• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2002.10a
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• pp.18-25
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• 2002

The pikromycin biosynthetic system in Streptomyces venezuleae is unique for its ability to produce two groups of antibiotics that include the 12-membered ring macrolides methymycin and neomethymycin, and the 14-membered ring macrolides narbomycin and pikromycin. The metabolic pathway also contains two post polyketide-modification enzymes, a glycosyltransferase and P450 hydroxylase that have unusually broad substrate specificities. In order to explore further the substrate flexibility of these enzymes a series of hybrid polyketide synthases were constructed and their metabolic products characterized. The plasmid-based replacement of the multifunctional protein subunits of the pikromycin PKS in S. venezuelae by the corresponding subunits from heterologous modular PKSs resulted in recombinant strains that produce both 12- and 14-membered ring macrolactones with predicted structural alterations. In all cases, novel macrolactones were produced and further modified by the DesVII glycosyltransferase and PikC hydroxylase leading to biologically active macrolide structures. These results demonstrate that hybrid PKSs in S. venezuelae can produce a multiplicity of new macrolactones that are modified further by the highly flexible DesVII glycosyltransferase and PikC hydroxylase tailoring enzymes. This work demonstrates the unique capacity of the S. venezuelae pikromycin pathway to expand the toolbox of combinatorial biosynthesis and to accelerate the creation of novel biologically active natural products. The polyketide backbone of rifamycin B is assembled through successive condensation and ${\beta}$-carbonyl processing of the extender units by the modular rifamycin PKS. The eighth module, in the RifD protein, contains nonfunctional DH domain and functional KR domain, which specify the reduction of the ${\beta}$-carbonyl group resulting in the C-21 bydroxyl of rifamycin B. A four amino acid substitution and one amino acid deletion were introduced in the putative NADPH binding motif in the proposed KR domain encoded by rifD. This strategy of mutation was based on the amino acid sequences of the corresponding motif of the KR domain of module 3 in the RifA protein, which is believed dysfunctional, so as to introduce a minimum alteration and retain the reading frame intact, yet ensure loss of function. The resulting strain produces linear polyketides, from tetraketide to octaketide, which are also produced by a rifD disrupted mutant as a consequence of premature termination of polyketide assembly. Much of the structural diversity within the polyketide superfamily of natural products is due to the ability of PKSs to vary the reduction level of every other alternate carbon atom in the backbone. Thus, the ability to introduce heterologous reductive segments such as ketoreductase (KR), dehydratase (DH), and enoylreductase (ER) into modules that naturally lack these activities would increase the power of the combinatorial biosynthetic toolbox. The dehydratase domain of module 7 of the rifamycin PKS, which is predicted to be nonfunctional in view of the sequence of the apparent active site, was replaced with its functional homolog from module 7 of rapamycin-producing polyketide synthase. The resulting mutant strain behaved like a rifC disrupted mutant, i.e., it accumulated the heptaketide intermediate and its precursors. This result points out a major difficulty we have encountered with all the Amycolatopsis mediterranei strain containing hybrid polyketide synthases: all the engineered strains prepared so far accumulate a plethora of products derived from the polyketide chain assembly intermediates as major products instead of just analogs of rifamycin B or its ansamycin precursors.

• Journal of Powder Materials
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• v.15 no.3
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• pp.182-187
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• 2008

Carbon was known to be one of effective additives which can improve the flux pinning of $MgB_2$ at high magnetic fields. In this study, glycerin $(C_3H_8O_3)$ was selected as a chemical carbon source for the improvement of critical current density of $MgB_2$. In order to replace some of boron atoms by carbon atoms, the boron powder was heat-treated with liquid glycerin. The glycerin-treated boron powder was mixed with an appropriate amount of magnesium powder to $MgB_2$ composition and the powder pallets were heat treated at $650^{\circ}C\;and\;900^{\circ}C$ for 30 min in a flowing argon gas. It was found that the superconducting transition temperature $(T_c)$ of $Mg(B_{1-x}C_x)_2$ prepared using glycerin-treated boron powder was 36.6 K, which is slightly smaller than $T_c$(37.1 K) of undoped $MgB_2$. The critical current density $(J_c)$ of $Mg(B_{1-x}C_x)_2$ was higher than that of undoped $MgB_2$ and the $T_c$ improvement effect was more remarkable at higher magnetic fields. The $T_c$, decrease and $J_c$ increase associated with the glycerin treatment for boron powder was explained in terms of the carbon substitution to boron site.

• Asian Journal of Turfgrass Science
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• v.24 no.2
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• pp.131-137
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• 2010

Recently, many domestic golf course have showed interest in introducing new cultivars into existing putting greens without reconstruction. But, any report and survey method for interseeding substitute ratio haven't been published. Therefore, this study was conducted to evaluate the effectiveness for interseeding by investigating how many interseeding plants were introduced into existing putting green by using molecular biologic tool. The Woo Jung Hills Country Club was one time introduced penn A4 into penncross putting green. The Lake Side Country Club was three times introduced CY-2 into penncross putting green. The interseeding substitute ratio using SCAR markers was 20% in the Woo Jung Hills Country Club and 35.5% in Lake Side Country Club. The interseeding putting green in Lake Side Country Club was investigated to three site, strong pressure, middle pressure and weak pressure. These substitute ratios were strong pressure 27.1%, middle pressure 37.3%, and weak pressure 47.2%. The data indicated that an interseeding substitute ratio was more increased three times than one time to introducing new cultivar into existing putting green. A long-term program should be made in order to exchange existing cultivar in putting green without reconstruction.

• Microbiology and Biotechnology Letters
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• v.40 no.2
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• pp.104-110
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• 2012

Glycosynthase is an active site nucleophile mutant enzyme, prepared from glycosidase, which is capable of synthesizing oligosaccharide derivatives without the hydrolysis of the product. Thermoacidophilic ${\alpha}$-glucosidase of Thermoplasma acidophilum (AglA) exhibits a transglycosylating activity yielding various glycosides. AglA was converted to glycosynthase by the substitution of the catalytic nucleophile Asp-408 residue into non-nucleophile glycine in order to increase its ability to synthesize various glycosides by transglycosylation. The glycosynthase mutant was purified by Ni-NTA chromatography and its glycoside-synthesizing activity was measured by using an external nucleophile, sodium formate buffer, providing maltose as a donor and p-nitrophenyl-${\alpha}$-D-glucopyranoside ($pNP{\alpha}G$) as an acceptor, respectively. In addition, $pNP{\alpha}G$ was examined for its feasibility to act as both a donor and an acceptor, and products were compared with those of the wildtype enzyme. The mutant enzyme was found to catalyze the formation of a specific product from $pNP{\alpha}G$ with a yield of 42.5% without further hydrolysis, while the wild-type enzyme produced two $pNP{\alpha}G$ products at low yields. The results demonstrate the possibility of satisfactory yields for the reactions in the presence of small amounts of acceptor, and demonstrate that the high activity of the mutant, at pHs below neutrality, was applicable in the transfer of glucose from the natural donor.

• Korean Journal of Construction Engineering and Management
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• v.12 no.1
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• pp.115-124
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• 2011

The Construction New Excellent Technology was introduced to promote the development of domestic construction technology and enhance national competitiveness, by inspiring the development desire of technology developers in 1989. Domestic Construction New Excellent Technology is first introduced in 1989, the total 596 cases were assigned to the new technology and it was used the construction site. Also, it was achieved the quantitative performance of thirty-hundred applications and was effected to the domestic construction industry. However, the systematic analysis about the introduction effects of Construction New Excellent Technology are none. Therefore, the quantitative analysis of economic effects in construction industry have to a justification in according to operation of Construction New Excellent Technology system and verification of superiority. And then, the Construction New Excellent Technology system needs a developing base in the future. In this study, The Economic Impacts of Construction New Excellent Technology were analyzed in quantitatively such as a cost savings effects, the employment effects, the effects on Value-added, the import substitution effects due to use of Construction New Excellent Technology. This study will be used promotion of economics and superiority in the Construction New Excellent Technology.

• Journal of the Korean Ceramic Society
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• v.33 no.11
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• pp.1292-1300
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• 1996

In-site X-ray diffraction measurements under electric field up to 20kV/ cm were carried out on PLZT (x/70/30) with x=7.5, 8.0, 8.5, and 10.5 All of PLZT belonged to cubic phases. At x=7.5, 8.0 and 8.5 PLZT behaved as an antiferroelectric under low electric fields up to 4-8 kV/cm. PLZT became ferroelectric at the higher electric fields. The high-temperature measurements on the dielectric constants of PLZT with x=7.5, 8.0 and 8.5 showed that they were similar to relaxor ferroelectrics and underwent a diffuse phase transition from antiferroelectrics to paraelectrics at 50-7$0^{\circ}C$. Their P-E hysteresis curves confirmed that they were antifer-roelectrics. The broad distribution of Curie points suggests that there is a significant disorder of cations and vacances in the crystal structure of those PLZT due to La-substitution. The variation of the lattice strain of PLZT(10.5/70/30) with electic field was very small and did not show any hysteresis confirming that it was paraelectric. The degree of the electric-induced strain variation decreased as La doping increased. In PLZT(7.5/70/30) the intensity of 110 reflection changes sensitively by applying electric field. Some domains with polarization parallel to [110] appeared to be developed in the field-induced ferroelectric phase of the PLZT.

• Journal of Gastric Cancer
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• v.7 no.4
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• pp.200-205
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• 2007

Purpose: The aim of this study was to analyze the post operative outcome of reconstruction with using the stomach after performing total pharyngolaryngoesophagectomy in patients with hypopharyngeal cancer or cervical esophageal cancer. Materials and Methods: We conducted a retrospective chart review of 23 patients who underwent gastric pull up for esophageal substitution at the Department of Surgery, Yonsei University College of Medicine, between January 1991 and December 2006. All the patients had transhiatal esophagectomy performed without thoracotomy. Results: There were seventeen males and six females with a median age of 58.1 years (range: 40-70 years). 19 cases were hypopharyngeal cancer, 13 cases had cancer in the pyriform sinus, 15 cases had cancer in the postcricoid area and one case had cancer in the glottic area. The rest were cervical esophageal cancers. The pathologic result was squamous cell carcinoma in all cases. The median total follow-up period was 33 months (range: 1-62 months) and there were two (8.6%) postoperative deaths: one was due to carotid rupture and the other was due to hepatic failure with liver metastasis. The complications were leakage in 1 patient (4.4%), pneumothorax in 1 patient (4.4%) and pneumonia in 1 patient (4.4%). Conclusion: The use of stomach for esophageal reconstruction has many benefits for treating hypopharyngeal cancer or cervical esophageal cancer, So, we made sure there was a sufficient length for the anastomosis after pharyngolaryngoesophagectomy and a rich blood supply from the stomach. There was a low incidence of the leakage at the anastomotic site, along with a low incidence of stenosis and bleeding.

• Journal of the Korean Society of Environmental Restoration Technology
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• v.18 no.2
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• pp.1-17
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• 2015

Ecological reservoir is a multifunctional space where provides the functions of retention, animal habitat and improvement of ecosystem health and landscape. The ecological reservoir of Anteo Eco Park located in Gwangmyeong-si has established to functions for water purification, maintenance of healthy aquatic ecosystem. Because the Anteo Eco Park is located in the site where nonpoint pollutant materials flow in, Anteo Eco Park has potential factors which aquatic ecosystem health deteriorates and damages the habitat of golden frog(Rana plancyi chosenica) which is restoration target species. Therefore, the purpose of this study is to suggest the plan to manage the variables which impede the right functions of aquatic ecosystem by understanding the causal loop diagram for the change of water quality environment and the interaction of predator-prey through system thinking. The results are as follows. First, the study showed that the individual number of golden frog which is an indicator species of Anteo Eco Park is threatened by snakeheaded fish, which is an upper predator. Therefore, balanced food chain should be hold to protect golden frog by capturing the snakeheaded fish which is individual number's density is high, and the monitoring management of the individual number for predator(snakeheaded fish)-prey(golden frog) should be performed. Second, the study represented that water pollution and carnification is caused by the sediment as the dead body of the large emergent vegetation in the winter cumulates as sediment. Ecological reservoir in Anteo Eco Park has been managed by eliminating the dead body of the large emergent vegetation, but the guideline for the proper density maintenance of vegetation community is additionally needed. Lastly, the study showed that aquatic ecosystem of Anteo Eco Park where is contaminated from the inflow of nonpoint pollutants affects the individual number's decline of golden frog and snakeheaded fish. Accordingly, the creation of a buffer area and a substitution wetland is needed in the periphery of the Anteo Eco Park to control the inflow of nonpoint pollutants including organic matters, nutrients and heavy metals. This study will be helpful that Anteo Eco Park improves the regional landscape and maintain healthy aquatic ecosystem space for the park visitors including local residents.

• Journal of Microbiology and Biotechnology
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• v.18 no.1
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• pp.15-22
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• 2008

Four series (S, M, R, and W) of Alternaria longipes isolates were obtained based on consecutive selection with Dimethachlon (Dim) and ultraviolet irradiation. These isolates were then characterized according to their tolerance to Dim, sensitivity to osmotic stress, and phenotypic properties. All the selected Dim-resistant isolates showed a higher osmosensitivity than the parental strains, and the last generation was more resistant than the first generation in the M, R, and W series. In addition, the changes in the Dim resistance and osmotic sensitivity were not found to be directly correlated, and no distinct morphologic characteristics were found among the resistant and sensitive isolates, with the exception of the resistant isolate K-11. Thus, to investigate the molecular basis of the fungicide resistance, a group III two-component histidine kinase (HK) gene, AlHK1, was cloned from nineteen A. longipes isolates. AlHK1p was found to be comprised of a six 92-amino-acid repeat domain (AARD), HK domain, and response regulator domain, similar to the Os-1p from Neurospora crassa. A comparison of the nucleotide sequences of the AlHK1 gene from the Dim-sensitive and -resistant isolates revealed that all the resistant isolates contained a single-point mutation in the AARD of AlHK1p, with the exception of isolate K-11, where the AlHK1p contained a deletion of 107 amino acids. Moreover, the AlHK1p mutations in the isolates of each respective series involved the same amino acid substitution at the same site, although the resistance levels differed significantly in each series. Therefore, these findings suggested that a mutation in the AARD of AlHK1p was not the sole factor responsible for A. longipes resistance to dicarboximide fungicides.

• Food Science of Animal Resources
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• v.24 no.4
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• pp.355-360
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• 2004

In Korean beef market, one of the major problems is mislabeling or fraudulent distribution of Holstein dairy meat or imported beef as domestic Hanwoo meat. Therefore, there has been a great need for a development of technology to identify beef breeds in meat and meat products. This study was carried out to develop the accurate and reliable method for the identification of beef breed using PCR-RFLP marker of MC1R, MGF and TYRPl genes affecting coat colors in cattle. A single base substitution (G\longrightarrowT transition) at the codon for amino acid position 104 of MC1R gene was identified between Hanwoo and Holstein and Angus breeds. The change at this position creates Msp I restriction site in Holstein and Angus, but not in Hanwoo. When the DNA amplified products (537 bp) was digested with Msp I, Hanwoo meat showed a single band of 537bp, while two fragments of 329bp and 208 bp were observed in Holstein meat and Angus breed, respectively. Thus, breed-specific RFLP marker in the MC1R gene can be used to distinguish between Hanwoo meat and Holstein and Angus meats. In the RFLP genotype of MGF gene, the frequency of r/r type was 75％ in Manwoo, whereas the frequency of R/R was 80％ in Hereford breed. Holstein and Angus breeds showed 100％ for R/r type. Therefore, Hanwoo meat showed significant difference in the MGF genotype frequencies compared with those of Holstein meat and imported beef cattle breeds. However, TYRP1 gene showed the same genotype in all breeds examined. Thus, this TYRP1 gene can not be used as a molecular marker for breed identification. As a consequence, we suggest that RFLP markers of the MC1R and MGF coat color genes could be used as DNA marker for identification of Hanwoo meat from Holstein and imported meats.