• Bulletin of the Korean Chemical Society
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• v.28 no.12
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• pp.2539-2542
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• 2007

• Journal of Microbiology and Biotechnology
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• v.18 no.1
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• pp.48-54
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• 2008

A putative ${\omega}$-aminotransferase gene, cc3143 (aptA), from Caulobacter crescentus was screened by bioinformatical tools and overexpressed in E. coli, and the substrate specificity of the ${\omega}$-aminotransferase was investigated. AptA showed high activity for short-chain ${\beta}$-amino acids. It showed the highest activity for 3-amino-n-butyric acid. It showed higher activity toward aromatic amines than aliphatic amines. The 3D model of the ${\omega}$-aminotransferase was constructed by homology modeling using a dialkylglycine decarboxylase (PDB ID: 1DGE) as a template. Then, the ${\omega}$-aminotransferase was rationally redesigned to increase the activity for 3-amino-3-phenylpropionic acid. The mutants N285A and V227G increased the relative activity for 3-amino-3-phenylpropionic acid to 3-amino-n-butyric acid by 11-fold and 3-fold, respectively, over that of wild type.

• Journal of Microbiology and Biotechnology
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• v.16 no.9
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• pp.1477-1480
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• 2006

The 63-amino-acid-encoding afsR2 is a global antibiotics-stimulating regulatory gene identified from the chromosome of Streptomyces lividans. To dissect a putative functional domain in afsR2, several afsR2-derivative deletion constructs were generated and screened for the loss of actinorhodin-stimulating capability. The afsR2-derivative construct missing a 50-bp C-terminal region significantly lost its actinorhodin-stimulating capability in S. lividans. In addition, site-directed mutagenesis on amino acid positions of #57-#61 in a 50-bp C-terminal region, some of which are conserved among known Sigma 70 family proteins, significantly changed the AfsR2's activity. These results imply that the C-terminal region of AfsR2 is functionally important for antibiotics-stimulating capability and the regulatory mechanism might be somehow related to the sigma-like domain present in the C-terminal of AfsR2.

• Journal of Acupuncture Research
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• v.17 no.3
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• pp.151-167
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• 2000

Objective : The aim of this study is to determine the antimutagenic effect and genetic safety of Buthus martensi Karsch aqua-acupuncture solution(BMKAS) against various chemical carcinogens. Method : Ames(Salmonella typhimurium) test and Rec assay(Bacillus subtilis) were used as indicators for DNA damage and antimutagenesis. Furthermore, the levels of umu operon expression by measuring the ${\beta}$-galactosidase activity wete monitored with the SOS umu test using S. typhimurium 1535 containing plasmid pSK1002. And the host-mediated assay was used to investigate the mutagenicity and antimutagenicity of BMKAS inducing various chemical carcinogens after the activation with in vivo metabolic systems. Results : From the results, BMKAS did not atfect DNA of S. typhimurium and B. subtilis strains and showed no mutagenicity at the all concentrations of tested solution. Furthermore BMKAS dose-dependently protected the mutagenecity by AF-2, 2-AA and B[a]P. These phenomena was also similar to that after metabolic activation of BMKAS in in vivo system. Conclusion : These results suggested that BMKAS did not show the mutagenicity and protected the mutagenesis against various chemical carcinogens by four different methods used in this study.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.66.2-66
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• 2003

A gain-of-function mutant, SHM-11 obtained through gamma-ray mutagenesis, is resistant to rice blast caused by Magnaporthe grisea while wild type Sanghaehyanghyella is highly susceptible to the same disease. The resistance in the mutant was not race-specific when we tested with four races (KJ-201, KI-1113a, KI-313, KI-409) of M. grisea. To identify genes involved disease resistance in the gain-of-function mutant, genes specifically expressed in the mutant were selected by suppression subtractive hybridization using cDNAS of blast-inoculated mutant and wild type as a tester and a driver, respectively, Random 200 clones from the subtracted library were selected and analyzed by DNA sequencing. The sequenced genes represented three major groups related with disease resistance； genes encoding PR proteins, genes probably for phytoalexin biosynthesis, and genes involved in disease resistance signal transduction. A gene encoding a putative receptor-like protein kinase was identified as highly expressed only in the gain-of-function mutant after blast infection. The role of the putative receptor-like protein kinase gene during blast resistance will be further studied.

• Journal of Plant Biotechnology
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• v.47 no.4
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• pp.324-329
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• 2020

Reed (Phragmites spp.) is a rhizomatous plant of the Poaceae family and is known as high tolerant plant to heavy metal contaminants. This plant is widely recognized as a Cd root-accumulator, but improved heavy metal tolerance and uptake capacity are still required for phytoremediation efficiency. To enhance capacity of hyperaccumulator plants, ethyl methane sulfonate (EMS) as chemical mutagen has been introduced and applied to remediation approaches. This study aimed to select EMS-mutagenized reeds representing high Cd resistance and large biomass and to investigate their ability of Cd accumulation. After 6 months cultivation of M₂ mutant reeds under Cd stress conditions (up to 1,500 µM), we discovered seven mutant individuals that showed good performances like survivorship, vitality, and high accumulation of Cd, particularly in their roots. Compared to wild type (WT) reeds as control, on average, dry weight of mutant type (MT) reeds was larger by 2 and 1.5 times in roots and shoots, respectively. In addition, these mutant plants accumulated 6 times more Cd, mostly in the roots. In particular, MT8 reeds showed the greatest ability to accumulate Cd. These results suggest that EMS mutagenesis could generate hyperaccumulator plants with enhanced Cd tolerance and biomass, thereby contributing to improvement of phytoremediation efficiency in Cd-contaminated soil or wastewater. Further studies should focus on identifying Cd tolerance mechanisms of such EMS-mutagenized plants, developing techniques for its biomass production, and investigating the practical potential of the EMS mutants for phytoremediation.

• Bulletin of the Korean Chemical Society
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• v.32 no.5
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• pp.1640-1644
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• 2011

Water-soluble mutant of intrinsically insoluble protein, crambin, was produced by mutagenesis based on the sequence analysis with homologous proteins. Thr1, Phe13, and Lys33 of crambin were substituted for Lys, Tyr, and Lys, respectively. The resultant mutant was soluble in aqueous buffer as well as in dodecylphosphocholine (DPC) micelle solution. The $^1H-^{15}N$ spectrum of the mutant crambin showed spectral similarity to that of the wild-type protein except for local regions proximal to the sites of mutation. Solution structure of water-soluble mutant crambin was determined in aqueous buffer by NMR spectroscopy. The structure was almost identical to the wild-type structure determined in non-aqueous solvent. Subtle difference in structure was very local and related to the change of the intra- and inter-protein hydrophobic interaction of crambin. The structural details for the enhanced solubility of crambin in aqueous solvent by the mutation were provided and discussed.

• Korean Journal of Plant Resources
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• v.19 no.2
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• pp.374-380
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• 2006

With the aim of inducing mutation in fern Cyrtomium caryoptideum var. coreanum, rhizome segments of In vitro-grown cultures were treated with chemical mutagens such as EMS, NMU and colchicine. Based on regeneration ratios, sensitivities for each treatments were assessed and also optimum treatment condition of each mutagens was explored. Optimum concentration for EMS treatment was considered to be 20 to 50mM and for NMU 5 to 10mM. NMU was found to be more effective in inducing chlorophyll and morphological variations than EMS. The RAPD were performed to check the genetic modification of phenotypical variants. As a result, polymorphic DNA band patterns between wild type and variants were observed by two 10-mer primers.

• Journal of Microbiology and Biotechnology
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• v.11 no.1
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• pp.118-123
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• 2001

The alginate lyase A1-III gene of Sphingomonas species A1 is composed of 1,077 nucleotides, encoding a protein (359 amino acids) with a molecular mass of 40,322 Da. Recombinant A1-III expressed in Escherichia coli exhibited the same full enzymatic activity as native A1-III. In order to identify the critical residue for activity, a site-directed mutation was introduced into the A1-III gene (H192A, His192->Ala). Recombinant A1-III (H192A) exhibited a significant decrease in enzyme activity (one-thirty thousandth of that of A1-III), without any conformational change, as detected by the CD spectra in the far UV region. Also, the chemical modification of wild-type A1-III with methyl 4-nitro benzene sulfonate resulted in a 40% decrease from the initial activity, whereas the same modification of A1-III (H192A) produced no change in the activity. The role of His192 on the catalytic process was also explored based on a model of A1-III docked with mannuronic acid into the active site.

• Bulletin of the Korean Chemical Society
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• v.26 no.6
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• pp.916-920
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• 2005

Acetohydroxy acid synthase catalyzes the first common step in the biosynthesis of branched chain amino acids. AHAS plays two distinct metabolic roles, and is designated as anabolic AHAS and catabolic AHAS, depending on its function. Anabolic AHAS is FAD-dependent, while its catabolic counterpart is not. In this work, a conserved motif was identified in the $\beta$-domain of anabolic AHASs, but not in catabolic AHAS ($_{372}RFDDR_{376}$). In order to determine the functions of this motif, we replaced the motif with the corresponding sequence in FAD-independent AHAS, SPVEY. None of these three mutants (SPV, SPVE, and SPVEY) was detected with bound FAD. However, two of these mutants (SPVE and SPVEY) were active at a low level of specific activity. Although they exhibited pyruvate- and ThDP- dependent characteristics, the activity of the two active mutants appears to be FAD-independent. The SPVEY mutant was completely insensitive to the three tested herbicides, even at extremely high concentrations and is also somewhat more thermolabile than the wild type enzyme. The data provided in this work suggest that the RFDDR motif is a possible determinant of the FAD-dependent and herbicide-resistant properties of tobacco AHAS. The SPVEY mutant appears to exhibit catabolic AHAS-like activity.