• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.31-32
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• 2005

Neoagaro-oligosaccharides are produced only by enzymatic degradation of agarose by ${\beta}-agarase.^{1)}$ Neoagaro-oligosaccharides inhibit the growth of bacteria, slow the rate of degradation of starch, are used as low-calorie additives to improve food quality, and have macrophage-stimulating activity. Furthermore, neoagarobiose is a rare reagent that has both moisturizing effect on skin and whitening effect on melanoma $cells.^{2)}$ An agar-degrading marine bacterium was isolated from the sea water at the northeast coast in Cheju island, Korea. The strain was gram negative, aerobic, and motile rod. The 16S rRNA of the strain had the closest match of 98% homology, with that from Agarivorans albus. On the basis of several phenotypic characters and a phylogenetic analysis, this strain was designated Agarivorans sp. JA-1. In solid agar plate, Agarivorans sp. JA-1 produced a diffusible agarase that caused agar softening around the colonies. Agarivorans sp. JA-1 was cultured for 36 hr in marine broth 2216 (Difco, USA) and the supernatant that containing an extracellular ${\beta}-agarase$ was prepared by centrifugation of culture media. The enzyme exhibited relatively strong activity at $40^{\circ}C$ and was stable up to $60^{\circ}C$. Using PCR primers derived from the ${\beta}-agarase$ gene of Vibrio sp., the gene encoding ${\beta}-agarase$ from Agarivorans sp. JA-1 was cloned and sequenced. The structural gene consists of 2931 bp encoding 976 amino acids with a predicted molecular weight of 107,360 Da. The deduced amino acid sequence showed 99% and 34% homology to $agaA^{2)}$ and $agaB^{2)}$ genes for ${\beta}-agarase$ from Vibrio sp., respectively. The expression plasmid for ${\beta}-agarase$ gene of Agarivorans sp. JA-1 is being constructed and the recombinant enzyme will be biochemically characterized.

• Journal of Microbiology and Biotechnology
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• v.22 no.12
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• pp.1705-1713
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• 2012

A gene encoding ${\beta}$-glucosidase was cloned from Weissella cibaria 37, an isolate from human feces. Sequence analysis showed that the gene could encode a protein of 415 amino acids in length, and the translated amino acid sequence showed homology (34-31%) with glycosyl hydrolase family 1 ${\beta}$-glucosidases. The gene was overexpressed in E. coli BL21(DE3) using pET26b(+) and a 50 kDa protein was overproduced, which matched well with the calculated size of the enzyme, 49,950.87 Da. Recombinant ${\beta}$-glucosidase was purified by using a his-tag affinity column. The purified ${\beta}$-glucosidase had an optimum pH and a temperature of 5.5 and $45^{\circ}C$, respectively. Among the metal ions (5mM concentration), $Ca^{2+}$ slightly increased the activity (108.2%) whereas $Cu^{2+}$ (46.1%) and $Zn^{2+}$ (56.7%) reduced the activity. Among the enzyme inhibitors (1 mM concentration), SDS was the strongest inhibitor (16.9%), followed by pepstatin A (45.2%). The $K_m$ and $V_{max}$ values of purified enzyme were 4.04 mM and 0.92 ${\mu}mol/min$, respectively, when assayed using pNPG (p-nitrophenyl-${\beta}$-D-glucopyranoside) as the substrate. The enzyme liberated reducing sugars from carboxymethyl cellulose (CMC).

• Journal of Pharmaceutical Investigation
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• v.39 no.1
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• pp.51-54
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• 2009

${\beta}ig$-h3, is a TGF-${\beta}$-induced gene product, extracellular matrix protein with 68 kDa MW(683 amino acids) and has been known for its possible roles in cell adhesion, spreading, migration and proliferation. To minimize a proteolytic degradation of ${\beta}ig$-h3, ${\beta}ig$-h3 incorporated chitosan sponge was prepared and its effects on fibroblast adhesion and migration were investigated. And its wound healing efficacy was evaluated in deep 2nd degree burn rabbit ear wound model. ${\beta}ig$-h3 enhanced fibroblast adhesion and proliferation. In histological observation, a significant over-proliferation of epidermal regeneration was observed in ${\beta}ig$-h3/chitosan dressing applied wound while epidermal regeneration was not proceeded yet in chitosan only treated wound. ${\beta}ig$-h3/sponge dressing could enhance epidermal regeneration.

• Korean Journal of Food Science and Technology
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• v.28 no.1
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• pp.19-27
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• 1996

Zanthoxylum piperitum and Z. schinifolium have been utilized not only as food stuffs, but also as medicinal plants in Korea. In this study, lipids, sugar, amino acids and other components of Zanthoxylum piperitum and Z. schinifolium peels and seeds were analyzed by HPLC and GLC. Four samples contained common fatty acids such as linoleic, linolenic, palmitic, oleic and stearic acid. The contents of unsaturated fatty acids were 87.1% and 64.8% in Z. schinifolium peels and seeds, 73.6% and 62.9% in Z. piperitum peels and seeds, respectively. Z. schinifolium peels contained only beta-sitosterol, whereas other three samples contained campesterol, stigmasterol and beta-sitosterol. In case of free amino acids, peels of both species showed higher contents of acids than seeds of both species. Glutamic acid, aspartic acid, arginine, valine, and leucine were found in all four samples. Essential oils consisted of limonene (30.1-66.8%), beta-phellandrene (4.8-13.3%), citronellal (1.5-22%) and cineol (1.6-3.9%). It is worthwhile to note that the content of citronellal in Z. schinifolium seeds was higher than that of the others.

• Journal of Life Science
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• v.28 no.2
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• pp.195-200
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• 2018

Beta-asarone is the well-known active ingredient of Rhizoma acori graminei. In this study, we investigated and compared the binding affinity of mosquito oviposition pheromone (MOP; (5R,6S)-6-acetoxy-5-hexadecanolide) and beta-asarone on the A domain of the mosquito odorant binding protein 1 (CquiOBP1) by in silico computational docking studies. The three-dimensional crystallographic structure of CquiOBP1 was obtained from the PDB database (PDB ID: 3OGN). In silico computational auto-docking analysis was performed using PyRx, Autodock Vina, Discovery Studio Version 4.5, and the NX-QuickPharm option based on scoring functions. The beta-asarone showed optimum binding affinity (docking energy) with CquiOBP1 as -6.40 kcal/mol as compared to the MOP (-6.00 kcal/mol). Among the interacting amino acids (LEU76, LEU80, ALA88, MET89, HIS111, TRP114, and TYR122), tryptophan 114 in the CquiOBP1 active site significantly interacted with both MOP and beta-asarone. Amino acids substitution (mutation) from non-polar groups to the polar (or charged) groups of the CquiOBP1 dramatically changed the X, Y, Z grid position and binding affinity of both ligands. These results significantly indicated that beta-asarone could be a more potent ligand to the CquiOBP1 than MOP. Therefore, the extract of Rhizoma acori graminei or beta-asarone can be applied to the fields of insecticidal and repellant biomaterial development.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.36-36
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• 2017

Out of twenty common protein amino acids, there are many kinds of non protein amino acids (NPAAs) that exist as secondary metabolites and exert ecological functions in plants. Mimosine (Mim), one of those NPAAs derived from L. leucocephala acts as an iron chelator and reversely block mammalian cell cycle at G1/S phases. Cysteine (Cys) is decisive for protein and glutathione that acts as an indispensable sulfur grantor for methionine and many other sulfur-containing secondary products. Cys biosynthesis includes consecutive two steps using two enzymes-serine acetyl transferase (SAT) and O-acetylserine (thiol)lyase (OASTL) and appeared in plant cytosol, chloroplast, and mitochondria. In the first step, the acetylation of the ${\beta}$-hydroxyl of L-serine by acetyl-CoA in the existence of SAT and finally, OASTL triggers ${\alpha}$, ${\beta}$-elimination of acetate from OAS and bind $H_2S$ to catalyze the synthesis of Cys. Mimosine synthase, one of the isozymes of the OASTLs, is able to synthesize Mim with 3-hydroxy-4-pyridone (3H4P) instead of $H_2S$ for Cys in the last step. Thus, the aim of this study was to clone and characterize the cytosolic (Cy) OASTL gene from L. leucocephala, express the recombinant OASTL in Escherichia coli, purify it, do enzyme kinetic analysis, perform docking dynamics simulation analysis between the receptor and the ligands and compare its performance between Cys and Mim synthesis. Cy-OASTL was obtained through both directional degenerate primers corresponding to conserved amino acid region among plant Cys synthase family and the purified protein was 34.3KDa. After cleaving the GST-tag, Cy-OASTL was observed to form mimosine with 3H4P and OAS. The optimum Cys and Mim reaction pH and temperature were 7.5 and $40^{\circ}C$, and 8.0 and $35^{\circ}C$ respectively. Michaelis constant (Km) values of OAS from Cys were higher than the OAS from Mim. Inter fragment interaction energy (IFIE) of substrate OAS-Cy-OASTL complex model showed that Lys, Thr81, Thr77 and Gln150 demonstrated higher attraction force for Cys but 3H4P-mimosine synthase-OAS intermediate complex showed that Gly230, Tyr227, Ala231, Gly228 and Gly232 might provide higher attraction energy for the Mim. It may be concluded that Cy-OASTL demonstrates a dual role in biosynthesis both Cys and Mim and extending the knowledge on the biochemical regulatory mechanism of mimosine and cysteine.

• Food Science and Preservation
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• v.19 no.4
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• pp.594-603
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• 2012

This study developed a high-utility type of vinegar from Rubus coreanus by optimizing its fermentation conditions. In the alcohol fermentation process, the optimal conditions for the maximization of the alcohol contents were an initial sugar concentration of 15 $^{\circ}Brix$, a temperature of $30^{\circ}C$ and 4 days. The optimal conditions for the acetic acid fermentation were 9 days of fermentation at $30^{\circ}C$ and 200 rpm, with 6% alcohol and 2% initial acidity. The sucrose, fructose, and glucose contents were 952.90, 491.01, and 386.62 mg%, respectively. The free organic acids were acetic, malic, succinic, malonic, oxalic, and lactic acids. The total free amino acid content was 104.33 ${\mu}g/mL$, with alanine, glutamic acid, ${\gamma}$-amino-N-butyric acid, and o-phospho-ethanolamine as the major amino acids. The K, Na, and Mg contents were 1,686.10, 172.50, and 69.33 ppm, respectively. The total phenolic and anthocyanin contents were 25.19 and 80.71 mg/100 mL, respectively. The DPPH- and $ABTS^{.+}$ radical scavenging activities were approximately 65 and 94%, respectively. Moreover, the vinegar's ${\beta}$-carotene bleaching activity and reducing power showed that it had strong anti-oxidant properties. These results show that Rubus coreanus vinegar has anti-oxidant properties and may be used as functional food.

• The Korean Journal of Food And Nutrition
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• v.31 no.6
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• pp.811-820
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• 2018

This study suggests a method of making cheese using soybeans for healthy food for people that are allergic to animal proteins, vegetarians, people on a diet, infants and toddlers. Additionally, the study was conducted to provide basic data that can contribute to the development of a quality control class of Korean cheese and underdeveloped cheese industry. Soybean cheeses have a high protein content and low fat. The free amino acids of soybean cheeses contained 11.48 mg of arginine per 100 g, 9.33 mg of glutamate, and leucine 4.91 mg, in that order. The free amino acids of Company A's milk cheese contained 20.95 mg of glutamate, 8.95 mg of proline and 8.02 mg of lysine per 100 g. In soybean cheeses, there were 2.21 mg of tryptophan and 0.73 mg of cysteine, which were not analyzed in the milk cheese of company A. The contents of the constituent amino acids was 1,070.22 mg of glutamate, 467.30 mg of aspartate and 446.30 mg of leucine in 100 grams of soybean cheeses per 100 grams. The milk cheese of Company A was 1,715.97 mg of glutamate, 798.72 mg of leucine and 685.31 mg of proline. The mineral contents of the soybean cheese were 120.29 mg/100 g of calcium, 0.92 mg/100 g of iron, 0.78 mg/100 g of zinc and 0.40 mg/100 g of selenium, respectively. The contents of vitamin $B_1$ and ${\beta}$-glucan in soybean cheese were higher than that of the milk cheese in Company A at 0.08 mg/100 g and 13.73 mg/g, respectively. Therefore, it is considered that the soy cheese is excellent in nutritional aspect and will contribute to health promotion. It is also suitable for people that are allergic to animal proteins, vegetarians, people on a diet and healthy foods for infants.

• Toxicological Research
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• v.6 no.1
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• pp.99-107
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• 1990

Dopamine beta-hydroxylase (DBH) is a neurotransmitter biosynthetic enzyme which catalyzes the conversion of dopamine to norepinephrine. Although structural studies of the mature form of this enzyme have been extensive, the culmination of these finding had been hightly controversial and contradictory. In this study, biochemical approaches were taken to characterize the structure of mature DBH. Soluble bovine DBH was purified from aderenal medulla. Three bands of 69 kDa, 72 kDa and 75 kDa which were physically separable and similar in structure were observed by SDS-PAGE. Furthermore, gas phase sequence analysis revealed that the 72 kDa band consists of two polypeptides which are present at equimolar concentrations and differed in that one had three extra amino acids at the N-terminus. Taken together, the soluble form of DBH exist in at least four forms, three identified by SDS-PAGE, one of which consists of two polypeptides as identified by N-terminal sequence analysis. The significance of these forms and their possible biosynthetic mechanisms are discussed.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.3
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• pp.432-444
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• 2001

In this manuscript, several effective feeding and management systems to reduce environmental pollution in swine production have been briefly introduced. It is logical that reducing the excretion of nutrients in manure should be the first step to reduce the environmental impact of pig production. it is evident that the excretion of nitrogen and phosphorus can be reduced when more digestible or available feedstuffs are used. Also, it is well known that proper feed processing can reduce anti nutritional factors (ANF) and improve nutrient digestibilities. Supplementation of effective feed additives can reduce excretion of nitrogen and phosphorus due to efficient feed utilization. These include enzymes (e.g., phytase), antibiotics, probiotics, organic acids and growth hormones ($\beta$-agonists and porcine somatotropin). One of the most effective ways to reduce pollutants from swine manure is to use synthetic amino acids in feed manufacturing. Many studies showed that reduction of 2 to 4% unit (U) of dietary protein with supplemental amino acid (AA) could dramatically reduce (15 to 20%) nitrogen excretion. Regarding feeding strategies, it has been recognized that phase feeding regimen could be used to reduce nitrogen and phosphorous excretion by feeding pigs in better agreement with age and physiological state. Feeding barrows and gilts separately, known as split sex feeding, can also decrease excretion of nitrogen and phosphorus. With the increasing concerns on the negative impact of animal production systems on the environment, animal nutritionists and producers should be aware that sustainability of animal agriculture is as important as high production performance. Therefore, some feeding and management strategies described in this manuscript will help to reduce environmental pollution in swine production. Proper combination of feeding regimen and environment-friendly diet formulation through nutritional approach will be more effective to reduce nutrient excretion in swine production system compared to single approach to do so.