• KSBB Journal
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• v.14 no.4
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• pp.440-444
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• 1999

For the effecient production of clavulanic acid., a mutant strain Streptomyces clavuligerus KK was selected from Streptomyces clavuligerus ATCC 27064 through mutation with NTG. S. clavuligerus ATCC 27064 produced about 200 mg/L of calvulanic acid when the medium was composed of 1%(W/V) glycerol, 1.5%(W/V) soybean flour, 0.1%(W/V) $KH_2PO_4$, 0.2%(V/V) soybean oil. A selected mutant, S. clavuligerus KK, produced about 1150 mg/L of clavulanic acid in the same medium. After the addition of $MgSO_4$ to the basal medium, S. clavuligerus KK produced about 1550 mg/L of clavulanic acid, with shows about 1.3 times higher than that produced in the basal medium. In order to select the proper bioreactor for the production of clavulanic acid, a batch culture was performed in an airlift, a bubble column and an stirred tank bioreactors. In an airlift bioreactor, about 1350 mg/L of clavulanic acid was produced, in a bubble column bioreactor, about 1550 mg/L, in a stirred tank bioreactor, about 2200 mg/L, respectively. The production of clavulanic acid in stirred tank bioreactor was about 50% higer than that by an airlift and a bubble column bioreactors. According to this result, the stirred tank bioreactor was selected as a proper bioreactor.

• Korean Chemical Engineering Research
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• v.53 no.2
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• pp.211-215
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• 2015

The hydrolysis of defatted rice bran using near-critical water was performed, and the feasibility of consequent hydrolyzate as a growth medium was investigated by the cultivation of Saccharomyces cerevisiae. The near-critical water hydrolysis was carried out through a series of batch experiments, and the contents of total carbohydrates, disaccharides, and monosaccharides, total organic carbon (TOC), total nitrogen (TN), pH of products were measured. The growth rate of Saccharomyces cerevisiae was measured with optical density. The yield of total carbohydrates, TOC, and TN increased with temperature below $240^{\circ}C$, however, decreased above $240^{\circ}C$. The decrease of yields above $240^{\circ}C$ was caused by the formation of organic acids, and it agreed with the change of pH of products. The yield of glucose was a maximum at $200^{\circ}C$ and it decreased dramatically at higher temperature. The growth rate of Saccharomyces cerevisiae cultivated in the hydrolyzate was similar with that in the commercial medium under certain conditions. The growth rate was correlated with the content of glucose in hydrolyzate.

• Korean Journal of Microbiology
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• v.46 no.2
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• pp.183-191
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• 2010

In this study, we isolated 179 bacterial strains using benzene, phenol, ethylbenzene, aniline, cumene, toluene as growth substrate from TCE contaminated soils and wastewaters. All the 179 strains were screened for TCE (30 mg/L) removal (growth substrate 0.2 g/L, $30^{\circ}C$, pH 7, cell biomass 1.0 g/L (w/v)) under aerobic condition for 21 days. EK2 strain using aniline showed the highest removal efficiency (74.4%) for TCE degradation. This strain was identified as Delftia acidovorans as the results of API kit, 16S rDNA sequence and fatty acid assay. In the batch culture, D. acidovorans EK2 showed the bio-degradation for TCE in the various TCE concentration (10 mg/L to 200 mg/L). However, D. acidovorans EK2 did not show the bio-degradation in the TCE 250 mg/L. D. acidovorans EK2 also show the removal efficiency (99.9%) for 12 days in the low concentration (1.0 mg/L). Optimal conditions to degrade TCE 200 mg/L were cell biomass 1.0 g/L (w/v), aniline 0.5 g/L, pH 7 and $30^{\circ}C$. Removal efficiency and removal rate by D. acidovorans EK2 strain was 71.0% and 94.7 nmol/h for 21 days under optimal conditions. Conclusion, we expect that D. acidovorans EK2 may contribute on the biological treatment in the contaminated soil or industrio us wastewater.

• Journal of Life Science
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• v.18 no.1
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• pp.52-57
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• 2008

Xylan is a major hemicellulose component of the cell walls of monocots and hardwood, representing up to 30% of the dry weight of these plants. To efficiently hydrolyze xylan, the endoxylanase gene from Bacillus sp. was expressed in B. subtilis DB431 by introducing the plasmid pJHKJ4. The total activity of the recombinant endoxylanase reached about 857 unit/ml by batch fermentation of B. subtilis DB431/pJHKJ4 in LB maltose medium. The majority (>92%) of endoxylanase was efficiently secreted into the culture medium. The recombinant endoxylanase hydrolyzed more the birchwood xylan efficiently than the other xylans. When 4 % concentration of xylan was used, the highest production of xylooligosaccharide was observed, and xylobiose and xylotriose were the major products. Optimal amount of enzyme and reaction time for producing xylooligosaccharide were found to be 10 unit and 1 hr, respectively. In addition, the temperature of $40^{\circ}C{\sim}50^{\circ}C$ gave the highest production of xylooligosaccharide. Consequently, the optimized conditions for the production of xylooligosaccharide through the hydrolysis of xylan were determined as follows: 10 unit endoxylanase, $50^{\circ}C$, 4% birchwood xylan, 1 hr reaction.

• KSBB Journal
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• v.10 no.4
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• pp.349-356
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• 1995

To produce PHA(polyhydroxyalkanoic acid) from microbr, dozens of microorganism have been screened from sewage sludge. Selected a strain HJ out of 50 strains of PHA producing bacteria has a capability of accumulating large amounts of PHB/HV copolymer when grown in batch culture with a single carbon source (glucose) that was not generally considered as precursor of hydroxyvalerate monomer unit. The strain HJ was identified as the genus Pseudomonas with respect to morphological, cultural, and biochemical characteristics. The optimal temperature and pH for cell growth were $37^{\circ}C$ and 7.0. The optimal medium compositions for cell growth were glucose 1% as a carbon source, (NH4) 2SO4 0.2% as a nitrogen source, K2HPO4 0.3%, and KH2PO4 0.45%. TO investigate she optimal condition for PHA production two-step cultivation method was employed. PHA production was inducted by deficiency of NH4+, SO4-2, Mg+2. Besides carbon source, deficiency of all nutrients stimulated PHA productivity but deficiency of NH4+ stimulated the most HV monomer content. The highest PHA production was C/N molar ratio 95.2. Pseudomonas sp. HJ was also able to pyoduc PHB/HV copolymer when cultivated on alkane, alkanoate, alcohol as carbon sources. The contents of PHA and she proportions of hydroxyvalerate monomer units varied depending on the carbon sources. Especially Pseudomonas sp. HJ was able to incorporate hydroxyvalerate into PHB/HV to level as high as from 49 to 74 mol% when grown in a medium containing hexadecane and propionate. The purified PHA was identified PHB/HV copolymer by HNMR analysis.

• KSBB Journal
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• v.19 no.6 s.89
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• pp.441-445
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• 2004

Cellular iron uptake was performed in the yeast Saccharomyces cerevisiae that transformed with human ferritin H- and L-chain genes. The recombinant yeasts were enriched in YEP medium supplemented with $2\%$ galactose for 3 days and the iron uptake was followed by incubating the cells with iron in 20 mM MOPS buffer (pH 6.5). The reactions were examined under different conditions including the iron compounds of Fe(II) and Fe(III), the concentration of iron, the concentration of cells and the reaction time. From our results, the recombinant yeast YGH2 producing H-chain ferritin showed higher cellular iron concentration at the cell concentration of 100 mg/ml than 200 mg/ml. Iron presented as Fe(II) rather than Fe(III) was taken up more efficiently. Iron uptake increased slightly when iron was added up to 14.3 mM Fe(II) and then its cellular iron concentration was $16.7{\pm}0.7\;{\mu}mol/g$ cell wet wt. In addition, the iron uptake reaction reached to maximum at about 2 hr incubation.

• KSBB Journal
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• v.20 no.1 s.90
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• pp.60-65
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• 2005

The objective of this work was to evaluate a solid-state fermentation process for the practical production of arachidonic acid(AA) by Mortierella alpina ATCC 32222. In the present investigation, batch culture kinetics for both submerged- and solid-state fermentations was carried out at $25^{\circ}C$ to identify the relationship between growth and arachidonic acid (AA) production. Glucose and yeast extract were used in submerged fermentations by using flasks, while rice bran was used as a sole raw material in the other type of fermentations by using a series of Petri dishes. It was evident that a mixed-growth associated pattern existed between the two variables, irrespective of modes of fermentations. The effect of carbon to nitrogen (CfN) ratio on AA production in solid-state fermentation was studied in the range of 6.5 - 20. As a result, an optimum condition was found to be 6.5. Supplementary carbon source was not necessary to meet the optimum C/N ratio. Unlike the Previous results obtained by other researchers, a supplement of sodium glutamate up to $4\%$ (w/w) to the rice bran medium did not have a positive effect on the AA productivity. However, an increase in AA productivity was obtained with the rice bran medium supplemented with sesame oil.

• Microbiology and Biotechnology Letters
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• v.42 no.2
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• pp.194-201
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• 2014

Clitocybin A is a novel anti-wrinkle cosmetic agent produced by the strain from a Korean native mushroom Clitocybe aurantiaca. In this study, fermentation, extraction, and purification conditions for a large scale production of clitocybin A were optimized, and its cytotoxicity and inhibition activity on the expression of matrix metalloproteinase-1 (MMP-1) were characterized. The mass production of anti-wrinkle agent was achieved according to the 300 L fermentation process with a fed-batch cultivation using the modified yeast-maltose (YM) broth, and a total of 12.5 kg of cell mass was obtained in a 120 L culture broth for 14 days. After extraction and purification, clitocybin A was identified by HPLC. The cytotoxicity of clitocybin A was examined by the MTT assay. When assayed at 100 and 200 ${\mu}g/ml$ concentrations, clitocybin A showed no cytotoxicity, demonstrating safety. The inhibition activity of clitocybin A on the expression of MMP-1 was examined against UV irradiation. Oleanolic acid (control group) showed a relatively low MMP-1 inhibiting activity (ca. 16.7%) at 10 ${\mu}g/ml$ and showed increased cytotoxicity at higher concentrations. In contrast, clitocybin A showed no cytotoxicity at 100 ${\mu}g/ml$, and exhibited a relatively high MMP-1-inhibiting activity (33.1%). These findings indicate that clitocybin A may be a safe and effective anti-wrinkle agent for use in functional cosmetics.

• Applied Biological Chemistry
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• v.38 no.3
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• pp.191-195
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• 1995

Production of a high viscosity exoploysaccharide, methylan, by Methylobacterium organophilum from methanol was carried out in fed-batch cultures and the rheological properties of methylan fermentation broth were studied. Bacterial biomass showed little influence on viscosity, but the accumulation of methylan caused the increase of viscosity. With proceeding fermention, the viscosity at the same concentration of methylan was significantly increased and methylan solution showed slightly higher pseudoplasticity. The composition changes of methylan were investigated at various fermentation times. Contents of total sugar, reducing sugar and methylan were decreased but contents of acids(pyruvic acid, uronic acid and acetic acid) were increased with the culture time. It was considered that the increased content of acids resulted in the increase of the hyrodynamic domain in the solution due to charge repulsion. Consequently, the solution viscosity increased in propotion to the acids contents of methylan. Cell growth and methylan production were severely decreased by the limitation of dissolved oxygen. However, the cellular activity for methylan production was almost constant regardless of the level of dissolved oxygen. As a result, the high speed of agitation increased the methylan production, the specific production rate of methylan, and the methylan yield of the cell.

• Korean Journal of Microbiology
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• v.34 no.4
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• pp.219-224
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• 1998

The cyclic metabolism of poly(3-hydroxyhutyrate-co-3-hydroxyvalerate) synthesized from glucose by Alcaligenes sp. SH-69 in the presence or absence of new carbon substrate was investigated. In batch culture, the content and weight average molecular weight of the copolymer already stored in the cell decreased rapidly when there was no other carbon source available. After the depletion of carbon source, the amount of high molecular weight copolymer decreased more rapidly than that of low molecular weight copolymer, and as a result, average molecular weight distribution shifted to the lower value. The addition of a mixture of glucose and levulinic acid when the initial carbon substrate, glucose, was nearly depleted supported the continual increase in cell mass and the accumulation of poly(3HB-co-3HV) with high molar fraction of 3HV. However, solvent fractionation of the polymer with acetone revealed the degradation of pre-existing polyhydroxyalkanoale (PHA) in parallel with the synthesis of PHA from new carbon substrate. Even though PHAs obtained from each substrate alone were the copolymer of 3HB and 3HV, it was found that the polymer accumulated in the cells grown by sequential feeding was mainly physical mixture of two poly(3HB-co-3HV) copolymers containing different molar fractions of 3HV.