• BMB Reports
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• v.30 no.5
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• pp.346-351
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• 1997

The maximum adsorption/desorption conditions and the adsorption mechanism of globular proteins to vaccine adjuvants were determined. The maximum adsorption ratio of protein to the $Al^{3+}$ content of aluminum oxyhydroxide and the optimal adsorption pH are 2:1 (${\mu}g:{\mu}g$) for bovine serum albumin (BSA) at pH 6.0 and 2.5:1 (${\mu}g:{\mu}g$) for immunoglobulin G (IgG) at pH 7.0, respectively. The maximum adsorption ratio onto aluminum phosphate gel was 1.5:1 (${\mu}g$ Protein:${\mu}g$ $Al^{3+}$) at pH 5.0 for both BSA and IgG. Adsorption of the native globular proteins, BSA and IgG, to aluminum oxyhydroxide and aluminum phosphate gel was reversible as a function of pH. Complete desorption of these proteins from aluminum phosphate gel was observed at alkaline pH, whereas only 80~90% removal from aluminum oxyhydroxide was achieved with alkaline pH and 50 mM phosphate buffer. We conclude that electrostatic and hydrogen bonding interactions between the native proteins and adjuvants are important binding mechanisms for adsorption, and that the surface charge of the protein and the colloid components control the maximum adsorption conditions.

• Journal of Life Science
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• v.18 no.7
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• pp.912-917
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• 2008

To investigate the biological activity of recombinant human granulocyte colony-stimulating factor (rec-hG-CSF) in mammalian cells, hG-CSF gene was cloned using the eDNA extracted from the human squamous carcinoma cell lines and rec-hG-CSF was produced in CHO cell lines. To analyze the biological activity in vivo, the rec-hG-CSF protein was injected into mice subcutaneously on days 0 and 2. Blood was withdrawn for white blood cell (WBC) determination 5 days after the first injection. WBC values were found to have increased significantly. A pEGFP-mUII-hG-CSF vector was transfected into somatic cell lines isolated from bovine fetal cells. The colony expressing EGFP signals was observed with a confocal microscope. These data suggest that the rec-hG-CSF produced in this study has potent activity in vivo. Thus, the results of this biological activity show that rec-hG-CSF can be enhanced considerably by genetic engineering that affects potential activity, including mutations, which add the oligosaccharide chain and construct double-fusion proteins. A pEGFP-mUII-hG-CSF vector can be utilized for the production of cloned transgenic livestock.

• KSBB Journal
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• v.10 no.5
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• pp.598-603
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• 1995

Batch and continuous cell retention cultures were carried out using tapioca hydrolysate. In batch culture, reducing sugar of about 180g/$\ell$ was almost consumed in about 36 hours, and the concentration of ethanol produced was about 84g/$\ell$ making the ethanol yield 0.48 g-ethanol/g-(reducing sugar). The final yeast concentration was 8.5${\times}$107 cells/ml(about 2.1g/$\ell$). In a total cell retention culture operated with a dilution rate of 0.18h-1, the yeast concentration, the residual reducing sugar concentration, the ethanol concentration, and the volumetric ethanol productivity were about 40g/$\ell$, about 15g/$\ell$, 81.4g/$\ell$, and 14.7g/$\ell$-h, respectively. In another cell retention culture operated with a dilution rate and a bleed ratio of 0.2h-1 and 0.14, respectively, the yeast concentration increased to 22g/$\ell$ and the ethanol concentration oscillated around 68g/$\ell$. The volumetric ethanol productivity was about 13.6g/$\ell$-h and the residual reducing sugar concentration about 12g/$\ell$ containing glucose of about 4.5g/$\ell$. According to the results of batch fermentation using the solid residue from hydrolysate filtration as the substrate, it seemed to have a certain value. Thus, development of an effective reactor system to produce ethanol from this solid residue is in need.

• Korean Journal of Plant Resources
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• v.21 no.5
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• pp.408-412
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• 2008

This research was designed to investigate the antioxidant activities and tyrosinase inhibitory effects of guava (Psidium guajava L.) leaf. Total phenol content was obtained from guava leaf extract of 19.0 (g/100g, D.W.). The crude extract exhibited significantly antioxidant activities (IC50value $102.5{\mu}g/ml$, free radical scavenging; $49.4{\mu}g/ml$, SOD like activity). The crude extract of guava leaf was fractionated into four partition layers; hexane (G-H), ethyl acetate (G-E), butanol (G-B) and water (G-W) layer. The extracts of G-E, G-B, G-W showed high radical scavenging activities of over 50% at $100{\mu}g/ml$. SOD like activities of G-E, G-B, G-W were revealed, as 81.8%, 84.7%, 65.3% at $100{\mu}g/ml$, while those of G-H did not showed the effectively. The crude extract of guava leaf showed high tyrosinase inhibitory effect as 60.8% at 1mg/ml, the measurement of G-E, G-B, G-W were 65.2%, 62.8%, 51.6% and that of G-H was not effective. These results indicate that useful bioactive substances exist in the guava leaf extracts, especially G-E, G-B. And the guava leaf has the potential of being developed into health related products.

• Journal of Nutrition and Health
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• v.8 no.1
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• pp.65-69
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• 1975

Human chorionic gonadotropin hormone(H.C.G.) is secreted from the villus tissues of the placenta and excreted in lange amount into the urine. Its isolation is chiefly made from the urine of a pregnant woman. Recently, Matsushima attempted isolation of H.C.G. directly from the placenta itself. In order to prepare H.C.G. from human placenta, general method of extractiag and purifying proteins was applied. Its way was as follow: Crude H.C.G. was extracted from placenta with pH 9.0 and pH 5.0 aqua ammonia, and purified with pH 8.0 ammonia and 50% ethanol at pH 4.8. The purified H.C.G. showed two moving bands on the anode by paper electrophoresis. On the other hand, the H.C.G. from pregnancy urine (Standard. Pharm. Co.) showed same two bands but their moving ratio were different. The purified H.C.G. showed gonadotropin effect when it was injected young fomale rats 40r/cc per day for 5 days and weighted the increased ovary weight.

• Development and Reproduction
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• v.14 no.1
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• pp.13-19
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• 2010

We developed a novel dicistronic system for the expression of target cDNA sequences in the milk of transgenic animals using goat beta-casein/hGH fusion construct, pGbc5.5hGH (Lee, 2006) and internal ribosome entry site (IRES) sequences of encephalomyocarditis virus (EMCV). Granulocyte colony-stimulating factor (hG-CSF) cDNA was linked to 3' untranslated region of hGH gene in the pGbc5.5hGH via EMCV IRES sequences. Transgenic mice were generated by microinjection and transgene expression was examined in the milk and mammary gland of transgenic mice at 10 days of lactation. Northern blot analysis showed that hGH gene and hG-CSF cDNA were transcribed as a single dicistronic mRNA. The hG-CSF and hGH proteins were independently translated from the dicistronic mRNA and secreted into the milk of transgenic mice. The highest concentration of hG-CSF and hGH in the milk of transgenic mice were $237{\mu}g/m{\ell}$ and $8,990{\mu}g/m{\ell}$, respectively. In contrast, another hG-CSF expression cassette, in which hG-CSF genomic sequences were inserted into a commercial milk-specific expression vector (pBC1), generated a lower level ($91{\mu}g/m{\ell}$) of hG-CSF expression in the milk of transgenic mice. These results demonstrated that the novel pGbc5.5hGH-based dicistronic construct could be useful for an efficient cDNA expression in the milk of transgenic animals.

• Journal of Plant Biology
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• v.24 no.4
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• pp.171-179
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• 1981

The membrane-bound ATPases were separated on sucrose gradient from corn roots and characterized by pH optima, sensitivity to monovalent salt, Km and Vmax. The pH optima for the activity of all the ATPases associated with 13, 000g pellet and 13, 000~80, 000g pellet were 5 and 9, respectively. The ATPases in Fractions B and C of the 13, 000 g pellet were more active at pH 5 than pH 9. While, in the case of Fractions D, E and F, they were reverse. The activities of the ATPase in Fractions A and C of the 13, 000~80, 000 g pellet were greater at pH 5 than pH 9. On the other hand, the ATPases in Fractions B, D, E, and F were more active at pH 9 than pH 5. The optimum concentraction of ATP for the assay was about 3 to 5 mM. The Km's for the membrane-bound ATPases in 13, 000g pellet and in 13, 000~80, 000 g pellet were 0.25 mM. While Vmax values for 13, 000g pellet were from 8.0 to 12.5 $\mu$M Pi/mg protein/hr. according to pH values, those for 13, 000~80, 000 g pellet were from 35.7 to 55.6 $\mu$M Pi/mg protein/hr. Activities of the membrane-bound ATPases in both 13, 000 g pellet and 13, 000~80, 000 g pellet were stimulated with increasing the concentration of $K^+$.

• Journal of Life Science
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• v.18 no.4
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• pp.494-502
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• 2008

The anaphylaxis shock reaction on the whole cells of H. pylori exhibited a symptom of slight illness for the first and second medication of causing antigen at an antigen concentration of WC (H) $60\;{\mu}g/100\;{\mu}l$ for WC (H) and no anaphylaxis shock symptom was observed at an antigen concentration of $20\;{\mu}g/100\;{\mu}l$ for WC (L). In the case of anaphylaxis shock reaction on the crude urease, no symptom was observed at an antigen concentration of $20\;{\mu}g/100\;{\mu}l$ for both urease (L) and urease (H). In the heterologous passive cutaneous anaphylaxis (PCA) test using a guinea pig-rat, no positive reaction was detected in all the medication groups of WC (H), WC (L), urease (H) and urease (L). In the skin sensitization test, it was observed that the best antigen concentration not causing skin disorder at each of $80\;{\mu}g/100\;{\mu}l$, $40\;{\mu}g/100\;{\mu}l$, $20\;{\mu}g/100\;{\mu}l$, and $20\;{\mu}g/100\;{\mu}l$ was $40\;{\mu}g/100\;{\mu}l$.

• Microbiology and Biotechnology Letters
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• v.8 no.3
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• pp.199-206
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• 1980

Some experiments on the citric acid production were carried out from soluble starch and molasses as raw materials. When soluble starch was used as substrate for the fermentation of citric acid by the strain M-80 which had assimilating ability of soluble starch in surface culture, the optimal culture media was 120g of soluble starch, 3.0g of (N $H_4$)$_2$S $O_4$, 2.0g of K $H_2$P $O_4$, 0.2g of MgS $O_4$.7$H_2O$, 1.5mg of F $e^{++}$, 1mg of Z $n^{++}$ and 20ml of methanol were added to 1 liter and optimal pH was 5.5. In about 8 days 61.8mg/ml of citric acid was produced. When treated molasses with potassium ferrocyanide was used as substrate for the fermentation of citric acid by the strain of M-315, the optimal condition in surface culture was 250g of molasses, 0.3g of N $H_4$N $O_3$, 0.05g of K $H_2$P $O_4$, 0.01g of MgS $O_4$.7$H_2O$, 0.5g of Potassium ferrocyanide and 30ml of methanol were added to 1.0 liter. On the other hand, the optimal condition in submersed culture was 250g of molasses, 0.3g of N $H_4$N $O_3$, 0.1g of K $H_2$P $O_4$, 0.01g of MgS $O_4$.7$H_2O$, 0.5g of potassium ferrocyanide, and 30m1 of methanol were added to 1.0 liter and optimal pH was all 5.0. After 9 days culture, 69.4mg/ ml, 39.6mg/ml of citric acid were separately produced in surface and submerged culture media.dia.

• Journal of Microbiology and Biotechnology
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• v.10 no.3
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• pp.321-326
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• 2000

To examine the feasibility of the long-term production of the human granulocyte colony stimulating factor (hG-CSF) using the $_{L}-arabinose$ promoter system of Escherichia coli, flask relay culture and cyclic fed-batch culture were performed. In the flask relay culture, it was found that the pismid was maintained stably up to about 170 generations in an uninduced condition, whereby the cells could also maintain the capability of expressing hG-CSF expression were maintained stably up to at least 100 generations. In contrast, in the cyclid fed-batch culture, segregational plasmid instability was observed within about 4 generations after induction, even though the cell growth and hG-CSF production reached their maximum balues, 78.0 g/l of dry cell weight and 7.0 g/l of hG-CSF, respectively. It would appear that, when compared to the flask relay culture, the high-cell density and high-level expression of hG-CSF in the cyclic fed-batch cultrure led to the segregational plasmid instability; in other words, a severe metabolic burden existe on the cells due to the high-level expression of hG-CSF. Accordingly, based on these long-term cultures, the segregational and structural plasmid instability was observed and a strategy to overcome such problems could be designed.