• Journal of Microbiology and Biotechnology
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• v.16 no.7
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• pp.1104-1110
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• 2006

Recombinant antibody-toxin is a bifunctional protein that binds and kills a target cell expressing a specific antigen on the surface of the cell, and its structure is chimeric, in which a toxin is fused to an antigen-binding domain such as scFv or Fab. Divalent antibody-toxin molecules showed higher cytotoxicities against cancer cell lines than monovalent molecules. However, the yields of the divalent molecules were very low. In this study, we introduced the CH2, CH3, or CH2-CH3 (=Fc) domain of antibody in the middle of the Fab-toxin between the hinge region of human IgG1 and the toxin domain to increase the yield. The covalently bonded dimer could be formed by three disulfide bridges from cysteine residues in the hinge region. The molecule with the CH3 domain showed about 3-fold higher dimerization yield than previously constructed Fab-toxin molecules, while maintaining the cytotoxic activity comparable to that of scFv-toxin. However, the introduction of CH2 or Fc domain to the same position showed little effect on the dimerization yield. We also observed that the introduction of the CH3 region made it possible to form noncovalently associated dimer molecules.

• Journal of Microbiology and Biotechnology
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• v.25 no.9
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• pp.1476-1484
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• 2015

Three endoxylanase-encoding genes from the moderately themophilic chemoorganotrophic bacterium Melioribacter roseus were cloned and expressed in Escherichia coli. Genes xyl2091 (Mros_2091) and xyl2495 (Mros_2495) encode GH10 family hydrolases, whereas xyl2090 (Mros_2090) represents the GH30 family. In addition to catalytic domains, Xyl2090 and Xyl2091 contain carbohydrate-binding modules that could facilitate their binding to xylans and Por sorting domains associated with the sorting of proteins from the periplasm to the outer membrane, where they are covalently attached. Recombinant endoxylanase Xyl2495 exhibited a high specific activity of 1,920 U/mg on birchwood xylan at 40℃. It is active at low temperatures, exhibiting more than 30% of the maximal activity even at 0℃. Endoxylanases Xyl2090 and Xyl2091 have lower specific activities but higher temperature optima at 80℃ and 65℃, respectively. Analysis of xylan hydrolysis products revealed that Xyl2090 generates xylo-oligosaccharides longer than xylopentaose. Xylose and xylobiose are the major products of xylan hydrolysis by the recombinant Xyl2091 and Xyl2495. No activity against cellulose was observed for all enzymes. The presence of three xylanases ensures efficient xylan hydrolysis by M. roseus. The highly processive "free" endoxylanase Xyl2495 could hydrolyze xylan under moderate temperatures. Xylan hydrolysis at elevated temperatures could be accomplished by concerted action of two cell-bound xylanases; Xyl2090 that probably degrades xylans to long xylo-oligosaccharides, and Xyl2091 hydrolyzing them to xylose and xylobiose. The new endoxylanases could be useful for saccharification of lignocellulosic biomass in biofuels production, bleaching of paper pulp, and obtaining low molecular weight xylooligosaccharides.

• Journal of Ginseng Research
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• v.17 no.2
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• pp.114-122
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• 1993

0.5 mg of natural ginsenoside mixture and 0.8 $\mu$Ci of synthesized 14C-ginsenosides were administered orally to a rat and killed at one hour after the ginsenoside administration and the liver was fractionated into nuclear fraction, mitrochondria microsomes and cytosol fraction. Radioactivity distribu lion in subcellular fractions of the liver showed that 32o1c of total radioactivity absorbed in the liver was in cytosol fraction but a significant portion of the radioactivity was also found in mitochondria (26.6%) and microsomal fraction (18.l%). 5.8% of the total radioactivity was recovered from the nuclear fraction as well. This suggested that ginsenosides might be distributed into all subcellular fractions. Activities of mitochondrial aldehyde dehydrogenase, lactate dehydrogenase and malate dehydrogenase of the liver of rat at two hours after the ginsenoside administraion were found appreciably stimulated, suggesting that the ginsenoside concentration in the liver might be around 10-5%, since optimum concentrations for most enzyme catalyzed reactions in vitro were known to be 10-6% 10-4%. A significant portion of the radioactivity recovered from subcellular fractions of the liver was found in protein fractions, suggesting that proteins might interact with ginsenosides. Examination of protein-ginsenoside interation by gel filtration, equilibrium dialysis and amonium sulfate precipitation technique suggesting that proteins and ginsenosides do not bound covalently but weakl\ulcorner combined. When purified ginsenoside Rbl and Rgl were incubated with rat liver cytosolic enzymes for 20 min, the above ginsenosides were hydrolyzed quickly, suggesting that ginsenosides might be rapidly hydrolyzed and metabolized in the liver. It was also observed in vitro that the ginsenosides such as Rbl and Rgl were easily hydrolyzed by rat liver cytosol preparation suggesting that absorbed ginsenosides might be quickly hydrolyzed and metabolized in the liver.

• Polymer(Korea)
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• v.33 no.5
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• pp.452-457
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• 2009

Hybrid nanomaterials consisting of multi-walled carbon nanotube(MWNT) and/or PEDOT of conductive polymer were prepared in this study. In the presence of catalyst and ligand, the MWNT-Br compound prepared by the successive surface treatment reaction was mixed with MMA to initiate the atom transfer radical polymerization process. PMMA was covalently linked to the surface of MWNT for the formation of MWNT/PMMA nanocomposites. The EDOT and oxidant were added in the aqueous emulsion of PS produced via a miniemulsion polymerization process and then it proceeded to carry out the oxidative chemical polymerization of EDOT for the preparation of PEDOT/PS nanoparticles with the core-shell structure. The aqueous dispersion of PEDOT:poly(styrene sulfonate) (PSS) was mixed with the silica particles treated with a silane compound and thus PEDOT:PSS-clad silica nanoparticles were prepared by the surface chemistry reaction. The hybrid nanomaterials were analyzed by using TEM, FE-SEM, TGA, EDX, UV, and FT-IR.

• Korean Journal of Veterinary Service
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• v.29 no.3
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• pp.277-285
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• 2006

Plasmids are covalently closed circular molecules of DNA that are stably inherited and replicate somewhat independently of the bacterial chromosome. Genes carried on plasmids can mediate a wide variety of important functions, including antibiotics (R plasmids) and heavy metals resistance, toxins production, cell penetration, iron chelation, complement resistance, and metabolic characteristics such as sucrose and lactose fermentation. Fifty strains of lactobacilli were isolated from 26 staters and 29 fermented milk products. They were classified 27 strains as Lactobacillus paracasei subsp paracasei, 11 stains as Lactococcus lactis subsp cremoris, 6 strains as L delbrueckii subsp lactis, 4 strains as L acidophius, and 2 strains as L delbrueckii subsp bulgaricus. All of these strains were examined for drug resistance and transferability of R plasmids. All of the isolates were sensitive to Am, C, CF, E, NB, P, T, and Te. But resistant to SXT 94% (47 strains), K 66% (33 strains), S 56% (28 strains), ENR 50% (25 strains), NOR 38% (19 strains) CIP 38% (19 strains), GM 16% (8 strains), and N 14% (7 strains), in order. And 32 different resistant patterns were found. The most frequently encountered patterns were CIP-ENR-K-NOR-S-SXT (5 strains). In vitro R plasmids transfer experiment, 57 antibiotic resistant strains which were not transfer to the recipient 2 Escherichia coli strains by conjugation, These results indicate that Lactobacillus in internal trade market' stater recognize R factor but transmissible R plasmid is not existed.

• Korean Journal of Materials Research
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• v.9 no.4
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• pp.355-361
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• 1999

We have synthesized the soluble polymide based side chain system with covalently attached NLO chromophore Disperse Red 1. The developed polymer is optically transparent in the range of optical communication wave-lengths. Its glass transition temperature$(T_g)$ and thermal decomposition temperature$(T_d)$ are $225^{\circ}C, 310^{\circ}C$ respectively. The poled film with the poling field of $100 V/\mu\textrm{m}$ shows that the dielectric constant is 3.37 at the 10-kHz frequency, the refractive indices of TM and TE modes are both 1.631 at $\lambda = 1300 nm$, and the Electro-optic coefficients are 4.6~9.2 pm/V at $\lambda = 1300 nm$. There are no decays of the EO coefficient in the poled polymer at $180^{\circ}C$ in one hour, and $90^{\circ}C$ in 500 hours.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.6
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• pp.576-581
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• 2005

Phytase improves the bioavailability of phytate phosphorus in plant foods to humans and animals, and reduces the phosphorus pollution of animal waste. We have engineered the cell surface of the yeast. Saccharomyces cerevisiae, by anchoring active fungal phytase on its cell wall, in order to apply it as a dietary supplement containing bioconversional functions in animal foods and a whole cell bio-catalyst for the treatment of waste. The phytase gene (phyA) of Aspergillus niger with a signal peptide of rice amylase 1A (Ramy1A) was fused with the gene encoding the C-terminal half (320 amino acid residues from the C-terminus) of yeast ${\alpha}-agglutinin$, a protein which is involved in mating and is covalently anchored to the cell wall. The resulting fusion construct was introduced into S. cerevisiae and expressed under the control of the constitutive glyceraldehydes-3-phosphate dehydrogenase (GPD) promoter. Phytase plate assay revealed that the surface-engineered cell exhibited a catalytically active opaque zone which was restricted to the margin of the colony. Additionally, the phytase activity was detected in the cell fraction, but was not detected in the culture medium when it was grown in liquid. These results indicate that the phytase was successfully anchored to the cell surface of yeast and was displayed as its active form. The amount of recombinant phytase on the surface of yeast cells was estimated to be 16,000 molecules per cell.

• Proceedings of the Korean Society of Toxicology Conference
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• 2002.05b
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• pp.20-45
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• 2002

In this study, we demonstrated the in vitro and in vivo formation of carcinogen-lipid adduct and its correlation with DNA or protein adducts. The lipids from serum or hepatocyte membranes of Spragu-Dawley rats. human serum, and standard major lipids were in vitro reacted with benzo[a]pyrene(BP) and BP metabolites. 7,8-Dihydroxy-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]-pyrene(BPDE-I), an ultimate carcinogenic form of BP, was covalently bound to triglyceride(TG). BPDE-I-TG adducts isolated by thin-layer chromatography (TLC) were further detected by high performance liquid chromatography(HPLC). TGs, including triolein, tripalmitin and tristearin, showed positive reactions with BPDE-I. However, cholesterol, phospholipids(Phosphatidylcholine, phosphatidyl-ethanolamine, phosphatidyl-inositol and sphingomyelin) and nonesterified fatty acids(palmitic acid, oleic acid, linoleic acid and stearic acid) did not react with BPDE-I. In addition, other BP metabolites (BP-phenols and -diols) did not react with TG, which TG appeared to be the most reactive lipid yet studied with respect to its ability to form an adduct with BPDE-I. There was a clear-cut dose-respect to its ability to form an adduct with BPDE-I-lipid adduct in vitro between TG and [1,3-3H]BPDE-I. In an animal study, BPDE-I-TG was also formed in the serum of rats orally treated with BP(25 mg/rat). Also, obvious correlations between [3H]BP related-biomolecule adducts (DNA, protein) or lipid damage and the BPDE-I-TG adduct were obtained in various tissues of mice i.p. treated with [3H]BP. These data suggest that TG can form an adduct with BPDE-I, as do other macromolecules (DNA, RNA, and protein). Therefore, a carcinogen-lipid adduct would be a useful biomarker for chemical carcinogenesis research and cancer risk assessment.

• Food Science of Animal Resources
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• v.23 no.1
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• pp.86-95
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• 2003

Potential mechanisms by which vitamin E improves oxidative stability of myoglobin are documented. The basis by which this lipid-soluble antioxidant, ${\alpha}$-tocopherol, protects water-soluble oxymyoglobin is beginning to be understood. Recent evidence suggests that ${\alpha}$-tocopherol delays the release of prooxidative products of lipid oxidation from biomembranes, which in turn delays oxymyoglobin oxidation and the concomitant loss of desirable beef color. ${\alpha}$, ${\beta}$-Unsaturated aldehydes are one class of lipid oxidation products that enhance oxymyoglobin oxidation in vitro and appear to act by covalently binding to the protein. If ${\alpha}$-tocopherol delays the formation of these reactive aldehydes, then this could inhibit the prooxidative effect of these oxidation products toward oxymyoglobin. Additionally, ${\alpha}$-tocopherol may exert part of its color-stabilizing effect in beef by enhancing the metmyoglobin reduction.

• Journal of the Korean Chemical Society
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• v.64 no.2
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• pp.74-78
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• 2020

A push-pull conjugated dye (DCMP) was covalently immobilized on a silanized glass surface to produce a high sensitivity pH sensor film for operating in the acidic region. A pH-sensitive sensor film was prepared by photo-initiating copolymerization of a modified DCMP (DCMA), 2-hydroxyethyl methacrylate (HEMA) and triethylene glycol dimethacrylate on the silanized glass surface. The absorbance of the sensor film increased with increasing pH between pH 2.0 and 5.0, and the fluorescence intensity of the film also increased about 50 times with increasing pH in the same pH range. The sensor film was reversible and reproducible under acidic conditions. The sensor film showed a relatively short response time between 20-50 seconds and high selectivity for proton in the presence of various metal ions.