• Microbiology and Biotechnology Letters
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• v.28 no.2
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• pp.105-110
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• 2000

Various environmental factors such as pH, temperature and initial glucose concentration were investigated for enhancing cell growth in fermentations of Phellinus linteus WI-OOl, a producer of polysaccarides with potent anticancer activities. Optimal pH and temperature were around 5.5 and $28^{\circ}C$, respectively. Relatively little variation of pH was observed ranging between 5.5 and 6.5 during the whole fermentation period. Maximum cell concentration and specific growth rate were investigated in the media containing initial glucose concentrations of 0.5%, 1 %, 2%, 3% and 4%. High initial glucose concentration enhanced biomass production but showed negative effect on specific growth rate. In bioreactor experiments with various feeding strategies, increases of 28% and 42% in final cell concentration were obtaind as compared to conventional batch process, by adopting pulse and continuous supplement of 2% glucose solution, respectively.

• KSBB Journal
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• v.15 no.5
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• pp.489-496
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• 2000

Several culture systems including batch, two-stage CSTR, semi-fed batch, and two-stage cyclic fed-batch were investigated for the efficient production of the Fab fraction of PDC-E2 specific human monoclonal antibody using high cell density recombinant E. coli. A two-phase batch system and a two-stage continuous system were examined to overcome plasmid instability problems, by separating the growth and the production stages. The cell density and productivity of the two-stage continuous culture was better than that of the two-phase batch fermentation. In the two-stage continuous culture system with DO-stat, the cell growth and the productivity were superior to those of the system without the DO control. Also, almost total plasmid stability was maintained in the two-stage continuous culture system. Modified M9 medium was selected as an optimum feeding medium for the fed-batch process, and the optimum C/N ratio determined to be 2:3. The optimum feeding rate was $0.6g/\ell/hr$ for a constant feeding strategy in semi-fed batch system. When the feeding medium was fed by pulsing, it was observed that more frequent pulsing resulted in improved cell growth. The linear feeding method was the most efficient of the various feeding methods tested. Finally, high cell density culture using a two-stage cyclic fed batch system with pH-stat was tried because the linear feeding method showed limitations in terms of obtaining high cell densities, and a cell density of $54 g/\ell$ was achieved. It was concluded that the two-stage cyclic fed batch system was the most efficient system for high cell density culture of the systems tested. However, productivity improvements were lower than expected due to the extremely high accumulations of acetate, although the low levels of residual glucose were maintained.

• Journal of Microbiology and Biotechnology
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• v.7 no.5
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• pp.323-328
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• 1997

The recombinant Escherichia coli BL21(DE3)pLysE : pTCGT1 was grown to overproduce Bacillus macerans cyclodextrin glucanotransferase (CGTase) able to synthesize ${\alpha}$-cyclodextrin (CD) with a selectivity of 67%. A number of batch fermentations were performed to test the possibility of using lactose as an inducer of the E. coli T7 promoter system. A mixture of isopropyl ${\beta}$-D-thiogalactoside (IPTG) and lactose (1 : 1) gave a maximum CGTase activity of 2.4 U/ml, which was higher than the value obtained with induction by IPTG alone. Fed-batch fermentations involving a glucose-controlled growth period followed by a gene-expression phase with mixtures of IPTG and lactose were employed to achieve high cell density and thereby increase total CGTase activity. Optimized fed-batch fermentation using the modified inducer (IPTG : lactose=1 : 3) and 100 g/l yeast extract solution in the gene-expression phase resulted in a maximum CGTase activity of 62.9 U/ml and a final cell mass of 53.5 g/l, corresponding to a 31-fold increase in CGTase activity and a 29-fold increase in cell mass compared with the control batch fermentation.

• Korean Journal of Microbiology
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• v.50 no.1
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• pp.84-86
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• 2014

Mass production of biocontrol agent is an essential step for its commercial use. Media composition and culture conditions for production of Bacillus amyloliquefaciens SKU-78, a potential biocontrol agent against bacterial wilts, were optimized by a flask culture. Low cost media combining nitrogen and carbon sources were tested. Maximum cell growth (> $2{\times}10^9$ CFU/ml) was obtained in a medium of 5% soy flour combined with 3% corn starch after 24 h cultivation. The optimum initial pH, temperature and shaking speed was 5.5, $30^{\circ}C$ and 150-250 rpm, respectively. Fermentation of SKU-78 was scaled up in 30 L fermenter and the profiles of cell density, pH, dissolved oxygen and spore formation were recorded. After 8 h lag phase, exponential growth occurred and reached at maximum viable cell number ($1.2{\times}10^{11}$ CFU/ml) after 20 h. The SKU-78 strain grown in a low cost medium exhibited the high suppression of bacterial wilts. The results indicate that SKU-78 strain can be produced in a low cost medium and provide a basis for scaling up to industrial level.

• Biotechnology and Bioprocess Engineering:BBE
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• v.5 no.4
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• pp.288-305
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• 2000

A review on the main aspects associated with yeast flocculation and its application in biotechnological processes is presented. This subject is addressed following three main aspects-the basics of yeast flocculation, the development of "new" flocculating yeast strains and bioreactor development. In what concerns the basics of yeast flocculation, the state of the art on the most relevant aspects of mechanism, physiology and genetics of yeast flocculation is reported. The construction of flocculating yeast strains includes not only the recombinant constitutive flocculent brewer's yeast, but also recombinant flocculent yeast for lactose metabolisation and ethanol production. Furthermore, recent work on the heterologous $\beta$-galactosidase production using a recombinant flocculent Saccharomyces cerevisiae is considered. As bioreactors using flocculating yeast cells have particular properties, mainly associated with a high solid phase hold-up, a section dedicated to its operation is presented. Aspects such as bioreactor productivity and culture stability as well as bioreactor hydrodynamics and mass transfer properties of flocculating cell cultures are considered. Finally, the paper concludes describing some of the applications of high cell density flocculating bioreactors and discussing potential new uses of these systems.e systems.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.651-654
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• 2000

A fermentation study of the recombinant Escherichia coli system has been tired to overproduce a recombinant Paragonimus westermani cysteine proteinase, rPwCP1. Using the modified LB media, main cultures and chemical induction with IPTG were carried out to examine the possibility of secretory protein overproduction at high cell density culture. As a result, the target protein of rPwCP1, purified by metal affinity chromatography and gel filtration, has been shown at 50.8 kDa on SDS-PAGE, and its final concentration turns out to be 350mg/L.

• KSBB Journal
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• v.8 no.5
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• pp.431-437
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• 1993

Active carbon which has the smallest bulk and wet density was found as the best support media among 4 different kinds of materials(celite, natural zeolite, Pusuk stone, active carbon) to make a proper fluidized-bed with small energy consumption. Its minimum and optimum fluidization velocity were found as 0.03cm/sec and 0.25cm/sec, respectively. As organic loading rate for methane fermentation was increased, CODcr removal efficiencies of all the media were decreased. But, CODcr, removal efficiencies of active carbon was maintained more than 90% in this experimental range of the organic loading rate. Larger amount of microorganism was adsorbed on the active carbon which has very high specific surface area. At the organic loading rate of 16g CODcr,/l day, its adsorbed cell mass was 157mg/g. Comparing natural zeolite with roast celite, adsorbed cell mass did not increase in proportion to specific surface area of the media. Even though roast celite has the same specific surface area as the Pusuk stone, its organic removal ability was superior to that of the Pusuk stone, which explains that the relatively great surface roughness and the positive surface charge are important for cell adsorption. It was concluded that the support media for anaerobic fluidized reactor should have small wet density and small fuidization velocity, if possible, in order to increase cell adsorption by reducing the fluid shear stress.

• Food Science and Preservation
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• v.20 no.6
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• pp.886-893
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• 2013

Several indigenous sulfite-resistant yeasts were isolated at the microbial succession stage of yeast flora during spontaneous fermentation of Campbell Early grapes using a YPD plate that contained 200 mg/L or 500 mg/L potassium metabisulfite. When they were applied to the wine fermentation using the Campbell Early grape and apple juices, strains S13 and D8 showed strong alcohol fermentation and good flavor production. They were identified as Saccharomyces cerevisiae in the phylogenetic analysis based on their ITS 1-5.8S-ITS II DNA sequences. The two yeast strains grew to a high cell density in the YPD media supplemented with 40%(w/v) glucose. They also grew rapidly in the YPD media at $40^{\circ}C$. While strain S13 showed some differences in cell density at the two temperatures, no marked difference was observed during the culture of strain D8. The strains grew relatively well at pH 5.0 and 9.0 compared with pH 7.0, which was the optimum pH for their growth. Especially, strain S13 cultivated in the YPD media at pH 9.0 grew to 93% of the growth of strain D8, which was obtained at pH 7.0.

• Journal of Microbiology and Biotechnology
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• v.10 no.4
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• pp.551-553
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• 2000

A strong constitutive $P_{JH}$ promoter from Bacillus was applied to overexpress the endoxylanase gene in B. subtilis. The expression plasmid, pHJKJ4, was designed to contain the $P_{JH}$ promoter and endoxylanase promoter ($P_B$), and introduced into B. subtilis DB104. Through batch fermentation of the trasformant cell on a maltose medium, endoxylanase was produced in a growth-associated manner as the predominant protein. The total activity reached about 600 unit/ml at the end of the cultivation, which corresponded to 698 mg endoxylanase protein/l with a specific activity of 860 unit/mg protein. It was also found that the segregational plasmid instability was less than 30% and most of the endoxylanase activity was detected in the culture medium. This result suggests that the secretory production of endoxylanase can be significantly enhanced with the use of the $P_{JH}$ promoter and high-cell density culture techniques, quantitatively as well as qualitatively.

• Microbiology and Biotechnology Letters
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• v.26 no.3
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• pp.257-263
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• 1998

Alcaligenes eutrophus was successfully recovered from high cell density broth by pre-treatment with Fe-based coagulants. An inorganic coagulant, Fe$_2$(SO$_4$)$_3$, and a polymerized coagulant, Ferix-3, were used. Good coagulation was observed in broad pH range of 3 to 13, the floe size was increased with increasing pH of culture broth. The optimum pH of fermentation broth for cell recovery was 10 to 13. The optimum coagulant dosages to recover cells with 95% cell recovery were increased with increasing cell concentration. Optimal coagulant dosage was lower when the polymerized coagulant was used rather than the inorganic coagulant. The coexistence of NH$_4$$\^$+/ was increased coagulant requirement, and the coagulant requirement was 0.066g Fe$_3$$\^$+//g NH$_4$$\^$+/.