• Journal of Korean Society of Environmental Engineers
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• v.29 no.1
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• pp.54-61
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• 2007

The influences of pH were investigated for anaerobic hydrogen gas production under the constant pH condition ranged from pH 3 to 10. Carbon dioxide and hydrogen gas were main components of the gas but methane was not detected in the produced gas when sucrose was added in enrichment medium. When the modified Gompartz equation was applied for the statistical analysis of experimental data, a hydrogen production potential and maximum gas production rate at pH 5 were 1,182 mL and 112.46 mL/g dry wt biomass/hr. The hydrogen conversion ratio was 22.56%. The butyrate/acetate ratios at pH 5 and pH 6 are 1.63 and 0.38. Higher butyrate/acetate ratio produced more hydrogen gas generation. The Haldane equation model was used to find the optimum pH and fitted well with the experimental data$(r^2=0.98)$. The optimum pH and specific hydrogen production were 5.5 and 119.61 mL/g VSS/h.

• Microbiology and Biotechnology Letters
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• v.32 no.1
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• pp.60-66
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• 2004

The cycloinulooligosaccharide fructanotransferase(CFTase) gene (cft) from Paenibacillus polymyxa was subcloned into the E. coli-yeast shuttle vector, pYES2.0 (GALI promoter). The constructed plasmid, pYGCFT (9.9 kb) was introduced into S. cerevisiae SEY2102 cell and then the yeast transformant was selected on the synthetic defined media lacking uracil Based on the cyclofructan(CF) spots on thin-layer chromatogram, the gene under the control of GALI promoter was successfully expressed in the yeast transformant. The recombinant CFTase was not secreted into the medium and was predominantly localized in the periplasmic space. CF was started to be produced after 3h of enzymatic reaction with inulin. The pH and temperature optimum for the CF production from inulin was pH 8.0 and 45$^{\circ}C$, respectively. Enzyme activity was stably maintained up to the pH of 10.0. The examination of the inulin sources revealed that a dahlia tuber and Jerusalem artichoke were the best for the production of CF.

• Korean Journal of Poultry Science
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• v.36 no.2
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• pp.177-186
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• 2009

It is well-known that unregulated over-expression of foreign gene may have unwanted physiological or toxic effects in transgenic animals. To circumvent these problems, we constructed retrovirus vector designed to express the foreign gene under the control of the tetracycline-inducible promoter. However, gene expressions in the tetracycline-inducible expression system (Tet system) are not completely regulated but a little leaky due to the inherent defects in conventional Tet-based systems. A more tightly controllable regulatory system can be achieved when the advanced versions ($rtTA2^SM2$) of rtTA and a minimal promoter in responsive components (pTRE-tight) are used in combination therein. In this study, we tried to produce human thrombopoietin (hTPO) from various target cells and transgenic chickens using the retrovirus vector combined with Tet system. hTPO is the primary regulator of platelet production and has an important role in the survival and expansion of hematopoietic stem cells. In a preliminary experiment in vitro, higher hTPO expression and tighter expression control were observed in chicken embryonic fibroblast (CEF) cells. We also measured the biological activity of the hTPO using Mo7e cells whose proliferation is dependant on hTPO. The biological activity of the recombinant hTPO from CEF was higher than both its commercial counterpart and hTPO from other target cells. The recombinant retrovirus was injected beneath the blastoderm of non-incubated chicken embryos (stage X). Out of 138 injected eggs, 15 chicks hatched after 21 days of incubation. Among them, 8 hatched chicks were hTPO positive. When the Go transgenic chicken was fed doxycycline (0.5 mg per 1 gram of feed), a tetracycline derivative, hTPO concentration of the transgenic chicken blood was 200 ng/mL. Germline transmission of the transgene was confirmed in sperm of the Go transgenic roosters. These results are informative to establish transgenic chickens as bioreactors for the mass production of commercially valuable and biological active human cytokine proteins.

• Microbiology and Biotechnology Letters
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• v.36 no.2
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• pp.128-134
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• 2008

In order to examine efficient L-threo-2,3-Dihydroxyphenylserine (L-threo-DOPS) synthesis process using whole cell biocatalyst, a thermostable L-threonine aldolase (L-TA), which cloned from Streptomyces coelicolor A3(2) and improved for stability, was expressed in a Corynebacterium glutamicum R strain. The constructed Corynebacterium expression vector, pCG-H44(1) successfully expressed L-TA in C. glutamicum R strain, but showed very low expression level. In order to improve the expression level, the expression vector named pCG-H44(2) was reconstructed by eliminating 1 nucleotide between SD sequence and start codon of L-TA. The pCG-H44(2) vector plasmid was able to overexpress L-TA approximately 3.2 times higher than pCG-H44(1) in C. glutamicum R strain (CGH-2). When the whole cell of CGH-2 was examined in a repeated batch system, L-threo-DOPS was successfully synthesized with a yield of 4.0 mg/ml and maintain synthesis rate constantly after 30 repeated batch reactions for 130 h.

• Journal of Life Science
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• v.21 no.12
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• pp.1726-1731
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• 2011

Human Stem Cell Factor (hSCF) is a cytokine that binds to the c-Kit receptor and plays an important role in hematopoiesis, spermatogenesis, and melanogenesis. To produce the human Stem Cell Factor (hSCF) recombinant protein, we constructed a germline transgenic silkworm using the piggyback vector. The expression of the hSCF gene was driven by the Drosophila heat shock protein 70 (dHsp70) promoter. 3XP3 promotor-driven EGFP was used as a marker which allowed us to rapidly distinguish the transgenic silkworm. A mixture of the donor and helper vector was micro-injected into 1,020 eggs of bivoltin silkworms, Keomokjam. We obtained approximately 22 G1 broods that were EGFP-positive. The expression of the hSCF gene in the transgenic silkworm was analyzed by SDS-PAGE and immunoblotting. Also, analysis of insertion sites into the silkworm genome using inverse PCR showed that exogenous DNA was inserted into the transgenic silkworm genome. These results show that successfully constructed transgenic silkworm expresses the hSCF recombinant protein.

• Applied Biological Chemistry
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• v.14 no.2
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• pp.131-135
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• 1971

1. L-Tyrosine-${\alpha}$-ketoglutaric transaminase and p-hydroxyphenylpyruvic acid oxidase are distributed in Aspergillus oryzae. 2. L-Tyrosine oxidation in extracts of acetone powder, cell free extract and culture liquid of Aspergillus oryzae cultivated in the shaking culture are considerably accelerated by the addition of ${\alpha}$-ketoglutaric acid and then formation of glutamic acid was identified by chromatography method. 3. The roles of ${\alpha}$-ketoglutaric acid and pyridoxal phosphate have been shown to be an amino group acceptor in a transamination reaction. 4. Enzyme systems of an extracts of acetone powder and cell free extract also rapidly oxidized L-tyrosine and p-hydroxyphenlpyruvic acid to homogentisic acid. 5. The optimum pH for L-tyrosine-${\alpha}$-ketoglutaric acid transaminase was pH values of 6.0 and 6.5, and that for p-hydroxyphenylpyruvic acid oxidase was at pH values of 7.5.

• Research in Plant Disease
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• v.18 no.3
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• pp.186-193
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• 2012

Biotransformation is an eco-friendly and efficient method for enhancing the bioavailability of biopesticide. To increase the antifungal activity of the crude extract of Cynanchum wilfordii roots against barely powdery mildew, we performed biotransformation of wilfoside C1G using ${\beta}$-glucosidase (cellobiase from Aspergillus niger). The mixture (G sample) of partially purified wilfoside C1G and cynauricuoside A (K1G) was treated with ${\beta}$-glucosidase to remove a glucopyranosyl moiety. The enzyme completely converted C1G to C1N and K1G to K1N. Optimal conditions for enzymatic biotransformation of G sample were determined to be 10% ethanol, 1,555 ${\mu}U$ ${\beta}$-glucosidase/ml, pH 5, and $45^{\circ}C$. In in vivo experiment, the G sample transformed by ${\beta}$-glucosidase showed stronger antifungal activity against barley powdery mildew than the non-treated G sample. These results suggest that ${\beta}$-glucosidase biotransformation can be applied to increase the antifungal activity of the crude extract of C. wilfordii roots against powdery mildews.

• Journal of the Korean Ceramic Society
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• v.40 no.6
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• pp.608-614
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• 2003

Well developed hydroxyapatite whiskers (length 5 ${\mu}{\textrm}{m}$, diameter 0.5 ${\mu}{\textrm}{m}$) have been synthesized by the hydrolysis reaction of $\alpha$-tricalcium phosphate ($\alpha$-Ca$_3$(PO$_4$)$_2$) under pH 9.1 at 9$0^{\circ}C$ for 6 h. The effect of reaction conditions (temperature, time, pH) on the conversion of $\alpha$-tricalcium phosphate to hydroxyapatite was examined. In addition, the hydroryapatite was characterized in terms of microstructure, composition and thermal stability using XRD, SEM, ICP, and TGA instruments.

• Journal of the Korea Organic Resources Recycling Association
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• v.18 no.1
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• pp.89-97
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• 2010

In this study, to investigate the effect of chemical pretreatment for livestock wastewater, laboratory scale test for ozonation and linked treatment of sewage were conducted. and the results were obtained as follows. The ozonation of livestock wastewater showed the COD removal rate per hour to be 17%, 78% and 62% at each pH 4, 7 and 10, respectively. With transformation of NBDCOD to biodegradable BDCOD by ozonation, the ratio of SCODcr/TCODcr was increased from 26% to 38%. Accordingly, pretreatment of livestock wastewater affected to the biological post treatment process to elevate removal efficiency by transformation of nonbiodegradable mass to biodegradable mass. As the results of linked treatment of pre-ozonated livestock wastewater and sewage in the MLE process, the treatment efficiencies of TCODcr 93.8%, T-N 74.3%, T-P 89.7%, SS 97.5% were earned at 100% of internal recycle rate. When the internal recycle rate was increased to 150%, the treatment efficiencies of TCODcr 94.5%, T-N 54.5%, T-P 70.8%, SS 98.5% were earned. Also the removal efficiencies of TCODcr 92.6%, T-N 83.1%, T-P 81.9%, SS 98.5% were earned as the internal recycle rate was increased to 200%. Especially, nitrogen removal efficiency in the linked treatment showed 74.3%, 54.5%, 83.1% at 100%, 150% and 200% of internal recycle ratio, respectively, which revealed the tendency of higher removal efficiency than that of sewage treatment.

• Journal of Plant Biotechnology
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• v.31 no.3
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• pp.231-237
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• 2004

Oxycarotenoids are oxygenated carotenoids that perform critical roles in plants. $\beta$-Carotene hydroxylase adds hydroxyl groups to the $\beta$-rings of carotenes and has been cloned from several bacteria and plants including Arabidopsis. This study was carried out to investigate the effect of $\beta$-carotene hydroxylase gene (Chyb) on the oxycarotenoids biosynthesis in the transgenic Arabidopsis. Construct of pGCHYB containing Chyb was established onto Gateway vector system (pENTR3C gateway vector and pH2GW7 destination vector). Arabidopsis thaliana (cv. Columbia) was transformed with Agrobacterium tumerfacience GV3101 harboring pGCHYB construct driven by 35S promoter and hygromycin resistant gene. Seven hundred bases paired PCR products, indicating the presence of Chyb gene, were found in the transformants by PCR analysis using Chyb primers. Hygromycin resistance assay showed that transgenes were stably inherited to next generation. The overexpression of the Chyb gene resulted in the decrease carotenoid content. Especially, astaxanthin unusual oxycarotenoid in wild type Arabidopsis was detected in the transgenic plants. This means that decreased carotenoids might be converted into astaxanthin metabolism with the aid of silent gene in the host.