• Korean Chemical Engineering Research
• /
• v.53 no.1
• /
• pp.121-126
• /
• 2015

The objective of this study is to evaluate the catalytic potential of the porous material prepared from water treatment sludge. The textural properties of the catalyst were studied using $N_2$ adsorption and desorption isotherms, scanning electron microscope, and X-ray diffraction. The pellet-type catalyst prepared using water treatment sludge is determined to be a material that contains mesopores as well as micropores. The specific surface area of the catalyst is $157m^2/g$. Acidic characteristics of the catalyst are analyzed by temperature-programmed desorption of ammonia and infrared spectroscopy of adsorbed pyridine. 2-Butanol dehydration reaction was carried out in a fixed bed catalytic reactor. Yields of 1-butene, trans-2-butene, and cis-2-butene at $350^{\circ}C$ were 25.6 wt%, 19.2 wt%, and 29.9 wt%, respectively. This catalytic activity of the catalyst based on water treatment sludge in 2-butanol dehydration is due to the acid sites composed of Bronsted acid sites and Lewis acid sites. It was confirmed that the catalyst based on water treatment sludge can be utilized to produce $C_4$ olefin through butanol dehydration.

• Journal of Hydrogen and New Energy
• /
• v.26 no.2
• /
• pp.170-178
• /
• 2015

In this study, we carry out a study on how to improve performance of vanadium redox flow battery (VRFB) through promoting reaction rate of rate determining vanadium reaction ($[VO]^{2+}/[VO_2]^+$). In order to do that, three different carbons like Vulcan (XC-72), CMK3 and MSU-F-C are adopted as the catalysts, while their catalytic activity and reaction reversibility are evaluated using half-cell tests. Their topological images are also measured by TEM. For estimation of the VRFB performance, multiple charge-discharge curves of VRFBs including the catalysts are measured by single cell tests. As a result of that, MSU-F-C shows relatively excellent catalytic activity and reaction reversibility as well as large surface area compared to those of Vulcan (XC-72) and CMK3. Also, in terms of the performance of VRFBs including the catalysts, VRFB including MSU-F-C indicates (i) low charging/discharging overpotentials and low internal resistance, (ii) high charge/discharge capacities and (iii) high energy efficiency. These VRFB performance data are well agreed with results on catalytic activity and reaction reversibility. The reason that MSU-F-C induces superior VRFB performances is attributed to (i) its large surface area and (ii) its hydrophilic surface functional groups that mainly consist of hydroxyl bonds that are supposed to play active surface site role for facilitaing $[VO]^{2+}/[VO_2]^+$ redox reaction. Based on the above results, it is found that adoption of MSU-F-C as catalyst for VRFB results in improvement in VRFB performance by promoting the languid $[VO]^{2+}/[VO_2]^+$ redox reaction.

• Applied Chemistry for Engineering
• /
• v.27 no.1
• /
• pp.92-100
• /
• 2016

Low-temperature conversion of nitrogen oxides using plasma-assisted hydrocarbon selective catalytic reduction of (HC-SCR) was investigated. Plasma was created in the catalyst-packed bed so that it could directly interact with the catalyst. The effect of the reaction temperature, the shape of catalyst, the concentration of n-heptane as a reducing agent, the oxygen content, the water vapor content and the energy density on $NO_x$ removal was examined. $NO_x$ conversion efficiencies achieved with the plasma-catalytic hybrid process at a temperature of $250^{\circ}C$ and an specific energy input (SIE) of $42J\;L^{-1}$ were 83% and 69% for one-dimensional Ag catalyst ($Ag\;(nanowire)/{\gamma}-Al_2O_3$) and spherical Ag catalyst ($Ag\;(sphere)/{\gamma}-Al_2O_3$), respectively, whereas that obtained with the catalyst-alone was considerably lower (about 30%) even with $Ag\;(nanowire)/{\gamma}-Al_2O_3$ under the same condition. The enhanced catalytic activity towards $NO_x$ conversion in the presence of plasma can be explained by the formation of more reactive $NO_2$ species and partially oxidized hydrocarbon intermediates from the oxidation of NO and n-heptane under plasma discharge. Increasing the SIE tended to improve $NO_x$ conversion efficiency, and so did the increase in the n-heptane concentration; however, a further increase in the n-heptane concentration beyond $C_1/NO_x$ ratio of 5 did not improve the $NO_x$ conversion efficiency any more. The increase in the humidity affected negatively the $NO_x$ conversion efficiency, resulting in lowering the $NO_x$ conversion efficiency at the higher water vapor content, because water molecules competed with $NO_x$ species for the same active site. The $NO_x$ conversion efficiency increased with increasing the oxygen content from 3 to 15%, in particular at low SIE values, because the formation of $NO_2$ and partially oxidized hydrocarbon intermediates was facilitated.

• Journal of Microbiology and Biotechnology
• /
• v.17 no.8
• /
• pp.1308-1315
• /
• 2007

Lipase B from Candida antarctica (CalB) displayed on the cell surface of H. polymorpha has been functionally improved for catalytic activity by molecular evolution. CalB was displayed on the cell surface by fusing to a cell-wall anchor motif (CwpF). A library of CalB mutants was constructed by in vivo recombination in H. polymorpha. Several mutants with increased whole-cell CalB activity were acquired from screening seven thousand transformants. The two independent mutants CalB 10 and CalB 14 showed an approximately 5 times greater whole-cell activity than the wild-type. When these mutants were made as a soluble form, CalB 10 showed 6 times greater activity and CalB 14 showed an 11 times greater activity compared with the wild-type. Sequence analyses of mutant CALB genes revealed amino acid substitutions of $Leu^{278}Pro$ in CalB10 and $Leu^{278}Pro/Leu^{219}Gln$ in CalB14. The substituted $Pro^{278}$ in both mutants was located near the proline site of the ${\alpha}$10 helix. This mutation was assumed to induce a conformational change in the ${\alpha}$10 helix and increased the $k_{cat}$ value of mutant CalB approximately 6 times. Site-directed mutagenized CalB, LQ ($Leu^{219}Gln$) was secreted into the culture supernatant at an amount of approximately 3 times more without an increase in the CalB transcript level, compared with the wild-type.

• BMB Reports
• /
• v.28 no.2
• /
• pp.112-117
• /
• 1995

Succinic semialdehyde reductase was inactivated by o-phthalaldehyde. The inactivation followed pseudo-first order kinetics, and the second-order rate constant for the inactivation process was 28 $M^{-1}s^{-1}$ at pH 7.4 and $25^{\circ}C$. The absorption spectrum ($\lambda_{max}$ 337 nm) and fluorescence excitation ($\lambda_{max}$ 340 nm) and fluorescence emission spectra ($\lambda_{max}$ 409 nm) were consistent with the formation of an isoindole derivative in the catalytic site between a cysteine and a lysine residue approximately about 3 $\AA$ apart. The substrate, succinic semialdehyde, did not protect enzymatic activity against inactivation, whereas the coenzyme NADPH protected against o-phthaladehyde induced inactivation of the enzyme. About 1 isoindole group per mol of the enzyme was formed following complete loss of enzymatic activity. These results suggest that the amino acid residues of the enzyme participating in a reaction with o-phthalaldehyde are cysteinyl and lysyl residues at or near the NADPH binding site.

• BMB Reports
• /
• v.32 no.5
• /
• pp.515-520
• /
• 1999

Incubation of two types of glutamate dehydrogenase isoproteins from bovine brain with the arginine-specific dicarbonyl reagent phenylglyoxal resulted in a biphasic loss of enzyme activity. Reaction of the glutamate dehydrogenase isoproteins with phenylglyoxal caused a rapid loss of 53~62% of the enzyme activities and modification of two residues of arginine per enzyme subunit. Prolonged incubation of the glutamate dehydrogenase isoproteins with phenylglyoxal resulted in the modification of an additional four residues of arginine per enzyme subunit without further loss of the residual activities. Partial protection against inactivation was provided by the coenzyme NADH or substrate 2-oxoglutarate. The most marked decrease in the rate of inactivation was observed by the combined addition of NADH and 2-oxoglutarate, suggesting that the first two modified arginine residues are in the vicinity of the catalytic site. However, inactivation of the glutamate dehydrogenase isoproteins by phenylglyoxal appears to be partial with approximately 40% activity remained after an extended reaction time with excess reagent, suggesting that the modified arginine residues may not be directly involved in catalysis. The lack of complete protection by substrates also suggest the possibility that the modified arginine residues are not directly involved at the active site, and the partial loss of activity by the modification of arginine residues may be due to a conformational change. There were no significant differences between the two glutamate dehydrogenase isoproteins in sensitivities to inactivation by phenylglyoxal, indicating that the microenvironmental structures of the glutamate dehydrogenase isoproteins are very similar to each other.

• Journal of Microbiology and Biotechnology
• /
• v.13 no.6
• /
• pp.969-975
• /
• 2003

Two conserved amino acid residues in the extra sugar-binding space near the catalytic site of Thermus maltogenic amylase (ThMA) were analyzed for their role in the hydrolysis and transglycosylation activity of the enzyme. Site-directed mutagenesis was carried out by replacing N33l with a lysine (N331K), E332 with a histidine (E332H), or by replacing both residues at the same time (N331K/E332H). The measured $K_m$ values indicated that affinities toward all substrates tested, including starch, pullulan, ${\beta}-cyclomaltodextrin$, and acarbose, were lower in all the mutants compared to that of wild-type ThMA, leading to reduced hydrolysis activity. In addition, the lower ratio of transglycosylation to hydrolysis in the mutants compared to that in the wild-type ThMA indicated that these mutants preferred hydrolysis to the transglycosylation reaction. These results demonstrated that the conserved dipeptide at 331 and 332 of ThMA is directly involved in the formation and accumulation of transfer products by accommodating acceptor sugar molecules.

• Bulletin of the Korean Chemical Society
• /
• v.32 no.8
• /
• pp.2549-2552
• /
• 2011

In this study, the kinetic properties of wild-type and C117D mutant H. influenzae MurA (Hi MurA), which catalyzes the first reaction in the biosynthetic pathway of the cell wall, were characterized. Purified recombinant Hi MurA was active at pH values ranging from pH 5.5 to pH 10, and its $K_m$ (UNAG), $K_m$ (PEP), and $k_{cat}$ values were measured to be 31 ${\mu}M$, 24 ${\mu}M$, and 210 $min^{-1}$, respectively. Hi MurA activity was effectively inhibited by fosfomycin with an $IC_{50}$ value of 60 ${\mu}M$. Hi MurA contains a cysteine residue (C117) at the loop region near the PEP binding, whereas MurA from fosfomycin resistant Mycobaterium tuberculosis or Chlamydia trachomatis contain an aspartate residue instead of the cysteine at the corresponding site. Aspartate substitution of Cys117 in Hi MurA shifted its optimum pH from 7.8 to 6.0. In addition, the $K_m$ values for UNAG and PEP were increased to 160 ${\mu}M$ and 150 ${\mu}M$, respectively, and the $k_{cat}$ value was significantly reduced to 41 $min^{-1}$. Furthermore, the C117D mutant form of Hi MurA was not inhibited by 1 mM fosfomycin. These results indicate that the Cys117 of Hi MurA is the binding site of fosfomycin and plays an important role in the fast turnover of the catalytic reaction.

• Bulletin of the Korean Chemical Society
• /
• v.26 no.2
• /
• pp.285-291
• /
• 2005

The nonstructural protein 5B (NS5B) of hepatitis C virus (HCV) is the viral RNA-dependent RNA polymerase (RdRp), which is the essential catalytic enzyme for the viral replication and is an appealing target for the development of new therapeutic agents against HCV infection. A small amount of serum from a single patient with hepatitis C was used to get the genome of a Korean HCV isolate. Sequence analysis of NS5B 1701 nucleotides showed the genotype of a Korean isolate to be subtype 1b. The soluble recombinant HCV NS5B polymerase lacking the C-terminal 24 amino acids was expressed and purified to homogeneity. With the highly purified NS5B protein, we established in vitro systems for RdRp activity to identify potential polymerase inhibitors. The rhodanine family compounds were found to be potent and specific inhibitors of NS5B from high throughput screening (HTS) assay utilizing the scintillation proximity assay (SPA) system. The binding mode of an inhibitor was analyzed by measuring various kinetic parameters. Lineweaver-Burk plots of the inhibitor suggested it binds not to the active site of NS5B polymerase, but to an allosteric site of the enzyme. The activity of NS5B in in vitro polymerase reactions with homopolymeric RNA requires interaction with multiple substrates that include a template/primer and ribonucleotide triphosphate. Steady-state kinetic parameter, such as Km, was determined for the ribonucleotide triphosphate. One of compounds found interacts directly with the viral polymerase and inhibits RNA synthesis in a manner noncompetitively with respect to UTP. Furthermore, we also investigated the ability of the compound to inhibit NS5B-directed viral RNA replication using the Huh7 cell-based HCV replicon system. The investigation is potentially very useful for the utility of such compounds as anti-hepatitic agents.

• Bulletin of the Korean Chemical Society
• /
• v.25 no.5
• /
• pp.721-725
• /
• 2004

Acetohydroxy acid synthase (AHAS) is one of several enzymes that require thiamine diphosphate and a divalent cation as essential cofactors. The active site contains several conserved ionizable groups and all of these appear to be important as judged by the fact that mutation diminishes or abolishes catalytic activity. Recently, we have shown [Yoon, M.-Y., Hwang, J.-H., Choi, M.-K., Baek, D.-K., Kim, J., Kim, Y.-T., Choi, J.-D. FEBS Letters 555 (2003), 185-191] that the activity is pH-dependent due to changes in $V_{max}$ and V/$K_m$. Data were consistent with a mechanism in which substrate was selectively catalyzed by the enzyme with an unprotonated base having a pK 6.48, and a protonated group having a pK of 8.25 for catalysis. Here, we have in detail studied the pH dependence of the kinetic parameters of the cofactors (ThDP, FAD, $Mg^{2+}$) in order to obtain information about the chemical mechanism in the active site. The $V_{max}$ of kinetic parameters for all cofactors was pH-dependent on the basic side. The pK of ThDP, FAD and $Mg^{2+}$ was 9.5, 9.3 and 10.1, respectively. The V/$K_m$ of kinetic parameters for all cofactors was pH-dependent on the acidic and on the basic side. The pK of ThDP, FAD and $Mg^{2+}$ was 6.2-6.4 on the acidic side and 9.0-9.1 on the basic side. The well-conserved histidine mutant (H392) did not affect the pH-dependence of the kinetic parameters. The data are discussed in terms of the acid-base chemical mechanism.