• Journal of Marine Bioscience and Biotechnology
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• v.1 no.3
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• pp.156-162
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• 2006

The halophilic enterobacteria, Enterobacteria cancerogenus, was isolated from the intestines of the fusiform fish (Trachurus japonicus) to yield a protein-like material termed PLM-f74. PLM-f74 was characterized by strong inhibition ratios to angiogenesis (82.8% at the concentration of $18.5{\mu}g/mL$) and elevated antioxidative capacities with low toxicity. The PLM-f74 is a glycoprotein comprised of saccharides and amino acids. PLM-f74 inhibited non-activated U937 monocytic cell adhesion to HUVECs activated with IL-$1{\beta}$ by 78.0%, and the adherence of U937 cells treated with the PLM-f74 and stimulated with IL-$1{\beta}$ to unstimulated HUVECs decreased by 102%. When both cell types were pretreated with PLM-f74, the adhesion of U937 cells to IL-$1{\beta}$ stimulated HUVECs was completely suppressed by 121% at a concentration of 18.5 ug/mL. PLM-f74 blocked signal pathways from VEGFR2, PI3K, ${\beta}$-catenin and VE-cadherin to NF-kB based on western bolt analysis. And also inhibited IL-1-stimulated HUVEC expression of the adhesion molecules, ICAM-1 by 40%, VCAM-1 by 60%, and E-selectin by 70% at the same concentration noted above. New anti-angiogenic and anti-cell adhesion materials showing elevated antioxidative capacities and non-toxicity may be expected from these results.

• Journal of the Korean Chemical Society
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• v.21 no.3
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• pp.171-179
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• 1977

Proton magnetic resonance spectra of five glycine-containing dipeptides glycyl-L-valine, L-valyl-glycine, glycyl-DL-alanine, glycyl-DL-serine and glycyl-L-aspartic acid in $D_2O$ were investigated as a function of pH at room temperature. From the analysis of the spectra, it was found that the chemical shift of the $C_{\alpha}H,\;C_{\beta}H\;and\;C_rH$protons varies with pH as a one-step titration curve, and that the spin-spin coupling constant remains almost unchanged. Two distinct values of the chemical shift for $C_{\alpha}H,\;C_{\beta}H\;or\;C_rH$protons of constituent amino acids in dipeptide solutions indicate the existence of two magnetically non-equivalent sites in solution. From this study, the structures of the five dipeptides have been confirmed by proton magnetic resonance spectra and it has been suggested that the structural change, conformation and sequence determination can be explored for oligopeptides by an analysis of proton magnetic resonance spectra.

• Journal of Microbiology and Biotechnology
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• v.16 no.10
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• pp.1544-1553
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• 2006

The halophilic enterobacteria, Enterobacteria cancerogenus, was isolated from the intestines of the fusiform fish (Trachurus japonicus) to yield a protein-like material termed PLM-f74. PLM-f74 was characterized by strong inhibition ratios to angiogenesis (82.8% at the concentration of $18.5{\mu}g/ml$) and elevated antioxidative capacities with low toxicity. The PLM-f74 is a glycoprotein comprised of saccharides and amino acids. PLM-f74 inhibited cell adhesion that non-activated U937 monocytic cell adhesion to HUVECs activated with $IL-1{\beta}$ by 78.0%, and the adherence of U937 cells treated with the PLM-f74 and stimulated with $IL-1{\beta}$ to unstimulated HUVECs decreased by 102%. When both cell types were pretreated with PLM-f74, the adhesion of U937 cells to $IL-1{\beta}$-stimulated HUVECs was completely suppressed by 121% at a concentration of $18.5{\mu}g/ml$. PLM-f74 blocked signal pathways from VEGFR2, PI3K, ${\beta}$-catenin, and VE-cadherin to NF-kB, based on western bolt analysis. It also inhibited IL-l-stimulated HUVEC expression of the adhesion molecules, ICAM-l by 40%, VCAM-l by 60%, and E-selectin by 70% at the same concentration noted above. New anti-angiogenic and anti-cell adhesion materials showing elevated antioxidative capacities, and non-toxicity may be expected from these results.

• Journal of Microbiology and Biotechnology
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• v.27 no.4
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• pp.775-784
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• 2017

A neutral xylanase (CcXyn) was identified from Coprinus cinereus. It has a single GH10 catalytic domain with a basic amino acid-rich extension (PVRRK) at the C-terminus. In this study, the wild-type (CcXyn) and C-terminus-truncated xylanase ($CcXyn-{\Delta}5C$) were heterologously expressed in Pichia pastoris and their characteristics were comparatively analyzed with aims to examine the effect of this extension on the enzyme function. The circular dichorism analysis indicated that both enzymes in general had a similar structure, but $CcXyn-{\Delta}5C$ contained less ${\alpha}-helices$ (42.9%) and more random coil contents (35.5%) than CcXyn (47.0% and 32.8%, respectively). Both enzymes had the same pH (7.0) and temperature ($45^{\circ}C$) optima, and similar substrate specificity on different xylans. They all hydrolyzed beechwood xylan primarily to xylobiose and xylotriose. The amounts of xylobiose and xylotriose accounted for 91.5% and 92.2% (w/w) of total xylooligosaccharides (XOS) generated from beechwood by CcXyn and $CcXyn-{\Delta}5C$, respectively. However, truncation of the C-terminal 5-amino-acids extension significantly improved the thermostability, SDS resistance, and pH stability at pH 6.0-9.0. Furthermore, $CcXyn-{\Delta}5C$ exhibited a much lower $K_m$ value than CcXyn (0.27 mg/ml vs 0.83 mg/ml), and therefore, the catalytic efficiency of $CcXyn-{\Delta}5C$ was 2.4-times higher than that of CcXyn. These properties make $CcXyn-{\Delta}5C$ a good model for the structure-function study of $({\alpha}/{\beta})_8$-barrel-folded enzymes and a promising candidate for various applications, especially in the detergent industry and XOS production.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.78.2-79
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• 2003

Pepper defensin ( CADEFl) clone was isolated from cDNA library constructed from pepper leaves infected with avirulent strain Bv5-4a of Xanthomonu campestris pv. vesicatoria. The deduced amino acid sequence of CADEFl is 82-64％ identical to that of other plant defensins. Putative protein encoded by CADEFl gene consists of 78 amino acids and 8 conserved cysteine residues to form four structure-stabilizing disulfide bridges. Transcription of the CADEF1 gene was earlier and stronger induced by X campestris pv. vesicatoria infection in the incompatible than in the compatible interaction. CADEF1 mRNA was constitutively expressed in stem, root and green fruit of pepper. Transcripts of CADEFl gene drastically accumulated in pepper leaf tissues treated With Salicylic acid (SA), methyl jasmonate (MeJA), abscisic acid (ABA), hydrogen Peroxide (H$_2$O$_2$), benzothiadiazole (BTH) and DL-${\beta}$-amino-n-butyric acid (BABA). In situ hybridization results revealed that CADEF1 mRNA was localized in the phloem areas of vascular bundles in leaf tissues treated with exogenous SA, MeJA and ABA. Strong accumulation of CADEF1 mRNA occurred in pepper leaves in response to wounding, high salinity and drought stress. These results suggest that bacterial pathogen infection, abiotic elicitors and some environmental stresses may play a significant role in signal transduction pathway for CADEF1 gene expression.

• BMB Reports
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• v.28 no.5
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• pp.375-381
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• 1995

Two forms of glucoamylase (GI and GII) from starch-grown Lipomyces kononenkoae CBS 5608 mutant were purified to apparent homogeneity by means of ultrafiltration, Sephacryl S-200 gel filtration and DEAE Sephadex A-50 chromatography. The apparent molecular weight was calculated as ca. 150 kDa for GI and ca. 128 kDa for GII, respectively. Both enzymes were glycoproteins with isoelectric points of 5.6 (GI) and 5.4 (GII). They had a pH optimun of 4.5 and were stable from pH 5 to 8. The temperature optimum for both enzymes was $60^{\circ}C$, but they were rapidly inactivated above $70^{\circ}C$. The $K_m$ values toward starch were estimated to be 6.57 mg per ml for GI and 4.52 mg per ml for GII, and the $V_{max}$ values were 16.28 ${\mu}M$ per mg for GI and 32.25 ${\mu}M$ per mg for GII, respectively. The $K_m$ and $V_{max}$ values of GII for ${\alpha}-$ or ${\beta}-cyclodextrin$ were estimated to be 0.15 mg per ml and 2.0 mg per ml, respectively ($K_m$) and 1.02 ${\mu}M$ per mg or 1.02 ${\mu}M$ per mg, respectively ($V_{max}$). Neither enzyme exhibited pullulanase activity but they released only glucose from starch or cyclodextrin. Amino acid analysis indicated that both glucoamylases were enriched in proline and acid amino acids. Glucoamylase GII strongly cross-reacted with a monoclonal antibody raised against GI enzymes, and the two enzymes shared very similar amino acid composition. Western blot analysis indicated that L. kononenkoae CBS 5608 mutant produced two forms of glucoamylase on starch, and that synthesis of them was subject to glucose repression.

• Journal of Radiation Industry
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• v.2 no.3
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• pp.121-128
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• 2008

Flavanone-3-hydroxylase (F3H) is one of the key enzymes for the biosynthesis of flavonals, anthocyanins, catechins and proanthocyanins. F3H catalyzes the $3{\beta}$-hydroxylation of (2S)-flavonones to form (2R, 3R)-dihydroflavonols. In this report, we isolated a full-length cDNA of RocF3H from black raspberry (Rubus occidentalis L.) using a reverse transcriptase-PCR and rapid amplification of the cDNA ends (RACE)-PCR. The full-length cDNA of RocF3H contains a 1,098 bp open reading frame (ORF) encoding a 365 amino acid protein with a calculated molecular weight of about 41.1 kDa and isoelectric point (pI) of 5.45. The genomic DNA analysis revealed that the RocF3H gene had three exons and two introns. Comparison of the deduced amino acid sequence of the RocF3H with other F3Hs revealed that the protein is highly homologous with various plant species. The conserved amino acids ligating the ferrous iron and the residues participating in the 2-oxoglutarate binding (R-X-S) were found in RocF3H at the similar positions to other F3Hs. Southern blot analysis indicated that RocF3H exist a multi-gene family. The isolation of RocF3H gene will be helpful to further study the role of F3H gene in the biosynthesis of flavonoids in R. occidnetalis.

• Animal Bioscience
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• v.34 no.12
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• pp.1974-1980
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• 2021

Objective: The objective of this study was to estimate standardized ileal digestibility (SID) of amino acids (AA) in growing pigs fed diets containing increasing levels of copra meal (CM) with β-mannanase supplementation. Methods: Twenty barrows (initial body weight: 34.43±0.11 kg) surgically fitted with T-cannulas at the distal ileum were individually housed in metabolism crates. Pigs were allotted to 5 dietary treatments in a completely randomized design with 4 replicates per treatment. The dietary treatments were: i) NC, negative control, corn-soybean meal (SBM) based diet, ii) PC, positive control, basal diet + 0.10% β-mannanase supplementation (800 IU/kg), iii) CM6, PC diet with 6% CM supplementation, iv) CM12, PC diet with 12% CM supplementation, and v) CM18, PC diet with 18% CM supplementation. A nitrogen-free diet was used to estimate basal endogenous losses of AA for SID calculation. All experimental diets contained 0.5% chromic oxide as an indigestible marker. Each period consisted of a 4-d diet adaptation period and a 3-d ileal digesta collection period. Results: There were no differences in apparent ileal digestibility (AID) and SID of all AA between the NC and PC treatments except that the PC treatment had lower AID and SID of glycine than the NC treatment (p<0.05). There were linear decreases in AID and SID of lysine (p<0.05) and aspartic acid (p = 0.06; tendency) with increasing levels of CM in the diets with β-mannanase. Conclusion: The β-mannanase supplementation had no effect on AA digestibility in pigs fed the corn-SBM based diet but increasing levels of CM reduced SID of lysine and aspartic acid.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.6
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• pp.733-738
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• 2012

Physiochemical characteristics of Luffa cylindrica (L.) Roem and antioxidant activities of its four extracts were analyzed. The chemical composition of L. Roem contained 93.69% moisture, 1.64% crude fiber, 0.52% crude protein, 0.27% crude fat, and 0.06% crude ash. The major minerals were potassium, phosphrous, and calcium. We also analyzed the major organic acids, acetic acid and succinic acid. In free amino acids, ${\beta}$-aminoisobutyric acid showed the highest concentration (100.74 mg/100 g), followed by phosphoethanolamine, urea, asparagine, and valine. Glutamic acid was the highest hydrolyzed amino acid with 1,039.99 mg/100 g followed by aspartic acid, lysine, and arginine. Four extracts from L. Roam, hot-water (LCH), cold-water (LCC), 80% ethanol (LCE), and methanol (LCM), were prepared. Total phenolic and flavonoid levels of LCE exhibited higher than three extracts. The antioxidant potential of extracts from L. Roem were investigated using DPPH, ABTS, and NBT assays. Of the four extracts, LCE had relatively high antioxidant capability on DPPH and superoxide anion radical scavenging activities (12.13%, 16.88%, and 26.61%). Based on the above results, it is suggested that an 80% ethanolic extract from L. Roem was a natural antioxidant material for health food and should be a good ingredient for functional food.

• Korean Journal of Food Science and Technology
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• v.44 no.4
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• pp.393-400
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• 2012

Compositional changes of the Korean chestnut (Castanea crenata) including Daebo, Tsukuba, Tanzawa and Okkwang were investigated in order to characterize them from different periods (immature, mature and storage period). Proximate compositions of mature cultivars were higher than that of the immature cultivars. Between minerals, K (263.0-420.6 mg/100 g) and P (45.8-69.6 mg/100 g) of Tanzawa were highest, and they gradually increased during storage. Tsukuba, in mature period, showed the highest contents of total essential amino acids and glutamic acids as savory amino acids. Major fatty acids were palmitic acid and linolenic acid in four cultivars. In addition, linoleic acids, as ${\omega}$-6 fatty acids, were increased during the storage period. Tanzawa, in the mature period, presented the highest levels of sucrose, however, tsukuba, in the storage period, showed relatively higher free sugar content than others. ${\beta}$-Carotene, as a provitamin A, of Tsukuba in the mature period was highly detected among them, and vitamin C of Tsukuba and Tanzawa was more plentiful than others.