• Journal of Microbiology and Biotechnology
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• v.30 no.1
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• pp.146-154
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• 2020

With the advantages of biocatalytic method, enzymes have been excavated for the synthesis of chiral amino acids by the reductive amination of ketones, offering a promising way of producing pharmaceutical intermediates. In this work, a robust phenylalanine dehydrogenase (PheDH) with wide substrate spectrum and high catalytic efficiency was constructed through rational design and active-site-targeted, site-specific mutagenesis by using the parent enzyme from Bacillus halodurans. Active sites with bonding substrate and amino acid residues surrounding the substrate binding pocket, 49L-50G-51G, 74M,77K, 122G-123T-124D-125M, 275N, 305L and 308V of the PheDH, were identified. Noticeably, the new mutant PheDH (E113D-N276L) showed approximately 6.06-fold increment of k_cat/Km in the oxidative deamination and more than 1.58-fold in the reductive amination compared to that of the wide type. Meanwhile, the PheDHs exhibit high capacity of accepting benzylic and aliphatic ketone substrates. The broad specificity, high catalytic efficiency and selectivity, along with excellent thermal stability, render these broad-spectrum enzymes ideal targets for further development with potential diagnostic reagent and pharmaceutical compounds applications.

• Applied Chemistry for Engineering
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• v.25 no.2
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• pp.198-203
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• 2014

In this study, steam reforming reaction and surface characteristics of Ni metal foam plate were investigated. Valence state of Ni could be changed by pretreatment, and metallic Ni species exposed on surface as a active site play important role in steam reforming reaction. Porous catalytic membrane also was prepared by mixing of Ni metal foam plate and Ni-YSZ catalyst to control the pore size and assign the catalytic function in Ni metal foam plate. In SEM analysis results, Pore size of Ni metal foam plate could be controlled and Ni-YSZ catalyst well dispersed on surface. Ni based porous catalytic membrane had a similar steam reforming activity regardless of space velocity.

• Bulletin of the Korean Chemical Society
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• v.22 no.3
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• pp.263-266
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• 2001

Isopropylation of phenol with 2-propanol has been carried out over Na-exchanged ZSM-5 zeolites to determine the effect of catalyst acidity on phenol conversion and product selectivity. The acid type and strength of the catalyst such as Lewis, weak and strong Bronsted acid sites are measured by pyridine adsorbed XPS and the catalytic properties are interpreted in terms of the acid properties. The active site and mechanism for the reaction are suggested based on evidence of study from the reactant adsorbed FT-IR.

• Korean Journal of Microbiology
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• v.29 no.4
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• pp.209-214
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• 1991

The PRD1 DNA polymerase is a small multi-functional enzyme containing conserved amino acid sequences shared by family B DNA polymerases. Thus the PRD1 DNA polymerase provides an useful model system with which to study structure-functional relationships of DNA polymerase molecules. In order to investigate the functional and structural roles of the highly conserved amino acid sequences, we have introduced three mutations into a conserved amino acid of the PRD1 DNA polymerase. Genetic complememtation study indicated that each mutation inactivated DNA polymerase catalytic activity.

• Journal of Life Science
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• v.10 no.1
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• pp.20-23
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• 2000

Mono-ADP-ribosylation, catalyzed by ADP-ribosyltransferases, is a post-translational modification of proteins in which the ADP-ribose moiety of NAD is transferred to an acceptor protein. Previously, we have identified and cloned a glycosylphosphatidylinositol-linked ADP-ribosyltransferase (Yac-1) from mouse lymphoma cells. Yac-1 enzyme contains three regions (region I,II,III) similar to those found in several bacterial toxins and vertebrate ADP-ribosyltransferases. Site-directed mutagenesis was performed to verify the role of Glu 233 in region III. Mutants E233Q, E233D and E233A were inactive for ADP-ribosyltransferase activity. Thus Glu 233 in Yac-1 is essential for enzyme activity, suggesting that Glu 233 in Glu-rich motif near the carboxy terminus plays a catalytic role in ADP-ribosyltransferase activity.

• Transactions of the Korean Society of Mechanical Engineers B
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• v.25 no.4
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• pp.583-592
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• 2001

The combustion characteristics of the hybrid catalytic(catalytic+thermal) combustor with a lean methane-air mixture on platinum catalyst were investigated numerically using a 2-D boundary layer model with detailed homogeneous and heterogeneous chemistries. for the more accurate calculations, the actual surface site density of monolith coated with platinum was decided by the comparison with experimental data. It was found that the homogeneous reactions in the monolith had little effect on the change of temperature profile, methane conversion rate and light off location. However, the radicals such as OH and CO were produced rapidly at exit by homogeneous reactions. The effect of operation conditions such as equivalence ratio, temperature, velocity, pressure and diameter of the monolith channel at the entrance were studied. In thermal combustor, the production of N$_2$O was more dominant than that of NO due to the relative importance of the reaction N$_2$+O(+M)→N$_2$O(+M). Finally the productions of CO and NOx by amount of methane addition were studied.

• Journal of the Korean Chemical Society
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• v.59 no.2
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• pp.183-187
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• 2015

• Journal of Life Science
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• v.21 no.3
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• pp.473-479
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• 2011

Adenylyl cyclase (AC) catalyzes the formation of cyclic AMP (cAMP) from ATP. The cAMP produced by AC serves as a secondary messenger in a variety of signal transduction pathways, and controls various cellular functions in many organisms. ACs can be grouped into six classes based on their primary amino acid sequences. Eukaryotes and mycobacteria contain only members of class III AC. The catalytic cyclase domains of class III AC are active as dimers: mammalian ACs, which are composed of a single polypeptide with two catalytic cyclase domains, form the active site as a result of intramolecular dimerization of the catalytic cyclase domains. In contrast, mycobacterial ACs function as homodimers, since their polypeptides contain a single catalytic cyclase domain. Six amino acids are required for the catalytic activity of class III AC - two aspartate residues, a lysine-aspartate pair and an arginine-asparagine pair. 16 ACs belonging to the class III were identified in Mycobacterium tuberculosis H37Rv, and their characteristics are reviewed.

• Journal of Microbiology and Biotechnology
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• v.16 no.6
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• pp.974-978
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• 2006

The cytochrome P450 monooxygenase from the pikromycin biosynthetic gene cluster in Streptomyces venezuelae, known as PikC, was observed to hydroxylate the unnatural substrate indole to indigo. Furthermore, the site-directed mutagenesis of PikC monooxygenase led to the mutant enzyme F171Q, in which Phe171 was altered to Gln, with enhanced activity for the hydroxylation of indole. From enzyme kinetic studies, F171Q showed an approximately five-fold higher catalytic efficiency compared with the wild-type PikC. Therefore, these results demonstrate the promising application of P450s originating from Streptomyces, normally involved in polyketide biosynthesis, to generate a diverse array of other industrially useful compounds.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1996.11a
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• pp.135-139
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• 1996

$p70^{s6k}$ lies on a $p21^{ras}$-independent signalling pathway and plays an important role in mitogenesis. Activation is associated with phosphorylation at multiple sites, four of which lie in an autoinhibitory region. The immunosuppressant rapamycin induces $p70^{s6k}$ inactivation through dephosphorylation of a second set of mitogen-induced sites. Here we identify these sites as $T_{229}$, $T_{389}$, and $S_{404}$. $T_{229}$ resides in the "T loop" of the catalytic domain, an essential phosphorylation site in other kinases. However, $p70^{s6k}$ inactivation by rapamycin most closely parallels $T_{389}$ dephosphorylation. Mutation of $T_{389}$ to alanine ablates kinase activity, whereas mutation to glutamic acid confers constitutive kinase activity and rapamycin resistance. indicating an essential role for phosphorylation at this site. $T_{389}$ resides in an unusual hydrophobic motif, not previously noted, between the catalytic and autoinhibitory domains. The importance of this site, and surrounding motif, is emphasized by its conservation in other kinases including homologues of $p70^{s6k}$ derived from such distantly related organisms as yeast and plant.