• Applied Chemistry for Engineering
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• v.30 no.5
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• pp.586-590
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• 2019

The aim of this study was to fabricate a dextran polyelectrolyte multi-layer on a bare metal stent (BMS) and to evaluate bio-physical properties of the layer. Diethylaminoethyl-dextran (DEAE-D) as a polycation and dextran sulfate (DS) as a polyanion were successively coated on the bare metal stent by a well-known layer-by-layer procedure. The morphology of the stent surface and its cell adhesion were studied after each coating step by scanning electron microscopy. The stent showed more blotched and slightly rougher morphology after dextran-DS coating. The contact angle of the DEAE-DS group ($39.5{\pm}0.15^{\circ}$) was significantly higher than that of the BMS group ($45.16{\pm}0.08^{\circ}$), indicating the improvement of hydrophilic. The SMC proliferation inhibition in the DEAE-DS-coated stent group ($20.9{\pm}0.04%$) was stronger than that in the control group ($21.7{\pm}0.10%$ in DS-coated group only). The DEAE-DS coating is desired for stent coating materials with biocompatibility and anti-restenosis effect.

• Journal of Microbiology and Biotechnology
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• v.9 no.2
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• pp.219-222
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• 1999

A simple sequence of membrane concentration and DEAE-Cellulose chromatography has been optimized to give a purified dextransucrase from Leuconostoc mesenteroides B-512FMCM with the highest specific activity (248.8 IU/mg protein) ever reported in high yield (overall 88.7%) for dextransucrase. When there was no sucrose in the dextransucrase and the dextran reaction digest, the dextransucrase hydrolyzed glucose from dextran. The glucose was transferred to the other glucoses from dextran and formed isomaltose and isomaltodextrin. The transglycosylation efficiency of glucose from dextran was much higher with acceptors. The dextransucrase can be used for the production of various kinds (or structures) of oligosaccharides using dextran and various acceptors with almost 100% theoretical yield.

• Asian-Australasian Journal of Animal Sciences
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• v.6 no.4
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• pp.581-589
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• 1993

A plasmid vector, $RSVLTR/{\beta}G2$, containing lacZ gene under the control of the RSVLTR promoter were transfected into chicken embryo fibroblasts by three different transfection methods. Calcium phosphate, lipsome and DEAE-dextran techniques were applied for transfection of chicken cells. A histochemical assay with X-gal was used as a simple method for screening transfected cells. Plasmid $RSVLTR/{\beta}G2$ was expressed proficiently in the chicken embryo fibroblast. Calcium phosphate-DNA precipitate transfection resulted in the highest efficiency for transient expression of $RSVLTR/{\beta}G2$. Transfected cells formed colonies on the 9th day of incubation indicating stable transformation of the inserted plasmid.

• The Korean Journal of Food And Nutrition
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• v.7 no.2
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• pp.119-123
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• 1994

Aspergillus sp. BY-54 which produced a strong dextran hydrolyzing enzyme was isolated from soil. Using this strain, the optimal cultural conditions, enzyme purification and characterization were studied. The results are as follows : The optimal concentration of dextran as carbon source was l%. and the optimum temperature and the initial pH for enzyme production was 3$0^{\circ}C$, and 7.0, respectively. Dextranase was purified by DEAE-cellulose column chromatography with a linear gradient increase in NaCl. Km value of dextranase was 0.222%, and several glucans containing various types of glucosidic linkages such as DEAE-sephadex, CM-sephadex and sephadex G-100 were almost digested to a large extent with this dextranase. The enzyme was strongly inhibited by sodium fluoride, KMnO4 and p-CMB, while KCN caused 20% of activation.

• The Korean Journal of Mycology
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• v.11 no.1
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• pp.23-26
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• 1983

Aspergillus ustus was cultured in the salts media contained dextran (2%). Then the cultured liquid media were filtrated and concentrated up to 10 folds by evaporation, and then purified by means of acetone precipitation, of a repeated chromatography on the columns of DEAE-Ccellulose, Biogel P-150, and Sephadex G-200. Total proteins in the initial culture filtrate were 38,500mg, but the final amounts of proteins were 172mg. The specific activity of the protein in the culture filtrate was $1,340\;{\mu}moles$ products per minute per mg protein, but the final specific activity of the protein was $2,448\;{\mu}\;moles$ products per minute per mg protein. The final yields remained about 30% of the initial.

• The Korean Journal of Zoology
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• v.34 no.2
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• pp.196-202
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• 1991

Juvenile honnone binding protein (JHBP) was identified in the last instar larval hemolymph of Lymantria dispar using dextran coated charcoal (DCC) binding assay and gel filtration. The p1 value of JHBP was estimated to be 5.3. JHBP was partially pudfied by polyethylene glycol(PEG) precipitation, DEAE-cellulose ion-exchange chromatography and gel filtration, and was confirmed by DCC binding assay.

• Journal of Microbiology and Biotechnology
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• v.10 no.5
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• pp.728-732
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• 2000

Propagation of Anagrapha falcifera nuclear polyhedrosis virus(AfNPV) was investigated using well-plates and split-flow air-lift bioreactors. In well-plate experiments, the effects of pH, cell density at a point of infection, serum concentration, DEAE-dextran, and lipid on virus propagation were all closely examined. The AfNPV titer in well-plates was optimal at pH 6.8 and $3{\times}10^6$ cells/$cm^2$. The virus titer was not dramatically affected when the fetal bovine serum concentration was reduced from 10% to 5%. The addition of cholesterol at AfNPV infection of Sf21 cells enhanced the virus titer, whereas the addition of DEAE-dextran did not improve the titer. The AfNPV titer ($3.8{\times}10^7$ $TCID_{50}/ml$) at optimized conditions for well-plate experiments was 2.5-fold higher than for the control. In bioreactor experiments, the AfNPV titer showed its maximum level at air flow rates of 20-40 ml/min. In a split-flow air-lift bioreactor, AfNPV titer ($2.3{\times}10^7\;TCID_{50}/ml$) was 1.5-fold higher than the control when the culture was at pH 6.8 and supplemented with 0.34 mM cholesterol.

• Journal of the Korean Chemical Society
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• v.43 no.2
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• pp.193-198
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• 1999

The transfection efficiency of plasmid DNA was inspected using multi-cationic polymer, 5, 10, 25 and 50KD polyethylenimine (PEI). The optimal neutralization ratio of PEI/DNA complexes by agarose assay was 1.5-2.0 (nmol/nmol) without much difference in molecular weight of PEI.In vitro transfection assay, most of PEI-mediated plasmid delivery was better compared to the naked DNA. Especially, 25KD PEI at optimal condition gave higher transfection rather than the standard assay of DEAE-dextran or Lipofectin. To enhance the cell targeting delivery, the liposome formulations were introduced using phospholipids. As a result, PC/PE liposomes increased 2-2.5 times of the transfection efficiency of PEI single or PC/PE single delivery, but not the case of 25KD PEI. Moreover, the DOTAP/PE-introduced PEI delivery reduced the transfection of DOTAP/PE single delivery. All these results proved that the PEI can be used not only good transfectants and but also good DNA condensing agents in neutral/anionic liposome for cell targeting delivery.

• KSBB Journal
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• v.19 no.5
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• pp.363-367
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• 2004

Leuconostoc mesenteroides NRRL B-1149 produces various glucoseyltransferases for the synthesis of dextran, levan and glucose-1-phosphate using sucrose as a substrate. A sucrose phosphorylase (1149SPase) was purified from L. mesenteroides NRRL B-1149 culture by using hollow fiber filtration (30 kDa cut off), Toyopearl DEAE 650 M column chromatography and following two times of DEAE-Sepharose column chromatographies. The specific activity of the purified 1149SPase was 25.7 (U/mg) with $16\%$ yield. The 1149SPase showed a molecular size of 56 kDa on denatured $10\%$ SDS-PAGE. The N-terminal amino acid sequence of the enzyme was MEIQNKAM. The optimum pH and temperature of this enzyme were 6.2~6.5 and 37^{circ}C, respectively. It had an apparent K_{m} of 6.0 mM and K_{cat} of 1.62/s for sucrose. 1149SPase crystal was formed by hanging drop diffusion technique using 20 mM calcium chloride dihydrate, 100 mM sodium acetate trihydrate pH 4.6 and $30\%$ 2-methyl-2,4-pentanediol as vaporizing and reservation solution. The 1149SPase catalyzes transferring of glucose from isomaltose or sucrose to salicin and salicyl alcohol by disproportionation reaction or acceptor reaction and synthesized two acceptor products, respectively.

• Journal of Microbiology and Biotechnology
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• v.11 no.4
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• pp.628-635
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• 2001

Recombinant E. coli DH5$\alpha$ harboring a dextransucrase gene (dsrB742) produced an extracellular dextransucrase in a 2% sucrose medium. The enzyme was purified by DEAE-Sepharose and Phenyl-Sepharose column chromatographies upto a 142.97-fold purification with a 11.11% recovery to near homogeneity. The enzyme had a calculated molecular mass of 168.6 kDa, which was in good agreement with the activity band of 170 kDa on a nondenaturing SDS-PAGE. An expression plasmid was constructed by inserting the dsrB742 into a pRSET expression vector. The activity after expression in E. coli BL21(DE3)pLysS increased about 6.7-fold compared to the extracellular dextransucrase from L. mesenteroides B-742CB. The expressed and purified enzyme from the clone showed similar biochemical properties (acceptor reaction, size of active dextransucrase, optimum pH, and temperature) to B-742CB dextransucrase, however, the ability to synthesize ${\alpha}$-(1$\rightarrow$3) branching decreased in comparison to that of L. mesenteroides B-742CB dextransucrase.