• Journal of Microbiology and Biotechnology
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• v.23 no.2
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• pp.205-210
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• 2013

Thiol-ene polymerization is a versatile tool for several applications. Here we report the preparation of epoxide groups containing thiol-ene photocurable polymeric support and the covalent immobilization of ${\alpha}$-amylase onto these polymeric materials. The morphology of the polymeric support was characterized by scanning electron microscopy (SEM), and energy dispersive spectroscopy (EDS) coupled with SEM was used to explore the chemical composition. The polymeric support and the immobilization of the enzyme were characterized by FTIR analysis. SEM-EDS and FTIR results showed that the enzyme was successfully covalently attached to the polymeric support. The immobilization efficiency and enzyme activity of ${\alpha}$-amylase were examined at various pH (5.0-8.0) and temperature ($30-80^{\circ}C$) values. The storage stability and reusability of immobilized ${\alpha}$-amylase were investigated. The immobilization yield was $276{\pm}1.6$ mg per gram of polymeric support. Enzyme assays demonstrated that the immobilized enzyme exhibited better thermostability than the free one. The storage stability and reusability were improved by the immobilization on this enzyme support. Free enzyme lost its activity completely within 15 days. On the other hand, the immobilized enzyme retained 86.7% of its activity after 30 days. These results confirm that ${\alpha}$-amylase was successfully immobilized and gained a more stable character compared with the free one.

• Molecules and Cells
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• v.39 no.1
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• pp.65-71
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• 2016

A challenge in the redox field is the elucidation of the molecular mechanisms, by which $H_2O_2$ mediates signal transduction in cells. This is relevant since redox pathways are disturbed in some pathologies. The transcription factor OxyR is the $H_2O_2$ sensor in bacteria, whereas Cys-based peroxidases are involved in the perception of this oxidant in eukaryotic cells. Three possible mechanisms may be involved in $H_2O_2$ signaling that are not mutually exclusive. In the simplest pathway, $H_2O_2$ signals through direct oxidation of the signaling protein, such as a phosphatase or a transcription factor. Although signaling proteins are frequently observed in the oxidized state in biological systems, in most cases their direct oxidation by $H_2O_2$ is too slow ($10^1M^{-1}s^{-1}$ range) to outcompete Cys-based peroxidases and glutathione. In some particular cellular compartments (such as vicinity of NADPH oxidases), it is possible that a signaling protein faces extremely high $H_2O_2$ concentrations, making the direct oxidation feasible. Alternatively, high $H_2O_2$ levels can hyperoxidize peroxiredoxins leading to local building up of $H_2O_2$ that then could oxidize a signaling protein (floodgate hypothesis). In a second model, $H_2O_2$ oxidizes Cys-based peroxidases that then through thiol-disulfide reshuffling would transmit the oxidized equivalents to the signaling protein. The third model of signaling is centered on the reducing substrate of Cys-based peroxidases that in most cases is thioredoxin. Is this model, peroxiredoxins would signal by modulating the thioredoxin redox status. More kinetic data is required to allow the identification of the complex network of thiol switches.

• Environmental Engineering Research
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• v.20 no.4
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• pp.342-346
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• 2015

Haloacetonitriles (HANs) are nitrogenous disinfection by-products (DBPs) that have been reported to have a higher toxicity than the other groups of DBPs. The adsorption process is mostly used to remove HANs in aqueous solutions. Functionalized composite materials tend to be effective adsorbents due to their hydrophobicity and specific adsorptive mechanism. In this study, the removal of dichloroacetonitrile (DCAN) from tap water by adsorption on thiol-functionalized mesoporous composites made from natural rubber (NR) and hexagonal mesoporous silica (HMS-SH) was investigated. Fourier-transform infrared spectroscopy (FTIR) results revealed that the thiol group of NR/HMS was covered with NR molecules. X-ray diffraction (XRD) analysis indicated an expansion of the hexagonal unit cell. Adsorption kinetic and isotherm models were used to determine the adsorption mechanisms and the experiments revealed that NR/HMS-SH had a higher DCAN adsorption capacity than powered activated carbon (PAC). NR/HMS-SH adsorption reached equilibrium after 12 hours and its adsorption kinetics fit well with a pseudo-second-order model. A linear model was found to fit well with the DCAN adsorption isotherm at a low concentration level.

• Journal of Environmental Health Sciences
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• v.11 no.2
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• pp.65-75
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• 1985

Microorganisms capable of utilizing diaminododecane containing amine groups diterminally were isolated from the soil by enrichment culture. One strain of these isolated strain, designated as EL-0112L, was selected for this study. The results of this study were as follows. 1. This isolated strain EL-0112L was identified as Microbacterium, from the results of morphological, cultural, and biochemical tests. This isolated strain was named temporarily Microbacterium sp. EL-0112L for convenience. 2. Microbacterium sp. EL-0112L was tested for ability to utilize different kinds of substitued alkanes containing cyan, amine, chloro, and thiol groups(monoterminally or diterminall substituted) as carbon source. Pentamethylenediamine, hexamethylenediamine, n-decane, laurylamine, and alkane derivatives containing cyan, chloro, and thiol groups were not utilized by Microbacterium sp. EL-0112L. 3. The alkane derivatives that did not serve as growth substrates were tested further in oxidation tests using resting cell preparation of Microbacterium sp. EL-0112 L. Alkane derivatives containing cyan, chloro, thiol groups, and n-decane were oxidized by Microbacterium sp. EL-0112 L. It is possible that this isolated strain is also able to degrade their substituted counterparts since they are structually similar to diaminododecane. The remarkable substrates that were being oxidized were dichlorodecane, and 1-dodecanethiol. Microbacterium sp. EL- 0112L could not oxidize pentamethylenediamine, and hexamethylenediamine. 4. The metabolic products formed from diaminododecane by Microbacterium sp. EL-0112 L were acid compound containing carboxyl group and not containing amine group. On the thin layer chromatography, Rf values of these metabolic products were different from that of the product formed by Corynebacterium sp. EL-0112L. These results suggested the specificity of diaminododecane as carbon source.

• BMB Reports
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• v.33 no.3
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• pp.234-241
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• 2000

A new type of the human TSA homologous gene was cloned from a HeLa cell cDNA and characterized. The gene product consists of 161 amino acids with a molecular mass of 16,900. The TSA homologous protein, as a new 6th member of the human TSA (hTSA VI), exerted a thioldependent peroxidase activity with the use of thioredoxin system as a physiological electron donor. The values of $V_{max}/K_m$ of hTSA VI for $H_2O_2$ and t-butyl hydroperoxide (t-BOOH) were calculated as $5.53{\times}10^{-2}$ and $3.70{\times}10^{-2}$, respectively. This implies that hTSA VI is a peroxidase, which reduces $H_2O_2$ and t-BOOH. The mutation of $Cys^{47}$ to serine resulted in a complete loss of the peroxidase activity. This suggests that $Cys^{47}$ acts as a primary site of catalysis. The analysis of the tryptic digest derived from hTSA VI revealed that the $Cys^{47}$ exists as a free thiol form. Taken together, these results suggest that the TSA homologous protein is a new type of the human family, which exerts thioredoxin-linked peroxidase activity toward $H_2O_2$ and alkyl hydroperoxide.

• Journal of Ecology and Environment
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• v.34 no.4
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• pp.393-400
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• 2011

The effects of the ectomycorrhizal fungus Pisolithus tinctorius and cadmium (Cd) on physiological properties and Cd uptake by Populus alba ${\times}$ glandulosa was investigated under greenhouse conditions. Cd treatment decreased the photosynthetic rate ($P_N$) of both non-mycorrhizal (NM) plants (16.3%) and ectomycorrhizal (ECM) plants (11.5%). In addition, the reduction in total dry weight by Cd treatment was greater in ECM plants (24.3%) than that in NM plants (17.6%). Mycorrhizal infection increased the $P_N$ and transpiration rate in both control and Cd-treated plants. Cd treatment increased superoxide dismutase (SOD) activity and decreased glutathione reductase activity, and the increase of SOD activity by Cd treatment was greater in NM plants (40.3%) than that in ECM plants (3.7%). Thiol content increased in both NM and ECM plants treated with Cd solution, and the increase in thiol content in NM plants (43.9%) was greater than that of ECM plants (15.6%). Cd uptake in the leaves, stems, and roots of ECM plants was 69.9%, 167.2% and 72.8%, respectively, higher than in the NM plants. However, the increase in Cd uptake ability of ECM plants resulted in a reduction in dry weight.

• BMB Reports
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• v.31 no.6
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• pp.572-577
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• 1998

A soluble protein from Saccharomyces cerevisiae provides protection against a thiol-containing oxidation system but not against an oxidation system without thiol. This 25-kDa protein acts as a peroxidase but requires the NADPH-dependent thioredoxin system or a thiol-containing intermediate, and was thus named thioredoxin peroxidase. The protective role of thioredoxin peroxidase against ionizing radiation, which generates reactive oxygen species harmful tocellular function, was investigated in wild-type and mutant yeast strains in which the tsa gene encoding thioredoxin peroxidase was disrupted by homologous recombination. Upon exposure to ionizing radiation, there was a distinct difference between these two strains in regard to viability and the level of protein carbonyl content, which is the indicative marker of oxidative damage to protein. Activities of other antioxidant enzymes, such as catalase, superoxide dismutase, glucose-6-phosphate dehydrogenase, and glutathione reductase were increased at 200-600 Gy of irradiation in wild-type cells. However, the activities of antioxidant enzymes were not significantly changed by ionizing radiation in thioredoxin peroxidase-deficient mutant cells. These results suggest that thioredoxin peroxidase acts as an antioxidant enzyme in cellular defense against ionizing radiation through the removal of reactive oxygen species as well as in the protection of antioxidant enzymes.

• Journal of Photoscience
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• v.4 no.3
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• pp.133-140
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• 1997

A system for studying oxidative hemolysis has been used by controling UV-irradiation and concentration of a novel molecular probe, N,N'-bis(2-hydroperoxy-2-methoxyethyl)-1,4,5,8-naphthalenetetra-carboxylic diimide (NP-III), which generates hydroxyl radical upon longer wavelength photoirradiation (> 350 nm). NP-III induces 25~30% of hemolysis at low concentration (50 $\mu$M) for 3h-irradiation of UVA. The simultaneous treatment of N-ethylmaleimide (NEM) with NP-IH completely hemotyzed erythrocytes under the same conditions as NP-III alone by both decreasing thiol group and increasing lipid peroxidation in erythrocyte membrane. However. thiol-reducing agents prevented the protein-crosslinking and lipid peroxidation on the NEM-synergistic hemolysis by partially scavenging hydroxyl radical and maintaining the thiol group of erythrocyte membrane in the reduced state. In addition, erythrocytes pretreated with 2,2,5,7,8-pentamethyl-6-hydroxychromane (PMC), vitamin E homologue was able to delay and decrease the lipid peroxidation when compared to cells pretreated with both NEM and PMC. We suggest that the presence of reduced thiols in inner membrane protein by GSH can prevent the protein-crosslinking and the lipid peroxidation, and eventually prevent the oxidative hemolysis of erythrocyte.

• Bulletin of the Korean Chemical Society
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• v.24 no.12
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• pp.1751-1756
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• 2003

An electrochemical study related to the electroreduction of 4-amino-6-methyl-3-thio-1,2,4-triazin-5-one(I), 6-methyl-3-thio-1,2,4-triazin-5-one(II), and 2,4-dimetoxy-6-methyl-1,3,5-triazine(III) in dimethylformamide at glassy carbon electrode has been performed. A variety of electrochemical techniques, such as differential pulse voltammetry (DPV), cyclic voltammetry (CV), chronoamperometry, and coulometry were employed to clarify the mechanism of the electrode process. The compounds I and II with thiol group exhibited similar redox behavior. Both displayed two cathodic peaks, whereas the third compound, III, without thiol group showed only one cathodic peak in the same potential range of the second peak of I and II. The results of this study suggest that in the first step the one electron reduction of thiol produced a disulfide derivative and in the second reduction step the azomethane in the triazine ring was reduced in two electron processes. A reduction mechanism for all three compounds is proposed on this basis. In addition, some numerical constants, such as diffusion constant, transfer coefficient, and rate constant of coupled chemical reaction in the first reduction peak were also reported.

• BMB Reports
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• v.29 no.6
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• pp.513-518
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• 1996

A soluble protein from Saccharomyces cerevisiae specifically provides protection against a thiolcontaining oxidation system but not against an oxidation system without thiol. This 25-kDa protein was thus named thiol-dependent protector protein (TPP). The role of TPP in the cellular defense against oxidative stress was investigated in Escherichia coli containing an expression vector with a yeast genomic DNA fragment that encodes TPP (strain YP) and a mutant in which the catalytically essential amino acid in the active site of TPP (Cys-47) has been replaced with alanine by site-directed mutagenesis (strain YPC47A). There was a distinct difference between these two strains in regard to viability, modulation of activities of superoxide dismutase and catalase, and the oxidative damage of DNA upon exposure to menadione. These results suggest that TPP may play a direct role in the cellular defense against oxidative stress by functioning as an antioxidant protein.