• KSBB Journal
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• v.5 no.3
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• pp.269-277
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• 1990

Methylobacillus sp. SK1, an obligate methylotroph, was cultivated in a fed-batch culture using DO as a methanol feeding indicator with a micro computer-aided control system. While 2.07g/l of cell density was obtained after 13 hr in the batch culture (initial methanol concentration: 1.0%(v/v)),45.3g/l of cell density was obtained after 17 hr by feeding methanol and metal ions in the fed-batch culture with oxygen supply. The high-density biomass was obtained in short cultuivation time by fed-batch culture with feedback control, and consequently the biomass productivity was significantly increased. It was mainly due to extension of logarithmic growth period by methanol feeding without methanol inhibition and intensive aeration without DO limitation with microcomputer-aided control system.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.233-236
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• 2000

Bacillus thuringiensis (Bt) is the most widely used microbial insecticide in the biological control market. Cultivation of the microorganism to high cell densities offers potential for enhancing the rate of formation as well as the concentration of the desired products In the fermentation broths in bioreactor. With this objective, we developed the new bioreactor incorporating ceramic membrane module for the retention of cell mass. Cell yield and spore formation of Bacillus thuringiensis was improved markedly by adopting this new bioreactor based on glucose -limited feeding operation. It was possible to grow the cell and the heat-resistant spore to above $1.2\;{\times}\;10^{10}\;CFU/ml$ density. With glucose-limited operation, we studied the growth behavior of Bacillus thuringiensis during the cell retention culture. Linear growth of Bacillus thuringiensis was observed under glucose-limited culture, which matched well with simple mathematical model of cell retention culture.

• Microbiology and Biotechnology Letters
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• v.22 no.4
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• pp.389-394
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• 1994

8 X 10$^{6}$(viable cells/ml) of maximum cell density and 9000(IU/ml) of $\gamma$-IFN production were obtained at 55(ml/hr) of a perfusion rate by cultivating HSF cells using a moving membrane aeration bioreactor. This system proves to be an efficient culture process by maintaning 90% of viable cells during the whole cultivation periods. The metabolic molar quotient of glucose to lactate was 0.81 for overall ranges of glucose consumed while the evolution of ammonia was not linearly related to the consumption of glutamine. Low molar conversion ratio was observed in low consumptions of glutamine and high molar conversion ratio in high comsumptions. It also shows that the glutamolysis plays important role in the steady state conditions by evolving larger quantities of ammonia than lactate. At the above of 50 rpm, which is the optimum agitation speed for this bioreactor, the cell growth was severely affected while the IFN production was less decrea- sed, maintaing 1.5 X 10$^{-3}$(IU/cell/day) specific IFN production rate. The cumulatvie $\gamma$-IFN production was 7.2 X 10$^{8}$(IU) for 70 days of the cultivation, which yields 1 X 10$^{7}$ (IU/day) of IFN production rate. Therefore, a commercial production of $\gamma$-IFN by this culture process can be achievable by maintaining the above IFN productivity in a scaled-up culture system.

• Journal of Microbiology and Biotechnology
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• v.12 no.5
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• pp.753-759
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• 2002

$\beta$-Cyclodextrin glucanotransferase ($\beta$-CGTase) was overproduced extracellularly using recombinant E. coli by transforming the plasmid pECGT harboring a secretive signal peptide. The $\beta$-CGTase gene of alkalophilic Bacillus firmus var alkalophilus was inserted into the high expression vector pET20b(+) containing a secretive pelB signal peptide, and then transformed into E. coli BL2l(DE3)pLysS. The optimum culture conditions fer the overproduction of $\beta$-CGTase were determined to be TB medium containing 0.5% (w/v) soluble starch at post-induction temperature of $25^{\circ}C$. A significant amount of $\beta$-CGTase, up to 5.83 U/ml, which was nine times higher than that in the parent strain B. firmus var. alkalophilus, was overproduced in the extracellular compartment. A pH-stat fed-batch cultivation of the recombinant E. coli was also performed to achieve the secretive overproduction of $\beta$-CGTase at a high cell density, resulting in production of up to 21.6 U/ml of $\beta$-CGTase.

• Proceedings of the Korean Vacuum Society Conference
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• 2015.08a
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• pp.231.1-231.1
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• 2015

Surface energy, being an important material parameter to control its interactions with the other surfaces plays a key role in bio-related application. Carbon films are found very promising due to their characteristics such as wear and corrosion resistant, high hardness, inert, low resistivity and biocompatibility. The present work deals with the deposition of carbon films using unbalanced facing target magnetron sputtering technique. The discharge characteristics were studied using optical emission spectroscopy and correlated with the film properties. Surface energy was investigated through contact angle measurement. The ID/IG ratio as calculated from Raman spectroscopy data increases with the increase in power density due to the higher number of sp2 clusters embedded in the amorphous matrix. The deposited films were smooth and homogeneous as observed by Atomic force microscopy having RMS roughness in the range of 1.74 to 2.25 nm. It is observed that electrical resistivity and surface energy varies in direct proportionality with operating pressure and has inverse relation with power density. The surface energy results clearly exhibited that these films can have promising applications in cell cultivation.

• Journal of Microbiology and Biotechnology
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• v.10 no.3
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• pp.367-374
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• 2000

The composition of dissolved gases and nutrients in a liquid medium were determined for establishment of the optimum conditions for in vitro culture of Helicobacter pylori. A microaerobic condition facored by the organism was prepared by adjusting the partial pressure of the gas, agitation speed, and viscosity of the medium. The gaseous concentrations were controlled by utilizing CampyPak Plus that reduced oxygen while augmenting carbon dioxide. Agitation of the broth facilitated the oxygen transfer to the cells, yet inhibited the growth at high rates. An increase of viscosity in the medium repressed the culture although this variable was relatively insignificant. The chemical constituents of the liquid broth were examined to establish an economic model for H. pylori cultivation. The microbe required a neutral pH for optimum growth, and yet was also able to proliferate in an acidic condition, presumably by releasing the acidity-modulating enzyme, urease. Cyclodextrin and casamino acid were investigated as growth enhancers in place of serum, while yeast extract unexpectedly inhibited the cells. A low concentration of glucose, the unique carbon source for the organism, increased the cell density, yet high concentrations resulted in an adverse effect. Under optimally dissolved gas conditions, the cell concentration in brucella broth supplemented with serum substitutes and glucose reached $1.6{\times}10^8$ viable cells/ml which was approximately 50% higher than that obtained in the liquid medium added with only cyclodextrin or serum.

• Microbiology and Biotechnology Letters
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• v.24 no.1
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• pp.126-131
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• 1996

HL-60 was cultivated to produce tPA (tissue-type plasminogen activator) and study the mechanism of cell death. Maximum cell density and tPA production were obtained as $5.27{\times}10^6$ cells/ml and 324ng/ml, respectively under perfusion cultivation. tPA production was enhanced to 420ng/ml in adding 160nM of phorbol ester. The cells were gradually differentiated to granulocytes rather than proliferation. By Fluorescent microscope, apoptosis was prevailed except the death phase and in high agitation speed, but necrosis was prevailed in thawed cells and during the latter periods of the cultivation. It was also proved that tPA was most produced in apoptosis. To obtain higher tPA productivity, the cells must be maintained in apoptosis, not necrosis phase when the cells were dying.

• Journal of Microbiology and Biotechnology
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• v.20 no.11
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• pp.1555-1562
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• 2010

Development of the strain and the fermentation process of Hansenula polymorpha was implemented for the production of ${\gamma}$-linolenic acid ($GLA,\;C18:3{\Delta}^{6,9,12}$), an n-6 polyunsaturated fatty acid (PUFA) that has been reported to possess a number of health benefits. The mutated ${\Delta}^6$-desaturase (S213A) gene of Mucor rouxii was expressed in H. polymorpha under the control of the methanol oxidase (MOX) promoter. Without the utilization of methanol, a high-cell-density culture of the yeast recombinant carrying the ${\Delta}^6$-desaturase gene was then achieved by fed-batch fermentation under glycerol-limited conditions. As a result, high levels of the ${\Delta}^6$-desaturated products, octadecadienoic acid ($C18:2{\Delta}^{6,9}$), GLA, and stearidonic acid ($C18:4{\Delta}^{6,9,12,15}$), were accumulated under the derepression conditions. The GLA production was also optimized by adjusting the specific growth rate. The results show that the specific growth rate affected both the lipid content and the fatty acid composition of the GLA-producing recombinant. Among the various specific growth rates tested, the highest GLA concentration of 697 mg/l was obtained in the culture with a specific growth rate of 0.08 /h. Interestingly, the fatty acid profile of the yeast recombinant bearing the Mucor ${\Delta}^6$-desaturase gene was similar to that of blackcurrant oil, with both containing similar proportions of n-3 and n-6 essential fatty acids.

• Journal of Microbiology and Biotechnology
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• v.1 no.2
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• pp.142-149
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• 1991

Gardenia callus was induced in MS medium containing $10{\;}{\mu}M$ of 2,4 diphenoxy acetic acid (2,4-D), $1{\;}{\mu}M$ kinetin, and 3% sucrose in the dark. $B_5$ medium was identified to be the most adequate medium for cell growth. Indole-3-acetic acid (IAA) was better growth regulator than 2,4-D not only for cell growth but slso for carotenoid production. Ligt also played a critical role on synthesis of carotenoid. Gardenia cells grown in $B_5$ medium could utilize a polysaccharide, soluble starch, as a carbon source. The cell growth was stimulated in $B_5$ medium fortified with 0.2% yeast extract. The optimum pH for cell growth was 5.7. High density cultures can be maintained by increasing inoculum size and medium concentration accordingly. Specific growth rate and mass doubling time were 0.095 $day^{-1}$ and 7.3 days, respectively. The cell immobilized in alginate tends to formulate more enlarged vacuoles containing yellow pigment compared with those of suspended cell. Carotenoid content of immobilized cell was about $264.4{\;}{\mu}g/g$ fresh weight (F.W.) corresponding twice of the content of suspended cell ($112.08{\;}{\mu}g/g$ F.W.). The color of gardenia cell was shifted from yellow to red when carbohydrase-secreting fungus, Trichoderma reesei, was co-cultivated with gardenia cells.

• Bulletin of the Korean Chemical Society
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• v.30 no.12
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• pp.2993-2998
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• 2009

An enzyme-linked immunosorbent assay (ELISA)-on-a-chip (EOC) biosensor combined with cell concentration technology based on immuno-magnetic separation (IMS) was investigated for use as a potential tool for early screening of Listeria monocytogenes (L. monocytogenes) in food products. The target analyte is a well-known pathogenic foodborne microorganism and outbreaks of the food poisoning typically occur due to contamination of normal food products. Thus, the aim of this study was to develop a rapid and reliable sensor that could be utilized on a daily basis to test food products for the presence of this pathogenic microorganism. The sensor was optimized to provide a high detection capability (e.g., 5.9 ${\times}\;10^3$ cells/mL) and, to eventually minimize cultivation time. The cell density was condensed using IMS prior to analysis. Since the concentration rate of IMS was greater than 100-fold, this combination resulted in a detection limit of 54 cells/mL. The EOC-IMS coupled analytical system was then applied to a real sample test of fish intestines. The system was able to detect L. monocytogenes at a concentration of 2.4 CFU/g after pre-enrichment for 6 h from the onset of cell cultivation. This may allow us to monitor the target analyte at a concentration less than 1 CFU/g within a 9 h-cultivation provided a doubling time of 40 min is typically maintained. Based on this estimation, the EOC-IMS system can screen and detect the presence of this microorganism in food products almost within working hours.