• Korean Journal of Microbiology
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• v.33 no.3
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• pp.175-180
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• 1997

In an artificial pH-gradient hatch culture system, the author analyzed the effects of acidification on the species composition of heterotrophic bacteria. As the result of this study, it was found that the numbers of total bacteria were not affected by acidification and that the population size of heterotrophic bacteria decreased as pH became lower. The heterotrophic bacteria isolated from all of the pH gradient were 12 genera and 22 species. and among them, gram negative and gram positive bacteria were 64% and 36%, respectivcly. As pH decreased, the distribution rate of gram negative bacteria increased while that of gram positive bacteria decreased. Regarding to distrihution rate of genuses in each pH gradient, 13 genuses appeared at pH 7 while only 5 genuses appeared at pH 3. which means that the diversity of genera decrease as pH decreased. As a result of cluster analysis, diversity indices 01 species had ranges from 1.13 to 2.37, and decreased as pH decreased. In order to evaluate the diversity of different size samples, we analyzed the expected number of species appearance according to pH by rarefaction method. The statistical significance of species diversity was verified by the fact that the number decreased at lower pH.

• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.4
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• pp.297-302
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• 2004

A chitinase-producing bacterium was isolated from seashore mud around Beobseongpo in Chunmam province through the use of a selective enrichment culture. The best chitinase producing strain was isolated and identified as Serratia marcescens KY from its characteristics. For effective production of chitinase, optimum pH, temperature, and agitation speed were investigated in flask cultures. The optimum pH using Serratia marcescens KY was between pH 6 and 7 and the chitinase produced was 37.9 unit/mL. On the other hand, the optimal pH of the Serratia marcescens ATCC 27117 was 7.5, and the produced amount of chitinase was 35.2 unit/mL. The optimal temperature for chitinase production for Serratia marcescens KY and Serratia marcescens ATCC 27117 was $30^{\circ}$. The cell growth pattern at different temperature was almost identical to the chitinase production. To investigate the optimal shaking speed under optimal culture, speeds were varied in the range of 0∼300 rpm. The maximum production of chitinase was carried at 200 rpm although the cell growth was the highest at 150 rpm. It indicates that oxygen adjustment is required for the maximum chitinase production. Using optimal conditions, batch cultures for comparing Serratia marcescens KY and Serratia marcescens ATCC 27117 were carried out in a 5 L fermentor. The oxygen consumption was increased with the increase of culture. Especially, at 120 h of culture Serratia marcescens KY and Serratia marcescens ATCC 27117 produced 38.3 unit/mL, and 33.5 unit/mL, respectively.

• Journal of Microbiology and Biotechnology
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• v.13 no.3
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• pp.451-456
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• 2003

An expression vector system was developed for the secretory production of recombinant Bacillus stearothermophilus L1 lipase in Saccharomyces cerevisiae. The mature L1 lipase gene was fused to ${\alpha}-amylase$ signal sequence from Aspergillus oryzae for the effective secretion into the culture broth and the expression was controlled under GAL10 (the gene coding UDP-galactose epimerase of S. cerevisiae) promoter. S. cerevisiae harboring the resulting plasmid successfully secreted L1 lipase into the culture broth. To examine an optimum condition for L1 lipase expression in the fed-batch culture, L1 lipase expression was induced at three different growth phases (early, mid, and late-exponential growth phases). Maximum product on of L1 lipase (1,254,000 U/l, corresponding to 0.65/1) was found when the culture was induced at an early growth phase. Secreted recombinant L1 lipase was purified only through CM-Sepharose chromatography, and the purified enzyme showed 1,963 U/mg of specific activity and thermoalkalophilic properties similar to those reported for the enzyme expressed in Escherichia coli.

• Journal of Microbiology and Biotechnology
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• v.17 no.3
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• pp.428-437
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• 2007

The potential for humic substances to serve as terminal electron acceptors in microbial respiration and the effects of humic substances on microbial azoreduction were investigated. The dissimilatory azoreducing microorganism Shewanella decolorationis S12 was able to conserve energy to support growth from electron transport to humics coupled to the oxidation of various organic substances or $H_2$. Batch experiments suggested that when the concentration of anthraquinone-2-sulfonate (AQS), a humics analog, was lower than 3 mmol/l, azoreduction of strain S12 was accelerated under anaerobic condition. However, there was obvious inhibition to azoreduction when the concentration of the AQS was higher than 5 mmol/l. Another humics analog, anthraquinone-2-sulfonate (AQDS), could still prominently accelerate azoreduction, even when the concentration was up to 12 mmol/l, but the rate of acceleration gradually decreased with the increasing concentration of the AQDS. Toxic experiments revealed that AQS can inhibit growth of strain S12 if the concentration past a critical one, but AQDS had no effect on the metabolism and growth of strain S12 although the concentration was up to 20 mmol/l. These results demonstrated that a low concentration of humic substances not only could serve as the terminal electron acceptors for conserving energy for growth, but also act as redox mediator shuttling electrons for the anaerobic azoreduction by S. decolorationis S12. However, a high concentration of humic substances could inhibit the bacterial azoreduction, resulting on the one hand from the toxic effect on cell metabolism and growth, and on the other hand from competion with azo dyes for electrons as electron acceptor.

• Journal of Microbiology and Biotechnology
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• v.24 no.6
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• pp.781-787
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• 2014

2-Ketogluconic acid production by Klebsiella pneumoniae is a pH-dependent process, strictly proceeding under acidic conditions. Unfortunately, cell growth is inhibited by acidic conditions, resulting in low productivity of 2-ketogluconic acid. To overcome this deficiency, a two-stage fermentation strategy was exploited in the current study. During the first stage, the culture was maintained at neutral pH, favoring cell growth. During the second stage, the culture pH was switched to acidic conditions favoring 2-ketogluconic acid accumulation. Culture parameters, including switching time, dissolved oxygen levels, pH, and temperature were optimized for the fed-batch fermentation. Characteristics of glucose dehydrogenase and gluconate dehydrogenase were revealed in vitro, and the optimal pHs of the two enzymes coincided with the optimum culture pH. Under optimum conditions, a total of 186 g/l 2-ketogluconic acid was produced at 26 h, and the conversion ratio was 0.98 mol/mol. This fermentation strategy has successfully overcome the mismatch between optimum parameters required for cell growth and 2-ketogluconic acid accumulation, and this result has the highest productivity and conversion ratio of 2-ketogluconic and produced by microorganism.

• Fisheries and Aquatic Sciences
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• v.2 no.2
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• pp.182-188
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• 1999

Production of extracellular polysaccharide by Monilinia fructigena in B-I medium containing cereals was higher than that in glucose medium. Productivities in B-I medium and glucose medium were 0.7g/l nd 0.2-03g/l respectively. The maximum content of polysaccharide occurred at the rising point from the lowest pH of culture. As the apparent viscosity of the polysaccharide solution increased, the flow Index (m) decreased, and the consistency Index (Kc) also increased. The polysaccharide solution was a typical pseudoplastic fluid. The mycelium was separated from the culture solution by $300\mu m$ mesh-filter and the polysaccharide was precipitated by adding 50% of ethanol (v/v). The amount of the polysaccharide removed from the filtrated solution was 0.45 g/l and the amount adhered to the mycelium was 0.25g/l. In experiments for investigating growth enhancement of rotifer (Brachionus plicatilis) by the polysaccharide, the dose of the polysaccharide was 1mg per 10,000 organisms of rotifer. Maximum specific growth rate of rotifer with feed consisting of sea Chlorella sp. and the polysaccharide was 1.095/day in the batch culture for 10 days. A semi-continuous culture was done for 30 days, the biomass of rotifer could be harvested twice. Maximum specific growth rate with sea Chlorella sp. and the polysaccharide was 0.734/day before the first harvest, and 1.685/day before the second harvest. Productivity was 38 $cells/ml\; \cdot\; day$ with sea Chlorella sp. and the polysaccharide.

• KSBB Journal
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• v.13 no.1
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• pp.58-64
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• 1998

An extracellular lipid-producing strain, Rhodotorula glutinis K-501, was isolated from soil. The extracellular lipid produced by the cell was found to have good and stable emulsifying activity. Factors affecting the extracellular lipid production were studied to optimize culture conditions for maximum production. Sucrose and ammonium sulfate were selected as best carbon and nitrogen sources, respectively because they gave the highest concentration of product. The optimum C/N ratio was 50. Optimum concentrations of $KH_2PO_4,\; Na_2HPO_4, \;and\; CaCl_2 $were 3.5, 1.0, 0.75, and 0.1g/L, respectively. The optimum temperature and initial pH were determined as $22^{\circ}C$ and 7.0, respectively. When the batch culture was conducted with a sucrose concentration of 60g/L under the optimized conditions, extracellular lipid was produced to a concentration of 8.1g/L with a high production yield of 51.9% based on dry cell weight.

• KSBB Journal
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• v.31 no.2
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• pp.105-112
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• 2016

Ferritin is known to regulate iron metabolism and maintain iron in a variety of the eukaryotic organisms. The region encoding the mature ferritin (0.47 kb, H-type) of Periserrula leucophryna was amplified using the designed primers including restriction enzyme site and termination codon and subcloned in frame to the pRBAS secretion vector containing the signal sequence, RBS, and promoter of amylase gene (E. coli-Bacillus shuttle vector), resulting in recombinant pRBAS-PLF vector. Recombinant ferritin (18 kDa) was correctly processed and secreted from Bacillus subtilis LKS strain harboring the pRBAS-PLF vector and quantitatively analyzed by SDS-PAGE and western blot, respectively. Secretion of the ferritin was optimized by culture conditions (host, medium, temperature, nitrogen source) in 3 L batch culture and 5 L jar fermenter. Finally. the ferritin was largely produced using 50 L fermenter as the following conditions; at $30^{\circ}C$, 150 rpm, 1 vvm in Bacillus subtilis LKS using PY medium. The secreted ferritin was maximally measured (approximately 177.6 ug/ml) when the cell density reached to 14.4 at $OD_{600}$ (20 h incubation). The iron binding activity was confirmed by Perls' staining in 7.5% non-denaturing gel, indicating that the multimeric ferritin (composed of 24 subunits) was formed in the culture broth after secretion. Biologically, the culture broth and powder type containing ferritin were tested for possibility as feed additive in chicken broiler. As a result, the ferritin stimulated the growth of chick broil and improved feed efficiency and production index.

• Journal of Microbiology and Biotechnology
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• v.14 no.1
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• pp.90-96
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• 2004

A synthetic defined medium, fortified with amino acids, was developed for the stable production of the kringle fragments of human apolipoprotein(a) (apo(a)), rhLK68. Using a complex rich medium containing yeast extract and a high-cell-density fed-batch culture, the expression level of rhLK68 reached 17% of the total cellular protein, which corresponded to $5\;g\;l^{-1}$ of the culture. To replace the complex media with chemically defined media, several amino acids that positively affect cell growth and gene expression were chosen by a statistical method. The various combinations of the selected amino acids were tested for its fortifying effect on a minimal defined medium. When glutamine only was added, the overall expression level of rhLK68 reached 93% of the complex rich medium increasing the specific expression level by 22.4% and decreasing the cell growth by 24%. Moreover, the addition of glutamine resulted in a 2-fold increase in the concentration of rhLK68 in the culture broth, compared with the minimal defined medium. The synthetic defined media developed in this study could be generally applied to high-cell-density cultures of the recombinant Escherichia coli BL21(DE3), especially for the production of therapeutic proteins that require a strict quality control of the culture media and fermentation processes.

• Journal of Korean Society on Water Environment
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• v.21 no.2
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• pp.118-124
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• 2005

The oxygen solubilized device(O.S.D) and standardized microorganism culture system is more efficient than physical and chemical purification techniques in closed water. This study was to determine how the O.S.D and standardized culture system is efficient in purification capacity in closed water based on the lab scale and pilot plant. In the batch test, inducing the quantitative results from pilot plant operation condition, removal efficiency of COD and TN were about 48.3% and 35% respectively, while SS and chlorophyll-a were 94.9% and 68.7%. The pilot plant results showed that suspended solid(SS) and chlorophyll-a removal efficiency were 60% and 59% respectively, due to coagulation characteristics by standardized culture. Total nitrogen(TN) and total phosphorus(TP)showed good effect for the purification of target pond water quality from field data. Additionally, released velocity was determined in control condition of $5.31mgPO{_4}^{3-}{\cdot}m^{-2}{\cdot}day^{-1}$ and $2486.8mgCOD{\cdot}m^{-2}{\cdot}day^{-1}$. Otherwise, phosphate and COD reflux in the aeration and microorganism condition was showed $-9.95mgPO{_4}^{3-}{\cdot}m^{-2}{\cdot}day^{-1}$ and $-397.88mgCOD{\cdot}m^{-2}{\cdot}day^{-1}$. This technology is the most effective not only removal of SS and chlorophyll-a but also control of phosphate and COD release which is very important phenomena in evaluating water quality in closed water like a reservoir and pond.