• Journal of Microbiology and Biotechnology
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• v.24 no.11
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• pp.1516-1524
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• 2014

Flavin adenine dinucleotide-dependent glucose dehydrogenase (FAD-GDH) can utilize a variety of external electron acceptors and also has stricter substrate specificity than any other glucose oxidoreductases, which makes it the ideal diagnostic enzyme in the field of glucose biosensors. A gene coding for a hypothetical protein, similar to glucose oxidase and derived from Aspergillus terreus NIH2624, was overexpressed in Pichia pastoris GS115 under the control of an AOX1 promoter with a level of 260,000 U/l in the culture supernatant after fed-batch cultivation for 84 h. After a three-step purification protocol that included isopropanol precipitation, affinity chromatography, and a second isopropanol precipitation, recombinant FAD-GDH was purified with a recovery of 65%. This is the first time that isopropanol precipitation has been used to concentrate a fermentation supernatant and exchange buffers after affinity chromatography purification. The purified FAD-GDH exhibited a broad and diffuse band between 83 and 150 kDa. The recombinant FAD-GDH was stable across a wide pH range (3.5 to 9.0) with maximum activity at pH 7.5 and $55^{\circ}C$. In addition, it displayed very high thermal stability, with a half-life of 82 min at $60^{\circ}C$. These characteristics indicate that FAD-GDH will be useful in the field of glucose biosensors.

• Journal of Applied Biological Chemistry
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• v.42 no.3
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• pp.113-118
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• 1999

The transgenic plant of Citrus species (Citrus aurantium L.) was constructed with a fatty acid desaturase gene using microprojectile bombardment transformation system. The DNA of a fatty acid desaturase gene, fad7, constructed in pBI121 was coated onto tungsten particles ($1.1{\mu}m$) and introduced into callus cells by bombarding with 1100 psi of helium pressure, 1/4 in of gap distance, 7.0 cm of target distance and 27 in Hg of chamber vacuum. The bombarded cells were selected on the medium containing kanamycin. The selected cells were successfully regenerated into plantlets via somatic embryogenesis on the media containing plant growth regulators. The results of polymerase chain reaction analysis of genomic DNAs from the putative transformants showed that the introduced DNAs of fad7 were present in both the selected callus cells and the regenerated plantlets.

• Bulletin of the Korean Chemical Society
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• v.26 no.2
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• pp.260-264
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• 2005

The nature of the active site of Tobacco acetohydroxyacid synthase (AHAS) in the substrate- and cofactorbinding was studied by kinetics and fluorescence spectroscopy. The substrate saturation curve does not follow Michaelis-Menten kinetics at different temperatures (7, 21 and 37 ${^{\circ}C}$), pH (6.5, 7.5 and 8.5) and buffers (Tris-HCl and MOPS). The concentration of one half of the maximum velocity ($S_{0.5}$) decreased in the following order: pyruvate $\gt$ ThDP $\approx$$Mg^{+2}$ $\gt$ FAD. However, the catalytic efficiency (K$_{cat}/S_{0.5}$) inversely decreased in the following order; FAD $\gt$ $Mg^{+2}$ $\approx$ThDP $\gt$ pyruvate, indicating that the cofactors by in decreasing order; FAD, $Mg^{+2}$, ThDP, affect the catalysis of AHAS. The dissociation constant ($K_d$) of the intrinsic tryptophan fluorescence decreased with the same tendency of the concentration of one half of the maximum velocity ($S_{0.5}$) decreasing order. This data provides evidence that the substrate and cofactor binding natures of the active site, as well as its activation characteristics, resemble those of other ThDP-dependent enzymes.

• Journal of Veterinary Science
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• v.19 no.6
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• pp.759-770
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• 2018

Fowl adenovirus (FAdV) is distributed worldwide and causes economic losses in the poultry industry. The objectives of this study were to determine the hexon and fiber gene changes in an attenuated FAdV isolate from Malaysia in specific pathogen-free chicken embryonated eggs (SPF CEE) and its infectivity in commercial broiler chickens. SPF CEE were inoculated with 0.1 mL FAdV inoculum via the chorioallantoic membrane (CAM) for 20 consecutive passages. The isolate at passage 20 (E20), with a virus titer of $10^{8.7}TCID_{50}/mL$ ($TCID_{50}$, 50% tissue culture infective dose), was inoculated (0.5 mL) into one-day-old commercial broiler chicks either via oral or intraperitoneal routes. The study demonstrated that 100% embryonic mortality was recorded from E2 to E20 with a delayed pattern at E17 onwards. The lesions were confined to the liver and CAM. Substitutions of amino acids in the L1 loop of hexon at positions 49 and 66, and in the knob of fiber at positions 318 and 322 were recorded in the E20 isolate. The isolate belongs to serotype 8b and is non-pathogenic to broiler chickens, but it is able to induce a FAdV antibody titer. It appears that molecular changes in the L1 loop of hexon and the knob of fiber are markers for FAdV infectivity.

• Journal of the Korean Society of Fisheries and Ocean Technology
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• v.57 no.3
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• pp.194-204
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• 2021

The research was analyzed the catch data of the five years (2016-2020) for six joint venture company tuna purse seiners in the Atlantic Ocean, with the aim of suggesting improvement measures for responsible and sustainable fishing according to changes of recommendation by International Commission for the Conservation of Atlantic Tuna (ICCAT) on the tropical tunas. In the last five years, the average catch of six tuna purse seiner gradually increased to 7,745 tons, 8,364 tons and 9,053 tons from 2016 to 2018, but decreased to 7,761 tons in 2019 and 6,214 tons in 2020. The reason for the decrease in fishing volume in 2019 and 2020 was the fluctuation of the formation of the cold water zone (22-23℃), and the total ICCAT convention area of FAD closure in January and February due to the expansion of the FAD closure area and poor free school catching during two months period, respectively. The analysis on fishing area showed that the percentage of fishing in the high sea was about 85% although the FAD closure area included the EEZ zone in coastal countries; the rise of the fishing license in coastal countries is also believed to be a factor. In order to overcome such situations and improve catching volume, it will be possible if excellent manpower is secured, school fishing is expanded, and the production of high value-added catch (purse seine special: PS).

• Korean Journal of Social Welfare
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• v.40
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• pp.5-37
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• 2000

The purpose of this study is to find out some characteristics of the family functions of single mothers to provide some useful data for the development of programs which may help single mothers to improve their family functions. The McMaster Family Assessment Device (FAD) which is composed of problem solving function (PS), communication function (CF), role function (RC), emotional reaction function (ER), emotional involvement (EI), behavioral control function (BC), and general function (GC) and McCubbins' FSI were administered to single mothers and their adolescent children and mothers and their adolescent children of ordinary families (control family). The single mothers' perception of their family function was lower than that of control family mothers in an the 7 categories of FAD. The perceptions of the family function of the single mothers' children were lower than those of control family mothers' children. There was on significant difference between the perceptions of single mothers and their children in PS, ER, EI, GC of their family. However, single mothers' perception in RC and BC was lower than that of their children, and the children's perception in CF were lower than that of their mothers. Single mothers who fall on the period of single motherhood was shorter than 7 years perceived their families' function as lower than that of single mothers' who fall on the periods of single motherhood longer than 7 years in all categories. Generally speaking, the single mothers' family functions were lower than that of control family. The different characteristics of each single mother's family functions should be considered in developing programs for single mothers.

• Applied Biological Chemistry
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• v.21 no.3
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• pp.137-143
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• 1978

Xanthine oxidase from bovine thyroid glands was found to contain FAD, molybdenum and iron in a ratio 1:0. 36:1. 6. The molecular weight of the thyroid enzyme was similar to that of the milk enzyme when estimated by gel filtration and polyacrylamide gel electrophoresis. The optimum pH for the enzyme activity was 7.8. The pH of the isoelectric point was determined to be 6.2 by electrofocusing. Sodium dodecyl sulfatepolyacrylamide gel electrophoresis experiment indicated that the enzyme was dissociated into subunits and that the molecular weight for the smallest subunit was 65,000 daltons. Absorption spectra were dissimilar between milk and thyroid xanthine oxidase.

• Applied Biological Chemistry
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• v.40 no.6
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• pp.501-507
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• 1997

Chloroplasts isolated from Stevia rebaudiana Bertoni leaves contained an enzyme activity which catalyzed hydroxylation of ent-kaurenoic acid (ent-kaur-16-en-19-oic acid; ent-KA) to steviol (ent-13-hydroxy kaur-16-en-19-oic acid), the diterpenoid carboxylic alcohol which is the aglycone of sweet stevioside-related glycosides. $[^(14)C]-methylated$ ent-KA was used to localize ent-KA hydroxylase. $[^(14)C]-methyl-KA$ was most actively was transformed into methyl-steviol in chloroplast. The enzymatic activity was found in stroma fraction but not in thylakoid membrane in Stevia rebaudiana Bertoni. However, ent-KA 13-hydroxylase activity was not detected in stroma fraction of either Spinacia oleracea and Solidago altissima. The reaction products using $[^(14)C]-methyl-KA$ were purified and identified on TLC autoradiogram. The hydroxylation of ent-KA from stromal protein to form steviol required NADPH and oxygen. FAD and riboflavin stimulated the enzyme activity 1.5-and 1.7-fold, respectively. It also turned out that the activity of this enzyme using methyl-KA as a substrate was 16.7% that of ent-KA. The purified ent-KA 13-hydroxylase did not act on t-cinnamic acid, 4-hydroxyphenyl acetic acid, choline and resorcinol, known as monooxygenase and hydroxylase substrates.

• Korean Journal of Microbiology
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• v.24 no.2
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• pp.133-140
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• 1986

Extracts of CO-autotrophically grown cells of Acinetobacter sp. 1 were shown to use thionin, methylene blue, or 2,6-dichlorophenol-indophenol, but not NAD, NADP, FAD, or FMN, as electron acceptors for the oxidation of CO under strictly anaerobic conditions. The CO dehydrogenase (CO-DH) in the thes bacterium was found to be an inducible enzyme. The enzyme activity was determined by an assay based on the CO-dependent reduction of thionin. Maximal reaction rates were found at pH 7.5 and $60^{\circ}C$, and the Arrhenius plot revealed an activation energy of 6.1 kcal/mol(25.5kJ/mol). THe $K_m$ m/ for CO was $154{\mu}M$. Known metalchelating agents tested had no effects on the CO-DH activity. No divalent cations tested affect the enzyme activity significantly escept $Cu^{2+}$ which suppressed the activity completely. The enzyme was inhibited by glucose and succinate. The same extracts catalyzed oxidation of hydrogen gas and formate with thionin as electron acceptor. The CO-DH of Acinetobacter sp. 1 was to have no immunological relationship with that of Pseudomonas carboxydohydrogena.

• Korean Journal of Poultry Science
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• v.38 no.3
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• pp.181-189
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• 2011

A 5-month (May to November in 2009) monitoring program for five viral pathogens in litter, such as avian influenza virus A (AIV), infectious bronchitis virus (IBV), infectious bursal disease virus (IBDV), fowl adenovirus (FAdV), and chicken infectious anemia virus (CIAV) was conducted in 62 flocks at 31 broiler farms (two flocks in each farm) in Korea in 2009. Viral pathogens were examined twice (before and at the end of the rearing period) at 31 broiler farms, and included fresh litter (n = 16) and recycled litter (n = 15) farms. Thirty-seven viruses (14 IBVs, 2 IBDVs, 9 FAdVs, and 12 CIAVs) were isolated from 75% (12/16) and 73% (11/15) of fresh litter and reused litter farms during the period, respectively, indicating no difference in viral contamination rate between farms using new and reused litter. Of these isolates, three (two CIAVs and one IBDV) were isolated from recycled litter samples collected before the rearing period at three broiler farms, whereas the others (n=34) were isolated from fresh and recycled litter samples collected at the end of the rearing period. When the performances, involving viability, body weight, and feed conversion ratio, were compared, no significant differences were found between farms using fresh and recycled litter during the period.