• Journal of Microbiology and Biotechnology
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• 제20권11호
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• pp.1534-1538
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• 2010

Fed-batch cultures of Hansenula polymorpha were studied to develop an efficient biosystem to produce recombinant human serum albumin (HSA). To comply with this purpose, we used a high-purity oxygen-supplying strategy to increase the viable cell density in a bioreactor and enhance the production of target protein. A mutant strain, H. polymorpha GOT7, was utilized in this study as a host strain in both 5-l and 30-l scale fermentors. To supply high-purity oxygen into a bioreactor, nearly 100% high-purity oxygen from a commercial bomb or higher than 93% oxygen available in situ from a pressure swing adsorption (PSA) oxygen generator was employed. Under the optimal fermentation of H. polymorpha with highpurity oxygen, the final cell densities and produced HSA concentrations were 24.6 g/l and 5.1 g/l in the 5-l fermentor, and 24.8 g/l and 4.5 g/l in the 30-l fermentor, respectively. These were about 2-10 times higher than those obtained in air-based fed-batch fermentations. The discrepancies between the 5-l and 30-l fermentors with air supply were presumably due to the higher contribution of surface aeration over submerged aeration in the 5-l fermentor. This study, therefore, proved the positive effect of high-purity oxygen in enhancing viable cell density as well as target recombinant protein production in microbial fermentations.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.845-848
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• 2001

For understanding physiology and metabolism under various culture conditions, combined analysis of transcriptome and proteome is attractable way. We have manufactured DNA microarray containing 2,850 genes including all functionally known and putative ones. In this study, we report analysis of transcriptome and proteome during the high cell density culture of E. coli by using DNA microarray and 2-DE. Fed-batch fermentation of E. coli was carried out by exponential feeding of nutrients until the maximum cell density reached 74 g dry cell weight/L (g DCW/L). Changes in transcriptome and proteome during the HCDC are analyzed qualitatively and quantitatively to provide their physiological and metabolic meanings.

• KSBB Journal
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• 제25권5호
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• pp.471-477
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• 2010

In order to improve bio-ethanol productivity by various cultivation methods in this paper, the culture modes using food wastes, such as batch culture, high-cell-density fermentation, SSF (simultaneous saccharification and fermentation) by fill & draw, continuous culture by fill & draw were performed and their productivities were compared. SSFs by fill & draw were performed by continuous decompression using 1 L evaporator system, and by 10 L bioreactor without decompression. In addition, the continuous cultures by fill & draw mode using SFW (saccharafied food wastes) medium were performed by changes of 40% culture broth with intervals of 12 h (0.03 $h^{-1}$), 6 h (0.07 $h^{-1}$), 3 h (0.13 $h^{-1}$). Consequently, productivities of bio-ethanol were 2.52 g/L-h and 1.30 g/L-h in batch culture and high- cell-density fermentation, respectively. The productivities of SSF by fill & draw showed 2.24 g/L-h and 2.03 g/L-h in continuous decompression with 1 L evaporator and 10 L bioreactor without decompression, respectively. Also, the productivities in continuous culture by fill & draw modes showed 2.02 g/L-h, 4.07 g/L-h and 6.25 g/L-h by medium change with intervals of 12 h, 6 h, and 3 h, respectively. In conclusion, the highest ethanol productivity was obtained in the continuous culture mode by fill & draw with dilution rate of 0.13 $h^{-1}$.

• KSBB Journal
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• 제6권3호
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• pp.231-240
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• 1991

Recently, developments of large scale and high density cell culture methods have been the objects of many researches, because the demand of various pharmaceutical products produced by animal cell culture has been rapidly increasing. The cell culture equipment should have the requirements such as sufficient oxygen transfer and mixing, low shear stress and surface tension, and small foaming. In order to develop a proper bioreactor meeting these requirements simultaneously, a perfluorocarbon having high solubility of oxygen was sprayed into the medium as an oxygen carrier instead of air. Also, a new impeller was developed and combined together with the perfluorocarbon spraying system so as to design a new bioreartor for cell cultivation. The new impeller had better characteristics of mixing and oxygen transfer than the paddle and cell-lift impellers based on the same, shear rate. But, it was observed that the volumetric oxygen transfer coefficient of the new bioreactor decreased with increasing cell density during E. coli fermentation.

• KSBB Journal
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• 제28권6호
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• pp.400-407
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• 2013

The growth characteristics and production of color pigments by Monascus strains were investigated during liquid culture, and production of monacolin K in red mold rice was carried out by solid state fermentation. Four different Monascus ruber strains were cultured in potato dextrose yeast extract broth (PDYB) media at $25^{\circ}C$ for 15 days. The high producing strain for red pigment was not corresponded to the strain for yellow pigment. Production of red pigment was high in the strain causing the fast pH change in culture broth. Production of monacolin K in red mold rice by solid state fermentation was influenced by a combination of wet cell weight and spore density in inoculum by liquid culture. Most strains showed the high production of monacolin K in red mold rice, when submerged fermentation was carried out for 5 days as inoculum for solid state fermentation. These results suggest that submerged fermentation period of inoculum have an effect on the production of monacolin K in red mold rice by solid state fermentation, and monacolin K in red mold rice could be increased by controlling the condition of submerged fermentation for inoculum.

• Journal of Microbiology and Biotechnology
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• 제4권4호
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• pp.316-321
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• 1994

We investigated the possibility of industrial application and economit process of high temperature fermentation by thermotolerant alcohol producing yeasts as previously reported. From the 20% glucose media, the RA-74-2 produced 11.8% (v/v) ethanol at $32^{\circ}C$ (0.5% inoculum) and 10.6% (v/v) ethanol at $40^{\circ}C$ (3% inoculum), respectively. Also, 11.3% (v/v) ethanol was produced for 96 hours in the temperature-gradient fermentation. These results suggest that the RA-74-2 could isuccessfully be applied to save the cooling water and energy in industrial scale without re-investment or modification of established fermentation systems. When potato starch was used as the substrate for the RA-74-2, high temperature fermentation above $40^{\circ}C$ was more appropriate for industrial utilization because organic nitrogen was not necessary to economical fermentation. As the naked barley media just prior to industrial inoculation, taken from the Poongkuk alcohol industry Co., were used, 9.6% (v/v) ethanol was produced at $40^{\circ}C$ for 48 hours in jar-fermentor scale (actually, 9.5-9.8% (v/v) ethanol was produced at 30~$32^{\circ}C$ for 100 hours in industrial scale). The ethanol productivity was increased by the high glucoamylase activity as well as the high metabolic ratio at $40^{\circ}C$ Therefore, if the thermotolerant yeast RA-74-2 would be used in industrial scale, we could obtain a high productivity and saving of the cooling water and energy. Meanwhile, the RA-912 produced 6%(v/v) ethanol in 10% glucose media at $45^{\circ}C$ and showed the less ethanol-tolerance compared with industrial strains. As the produced alcohol was recovered by the vacuum evaporator at $45^{\circ}C$ in 15% glucose media, the final fermentation ratio was enhanced (76% of theoretical yields). This suggest that a hyperproductive process could be achieved by a continuous input of the substrate and continuous recovery of the product under vacuum in high cell-density culture.

• Journal of Microbiology and Biotechnology
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• 제20권11호
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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.

• 상하수도학회지
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• 제20권2호
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• pp.225-234
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• 2006

Characteristics of electricity production from major fermentation products (acetate, propionate and butyrate) were evaluated in a microbial fuel cell (MFC). For each substrate, batch and continuous experiments were performed. The batch test result indicated that coulombic efficiency depended on the resistance connected in MFC circuit. With acetate, coulombic efficiency were 87% at $20{\Omega}$, but decreaced to 45% at$220{\Omega}$. In continuous tests, maximum power densities obtained was 220 Q with acetate. The maximum power densities of butyrate, acetate and propionate were 6.8, 6.1, and $5.2mW/m^2$, respectively. Propionate and butyrate were converted into acetate producing high currents. $H_2$ produced during butyrate and propionate probably used to produce electricity. In conclusion, butyrate conversion into acetate was faster than that of propionate with higher electricity production. If the production of propionate is inhibited during fermentation, anaerobically fermented liguor may be effectively applied for MFC.

• KSBB Journal
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• 제5권3호
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• pp.307-313
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• 1990

배양기와 16비트 마이크로컴퓨터를 접속하여 호기성 미생물의 고농도 유가배양시스템을 구성하였다. 기질공급을 위한 제어변수로는 용존산소(DO)를 이용하였다. 용존산소측정기의 출력신호를 컴퓨터에 입력시켜 측정한 DO값에 근거하여 교반모터의 교반속도와 산소유량을 제어함으로써 배양액의 DO를 일정하게 유지하였으며, 연동펌프의 제어에 의해 기질공급을 안정하게 수행하였다. 기질공급의 제어와 DO의 제어를 한가지의 하아드웨어 및 소프트 웨어로 수행할 수 있었다. 제어시스템을 이용하여 메탄올이용균인 Methylobacillus sp. SKI을 유가배양한 결과 배양액의 DO제어와 메탄올 공급의 제어가 무리없이 수행되었으며, 배양 10시간만에 16.53g/l의 균체농도에 도달하였다.

• Food Science and Biotechnology
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• 제17권5호
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• pp.990-995
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• 2008

A thermostable $\alpha$-L-arabinofuranosidases ($\alpha$-L-AFase) is an industrially important enzyme for recovery of L-arabinose from hemicellulose. The recombinant $\alpha$-L-AFase from Thermotoga maritima was expressed in Escherichia coli by using a constitutive pHCE or an inducible pRSET vectors. In batch fermentation, the constitutive expression system resulted in slightly faster growth rate (0.78 vs. 0.74/hr) but lower enzyme activity (2,553 vs. 3,723 units/L) than those of the induction system. When fed-batch fermentation was performed, biomass and enzyme activity reached the highest levels of 36 g/L and 9,152 units/L, respectively. The fed batch cultures performed superior results than batch culture in terms of biomass yield (4.62-5.42 folds) and enzyme synthesis (3.39-4.00 folds). In addition, the fed-batch induction strategy at high cell density resulted in the best productivity in cell growth as well as enzyme activity rather than the induction method at low cell density or the constitutive expression.