• Journal of Microbiology and Biotechnology
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• v.28 no.9
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• pp.1502-1510
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• 2018

Organic solvent-tolerant (OST) enzymes are widely applied in various industries for their activity and stability in organic solvents, for their higher substrate solubility, and for their greater stero-selectivity. However, the criteria for identifying OST enzymes largely remain undefined. In this study, we compared the amino acid composition of 19 OST esterases with that of 19 non OST esterases. OST esterases have increased the ratio of Ala and Arg residues and decreased the ratio of Asn, Ile, Tyr, Lys, and Phe residues. Based on our amino acid composition analysis, we cloned a carboxylesterase (EstSP2) from a psychrophilic bacterium, Sphingomonas glacialis PAMC 26605, and characterized its recombinant protein. EstSP2 is a substrate specific to p-nitrophenyl acetate and hydrolyzed aspirin, with optimal activity at $40^{\circ}C$; at $4^{\circ}C$, the activity is approximately 50% of its maximum. As expected, EstSP2 showed tolerance in up to 40% concentration of polar organic solvents, including dimethyl sulfoxide, methanol, and ethanol. The results of this study suggest that selecting OST esterases based on their amino acid composition could be a novel approach to identifying OST esterases produced from bacterial genomes.

• Korean Journal of Environmental Agriculture
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• v.8 no.1
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• pp.47-54
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• 1989

This study was conducted to investigate insecticide toxicities to a honeybee, Apis cerana F. being raised in Korea and its detoxifying enzyme activities. In order to determine the appropriate usage of insecticides, median effective dose and detoxifying enzyme activities to seven insecticides were observed. Various detoxifying enzymes, including microsomal oxidases, glutathione S-transferases, esterases, and DDT-dehydrochlorinase were assayed in the midguts of adult worker bees as the enzyme source. Of the insecticides used, $LC_{50}$ value in DDT treatment was the highest as 19ppm, and that in EPN treatment was the lowest as 0.75ppm. Sublethal exposures of honeybees to various insecticides had some effects on microsomal enzyme activities. Aldrin epoxidase activity was inhibited by malathion and demeton S-methyl treatment. N-demethylase activity was induced by carbaryl treatment. Of the glutathione S-transferases, aryltransferase(DCNB conjugation) activity was significantly induced by diazinon, and moderately induced by malathion. Of the esterases, ${\alpha}-NA$ esterase activity was moderately inhibited by malathion and permethrin. Carboxylesterase and acetylcholinesterase activity were not affected by the sublethal exposure of honeybee to the insecticides. Sublethal exposure of honeybee to the insecticides had no effect on DDT- dehydrochlorinase activity, except carbaryl, malathion and demeton S-methyl were inhibited.

• Environmental Analysis Health and Toxicology
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• v.7 no.1_2
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• pp.17-34
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• 1992

The object of this study is to investigate the toxicity of S-biol (d-allethron-d-transchrysanthemate) and the mode of action between other synthetic pyrethroid insecticides and S-biol in rats. Rats were treated daily with S-biol (25 mg/kg, 50 mg/kg, 100mg/kg) by oral administration for 5 weeks. The experimental results were summerized as follows: Biochemical parameters such as LDH and Glucose in serum were much more increased in control groups. No significant hematological change excepts for MCHC in rats treated with S-biol 100 mg/kg were observed in all groups compared to control groups. In animals treated with S-biol for 4∼5 weeks, the levels of cytochrome P-450 in the liver were significantly increased. In renal microsomal fractions, however, no significant changes of cytochrome P-450 contents were observed. The activitis of ATPase in groups treated with S-biol (50 mg/kg, 100 mg/kg) were decreased compared to those in other groups. TBA values and activities of glucose-6-phosphatase in the liver were increased a little in the groups treated with S-biol (100 mg/kg) for 5 weeks. The activities of cholinesterase in hepatic and serum were not significantly changed in all groups but slightly decreased in animals treated with high dose of S-biol (100 mg/kg). The activities of carboxylesterase in serum and in the liver were slightly increased but not significantly changed.

• Journal of agriculture & life science
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• v.46 no.3
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• pp.109-118
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• 2012

The gene encoding an esterase enzyme was cloned from a metagenomic library of cow rumen bacteria. The esterase gene (est2R) was 2,120 bp in length, encoding a protein of 516 amino acid residues with a calculated molecular weight of 57,286 Da. The molecular weight of the enzyme was estimated to be 57,000 Da by SDS-PAGE. Est2R shared 35.6% amino acid identity with esterase (CAH19079) of uncultured prokaryote. The Est2R was most active at $20-40^{\circ}C$, and showed optimum at $30^{\circ}C$ and pH 8.0. The most activity of Est2R for the different chain length of p-nitrophenyl ester group as substrate was p-nitrophenyl acetate. Moreover, the enzyme was found to be most active without organic solvent, followed by 98% active with ethanol, and the enzyme activity was highly affected by the acetonitrile. The enzyme was significantly inhibited by $Zn^{2+}$ but stimulated by $Ca^{2+}$. So, novel esterase gene est2R is likely to obtain from cow rumen metagenome and supposed to use for industrial purpose.

• The Korean Journal of Zoology
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• v.33 no.3
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• pp.337-345
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• 1990

The properties of three esterases (E2, E6 and E11) which were previously purified from Pieris rapae L. were determined and physiological role of E6 was inferred using immunological methods. Based on inhibitor study, all of the purified esterases were found to be carboxylesterases (EC 3.1.1.1). The Km values for E2, E6 and E11 were determined to be 6.89 X 10-$^4$M. 3.19 $\times$ l0-$^4$M and 3.69 X 10-$^4$M, respectively. The molecular weights of E2, E6 and E11 were estimated to be 42 KD, 81 KD and 174 KD, respectively. The isoelectric points of E2, E6 and E11 were estimated to be pH 5.54, pH 5.89 and pH 6.50, respectively. The concentration of E6 during development was highest at the late 5th instar larval stage and that according to organs at the same stage was highest in midgut. These results suggest that E6 might be a hydrolase involved in the digestion of dietary lipids.

• Journal of Applied Biological Chemistry
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• v.62 no.1
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• pp.73-79
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• 2019

A novel esterase gene (est7S) was cloned from an enriched metagenomic library derived from an oil-spill area. The gene encoded a protein of 505 amino acids, and the molecular mass of the Est7S was estimated to be 54,512 Da with no signal peptide. Est7S showed the highest identity of 40% to an esterase from a sludge metagenome compared to the characterized enzymes with their properties, although it showed 99% identity to a carboxylesterase in the genome sequence of Alcanivorax borkumensis SK2. Est7S had catalytic triad residues, Ser183, Glu312, and His420, and the GESAG motif in most family VII lipolytic enzymes. Est7S was purified from the crude extract of clone SM7 using Sephacryl S-200 HR and HiTrap Q column chromatographies. The purified Est7S was optimally active at $50^{\circ}C$ and pH 10.0. Est7S showed a high specific activity of 366.7 U/mg protein. It preferred short length esters, particularly p-nitrophenyl acetate, efficiently hydrolyzed R- and S-enantiomers of methyl-3-hydroxy-2-methylpropionate, and glyceryl tributyrate. These properties of Est7S may provide potential merits in biotechnological applications such as detergent and paper processing under alkaline conditions.

• Korean journal of applied entomology
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• v.32 no.3
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• pp.348-353
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• 1993

The fluctuation of insecticide resistance in the green peach aphid (GPA) in fields was investigated by caboxy1 esterase (CE) activity index analysis. Of the GP A Populations occurred on the red pepper seedlings, aphids in the untreaLed and treaLed with acephate plots showed 40 and 78 resistance percent (RP), respectively. Aphids in the untreated kale field showed the RP value 24 in July, contrast to 83 in October. Mean RPs of aphids from 18 localities were 50 + 14 in summer and B2+ 10 in late fall, indicating a seasonal fluctuation of Lhe CE activity.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.1
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• pp.138-148
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• 2018

Objective: Fusarium mycotoxins deoxynivalenol (DON) and zearalenone (ZEN), common contaminants in the feed of farm animals, cause immune function impairment and organ inflammation. Consequently, the main objective of this study was to elucidate DON and ZEN effects on the mRNA expression of pro-inflammatory cytokines and other immune related genes in the kidneys of piglets. Methods: Fifteen 6-week-old piglets were randomly assigned to three dietary treatments for 4 weeks: control diet, and diets contaminated with either 8 mg DON/kg feed or 0.8 mg ZEN/kg feed. Kidney samples were collected after treatment, and RNA-seq was used to investigate the effects on immune-related genes and gene networks. Results: A total of 186 differentially expressed genes (DEGs) were screened (120 upregulated and 66 downregulated). Gene ontology analysis revealed that the immune response, and cellular and metabolic processes were significantly controlled by these DEGs. The inflammatory stimulation might be an effect of the following enriched Kyoto encyclopedia of genes and genomes pathway analysis found related to immune and disease responses: cytokine-cytokine receptor interaction, chemokine signaling pathway, toll-like receptor signaling pathway, systemic lupus erythematosus (SLE), tuberculosis, Epstein-Barr virus infection, and chemical carcinogenesis. The effects of DON and ZEN on genome-wide expression were assessed, and it was found that the DEGs associated with inflammatory cytokines (interleukin 10 receptor, beta, chemokine [C-X-C motif] ligand 9, CXCL10, chemokine [C-C motif] ligand 4), proliferation (insulin like growth factor binding protein 4, IgG heavy chain, receptor-type tyrosine-protein phosphatase C, cytochrome P450 1A1, ATP-binding cassette sub-family 8), and other immune response networks (lysozyme, complement component 4 binding protein alpha, oligoadenylate synthetase 2, signaling lymphocytic activation molecule-9, ${\alpha}$-aminoadipic semialdehyde dehydrogenase, Ig lambda chain c region, pyruvate dehydrogenase kinase, isozyme 4, carboxylesterase 1), were suppressed by DON and ZEN. Conclusion: In summary, our results indicate that high concentrations of DON and ZEN suppress the inflammatory response in kidneys, leading to potential effects on immune homeostasis.