• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.12
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• pp.1647-1653
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• 2008

This study used response surface methodology (RSM) in an effort to optimize the water extraction conditions of Hypsizigus marmoreus in order to increase cytotoxicity activity of the extract. A central composite design was applied to investigate the effects of independent variables, which included the extraction temperature ($X_1$), extraction time ($X_2$), the ratio of solvent to sample ($X_3$) on dependent variables of the extracts, including extraction yield ($Y_1$) and protein content ($Y_2$). The estimated optimal conditions were as follows: $51.3^{\circ}C$ extraction temperature, 8.2 hrs extraction time, and 46.7 mL/g of solvent per sample. The extract (CE) was extracted at optimal condition and crude polysaccharides (CPS) were obtained from CE by ethanol precipitation, dialysis, and freeze drying. Neutral (NPS) and acidic (APS) fraction of polysaccharides were seperated from CPS by ion chromatography. The growth inhibitory effects of the APS (0.5 mg/mL) on AGS human cancer cells were 73.97%. CPS showed the highest growth inhibitory effects on HepG2 human cancer cell at 0.5 mg/mL. However all fraction polysaccharides from Hypsizigus marmoreus showed lower than 20% growth inhibition on SW480 human cancer cell.

• Korean Journal of Food Science and Technology
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• v.15 no.1
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• pp.56-61
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• 1983

Direct production of biomass from starch using amylolytic yeast, Sporobolomyces holsaticus FRI Y-5 was studied with varying the ratios between carbon and other nutrient sources in the medium. It was investigated under condition of constant C/P and C/S ratio to influence the initial concentration of starch $(S_o)$ and C/N ratio on its growth which is described as the specific growth rate $({\mu})$, cell yield (Y), the maximum concentration of cell $(X_m)$, and productivity (P). They were very dependent on both $S_o$ and C/N ratio. The form of the relationship between and ${\mu}$ and $S_o$ was observed to be similar to saturation kinetics at C/N = 100 but presented substrate inhibition at other C/N ratios. As $S_o$ was changed from 22.5 to 90 g/l, Y was observed to vary with C/N ratios but seemed to decrease as a wholes. $X_m$ was linearly related to $S_o$ at more than C/N = 50 but at less than C/N = 10 substrate inhibition was presented. P increased suddenly to $S_o$ = 45 g/l and then changed decreasingly at less than C/N = 50, but at more than C/N = 100 it changed increasingly. The effect of C/P ratio and C/S ratio on the yeast growth was also investigated at constant $S_o$ and C/N ratio. ${\mu}$ was dependent on C/P and C/S ratios, but Y, independent on them. But $X_m$ was reliant upon C/P ratio but not upon C/S ratio.

• Journal of Plant Biotechnology
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• v.33 no.3
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• pp.195-207
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• 2006

Stem cells are the primordial, initial cells which usually divide asymmetrically giving rise to on the one hand self-renewals and on the other hand progenitor cells with potential for differentiation. Zygote (fertilized egg), with totipotency, deserves the top-ranking stem cell - he totipotent stem cell (TSC). Both the ICM (inner cell mass) taken from the 6 days-old human blastocyst and ESC (embryonic stem cell) derived from the in vitro cultured ICM have slightly less potency for differentiation than the zygote, and are termed pluripotent stem cells. Stem cells in the tissues and organs of fetus, infant, and adult have highly reduced potency and committed to produce only progenitor cells for particular tissues. These tissue-specific stem cells are called multipotent stem cells. These tissue-specific/committed multipotent stem cells, when placed in altered environment other than their original niche, can yield cells characteristic of the altered environment. These findings are certainly of potential interest from the clinical, therapeutic perspective. The controversial terminology 'somatic stem cell plasticity' coined by the stem cell community seems to have been proved true. Followings are some of the recent knowledges related to the stem cell. Just as the tissues of our body have their own multipotent stem cells, cancerous tumor has undifferentiated cells known as cancer stem cell (CSC). Each time CSC cleaves, it makes two daughter cells with different fate. One is endowed with immortality, the remarkable ability to divide indefinitely, while the other progeny cell divides occasionally but lives forever. In the cancer tumor, CSC is minority being as few as 3-5% of the tumor mass but it is the culprit behind the tumor-malignancy, metastasis, and recurrence of cancer. CSC is like a master print. As long as the original exists, copies can be made and the disease can persist. If the CSC is destroyed, cancer tumor can't grow. In the decades-long cancer therapy, efforts were focused on the reducing of the bulk of cancerous growth. How cancer therapy is changing to destroy the origin of tumor, the CSC. The next generation of treatments should be to recognize and target the root cause of cancerous growth, the CSC, rather than the reducing of the bulk of tumor, Now the strategy is to find a way to identify and isolate the stem cells. The surfaces of normal as well as the cancer stem cells are studded with proteins. In leukaemia stem cell, for example, protein CD 34 is identified. In the new treatment of cancer disease it is needed to look for protein unique to the CSC. Blocking the stem cell's source of nutrients might be another effective strategy. The mystery of sternness of stem cells has begun to be deciphered. ESC can replicate indefinitely and yet retains the potential to turn into any kind of differentiated cells. Polycomb group protein such as Suz 12 repress most of the regulatory genes which, activated, are turned to be developmental genes. These protein molecules keep the ESC in an undifferentiated state. Many of the regulator genes silenced by polycomb proteins are also occupied by such ESC transcription factors as Oct 4, Sox 2, and Nanog. Both polycomb and transcription factor proteins seem to cooperate to keep the ESC in an undifferentiated state, pluripotent, and self-renewable. A normal prion protein (PrP) is found throughout the body from blood to the brain. Prion diseases such as mad cow disease (bovine spongiform encephalopathy) are caused when a normal prion protein misfolds to give rise to PrP$^{SC}$ and assault brain tissue. Why has human body kept such a deadly and enigmatic protein? Although our body has preserved the prion protein, prion diseases are of rare occurrence. Deadly prion diseases have been intensively studied, but normal prion problems are not. Very few facts on the benefit of prion proteins have been known so far. It was found that PrP was hugely expressed on the stem cell surface of bone marrow and on the cells of neural progenitor, PrP seems to have some function in cell maturation and facilitate the division of stem cells and their self-renewal. PrP also might help guide the decision of neural progenitor cell to become a neuron.

• KSBB Journal
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• v.21 no.6 s.101
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• pp.479-483
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• 2006

[ ${\beta}-Glucan$ ], one of the cell wall components, is most plentiful polysaccharides in cell wall and has several advantages in immune system. In yeast ${\beta}-glucan$ is mainly contained in the yeast cell wall, and thus it is important to produce high levels of cell mass for the mass production of yeast ${\beta}-glucan$. The best carbon and nitrogen sources on cell mass production were high fructose syrup and yeast extract. Response surface methodology (RSM) was very potential tool for the optimization of process factor and medium component. It was applied to estimate the effects of medium components on the production of cell mass. Optimal concentrations of high fructose syrup and yeast extract by response surface methodology were 8.0% (v/v) and 5.2% (w/v), respectively and the cell mass predicted was $17.0\;g/{\ell}$ at 20 h of cultivation.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.30 no.2
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• pp.100-107
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• 2004

Schwann cell, one of important components of peripheral nervous system, interact with neurons to mutually support the growth and replication of embryonal nerves and to maintain the different functions of adult nerves. The Ara-C, known as an antimitotic agent, have been used to have high effectiveness in eliminating fibroblasts during Schwann cell culture period. This enrichment effect is also known to be cummulative with each successive pulse of Ara-C applied and is due to a progressive loss of fibroblasts. But the cytotoxicity by Ara-C is also cummulative and noticeable over the period. To determine the most effective application time and interval of Ara-C in the Schwann cell culture, we observed the Schwann cell purity and density with the Ara-C treatment in plain and three-dimensional culture from dorsal root ganglion of new born rat. By culturing dispersed dorsal root ganglia, we can repeatedly generate homogenous Schwann cells, and cellular morphology and cell count with mean percentages were evaluated in the plain culture dishes and in the immunostainings of S-100 and GFAP in the three-dimensional culture. The Ara-C treated cultures showed a higher Schwann cell percentage ($31.0%{\pm}8.09%$ in P4 group to $65.5%{\pm}24.08%$ in P2 group), compared with that obtained in the abscence of Ara-C ($17.6%{\pm}6.03%$) in the plain culture after 2 weeks. And in the three-dimensional culture, S-100 positive cells increased to $56.22%{\pm}0.67%$ and GFAP positive cells to $66.46%{\pm}1.83%$ in G2 group (p<0.05), higher yield than other groups with Ara-C application. Therefore, we concluded that the Ara-C treatment is effective for the proliferation of Schwann cells contrast to the fibroblasts in vitro culture, and the first application after 24 hours from cell harvesting and subsequent 2 pulse treatment (P2 group in plain culture and G2 group in three-dimensional culture) was more effective than other application protocols.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.32 no.1
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• pp.42-51
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• 2006

Schwann cell, one of important components of peripheral nervous system, interact with neurons to mutually support the growth and replication of embryonal nerves and to maintain the different functions of adult nerves. The Ara-C, known as an antimitotic agent, have been used to have high effectiveness in eliminating fibroblasts during Schwann cell culture period. This enrichment effect is also known to be cummulative with each successive pulse of Ara-C applied and is due to a progressive loss of fibroblasts. But the cytotoxicity by Ara-C is also cummulative and noticeable over the period. To determine the most effective application time and interval of Ara-C in the Schwann cell culture, we observed the Schwann cell purity and density with the Ara-C treatment in plain and three-dimensional culture from dorsal root ganglion of new born rat. By culturing dispersed dorsal root ganglia, we can repeatedly generate homogenous Schwann cells, and cellular morphology and cell count with mean percentages were evaluated in the plain culture dishes and in the immunostainings of S-100 and GFAP in the three-dimensional culture. The Ara-C treated cultures showed a higher Schwann cell percentage (31.0%${\pm}$8.09% in P4 group to 65.5%${\pm}$24.08% in P2 group), compared with that obtained in the abscence of Ara-C (17.6%${\pm}$6.03%) in the plain culture after 2 weeks. And in the three-dimensional culture, S-100 positive cells increased to 56.22%${\pm}$0.67% and GFAP positive cells to 66.46%${\pm}$1.83% in G2 group (p<0.05), higher yield than other groups with Ara-C application. Therefore, we concluded that the Ara-C treatment is effective for the proliferation of Schwann cells contrast to the fibroblasts in vitro culture, and the first application after 24 hours from cell harvesting and subsequent 2 pulse treatment (P2 group in plain culture and G2 group in three-dimensional culture) was more effective than other application protocols.

• 한국생물공학회:학술대회논문집
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• 2000.04a
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• pp.53-56
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• 2000

Two-stage fed-batch culture was peformed to improve the volumetric productivity of erythritol. In the growth phase dissolved oxygen was maintained to 20% and the feed medium was automatically supplied to the fermenter by pH-stat mode. The cell yield was 0.76 g-cell/g-glucose. In two-stage fed-batch culture, 41% of total erythritol conversion yield with 187 g/L of erythritol concentration and 2.79 g/L-h of maximum erythritol Productivity were obtained when 400 g/L of glucose was directly added in the form of non-sterile powder at production phase. The erythritol productivity increased in parallel with cell mass. The metabolic shift in the biosynthetic pathway of erythritol was caused by dissolved oxygen concentration. The production of gluconic acid was observed when the dissolved oxygen in the medium was maintained over 40% during the production phase, whereas the dissolved oxygen concentration lower than 40% caused the production of citric acid. But the butyric acid was produced independently with dissolved oxygen concentration in the medium. The production of organic acids such as gluconic acid, citric acid, and butyric acid was decreased by addition of mineral salts.

• Korean Chemical Engineering Research
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• v.55 no.3
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• pp.369-378
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• 2017

Vascular endotherial growth factor (VEGF) is a potent mitogen that stimulates vascular permeability and angiogenesis and has a potential in therapeutic applications. An industrial production method that provides high yield as well as purity is needed. Researches for various factors of mild solubilization with combination of ubiquitin fusion protein to increase solubility were carried out as well as by changing pH and denaturant concentration. Usage of pET28-a bacteral expression vector in BL21 (DE3) host cell was capable of producing approximately 14 g/L VEGF fusion protein in 20L fermentor. A purification process consisting of four chromatography steps including refolding and digestion with UBP1 resulted in mild solublization under the conditions of 2M urea and pH 10.0 due to ubiquitin fusion tag protein that increases in solubility of target protein VEGF. High yield of refolding and dimerization could be obtained between two step Ni-affinity chromatography. Multimeric and misfolded proteins and endotoxin were removed by DEAE anion exchange chromatography. Final monomers were removed from dimers by gel filtration chromatography. Characterization analysis of purified dimeric VEGF was performed using SDS-PAGE and RP-HPLC with a purity of 97%.

• Polymer(Korea)
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• v.34 no.6
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• pp.522-526
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• 2010

Bacterial cellulose is produced by the bacterium Gluconacetobacter xylinus, which forms a nanofibrous pellicle in its culture medium. We studied properties of the bacterial cellulose such as crystallinity, viscosity, morphology, and mechanical properties according to the carbon source. Static cultures of Gluconacetobacter sp. V6 were performed in three kinds of media: standard Hestrin-Schramm medium, and modified medium with either glycerol or molasses as carbon sources. Cell growth and cellulose yield were increased in the glycerol and molasses media. The culture in the glycerol medium improved the physical properties of cellulose such as crystallinity, intrinsic viscosity, and breaking stress. However, the culture in the molasses medium decreased crystallinity, crystallite size, and intrinsic viscosity of cellulose. In summary, the cellulose yield was remarkably improved in the molasses medium, but with inferior structural properties.

• Microbiology and Biotechnology Letters
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• v.10 no.2
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• pp.95-100
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• 1982

Torulopsis candida FRI YA-15, a selected yeast, was cultivated in alcohol distiller's waste-filtrate of dried sweet potato for microbial protein production and BOD reduction. The General composition of waste-filterate was BOD$_{5}$ 15700 ppm, COD 36800 ppm, reducing sugar 3300 ppm, total nitrogen 910 ppm, total solids 51800 ppm and ash 390 ppm. The pH of waste was 3.85. The yield to the medium of T. candida cultivated in shake-flask at $25^{\circ}C$ for 48 hrs was 3.38g/$\ell$ and effectiveness in reducing BOD$_{5}$ and COD of waste was 38.9% and 31.8%, respectively. In batch cultivation using 3 $\ell$-jar fermenter, maximum yield to the medium reached 3.2g/$\ell$after 28 hrs cultivation under the condition of temperature 35$^{\circ}C$, initial pH 4.0, aeration rate 2vvm, agitation speed 100rpm. Dry yeast was composed of crude protein 47.98% and ash 5.23%.