in order.

• International Journal of Industrial Entomology and Biomaterials
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• v.9 no.1
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• pp.29-33
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• 2004

Low molecular weight silk fibroin (LMSF), which was prepared by hydrolysis of silk fibroin using high-temperature and high-pressure method, was found to inhibit the oxidation of L-3,4,-dihydroxyphenylalanine (L-DOPA) catalyzed by mushroom tyrosinase (EC 1.14.18.1). LMSF contained mostly free amino acids such as L-glycine, L-alanine, and L-serine and oligopeptides, mainly glycine-alanine dimer. As a result of analyzing the inhibition kinetics from Lineweaver-Burk plots, L-glycine and glycine-alanine dimer showed noncompetitive behavior while uncompetitive behavior was observed in L-alanine, and L-serine. When weight percent concentration of ${ID_50}$ was compared, L-glycine was most effective on the inhibition and LMSF was also good enough for the inhibition effect of tyrosinase activity. LMSF showed a mixed-type inhibition and the inhibitory mechanism of LMSF might be caused by free amino acids and oligopeptides. As a result of spectroscopic observation with time, initial rate of increase of DOPAchrome decreased remarkably and the time to reach maximum absorbance increased as an increase of the concentration of L-glycine, meaning that L-glycine made itself mainly responsible for the formation of chelate with ${Cu^2+}$ in tyrosinase. However, in case of L-alanine, L-serine, and especially glycine-alanine dimmer, the production of DOPAchrome after an arrival at maximum absorbance decreased, indicating the production of adducts through the reaction with DOPAquinone.

• Bulletin of the Korean Chemical Society
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• v.28 no.12
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• pp.2283-2287
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• 2007

Amyloid fibril formation of amyloid β/A4 protein (Aβ) is critical to understand the pathological mechanism of Alzheimer's disease and develop controlling strategy toward the neurodegenerative disease. For this purpose, dequalinium (DQ) has been employed as a specific modifier for Aβ aggregation and its subsequent cytotoxicity. In the presence of DQ, the final thioflavin-T binding fluorescence of Aβ aggregates decreased significantly. It was the altered morphology of Aβ aggregates in a form of the bundles of the fibrils, distinctive from normal single-stranded amyloid fibrils, and the resulting reduced β-sheet content that were responsible for the decreased fluorescence. The morphological transition of Aβ aggregates assessed with atomic force microscope indicated that the bundle structure observed with DQ appeared to be resulted from the initial multimeric seed structure rather than lateral association of preformed single-stranded fibrils. Investigation of the seeding effect of the DQ-induced Aβ aggregates clearly demonstrated that the seed structure has determined the final morphology of Aβ aggregates as well as the aggregative kinetics by shortening the lag phase. In addition, the cytotoxicity was also varied depending on the final morphology of the aggregates. Taken together, DQ has been considered to be a useful chemical probe to control the cytotoxicity of the amyloid fibrils by influencing the seed structures which turned out to be central to develop therapeutic strategy by inducing the amyloid fibrils in different shapes with varied toxicities.

• Biomolecules & Therapeutics
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• v.20 no.6
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• pp.562-568
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• 2012

Cytochrome P450 BM3 (CYP102A1) from Bacillus megaterium is a self-sufficient monooxygenase that consists of a heme domain and FAD/FMN-containing reductase domain (BMR). In this report, the reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) by BMR was evaluated as a method for monitoring BMR activity. The electron transfer proceeds from NADPH to BMR and then to BMR substrates, MTT and CTC. MTT and CTC are monotetrazolium salts that form formazans upon reduction. The reduction of MTT and CTC followed classical Michaelis-Menten kinetics ($k_{cat}=4120\;min^{-1}$, $K_m=77{\mu}M$ for MTT and $k_{cat}=6580\;min^{-1}$, $K_m=51{\mu}M$ for CTC). Our continuous assay using MTT and CTC allows the simple, rapid measurement of BMR activity. The BMR was able to metabolize mitomycin C and doxorubicin, which are anticancer drug substrates for CPR, producing the same metabolites as those produced by CPR. Moreover, the BMR was able to interact with CYP1A2 and transfer electrons to promote the oxidation reactions of substrates by CYP1A2 and CYP2E1 in humans. The results of this study suggest the possibility of the utilization of BMR as a surrogate for mammalian CPR.

• Korean Chemical Engineering Research
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• v.44 no.6
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• pp.636-643
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• 2006

Supercritical water oxidation of DMMP using continuous flow reactor was studied at temperature ranging from 440 to $540^{\circ}C$ and a fixed pressure of 242 bar. The range of residence times in the reactor was from 10 to 26 s, and oxygen excess value varied from -40 to 200%. Destruction efficiencies (DE) of DMMP were greater than 99.7% at $540^{\circ}C$, and increased as the DMMP concentrations were increased. DE of DMMP were significantly affected by oxygen concentration under stoichiometric amount, but showed little difference over stoichiometric amount. On the basis of 30 data with conversions greater than 85%, kinetic correlations for the DE of DMMP were developed. The pre-exponential factor was $(1.10{\pm}0.76){\times}10^6$, and the activation energy was $90.66{\pm}3.87kJ/mol$, and the reaction orders for DMMP and oxygen were $1.02{\pm}0.03$, $0.32{\pm}0.03$, respectively. The model predictions agreed well with the experimental data.

• Journal of Applied Biological Chemistry
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• v.42 no.1
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• pp.12-18
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• 1999

Sucrose synthase (EC 2.4.1.13) has been purified from the plant cytosolic fraction of chickpea (Cicer arietinum L. cv. Amethyst) nodules. The native enzyme had a molecular mass of $356{\pm}15kD$. The subunit molecular mass was $87{\pm}2kD$, and a tetrameric structure is proposed for sucrose synthase of chickpea nodule. Optimum activities in the sucrose cleavage and synthesis directions were at pH 6.5 and 9.0, respectively. The purified enzyme displayed typical hyperbolic kinetics with substrates in cleavage and synthesis reactions. Chickpea nodules sucrose synthase had a high affinity for UDP ($K_m$, $8.0{\mu}M$) and relatively low affinities for ADP ($K_m$, 0.23 mM), CDP ($K_m$, 0.87 mM), and GDP ($K_m$, 1.51 mM). The $K_m$ for sucrose was 29.4 mM. In the synthesis reaction, UDP-glucose ($K_m$, $24.1{\mu}M$) was a more effective glucosyl donor than ADP-glucose ($K_m$, 2.7 mM), and the $K_m$ for fructose was 5.4 mM. Divalent cations, such as $Ca^{2+}$, $Mg^{2+}$, and $Mn^{2+}$, stimulated the enzyme activity in both the cleavage and synthesis directions, and the enzyme was very sensitive to inhibition by $HgCl_2$ and $CuSO_4$.