• BMB Reports
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• v.37 no.4
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• pp.408-415
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• 2004

The expression of the Paenibacillus sp. A11 cyclodextrinase (CDase) gene using the pUC 18 vector in Escherichia coli JM 109 resulted in the formation of an insoluble CDase protein in the cell debris in addition to a soluble CDase protein in the cytoplasm. Unlike the expression in Paenibacillus sp. A11, CDase was primarily observed in cytoplasm. However, by adding 0.5 M sorbitol as an osmolyte, the formation of insoluble CDase was prevented while a three-fold increase in cytoplasmic CDase activity was achieved after a 24 h-induction. The recombinant CDase protein was purified to approximately 14-fold with a 31% recovery to a specific activity of 141 units/mg protein by 40-60% ammonium sulfate precipitation, DEAE-Toyopearl 650 M, and Phenyl Sepharose CL-4B chromatography. It was homogeneous by non-denaturing and SDS-PAGE. The enzyme was a single polypeptide with a molecular weight of 80 kDa, as determined by gel filtration and SDS-PAGE. It showed the highest activity at pH 7.0 and $40^{\circ}C$. The catalytic efficiency ($k_{cat}/K_m$) values for $\alpha$-, $\beta$-, and $\gamma$-CD were $3.0{\times}10^5$, $8.8{\times}10^5$, and $5.5{\times}10^5\;M^{-1}\;min^{-1}$, respectively. The enzyme hydrolyzed CDs and linear maltooligosaccharides to yield maltose and glucose with less amounts of maltotriose and maltotetraose. The rates of hydrolysis for polysaccharides, soluble starch, and pullulan were very low. The cloned CDase was strongly inactivated by N-bromosuccinimide and diethylpyrocarbonate, but activated by dithiothreitol. A comparison of the biochemical properties of the CDases from Paenibacillus sp. A11 and E. coli transformant (pJK 555) indicates that they were almost identical.

• Journal of Microbiology and Biotechnology
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• v.21 no.2
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• pp.183-186
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• 2011

Biochemical methods were selected to evaluate the role of exopolymeric substances in the stability of biofilms used in bioremediation processes. Biofilms of Thiomonas arsenivorans formed on pozzolana were thus treated with pronase (protein target), lectins (Con A or PNA), calcofluor or periodic acid (polysaccharides target), DNase (DNA target), and lipase (triglycerides target). Neither protease nor DNase treatments had any effect on bacterial adhesion. Lectins and calcofluor treatments mainly affected young biofilms. Lipase treatment had a noticeable effect on biofilm stability whatever the biofilm age. Results suggest that it would be an increased resistance of mature biofilms that protects them from external attacks.

• Journal of Plant Biology
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• v.29 no.4
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• pp.263-284
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• 1986

Effects of putrescine, spermidine and spermine on the activity of D-glucose-6-phosphate cyclohydrolase in the Daucus carota L. protoplast cultured for 4 days and effects of polyamines on the incorporation of D-[u-14C]-glucose treated to protoplasts in culture-medium were investigated. The activity of D-glucose-6-phosphate cyclohydrolase was increased by polyamines and among them spermine was the most effective. Polyamiens increased protein synthesis and this due to the increasing effect of the polyamines on the synthesis of glycoprotein which is one of cell wall components. The synthesis of cell polysaccharides, such sa pectic substances, hemicelluloses and cellulose was increased by polyamines, which stimulated synthesis of pectin substances, and hemicellulose more greatly than that of cellulose, and spermidine was the most effective. In the light of the above results it seems that the polyamines increase cell wall regeneration by the stimulation of enzyme activities which synthesize cell wall components.

• Archives of Pharmacal Research
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• v.8 no.1
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• pp.31-38
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• 1985

Polysaccharide fractions were prepared from Forsythia Corea by several different extraction schedules. The fractions obtained were designated as ForCo A, ForCo B and ForCo C respectively. ForCo C was further purified through Sephadex G 200 column chromatography and obtianed two subfractions (Coreana A, Coreana B). ForCo showed marked antitumor activity against sarooma 180 and its activity was dose-dependent. Coreana A, purified from ForCo C, showed somewhat higher antitumor activity. ForCo C increased the number of circulating leucocytes and peritoneal exudate cells, but didn't show any significant effects on the phagocytic activity and total serum protein level. Chemical analysis showed that ForCo C was composed of glucose, galactose, xylose and arabinose. A close relationship between antitumor activities and polysaccharide contents waw observed. These results indicate strongly that antitumor active principle of Forsythia Corea is polysaccharide.

• Archives of Pharmacal Research
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• v.2 no.1
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• pp.9-16
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• 1979

3-Amino-2-phenylthietane derivatives were considered as a useful tool to elusidate the mechanism of inhibiton of MAO by tranylcypromine-type inhitors. The synthesis of 3-benzoylamino-2-phenylthieetane, 3-amino-2-phenylthietane, and 3-N, N-dimentylamino-2-p-nitrophenythietane was attempted using the reaction between 1, 3 dihalogeno alkanes with alkali sulfide. When 1-pheny1-1, 3-dihalo-2-benzolaminopropane was treated with sodium sulfide, 2-pheny 1-4 benzylidene-2-oxazoline was isolated, indicating the case of elimination reaction compared to ring formation. The reaction of 1-p-nitropheny1-1, 3-dichloro-2-N, N-dimethylaminopropane with sodium sulfide gave bis (1-p-nitropheny1-2-N, N-dimethylamino-3-chloropropane)sulfide. The mechanism of reaction was discussed.

• Korean Journal of Pharmacognosy
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• v.15 no.1
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• pp.30-38
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• 1984

Yam, Dioscorea batatas Decne (Dioscoreaceae) is a plant growing in Korea. Dioscoreae Rhizoma in the markets are classified into natural and cultural ones. These studies were conducted to investigate the differencies between natural and cultural Yams by characteristics of morphology, cell contents, chemical components and physical properties. More mucilage cells which contain calcium-oxalate raphides were contained in cultural Yam than natural Yam. However, the amount of crude protein, crude lipid and mucilage viscosity, swelling, gelatinization of polysaccharides were greater in natural Yam than the other. It is suggested that natural Yam is excellent as nutrition agent in quality and worthwhile as a resource of crude drugs.

• Journal of the Korean Society of Food Science and Nutrition
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• v.24 no.1
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• pp.147-153
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• 1995

The cultured mycelial cells of Ganoderma lucidum was extracted by alkali, and then neutralized by acid. The extract was passed through the column of DEAE cellulose for more purification. The neutral fraction was concentrated and precipitated with 95% ethanol. The precipitate was lyophilized and then PSG(polysaccharides) was obtained. PSG was composed of 82.2% polysaccharide, 0.7% protein and 17.1% uronic acid. Sugar conjugates of its hydrolysates were produced using with fluorescent compound(7-amino-1,3-naphthalene disulfonic aicd : 7-AGA), and then fluorescent labeled sugar conjugates were separated by reverse phase high perfomance liquid chromatography. Hydrolysates of PSG were composed of sixteen amino acids and 95.7% glucose, 2.7% xylose, 1.6% fucose and tract amount of galactose and mannose. The immunomodulating effects of PSG on macrophage were perfomed using murine macrophage cell line ATCC TIB 71 cells. PSG augumented the phagocytic activity of TIB 71 cells against fluorescent latex beads.

• Journal of the Korean Society of Food Science and Nutrition
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• v.26 no.6
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• pp.1102-1108
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• 1997

Proteins and polysaccharides confer distinct functional properties in food systems. This research was attempted to improve emulsion properties of casein by protein-polysaccharide conjugation, in which alginates with various molecular weights were employed as polysaccharide sources. Casein-alginate mixtures were conjugated by the amino-carbonyl or Maillard reaction at 6$0^{\circ}C$ and 79% relative humidity. The resulting casein-alginate conjugates were tested for their emulsion activity and emulsion stabilizing properties. In general, the emulsion stability of casein-alginate mixture greatly increased due to the amino-carbonyl reaction between casein and alginates, whose magnitude depended on the molecular weight of alginate, weight ratio of casein to alginate and incubation time. It was also found that thermal stability and pH stability were markedly improved by the casein-alginate conjugation.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.11
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• pp.1634-1641
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• 2005

Generally, dietary fibre (DF) includes lignin, non-starch polysaccharides (NSP) and resistant starch (RS). In monogastric species, low levels of dietary fibre in the diet are associated with various diseases and high levels reduce nutrient digestibilities. In this study, the effects of different types and levels of NSP (soluble: $\beta$-glucan, insoluble cellulose) and resistant starch on mucin secretion and endogenous nitrogen and amino acid losses in pigs were investigated. A total of 25 five-week-old weaner pigs (9.5 kg${\pm}$1.5 kg), were randomly allocated to each of five experimental diets. Different levels of purified barley $\beta$-glucan (BG) extract (5 or 10% of $Glucagel^{(R)}$ $\beta$-glucan, providing 4 or 8% of BG in the diet), and resistant starch (RS) (8.3 or 16.6% of Hi-$Maize^{TM}$, providing 5 or 10% RS in the diet) were substituted for wheat starch in a purified diet in which enzymatically-hydrolysed casein was the sole source of protein. The diets were fed for 21 days. No statistically significant difference between treatments (p>0.05) was observed for growth performance and organs weights. No difference in ileal starch digestibility was observed between pigs on the cellulose or $\beta$-glucan diets. However, as the level of resistant starch in the diet increased the ileal starch digestibility decreased (p<0.05). The inclusion of resistant starch in the diet (5 or 10%) did not increase mucin production when compared with the cellulose-only diet. However, as the level of beta-glucan in the diet increased, both crude mucin in the digesta dry matter and per kg dry matter intake increased (p<0.05). Pigs fed the diet containing 8% of beta-glucan had higher endogenous loss flow than those fed the diets including 5 or 10% of resistant starch or 4% of $\beta$-glucan. In conclusion, dietary inclusion of resistant starch increased the level of starch reaching the large intestine without any effect on mucin secretion, or endogenous nitrogen or amino acid losses content in the small intestine. The addition of $\beta$-glucan to a diet containing cellulose increases both mucin secretion and endogenous amino acid and nitrogen losses in the small intestine.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.31 no.3
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• pp.447-451
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• 1998

Functional properties such as anti-blood coagulation, angiotensin converting enzyme (ACE) inhibitory activity, fat binding capacity, foaming properties, emulsifying properties and chemical components of sulfated polysaccharides isolated from ascidian tunics were investigated. The sulfated polysaccharide mainly consisted of sulfate, uronic acid, protein, and chondroitin sulfate, among which chondroitin sulfate showed higher concentration while sulfate and uronic acid did lower. Compositional menosaccharides were arabinose, xylose, glucose, galactose, glucuronic acid, N-acetylgalactosamine and N-acetyglucosamine. Especially, galactose content was dominant among them. And emulsifiability and foaminess of the sulfated polysaccharide was higher than the control group. Anti-blood coagulation of sulfated polysaccharide showed with respect to APTT (Activated partial thromboplastin time). ACE inhibitory activity showed about $16.7\%$.