• Korean Journal of Food Science and Technology
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• v.29 no.5
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• pp.1028-1032
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• 1997

The strain No. 33, which produces cellulose-degrading enzyme, was isolated from soil. Yellow halo was identified when the culture supernatant of the strain was loaded onto agar plate containing 2.0% CMC using paper disc method. From scanning electron microscopic observation, the morphology of the stain was rod-shaped. For identification, several biochemical characteristics were tested, and this strain was identified to Bacillus sp. So, we named this strain as Bacillus sp. No. 33. The maximal growth was observed when the stain was cultured in the medium containing 1.0% glucose, 3.0% yeast extract, 0.5% $KH_2PO_4$, 0.02% $MgSO_4{\cdot}7H_2O$, pH 7.0 at $30^{\circ}C$ for 39 hours with shaking. The maximal enzyme production was accomplished using the medium containing 4.0% CMC, 2.0% yeast extract, 0.5% $KH_2PO_4$, 0.04% $MgSO_4{\cdot}7H_2O$, pH 7.0 at $30^{\circ}C$ for 42 hours with shaking.

• The Korean Journal of Mycology
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• v.27 no.2 s.89
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• pp.100-106
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• 1999

The polysaccharides, intracellular and extracellular, extracted from the liquid culture of the Agrocybe cylindracea were purified and characterized. The mycellial cellular productivity of Agrocybe cylindracea was proved to be almost 2 folds in the shaking culture compared to the standing culture. These polysaccharides were purified by the DEAE-cellulose ion exchange chromatography and the Sepharose 2B size exclusive gel filteration. The two purified fractions of extracellular polysaccharides, ACEPDG and ACEPAG, contained 75.8% and 65.4% total sugar respectively. The total sugar content of ACIPDG and ACIPAG, the two purified fractions of intracellular polysaccharides, were 89.2% and 54.2% respectively. The molecular weights range of all the substances were estimated to be above 100,000, from 300KDa of ACEPDG to 600 KDa of ACIPAG. The results of sugar analysis by HPLC showed that the sugar part of ACEPDG was consisted of glucose and inositol. The ACIPDG, ACEPAG and ACIPAG contained three kinds of monosaccharides, glucose, fructose and inositol.

• Journal of Microbiology and Biotechnology
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• v.25 no.3
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• pp.366-374
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• 2015

Herein, we established a repeated-batch process for ethanol production from glycerol by immobilized Pachysolen tannophilus. The aim of this study was to develop a more practical and applicable ethanol production process for biofuel. In particular, using industrial-grade medium ingredients, the microaeration rate was optimized for maximization of the ethanol production, and the relevant metabolic parameters were then analyzed. The microaeration rate of 0.11 vvm, which is far lower than those occurring in a shaking flask culture, was found to be the optimal value for ethanol production from glycerol. In addition, it was found that, among those tested, Celite was a more appropriate carrier for the immobilization of P. tannophilus to induce production of ethanol from glycerol. Finally, through a repeated-batch culture, the ethanol yield (Y_e/g) of 0.126 ± 0.017 g-ethanol/g-glycerol (n = 4) was obtained, and this value was remarkably comparable with a previous report. In the future, it is expected that the results of this study will be applied for the development of a more practical and profitable long-term ethanol production process, thanks to the industrial-grade medium preparation, simple immobilization method, and easy repeated-batch operation.

• Korean Journal of Plant Tissue Culture
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• v.28 no.3
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• pp.153-157
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• 2001

Protoplasts were isolated from hypocotyls, cotyledons, and young leaves of Chinese cabbage grown under in vitro environmental condition. An enzyme mixture of 1% Cellulysin and 0.5% Macerozyme in combination with 0.4 M mannitol was most effective condition for protoplast isolation. The highest yield of protoplasts, 7.6$\times$10$^{5}$ protoplast/g of fresh weight, was obtained from the treatment of leaves for 12~16 hours at 27~28$^{\circ}C$ with shaking at 30 rpm. The most suitable medium for an initial cell division was K8p basal medium supplemented with 5 mg/L 2,4-D and 2 mg/L kinetin. Within 7~10 days, protoplasts derived from hypocotyl and cotyledon tissues formed cell colonies. When the cells were grown at the size of 8~10 cells, they were embedded into semi-solid medium containing 0.2% agarose. Calli derived from protoplast culture were transferred to the 100 different types of plant regeneration media, but no completely regenerated plants from inbred lines of Chinese cabbage used for this study wore obtained, though frequent rooting took place in several media tested.

• Korean Journal of Microbiology
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• v.46 no.4
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• pp.389-396
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• 2010

Insecticidal crystalline toxin producing Bacillus thuringiensis BT17 strain was isolated and identified as B. thuringiensis serovar colmeri by 16S rRNA analysis. BT17 strain produced crystalline ${\delta}$-endotoxin against to Lepidopteran larvae effectively on the culture broth of soybean meal and skim milk, $30^{\circ}C$ and 36 h shaking culture of 280 rpm. The maximum colony forming unit achieved when the culture was continued for 24 h, but the number of crystals increased until 36 h in the 200 L fermentor. Liquid type of biological insecticide product was made, and after 3 months storage in $20^{\circ}C$ the number of crystals was increased up to twice than beginning. Biocontrol effect of BT17 insecticide product was better in Plutella xylostella than in Spodoptera exigua, and the toxicity to animals was negligible.

• KSBB Journal
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• v.19 no.3
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• pp.192-199
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• 2004

In this work the production of glutathione (GSH) by yeast Saccharomyces cerevisiae and the monitoring of the process were studied. In shaking culture the production of GSH was high at initial pH value of 4 and at temperature of 30$^{\circ}C$. But when L-cysteine was added to the culture medium at the beginning of the cultivation, the productivity of GSH was low. In case 0,5% (v/v) of L-cysteine, glycine and glutamic acid were introduced to the culture medium in the exponential cell growth phase, high concentration of GSH (about 90 mg/L) was produced in the bioreactor. A fed-batch operation with stepwise glucose feeding strategy allowed to produce 102 mg/L of GSH. The cultivation processes were on-line monitored by a 2-dimensional fluorescence sensor. A few off-line data such as cell growth, cystein concentration, phosphate concentration and GSH productivity could be well correlated to the fluorescence intensity of some combinations of excitation and emission wavelengths.

• The Korean Journal of Mycology
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• v.12 no.4
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• pp.129-132
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• 1984

To find antitumor constituents in Korean basidiomycetes, the mycelia of Lentinus edodes DMC7, which had shown a good mycelial growth in shakeflasks, were cultured at $27^{\circ}C$ on an orbital shaking incubator at 180 rev/min for 12 days. The medium was composed of glucose (50g/l), yeast extract (9g/l), peptone (9g/l), and seven inorganic salts. A water soluble macromolecular fraction, LF-3, was obtained from the culure filtrate by fractionation with ethanol and dialysis using a Visking tube. When LF-3 was administered i.p. at 50mg/kg/day once daily for 10 consecutive days to female ICR mice which were implanted s.c. with sarcoma 180 $(10^6\;cells/mouse)$, it exerted a highly significant antitumor activity, with the tumor inhibition ratio of 53. 1%.

• Applied Biological Chemistry
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• v.49 no.3
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• pp.165-169
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• 2006

A Citrobacter sp. MB 2, azo dyes decolorizing bacterium, was isolated from the wastewater and soil and identified as Citrobacter sp.. It was examined that optimum conditions for culture media were 0.5% of sucrose, 1.0% of yeast extract, 0.1% of $K_2HPO_4$, 0.1% of $NaHCO_3$ per distilled water. The best efficient condition of culture was obtained at pH 7.0, $30^{\circ}C$ and aerobic shaking culture. The number of Citrobacter sp. MB2 in optimum medium was increased more than 7 fold compared to basal medium and 50 fold compared to nutrient broth. This strain was exhibited strong resistance against metal salts and antibiotics (ampicillin and penicillin G).

• Journal of Microbiology and Biotechnology
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• v.13 no.6
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• pp.960-968
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• 2003

A bacterial isolate showing a strong endochitosanase activity was isolated from soil and then characterized. The isolate was identified and designated as Bacillus cereus P16, based on morphological and biochemical properties, assimilation tests, cellular fatty acids pattern, along with 16S rRNA gene sequence. The optimized medium for producing extracellular chitosanase in a batch culture contained 1% tryptone, 0.5% chitosan, and 1% NaCl (pH 7.0). Powder chitosan and tryptone served the best as carbon and nitrogen sources, respectively, for the chitosanase production. Chitosanase activity was the highest when culture was completed at $37^{\circ}C$ among various temperatures ($20-42^{\circ}C$) tested in a shaking incubator (200 rpm). The levels of chitosanase activity in the culture fluid were 2.0 U/ml and 3.8 U/ml, respectively, when incubated in a flask for 60 h and in a jar fermenter for 24 h. The culture supernatant showed a strong liquefying activity on the soluble chitosan. The viscosity of 1% chitosan solution, that was incubated with the culture supernatant, was rapidly decreased, suggesting the secretion of endochitosanolytic enzymes by P16. The culture fluid revealed six endo-type chitosanase isozymes, two major (38 and 45 kD), and four minor (54, 65, 82, and 96 kD) forms by staining profile. The crude enzymes were very stable, and full activity was maintained for 4 weeks at $4^{\circ}C\;or\;-20^{\circ}C$ in the culture supernatant, suggesting a highly desirable stability rate for making an industrial application of the crude enzymes. The supernatant also cleaved the insoluble chitosan powder, but the hydrolysis rate was much lower. The enzymic degradation products of chitosan contained $(GlcN)_n$ (n=2-8). The concentration of chitosan in the reaction mixture of the crude enzyme affected the chitooligosaccharides composition of the hydrolysis products. When the higher concentration of chitosan was used, the higher degree of polymerized chitooligosaccharides were produced. By comparison with other commercial chitosanase preparations, P16 was indeed found to be a valuable enzyme source for industrial production of chitooligosaccharides from chitosan.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.69-69
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• 2001

The cell cycle phase in which donor nuclei exist prior to nuclear transfer is an important factor governing developmental rates of reconstituted embryos. It was suggested that quiescent G0 and cycling G1 cells could support normal development of reconstituted embryos. In a quest of optimized donor nuclei treatment prior to nuclear transfer, this study was undertaken to examine the cell cycle characteristics of bovine fetal and adult somatic cells when cultured under a variety of culture treatments and the cell cycle change with the lapse of time after trypsinization. This was archived by measuring the DNA content of cells using flow cytometry, Cultured fetal fibroblast cells, adult skin and muscle cells, and cumulus cells were divided by 3 culture treatments; 1) grown to 60-70% confluency (cycling), 2) serum starved culture, 3) culture to confluency. Trypsinized cells were fixed by 70% ethanol and stained with propidium iodide. For one experiment, trypsinized cells were resuspended in DMEM＋10% FBS and incubated for 1.5, 3 and 6 h with occasional shaking before ethanol fixation. Cell cycle phases were determined by flow cytometry enabling calculation of percentages of G0＋G1, S and G2＋M. The majority of cells were in G0＋Gl stage regardless of origin of cells. Cultures that were serum starved or cultured to confluency contained significantly (P<0.05) higher percentages of cells in G0＋G1 (89.5-95.4%). For every cell lines and culture treatments, percentages of cells in existing in G0＋G1 increased with decreasing of the cell size from large to small. In the serum starved and confluency groups, about 98% of small cells were in G0＋G1 Serum starved culture contained higher percentages of small-sized cells (38.5-66.9%) than cycling and confluent cultures regardless of cell lines (P<0.05). After trypsinization of fetal fibroblast and adult skin cells that were serum starved and cultured to confluency, the percentages of cells in G0＋G1 significantly increased by incubation for 1.5(95.7-99.5%) and 3.0 h (95.9-98.6%). The results suggest that the efficient synchronization of bovine somatic cells in G0＋G1 for nuclear transfer can be established by incubation for a limited time period after trypsinization of serum starved or confluent cells.