• KSBB Journal
• /
• v.10 no.3
• /
• pp.335-342
• /
• 1995

Solid and liquid phase peroxidases were extracted from Chinese cabbage roots by using commercial juicer in order to use peroxidases from agricultural waste for industrial applications. Since peroxidases are distributed into 66% in liquid (juice) and 34% in solid phase (pulp), enzymes from both phases were applied to investigate the enzymatic removal of phenol from waste water. After contacting 150 ppm Phenol solution with liquid phase enzyme (1,800 unit/$\ell$) for 3 hours in a batch stirred reactor, 96% of phenol could be removed through polymerization and precipitation. Also, phenol could be removed from initial 120ppm to 5ppm by applying solid phase enzyme in an air lift reactor ($600 unit/\ell$). Almost equivalent efficiencies of phenol removal were observed between two systems, even though only one third of the enzymes in batch stirred reactor was applied in airlift reactor. The possible reason for this phenomenon is because peroxidases exist as immobilized forms in solid phase.

• Journal of Microbiology and Biotechnology
• /
• v.19 no.10
• /
• pp.1201-1205
• /
• 2009

Corn starches with different amylose contents were enzymatically modified using Thermus aquaticus 4-$\alpha$-glucanotransferase ($TA{\alpha}GTase$). Upon the enzyme treatment, the chain-length distributions of isoamylolytically debranched products became broader [degree of polymerization (DP): 3-40] than those of untreated corn starches. In addition, a variety of cycloamyloses (CAs) with different sizes were formed by the glucanotransfer activity of $TA{\alpha}GTase$. CAs with DP 5-40 were detectable in all of the $TA{\alpha}GTase$-treated corn starches. From the results of high-performance anion-exchange chromatography and high-performance size-exclusion chromatography analyses, it was suggested that the amount of CAs produced by the enzyme treatment increased as the amylose content of the starches increased. Thus, we concluded that the extent of modification of starch molecules was enhanced in proportion to amylose content by the transfer activity of $TA{\alpha}GTase$. This finding could be useful for developing an efficient process of CA production using this enzyme.

• BMB Reports
• /
• v.31 no.1
• /
• pp.25-30
• /
• 1998

Kinetic studies of L-Alanine dehydrogenase from Bacillus subtilis-catalyzed reactions in the presence of $Zn^{2+}$ were carried out. The substrate (L-alanine) saturation curve is hyperbolic in the absence of the metal ion but it becomes sigmoidal when $Zn^{2+}$ is added to the reaction mixture indicating the positive cooperative binding of the substrate in the presence of zinc ion. The cooperativity of substrate binding depends on the xinc ion concentration: the Hill coefficients ($n_H$) varied from 1.0 to 1.95 when the zinc ion concentration varied from 0 to $60\;{\mu}m$. The inhibition of AlaDH by $Zn^{2+}$ is reversible and noncompetitive with respect to $NAD^+$ ($K_i\;=\;5.28{\times}10^{-5}\;M$). $Zn^{2+}$ itself binds to AlaDH with positive cooperativity and the cooperativity is independent of substrate concentration. The Hill coefficients of substrate biding in the presence of $Zn^{2+}$ are not affected by the enzyme concentration indicating that $Zn^{2+}$ binding does not change the polymerization-depolymerization equilibria of the enzyme. Among other metal ions, $Zn^{2+}$ appears to be a specific reversible inhibitor inducing conformational change through the intersubunit interaction. These results indicate that $Zn^{2+}$ is an allosteric competitive inhibitor and substrate being a non-cooperative per se, excludes the $Zn^{2+}$ from its binding site and thus exhibits positive cooperativity. The allosteric mechanism of AlaDh from Bacillus subtilis is consistent with both MWC and Koshland's allosteric model.

• Archives of Pharmacal Research
• /
• v.10 no.1
• /
• pp.42-49
• /
• 1987

This study is to evaluate the potential use of aclarubicin-loaded gelatin microspheres as an intravascular biodegradable drug delivery system for the regional cancer therapy. The diameter of the microspheres prepared by water in oil emulsion polymerization could be controlled by adjusting the stirring rate in the range of 10-50 $\mu$m : D(in $\mu$m) = -73.8 log (rpm) + 262.7. The addition of proteolytic enzyme increased the in vitro aclarubicin release but it did not change the amount of the initial burst release which reached about 45%. Microspheres injected intravenously into the mouse tail vein embolized only to the lung when observed by fluorescence microscopy. From histological examination following injection of gelatin microspheres into mouse femoral muscle, mild inflammation was observed from the appearance of neutrophils after 2 days and rapid repair process was confirmed thereafter. Biodegradation process of gelatin microspheres lodged on the pulmonary capillary bed was followed up by microscopic observation; degradation was taking place by about 36 hrs, followed by severe damage on the spheerical shape and microspheres was no longer found 10 days after injection.

• Food Science and Biotechnology
• /
• v.17 no.2
• /
• pp.228-233
• /
• 2008

Considerable progress has been made in functionalization of the soy isoflavones through enzymatic modification of daidzin, genistin, and glycitin. After hydrolysis of $\beta$-glucosides into their corresponding aglycones, these compounds were structurally modified via biotransformations such as regioselective hydroxylation, enantioselective reduction, regioselective methylation, and polymerization. These reactions often resulted in an increase of the biological activities (e.g., anti oxidative activity, antiproliferative activity) and/or improvement of the physico-chemcial properties (e.g., water solubility, bioavailability). This review briefly summarizes on-going research activities on the biofunctionalization of the soy isoflavones.

• Macromolecular Research
• /
• v.15 no.1
• /
• pp.17-21
• /
• 2007

A series of new, vinyl sulfone-containing monomers was prepared by p-aminophenyl vinyl sulfone (1) with various acid chlorides such as adipoly chloride, terephthaloyl chloride and isophthaloyl chloride. Finally, tertiary amine-containing poly(amide-sulfone)s were prepared by the Michael-type addition polymerization using N,N-dimethylethylenediamine (DMEDA), p-phenylenediamine and piperazin. To evaluate the relevant properties for permselective membranes and enzyme substrate, various physical properties such as molecular weight, solubility, quaternization behavior and thermal properties were examined.

• BMB Reports
• /
• v.31 no.6
• /
• pp.585-589
• /
• 1998

The herpes simplex virus type-1 (HSV-1)-encoded DNA polymerase consists of two subunits, the products of the UL30 and UL42 genes. UL30 and UL42 were coexpressed in Sf9 cells infected with recombinant baculoviruses carrying the two genes. The UL30 and UL42 gene products remained tightly associated throughout the purification, which led to a near homogeneous heterodimer composed of the DNA polymerase and UL42 protein. The DNA polymerase-UL42 protein heterodimer, purified from the recombinant baculovirus-infected Sf9 cells, showed the same high degree of processivity of deoxynucleotide polymerization as the enzyme purified from the HSV-1 infected primate cells. Like the latter, it contained a 3'-5' exonuclease activity that specifically hydrolyzes an incorrectly matched nucleotide at the 3' terminus of a primer, thereby contributing to the fidelity of DNA replication.

• Journal of Environmental Health Sciences
• /
• v.31 no.4 s.85
• /
• pp.241-245
• /
• 2005

Laccase catalyzes the oxidation and polymerization of aromatic compounds in the presence of molecular oxygen. Studies were conducted to characterize the use of polyethylene glycol (PEG) as an additive to keep up the enzymatic stability. The enzymatic activities highly remained and bisphenol A (BPA) was rapidly converted in the presence of 5 mg/l of PEC. These effects were accomplished with PEG of molecular weight 3,350. A linear relationship was found between the quantity of BPA to be converted $(10-120\;{\mu}M)$ and the optimum dose of PEC required for greater than $95\%$ conversion. This result suggests that it is the interaction between the PEG and the reaction products. In the optimum dose of PEG, the aeration of reaction mixture neither enhanced the conversion of BPA nor retarded the inactivation of the enzyme.

• International Journal of Oral Biology
• /
• v.37 no.4
• /
• pp.181-188
• /
• 2012

As the demand for large-scale analysis of gene expression using DNA arrays increases, the importance of the surface characterization of DNA arrays has emerged. We compared the efficiency of molecular biological applications on solid-phases with different surface polarities to identify the most optimal conditions. We employed thiol-gold reactions for DNA immobilization on solid surfaces. The surface polarity was controlled by creating a self-assembled monolayer (SAM) of mercaptohexanol or hepthanethiol, which create hydrophilic or hydrophobic surface properties, respectively. A hydrophilic environment was found to be much more favorable to solid-phase molecular biological manipulations. A SAM of mercaptoethanol had the highest affinity to DNA molecules in our experimetns and it showed greater efficiency in terms of DNA hybridization and polymerization. The optimal DNA concentration for immobilization was found to be 0.5 ${\mu}M$. The optimal reaction time for both thiolated DNA and matrix molecules was 10 min and for the polymerase reaction time was 150 min. Under these optimized conditions, molecular biology techniques including DNA hybridization, ligation, polymerization, PCR and multiplex PCR were shown to be feasible in solid-state conditions. We demonstrated from our present analysis the importance of surface polarity in solid-phase molecular biological applications. A hydrophilic SAM generated a far more favorable environment than hydrophobic SAM for solid-state molecular techniques. Our findings suggest that the conditions and methods identified here could be used for DNA-DNA hybridization applications such as DNA chips and for the further development of solid-phase genetic engineering applications that involve DNA-enzyme interactions.

• Korean Journal of Food Science and Technology
• /
• v.31 no.5
• /
• pp.1262-1267
• /
• 1999

Optimum reaction conditions for gel formation of rapeseed, Brassica napus, protein catalyzed by microbial TGase(transglutaminase) were evaluated by measuring breaking strength and deformation of gel. The polymerization of the protein gel was ascertained by SDS-PAGE and content of GL crosslinking$[{\varepsilon}-({\gamma}-glutamyl)lysine]$. In the reaction between rapeseed protein and TGase at $45^{\circ}C$ for 60 min, the breaking strength and deformation of the gel was the maximum at the ratio of 1 : 40 of enzyme to substrate. 10%(w/v) of rapeseed protein concentrate was optimum for gel production. The maximum breaking strength and deformation was shown at $45^{\circ}C$ The breaking strength increased linearly up to 90 min of the reaction time and remained unchanged. The breaking strength and deformation by TGase treatment was pH dependent and pH 7 was optimum for 10% rapeseed protein solution. SDS-PAGE analysis indicated that new band of highmolecular polymers were formed by the enzyme reaction, with disappearing the original bands of rapeseed protein. According to HPLC analysis. the content of GL crosslinking was increased from 0 to $7.14\;{\mu}mol/g$ gel for 90 min of the reaction time.