• Korean Journal of Environmental Biology
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• v.20 no.2
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• pp.173-179
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• 2002

N-nitrosoethylurea (NEU) -induced changes of lipid peroxide content, aldehyde metabolic enzyme activities and antioxidant enzyme activities were examined in cultured rat liver cell. Aldehyde metabolic enzymes tested in this investigation were alcohol dehydrogenase and aldehyde dehydrogenase. Several antioxidant enzymes tested were glutathione transferase, superoxide dismutase, glutathione reductase and catalase. When the cell was exposed with various concentrations of NEU, lipid peroxide content increased about 2.5 fold with 6.25 mM NEU. Maximun 2.3 times higher alcohol dehydrogenase activity was found after NEU treatment. About 2 times higher aldehyde dehydrogenase activity could also be observed. Only slight increases of glutathione transferase and catalase activities occurred with NEU treatment. In addition mnximun 1.5 times higher superoxide dismutase activities and 3 times higher glutathione reductase activities were found after NEU treatment. Therefore, it is likely that the increases of superoxide dismutase and glutathione reductase could contribute in a antioxidative process against NEU toxicity.

• Journal of Life Science
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• v.16 no.3 s.76
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• pp.375-381
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• 2006

Aldehyde reductase was purified to electrophoretic homogeneity from Saccharomyces cerevisiae, and then enzymatic reduction of substituted carbonyl compounds was carried out by using the purified aldehyde reductase as a biocatalyst. Under preparative scale reaction renditions, the enzymatic reduction proceeded in high chemical yield with excellent chemoselectivity. The enzymatic reduction product was identified by TLC, GC, Mass, NMR and FT-IR. Benzoic acid, an inhibitor of aldehyde reductase, also potently inhibited the reduction of substituded carbonyl compounds. This enzyme exhibited a broad substrate specificity , and can utilize both NADH and NADPH as cofactors. The enzyme was strongly inhibited by benzoic acid and quercetin. The apparent Km for 4-cyanobenzaldehyde and 3-nitrobenzamide were 4.894 mM and 0.305 mM, respectively.

• Journal of Microbiology and Biotechnology
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• v.23 no.12
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• pp.1699-1707
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• 2013

During the fermentative production of 1,3-propanediol under high substrate concentrations, accumulation of intracellular 3-hydroxypropionaldehyde will cause premature cessation of cell growth and glycerol consumption. Discovery of oxidoreductases that can convert 3-hydroxypropionaldehyde to 1,3-propanediol using NADPH as cofactor could serve as a solution to this problem. In this paper, the yqhD gene from Klebsiella pneumoniae DSM2026, which was found encoding an aldehyde reductase (KpAR), was cloned and characterized. KpAR showed broad substrate specificity under physiological direction, whereas no catalytic activity was detected in the oxidation direction, and both NADPH and NADH can be utilized as cofactors. The cofactor binding mechanism was then investigated employing homology modeling and molecular dynamics simulations. Hydrogen-bond analysis showed that the hydrogen-bond interactions between KpAR and NADPH are much stronger than that for NADH. Free-energy decomposition dedicated that residues Gly37 to Val41 contribute most to the cofactor preference through polar interactions. In conclusion, this work provides a novel aldehyde reductase that has potential applications in the development of novel genetically engineered strains in the 1,3-propanediol industry, and gives a better understanding of the mechanisms involved in cofactor binding.

• Proceedings of the Korean Society of Life Science Conference
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• 2002.09a
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• pp.74-74
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• 2002

• Proceedings of the Microbiological Society of Korea Conference
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• 2008.05a
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• pp.162-162
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• 2008

• Proceedings of the Korean Society of Applied Pharmacology
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• 1995.04a
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• pp.72-72
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• 1995

Monoclonal antibodies against bovine brain succinic semialdehyde reductase were produced and characterized. A total of nine monoclonal antibodies recognizing different epitopes of the enzyme were obtained, of which two inhibited the enzyme activity and three stained cytosol of rat spinal cord neurons as observed by indirect immunofluorescence microscopy. When unfractionated total proteins of bovine brain homogenate were seperated by gel electrophoresis and immunoblotted, the antibodies specifically recognized a single protein band of 34 kDa, which comigrates with purified bovine succinic semialdehyde reducatase Using the antisuccinic semialdehyde reductase antibodies as probes, we investigated the cross-reactivites of brain succinic semialdehyde reductases from some mammalian and an avian species. The immunoreactive bands on Western blots appeared to be the same in molecular mass-34 kDa-in all animal species tested, including humans. The result indicated that brain succinic semialdehyde reductase is distinct from other aldehyde reductases and that mammalian brains contain only one succinic semialdehyde reductase. Moreover, the enzymes among the species are imunologically very similar, although some properties of the enzymes reported previously were different from one another.

• Journal of Plant Biotechnology
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• v.1 no.1
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• pp.61-68
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• 1999

Rapid accumulation of reactive oxygen species (ROS) and their toxic reaction products with lipids and proteins significantly contributes to the damage of crop plants under biotic and abiotic stresses. We have identified several stress activated alfalfa genes, including the gene of the alfalfa ferritin and a novel NADPH-dependent aldose/aldehyde reductase enzyme. Transgenic tobacco plants that synthesize alfalfa ferritin in vegetative tissues-either in its processed form in chloroplast or in the cytoplasmic non-processed form-retained photosynthetic function upon free radical toxicity generated by paraquat treatment and exhibited tolerance to necrotic damage caused by viral and fungal infections. We propose that by sequestering intracellular iron involved in generation of the very reactive hydroxyl radicals through a Fenton reaction, ferritin protects plant cells from oxidative damage. Our preliminary results with the other stress-inducable alfalfa gene (a NADPH-dependent aldo-keto reductase) indicate, that the encoded enzyme may play role in the stress response of the plant cells. These studies reveal new pathways in plants that can contribute to the increased stress resistance with a potential use in crop improvement.

• Microbiology and Biotechnology Letters
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• v.48 no.4
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• pp.506-514
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• 2020

4-Hydroxybenzyl alcohol (4-HB alcohol) is one of the major active components of Gastrodia elata Blume, with beneficial effects on neurological disorders such as headache, convulsive behavior, and dizziness. Here, we developed a metabolically engineered Corynebacterium glutamicum strain able to produce 4-HB alcohol from 4-hydroxybenzoate (4-HBA). First, the strain APS963 was obtained from the APS809 strain via the insertion of aroK from Methanocaldococcus jannaschii into the NCgl2922-deleted locus. As carboxylic acid reductase from Nocardia iowensis catalyzes the reduction of 4HBA to 4-hydroxybenzaldehyde (4-HB aldehyde), we then introduced a codon-optimized car gene into the genome of APS963, generating the GAS177 strain. Then, we deleted creG coding for a putative short-chain dehydrogenase and inserted ubiCpr encoding a product-resistant chorismate-pyruvate lyase into the pcaHG-deleted locus. The resulting engineered GAS355 strain accumulated 2.3 g/l 4-HB alcohol with 0.32 g/l 4-HBA and 0.3 g/l 4-HB aldehyde as byproducts from 8% glucose after 48 h of culture.

• The Korean Journal of Physiology and Pharmacology
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• v.5 no.3
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• pp.271-278
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• 2001

Oxidative stress and methylglyoxal (MG), a reactive dicarbonyl metabolites produced by enzymatic and non-enzymatic reaction of normal metabolism, induced aldose reductase (AR) expression in rat aortic smooth muscle cells (SMC). AR expression was induced in a time-dependent manner and reached at a maximum of 4.5-fold in 12 h of MG treatment. This effect of MG was completely abolished by cyclohemide and actinomycin D treatment suggesting AR was synthesized by de novo pathway. Pretreatment of the SMC with N-acetyl-L-cysteine significantly down-regulated the MG-induced AR mRNA. Furthermore, DL-Buthionine-(S,R)-sulfoximine, a reagent which depletes intracellular glutathione levels, increased the levels of MG-induced AR mRNA. These results indicated that MG induces AR mRNA by increasing the intracellular peroxide levels. Aminoguanidine, a scanvenger of dicarbonyl, significantly down-regulated the MG-induced AR mRNA. In addition, the inhibition of AR activities with statil, an AR inhibitor, enhanced the cytotoxic effect of MG on SMC under normal glucose, suggesting a protective role of AR against MG-induced cell damages. These results imply that the induction of AR by MG may contribute to an important cellular detoxification of reactive aldehyde compounds generated under oxidative stress in extrahepatic tissues.

• BMB Reports
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• v.32 no.3
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• pp.254-258
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• 1999

The NADPH-dependent succinic semialdehyde reductase is one of the key enzymes in the brain GABA shunt, and it catalyzes the formation of the neuromodulator $\gamma$-hydroxybutyrate from succinic semi aldehyde. This enzyme was inactivated by diethylpyrocarbonate (DEP) with the second-order rate constant of $1.1{\times}10^3\;M^{-1}min^{-1}$ at pH 7.0, $25^{\circ}C$, showing a concomitant increase in absorbance at 242 nm due to the formation of N-carbethoxyhistidyl derivatives. Complete inactivation of succinic semialdehyde reductase required the modification of five histidyl residues per molecule of enzyme. However, only one residue was calculated to be essential for enzyme activity by a statistical analysis of the residual enzyme activity. The inactivation of the enzyme by DEP was prevented by preincubation of the enzyme with the coenzyme NADPH but not with the substrate succinic semialdehyde. These results suggest that an essential histidyl residue involved in the catalytic activity is located at or near the coenzyme binding site of the brain succinic semialdehyde reductase.