• KSBB Journal
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• v.11 no.3
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• pp.303-310
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• 1996

The marine microalga Isochrysis galbana Parke was studied to optimize its growth conditions in flask culture. Important medium components studied include nitrogen source, buffer, trace elements and vitamins. Environmental conditions include pH, temperature, light intensity, mixing extent and working volume. The medium prepared from natural sea-waters gave a higher final cell density than the medium prepared from synthetic sea-water Nitrate was a better source than ammonium. In the range of 0.4∼2mM, the final cell density was proportional to the initial nitrate concentration and the cell yield was estimated to be 8.5g dry cell wt/g N. For phosphate, optimal growth was observed in 0.1∼1.0mM but a considerable variation in pH was resulted. The addition of Tris at 5mM or 7mM could stabilize the medium pH, but this significantly reduced both growth rate and final cell density, The effect of trace elements and vitamins was negligible. Optimal temperature and initial pH were $20^{\circ}C$ and 8. When the intensity of incident light was varied in the range of 400∼2100 lux, the growth rate increased from 10mL to 70mL, the final cell density decreased although the initial growth rate did not change. Optimal agitation speed was 100rpm when working volume was 30mL. With optimal conditions, the maximum specific growth rate obtained was 0.021hr-1 and the final cell density was 1.1g/L.

• Korean Journal of Environmental Biology
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• v.29 no.4
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• pp.321-325
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• 2011

The mutant strain M6 derived from Acetobacter sp. A9, which produces high levels of the bacterial cellulose derived by random mutagenesis with N-methyl-N'-nitro-N-nitrosoguanidine or UV treatment, was selected by a Hestrin and Schramm medium (HSB) plate assay. The characterization of the cellulose production was studied in flask culture to improve the productivity of bacterial cellulose by $Acetobacter$ sp. A9 and mutant strain M6. The yield of cellulose production was superior to mutant M6 than $Acetobacter$ sp. A9. Cellulose was produced 0.12 g $L^{-1}$ by $Acetobacter$ sp. A9 at HS medium and the mutant M6 produced the cellulose 6.95 g $L^{-1}$at HS medium. Strain M6 produced less amount of gluconic acid than A9, thus showing that cellulose production is negatively relted with the gluconic acid production.

• Microbiology and Biotechnology Letters
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• v.2 no.2
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• pp.83-88
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• 1974

The water pollution by waste water is one of the important issue and the short of animal feed is too, in Korea. So, this experiment is accomplished to treat alcohol distillers' waste by micro-organisms and planning to produce yeasts, which can be used as animal feed, pharmacy and condiments. 1. The raw material, alcohol distillers' waste, of this experiment consists of insoluble solids (residue) and filterate (supernatant). The residue contains 33.08％ of crude protein, 19.96％ of total sugar. and 2.06％ of ash, respectively. On the other hand the flterate through the Toyo filter paper No. 5C, contains 2.48％ of crude protein, 1.54％ of reducing sugar, and 0.43％ of ash, respectively. 2. Optimum pH of the basal medium for the growth of Saccharomyces cerevisiae YF-1 is 4.0. Optimum culture condition of this is as follows : when 0.43g of urea, 0.43g of potassium phosphate monobasic, and 0.21g of magnesium sulfate are added to the 100m1 of basal medium. Optimum temperature and optimum incubation time are 30$^{\circ}C$ and 24-28 hrs. 3. Under these conditions, the maximum yield of dry yeast is 1.38％ to the medium. 4. The composition of dry yeast, produced under these conditions, is as follows: crude protein, 56.96％, lipid, 1.30％. total sugar, 6.53％, and ash 9.62％.

• Journal of Chest Surgery
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• v.29 no.1
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• pp.7-13
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• 1996

The conventional glutaraldehyde (GA) fixation method of tissue valves is considered to be responsible for accelerated valve degeneration. The release of toxic GA from the valve tissue is believed to limit endothelial cell (EC) ingrowth. Removal of toxic GA by reaction with L-glutamic acid and storage in a Paraben solution may offer good EC growth. To investigate the conditions for endothelialization of tissue valves, the growth properties of ECs on the conventionally and alternatively treated pericardial tissue were compared. Conventional preparation included zero-pressure fixation for 72 hours in phosphated-buffered saline (PBS) solution containing 0.5% GA at 4$^{\circ}C$ and storage into PBS containing 0.2% GA(group I). Alternatively treated pericardial tissues were divided into three postfixation treatment groups : (1) storage in PBS solution containing Paraben(group II), (2) treatment with PBS containing 8$^{\circ}C$ L-glutamic acid(PH 7.35) and storage in PBS solution containing Paraben (g oup III), (3) treatment with L-glutamic acid dissolved in distilled water (PH 3.5) (group IV). Pericardial tissue were transferred into the 24-well plate after storage for 4 weeks. ECs were harvested enzymatically from the bovine pulmonary artery and grown to confluence on culture flask surfaces. Detached ECs by trypsin were incubated into the each well of the 24-well plate including test pericardial tissues. Cells were detached by trypsin, 1, 2, 3, 5, 7 days after incubation and counted on the hemacytometer. Cell viability test was performed by frypan-blue exclusion method. Acute cell death in the group I were found even after prolonged washing. The group II showed prolonged cell survival compared with the group I. Both group III and group IV showed better cell growth than group II. There was no statistically significant difference between group III and group IV method in terms of EC growth. This results suggest that treatment by L-glutamic ac id and storage in a Paraben solution be a promising approach for improvement of durability of GA-treated tissue valves.

• Microbiology and Biotechnology Letters
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• v.26 no.5
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• pp.406-412
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• 1998

The endoglucanase gene, egl6, of Trichoderma sp. was connected with the yeast ADH1 promoter, and the resultant plasmid, pVT-C4, was introduced into three S. cerevisiae host strains (YNN27, 2805, and SEY2102). Among each 80 transformants, the cell growth and expression level of endoglucanase were compared in test-tube cultivation, and three respective transformants for each host cells showing the highest expression level and cell growth were selected. When three recombinant yeast cells were batchwise cultivated for 48 hr in flask, the total activities of endoglucanase expressed were about 1140 unit/l with 2805/pVT-C4, 1020 unit/l with SEY2102/pVT-C4, and 590 unit/l with YNN27/pVT-C4. Irrespective of host strain, about 80% of the expressed endoglucanase was detected in the extracellular medium. In addition, it was also found that the recombinant enzyme was secreted into the culture medium as two major forms of lightly and heavily glycosylated proteins.

• Applied Biological Chemistry
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• v.40 no.4
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• pp.329-333
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• 1997

Phosphate solubilizing microorganisms (1,000 bacteria and 200 fungi) were isolated from soil around Kyungnam and Kyungbook regions using potato dextrose agar-calcium phosphate medium. A fungus with the greatest phosphate solubilizing activity was selected and identified to Penicillium sp. GL-101, based on the morphological characteristics of conidiophore and conidia; flask shape of phialide, simple branching type of conidiophore, and columnar shape of conidial head, in malt extract agar and potato dextrose agar media. The optimum temperature and initial pH to solubilize rock phosphate in potato dextrose broth-rock phosphate medium were $25^{\circ}C$ and pH 7.5, respectively. In these optimum conditions, phosphate solubilizing activities of Penicillium sp. GL-101 against four types of insoluble phosphate: tricalcium-phosphate, aluminium phosphate, hydroxyapatite and rock phosphate, were quantitatively determined. As results, this fungus highly discharged free phosphates to the culture broth with the concentrations of 1,152 ppm against tricalcium-phosphate, 565 ppm against rock phosphate, 292 ppm against aluminium phosphate, and 217 ppm against hydroxyapatite, respectively.

• KSBB Journal
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• v.11 no.1
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• pp.77-85
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• 1996

The optimal initial pH for the ethanol production by Saccharomyces K35 was found to be 5.0, and about 80% of yield was obtained when 200g/$\ell$ of glucose was used as a substrate, which showed sugar tolerant. As the additives and cross-linking agent, the addition of 1.67%(w/v) Celite R-634 together with 0.33%(v/v) of glutaraldehyde(ACG bead) resulted in better stability, ethanol productivity and cell viability than Ca-alginate bead. Also, ACG bead seemed to be more resistant to phosphate ion than Ca-alginate bead, considering outgrowing cell concentration in the media. Scanning electron microscopic observation depicted that the surface of ACG bead was almost similar to the original state but not for Ca-alginate bead. When repealpd-batch culture was performed with Ca-alginate bead for 60 days in a 500m1 Erlenmeyer flask, ethanol and cell concentration were maintained about 138g/$\ell$-gel and 29~30g/$\ell$-gel, respectively, up to 40 days(7th run number), and then both were rapidly decreased. In the case of ACG bead, ethanol and cell concentration were maintained about 130~150g/$\ell$-gel and 32~35g/$\ell$-gel, respectively, up to 60days(10th run number). Cell viability was maintained about 70%, and outgrowing cell concentration was below 5.8% of total cell concentration.

• The Korean Journal of Mycology
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• v.24 no.4 s.79
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• pp.246-254
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• 1996

Since the mid of 1980's, cultivation area and production of Ganoderma lucidum have been increased annually in Korea. However, the presence of a fungal disease has become a major limiting factor in the cultivation of Ganoderma lucidum, causing a serious economic loss. The present study was carried out to isolate and identify the pathogenic fungus to Ganoderma lucidum. Several fungi isolated from the wood logs showing typical symptoms were tested whether they are pathogenic to Ganoderma lucidum or not by cross-pairing culture method, flask inoculation method, and wood log inoculation method. The pathogenic fungus produced ascomata. Mature ascomata was spherical, dark, thick-walled, $45{\sim}95\;{\mu}m$ diameter. Asci were thin-walled, evanescent when mature, disintegrate early. Ascospores were spherical, hyaline, glaborous, thick-walled, refractive, $3.6{\sim}4.3\;{\mu}m$ in size. Conidiophores soon became abundantly septate and broke up into arthrospores, which are cylindrical, $3{\sim}6\;{\mu}m$ long and $3{\sim}4\;{\mu}m$ wide. Based on the observations under dissecting microscope, light microscope and scanning electron microscope, teleomorph and anamorph of the pathogenic fungus were identified as Xylogone sphaerospora Von Arx & Nilsson and Sporendonema purpurascens (Bonordon) Mason & Hughes, respectively. X. sphaerospora is first reported as a pathogenic fungus of Ganoderma lucidum.

• Microbiology and Biotechnology Letters
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• v.26 no.1
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• pp.68-75
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• 1998

The overproduction and high level secretion of Glucose Oxidase (GOD) from A. niger in S. cerevisiae was carried out by cloning GOD gene. For this purpose, using two different strong promoters (ADH1 promoter, GAL10 promoter) and signal sequences (${alpha}$-MF signal sequence of S. cerevisiae and ${alpha}$-amylase signal sequence of A. oryzae) and GAL7- and GOD terminator, four expression vectors were constructed. All the expression vectors were transformed in S. cerevisiae 2805 using auxotroph method. By the flask culture, transformants of pGAL expression vector series containing GAL 10 promotor showed much higher GOD productivity than transformants of pADH expression vector series containing ADH1 promoter Transformants of pGALGO2 containing GAL10 promotor and ${alpha}$-amylase signal sequence has shown the best productivity of GOD ($GOD_{total}$: 10.3 unit/mL, $GOD_{ex}$: 8.7 unit/mL) at 115 hr. This value was three fold higher than that of pGALGO1 containing GAL 10 promotor and ${alpha}$-MF signal sequence, even if the same promotor was involved. Through the ${alpha}$-amylase signal sequence of A. oryzae, GOD was secreted much more than the case of ${alpha}$-MF signal sequence from S. cerevisiae. These results suggest that signal sequence may play a important roles in not only the secretion but also the overproduction of foreign protein. Secretion rate of GOD in pGALGO1 and pGALGO2 was 89% and 84%, respectively, Because of the overglycosylation in S. cerevisiae the molecular weight of recombinant GOD in S. cerevisiae was much larger (250 kDa) than that of nature GOD in A. niger (170 kDa).

• KSBB Journal
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• v.24 no.3
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• pp.321-326
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• 2009

This study was to optimize the medium components for astaxanthin production in Paracoccus sp. through surface response methodology. A screening test was first conducted on 5 medium components using a Plackett-Burman design, from which $MgSO_4$ and yeast extract were identified as the significant factors affecting astaxanthin production. These significant factors were optimized by central composite design of experiments and response surface methodology, as 2.83 g/L $MgSO_4$ and 7.02 g/L yeast extract, respectively. The expected astaxanthin concentration with these optimized medium compositions were 0.925 mg/L. In flask culture, the experimentally obtained concentration of astaxantin was 1.021 mg/L, where it had been 0.4 mg/L before optimization.