• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.99-102
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• 2001

Recombinant Saccharomyces cerevisiae strains harboring various copy numbers of hirudin gene were developed to study dependency of hirudin expression level on its gene copy number. A linear relationship between the copy number of hirudin expression cassette and hirudin expression level was observed up to 10 copies. A double <5-integration vector truncated wi 디 1 the unnecessary bacterial genes before yeast transformation showed a four-fold increase in transformation efficiency and a 1.3-fold enhancement in hirudin expression level compared with a single <5 system. Gratuitous hirudin expression strain was developed by disrupting the GALl gene of S. cerevisiae. Glucose that was fed in a limited manner effectively supported cell growth and hi겨din expression by the gratuitous strain. Effects of methanol concentrations on hirudin production in recombinant Hansenula polymorpha were investigated in continuous and fed-batch cultures. At a steady-state of continuous culture, an optimum methanol concentration of 1.7 g/L was determined at a dilution rate of 0.18 $h^{-1}$ with 1.8 mg/L ${\cdot}$ h hirudin productivity.

• Annals of Clinical Microbiology
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• v.19 no.1
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• pp.20-23
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• 2016

In the RNA-based study, it is important to extract high-quality RNA. However, RNA extraction from Mycobacterium tuberculosis is problematic due to its thick, waxy cell wall rich in mycolic acid, which renders the cells resistant to lysis. Using TRIzol reagent and several powerful bead-beating steps, a high quantity of RNA was obtained.

• Applied Chemistry for Engineering
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• v.30 no.5
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• pp.569-579
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• 2019

In this study, the effect anti-oxidant, anti-inflammatory, and liver protective activity was investigated via quick ultrasonic disintegration of pine pollen using a probe sonicator (PS) followed by the extraction with water, 70% ethanol, and 100% ethanol. The anti-inflammatory effect was studied by measuring the production of nitric oxide (NO) and cytokine in RAW264.7 cells induced with lipopolysaccharides (LPS). The cell toxicity was also checked with an 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and the experiment was conducted using non-toxic $100{\mu}g/mL$. The NO inhibition rate was highest in the 70% ethanol PS group at $85.99{\pm}0.12%$. Also an excellent efficiency was obtained from the results of interlukin-1 beta ($IL-1{\beta}$) and tumor necrosis factor alpha ($TNF-{\alpha}$), which is related to inflammation-related cytokine, with the respective inhibition rates of 63 and 22%. To examine liver protective activity, HepG2 cells were treated with Taclin, and the generation of glutamic oxaloacetic transaminase (GOT) and lactate dehydrogenase (LDH) was measured in the culture solution. From GOT and LDH generation results, the inhibition rates in the 70% ethanol PS group were 28% and 13%, respectively, which was higher compared to that of using negative control group. Our results suggest that pine pollen extracted in 70% ethanol using PS may be used to develop food products that have anti-aging, anti-inflammatory, and liver protective effects.

• Parasites, Hosts and Diseases
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• v.33 no.4
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• pp.357-364
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• 1995

The protozoan parasite, Entcmoeba histoIWticc, is one of major causative agents of intestinal disease all over the world. In acute experimental infection, the early host response to 5. histoIHtica is characterized by an infiltration of neutrophils. However, the chemotactic signal for this response is not well known. Based on the (jading that human epithelial cells produce the potent neutrophil chemoattractant and activator, interleukin-8 (IL-8), IL-8 gene expression was examined thoroughly in human colon epithelial cells exposed to 5. histolvtica trophozoites. Cellular RNAs were extracted from HT-29 or Caco-2 human colon epithelial cells exposed to 5. histoLvtica trophozoites for 30 minutes, 1 and 3 hours. IL-8 mRNA transcripts were measured by reverse transcriptional polprnerase chain reaction (RT-PCR) using synthetic standard RNA. The number of IL-8 mRNA molecules increased from 30 minutes to 3 hours of exposure period, reaching 3.1 H 107 molecules/ug of total RNA. Expression pattern of IL-8 mRNA transcripts was parallel to the amounts of IL-8 protein measured by enzyme-linked immunosorbent assay (ELISA) . Lysates of 5. histoIVtica also induced expression of mRNA for IL-8 in colon epithelial cells. These results sugf:esc that acute inflammatory reaction by 5. histoIVticc may be initially triggered by proinflammatory cytokines such as IL-8 secreted from epithelial cells of the colon.

• Microbiology and Biotechnology Letters
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• v.35 no.4
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• pp.325-333
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• 2007

One bacterium with high proteinase production and spore-forming ability was isolated from korean traditional soybean paste(doenjang). The isolated strain was identified as Bacillus subtilis, based on gram-staining, biochemical properties and l6S rRNA gene sequencing, and designated as B. subtilis DJI. Its growth rate was very fast, and it reached its stationary phase within 9 h, and then started to form spores. Bacterial-koji and doenjang were prepared using B. subtilis DJI. Chemical components of the doenjang were determined after 2 months of aging period: amino nitrogen 507 mg%, crude protein 14.3%, crude fat 4.8% and water 54.9%. The composition of total and free amino acids and their ratios of doenjang were changed during the aging period. Among total amino acids in DJI doenjang, glutamic acid, aspartic acid, leucine and arginine were the major amino acids. The fibrinolytic activities of DJI doenjang and traditional doenjangs were 909.7 units/ml and $363.3{\sim}618.6\;units/ml$, respectively. Flavor compounds of DJI doenjang and traditional doenjang were extracted by SDE(simultaneous steam distillation and extraction), and analyzed by GC/MS; DJI doenjang possessed the typically favorable flavor compounds in traditional korean doenjang, with reduced off-flavor compounds.

• Journal of the Korea Organic Resources Recycling Association
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• v.30 no.3
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• pp.23-28
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• 2022

In order to investigate the degree of sewage sludge dewaterability using Fe(II)/Na₂S₂O₈, STTF, SCST, water content, TS, VS, TB-EPS as carbohydrate and Protein were measured. The dosage of Na₂S₂O₈ was varied from 0.4 to 0.7 mmol/gVS and molar ratio of Fe(II)/Na₂S₂O₈ was varied from 0.5 to 0.7 mol/mol. According to the increase of the dosage of Na₂S₂O₈ and Fe(II)/Na₂S₂O₈ molar ratio, STTF and SCST increased from 1.00 to 15.00 and 4.51, respectively. Water content decreased to 82.6%. TB-EPS as carbohydrate and protein decreasing rate also increased to 37.16% and 57.34%, respectively. Especially, Na₂S₂O₈ 0.6 mmol/gVS and Fe(II)/Na₂S₂O₈ 0.6 mol/mol condition, water content dercreased to 83.1%, STTF and SCST increased to 13.64 and 4.19 which showed the cost effective improvement of dewaterability. It is considered that SO₄^- radical generated by Fe(II)/Na₂S₂O₈ degraded EPS and converted bound water to free water.

• Korean Journal of Food Science and Technology
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• v.22 no.2
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• pp.129-133
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• 1990

The strain of Nuruk yeat No. 15 (NY-15) which ferments lactose in milk was Isolated from Nuruk and identified as Saccharomyces marxianus according to the API 20C profile index. The lactose hydrolysing ability of NY-15 was similar to that Saccharomyces fragilis ATCC 8583 which has ${\beta}-galactosidase$ activity. Its optimum growth temperature, pH and time for the production of maximum enzyme activity showed $28^{\circ}C$, 4.5 and 28hr, respectively. Galactose as well as sucrose as carbon sources, and urea as nitrogen source Increased the production of enzyme. In order to test the production of alcohol, NY-15 was inoculated in whey medium and whey medium added with sugar. In the former, NY-15 produced 2% alcohol and in the latter, it showed 12% alcohol production. The optimal medium pH for lactose hydrolysis of NY-15 is 4.5, whereas that of Saccharomyces fragilis ATCC 8583 is 3.5

• KSBB Journal
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• v.17 no.2
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• pp.176-181
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• 2002

Astaxanthin (3,3＇-dihydroxy-${\beta}$, ${\beta}$-carotene-4-4＇-dione), a natural pigment of pink to red color, is widely distributed in nature particularly in the skin layer of salmonoids and the crust of shrimp, lobster, etc. Recently, it was produced from the yeast culture of Phaffia rhodozyma. Because of its high thermal stability and antioxidant functionality, its applications can be extended into food, cosmetics, and pharmaceutical ingredient beyond the traditional feed additive. Because of its very high lipophilicity, astaxanthin has been extracted traditionally by strong organic solvents such as chloroform, petroleum ether, acetone, etc. In this study, we developed a surfactant-based solubillization system for astaxanthin, and used it to extract astaxanthin from disrupted yeast cells. Among Tween 20, Triton X-100 and SDS, Tween 20 was identified as the most suitable surfactant in terms of extraction capacity and safety. The ethylene oxide group of Tween 20 was identified as the most significant factor to increase the HLB value that determined the extraction capacity. The effects of micelle formation condition, such as the molar ratio of astaxanthin and Tween 20, pH, and ionic strength were also investigated. pH and ionic strength showed no significant effects. The optimal molar ratio between astaxanthin and Tween 20 was 1 : 12. Antioxidant activity of astaxanthin was higher than ${\beta}$-carotene and ${\alpha}$-tocopherol. Astaxanthin in the crude extract from the yeast cell was more resistant to air and/or light degradation than pure astaxanthin, probably because of the presence of other carotenoids and lipids.

• Applied Chemistry for Engineering
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• v.18 no.5
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• pp.501-505
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• 2007

Organophosphorus hydrolase (OPH) expressed from recombinant Escherichia coli was used to biodegrade organophosphate insecticide coumaphos which has a very high toxicity in mammalian cells. To improve the productivity of OPH, the effects of nonionic surfactants (Tween 20, PEG 1000) and organic solvents, such as glycerol, propanol, and ethanol, were investigated in the strain culture. The maximum OPH was produced when the 0.25% of Tween 20 and 0.5% of glycerol were added to the medium. As the OPH obtained from disrupt-cell process by ultrasound treatment was used, the biodegradation efficiencies of 0.2, 0.5, 1.0 and 2.0 mM coumaphos were 100, 88, 84 and 78%, respectively. A novel method developed in this study could be applied to the biodetoxification technology in the contaminated region with various coumaphos concentration.

• KSBB Journal
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• v.18 no.6
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• pp.500-505
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• 2003

‘LK (lipoprotein kringle) 68’is a polypeptide of a modified ansiostatin consisting of three kringle structures that might be clinically useful as a potential cancer therapeutics. It can be produced by overexpressing it as inclusion body in recombinant E. coli. In this study, solid-phase refolding processes using packed bed adsorption (PBA) and expanded bed adsorption (EBA) column were carried out to compare their refolding yields with that of the conventional, solution-phase refolding process, For the solution-phase and the PBA-mediated processes employing Q-Sepharose, washed inclusion body was used as the starting material, whereas both washed inclusion body and E. coli homogenate were used for the EBA-mediated process employing streamline DEAE. On the final recovery LK68 per unit mass of wet cell basis, the EBA- and PBA-mediated processes showed about 2.7- and 1.5-fold higher yields, respectively, than the solution-phase refolding method. The solid-phase refolded LK68 demonstrated the same Iysine binding bioactivity and the retention time in the RP-and SEC-HPLC as those of the native protein.