• KSBB Journal
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• 제17권1호
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• pp.93-99
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• 2002

C. tropicalis ATCC 20336을 이용하여 높은 수율과 생산성으로 xylitol을 생산하기 위하여 glucose 배지에서 짧은 시간에 고농도로 세포를 배양한 후에 xylose를 첨가하여 xylitol로 전환하는 2단계 유가식 발효공정에 관한 연구를 수행하였다. Cell growth-stage에서의 배지공급 방법으로는 glucose가 모두 고갈된 후에 pH가 5.7 이상으로 올라가는 것을 신호로 glucose를 첨가하는 pH-stat 방법으로 18.5시간 배양에 의하여 67.9 g/L의 세포 농도를 얻었으며, ethanol 생성은 1.0 g/L로 매우 낮았다. Production-stage에서 최적의 초기 xylose 농도는 150 g/L이었으며, 이때에 2 g/L ($NH_4)_2SO_4$, 1 g/L $K_2HPO_4$, 0.2 g/L $MgSO_47H_2$O로 구성된 mineral salt를 2회 첨가에 의하여 xylitol의 생성이 향상되었다 그러나 0.2 vvm에서 1.0 vvm 사이의 통기조건에서는 xylitol의 생산에 큰 영향을 미치지 못하였다. 반면에 production-stage에서 yeast extract를 10 g/L가 되도록 첨가한 경우에 xylitol 수율과 생산성이 큰 폭으로 증가하였다. 즉 xylose 농도가 약 150 g/L이 되도록 232.5 g의 xylose를 2회 첨가한 경우에 배양액의 최종 부피는 1.67 L이 되었고, 이때의 xylitol 농도는 193 g/L이었으며, 수율은 70%이고 생산성은 3.55 g/L.h이었다.

• ALGAE
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• 제21권1호
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• pp.133-139
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• 2006

Outdoor cultivation of cyanobacterium Spirulina platensis was carried out for 40 days in a batch mode. A half concentration of the SOT based on the underground water was used as culture medium. Working volume was 5.7 tons with 0.2 m depth. During cultivation, mean water temperature, DO and light intensity were all in proper conditions for the S. platensis growth. The adjustment of pH to over 10 with Na2CO3 and addition of the 1.5% natural salt were very effective to delete contaminant organisms, Chlamydomonas moewusii and Chlorella minutissima occurred one after the other in the culture. The mean productivity of the biomass based on the dry cell weight from 14 to 25 days, after the contaminants were deleted, was 7.8 g ·m–2· d–1, which was relatively high productivity in that a half concentration of the SOT was used for the culture. Underground water used in the culture minimized contaminants invasion and addition of the 1.5% natural salt was effective to delete contaminants as well as acted as mineral supplement in outdoor cultivation of S. platensis. Harvesting using the floating activity of S. platensis was effective from mass floating in day time after overnight without agitation and illumination.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.758-761
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• 2001

In this study, the nar promoter of E. coli was characterized to see whether the nar promoter cloned onto pBBR122 can be used as an expression promoter of gram negative microbes. For this purpose, a plasmid with lacZ gene expressing ${\beta}-galactosidase$ instead of the structural genes of nar operon in a gram negative host strain(Agrobacterium.tumefaciens) was used to simplify an assay of induction of the nar promoter. The following effects were investigated to find optimal conditions: methods of inducing the nar promoter, optimal nitrate concentration, maximally inducing the nar promoter, the amount of expressed ${\beta}-galactosidase$ and induction ratio(specific ${\beta}-galactosidase$ activity after maximal induction/specific ${\beta}-galactosidase$ activity before induction). The following results were obtained from the experiments: the growth of Agrobacterium with E.coli nar promoter was not much affected by nitrate concentration in the shake-flask; induction of nar promoter was optimal when Agrobacterium was grown in the presence of 1% nitrate ion at the beginning of culture and when overnight culture was completely grown in the shake-flask before being transferred to other shake-flask; the amount of ${\beta}-galactosidase$ per cell and per medium volume was maximal when Agrobacterium was grown under aerobic condition to $OD_{600}$ of 1.7; then the nar promoter was induced under microaerobic and anaerobic condition made by lowering dissolved oxygen level(DO). After 2-3h of induction in the YEP medium selected as a main culture medium, the specific ${\beta}-galactosidase$ activity became about 17,000 Miller units in the fermentor cluture.

• KSBB Journal
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• 제15권6호
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• pp.573-577
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• 2000

부패된 사과로부터 새롭게 분리된 Acetobacte sp. A9에 의 한 Be 생산조건을 정치배양에 의하여 검토하였다. 본 균주는 2 $25-30^{\circ}C$에서 BC를 생산할 수 있었으며, $30^{\circ}C$에서 최대 생산 능을 나타내었다. 또한 본 균주는 배지의 초기 pH 6.5-8.0에 서 BC를 생산할 수 있었으며, pH 6.5에서 최대 생산능을 나타내었다. BC 생산올 위한 최적 배지조건은 glucose 1.0%, y yeast extract 1.0%, polypeptone 0.7%, acetic acid 0.15% 및 succinic acid 0.02 %였다. Acetobacter sp. A9에 의한 BC 생산 은 "growth-associated type"이었다. 최적 배양조건하에서 배양 6일만에 최대 $24.1 g/m^2$ 의 BC가 생산되어 기본배지보다 약 2배의 생산성 향상이 이루어졌다.. BC 생산능은 배지의 표면 적보다는 배지량과 배지의 깊이에 크게 영향올 받았다.

• KSBB Journal
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• 제30권6호
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• pp.307-312
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• 2015

Transgenic rice cells using RAmy3D promoter can provide high productivity, and the production of recombinant protein is induced by sugar starvation. In this system, productivity was reduced during the scale-up processes. To ensure the influences of shear stress and oxygen transfer rate, working volume and mixing performances were investigated under various agitation speeds and working volumes. In addition, inoculation methods including suspended cells and filtered cells were compared. Working volumes and shaking speeds were 300, 450 mL and 80, 120 rpm, respectively. Hydrodynamic environment of each condition was measured numerically like mixing time and $k_La$. Good mixing performance and high shear stress were measured at high agitation speed and low volume. The highest level of hCTLA4Ig was 30.7 mg/L at 120 rpm, 300 mL. When conditioned medium was used for inoculation, increased cell growth was noticed during the day 0~4 and decreased slower than filtered cells. Compared with filtered cells, the maximum hCTLA4Ig level reached 37.8 mg/L at 120 rpm, 300 mL and lower protease activity level was observed. In conclusion mixing performance is critical factor for productivity and conditioned medium can have a positive effect on damaged cells caused by hydrodynamic shear stress.

• 한국가축번식학회지
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• 제8권1호
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• pp.29-35
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• 1984

These experiments were carried out to obtain basic information necessary for in vitro culture of aggregated mouse embryos. Inbred ICR mice were used to obtain embryos. The zona pellucida was removed by placing the embryos in Whittingham's medium containing 0.5% protease for about 5-10minutes at 37$^{\circ}C$. Total 263 pairs of 2-, 4- and 8-cell zona free mouse embryos were subjected to aggregation by physical pressure and cultured in Whittingham's medium under the gas phase of 5% CO2 in air at 37$^{\circ}C$ for 24 to 60 hours. The results obtained in these experiments were summarized as follows: 1. Time needed for fusion of 2-, 4- and 8-cell embryos were 0-3, 0-3 and 0-3 hours, respectively and average time needed for in vitro development of 2-, 4- and 8-cell embryos after aggregation to morula and blastocyst were 42, 30 and 13.5 hours, and 51, 39 and 27 hours, respectively. 2. Of total 263 pairs of naked embryos, 227 were firmly aggregated together and the rats of aggregation in 2-, 4- and 8-cell embryos were 71.8, 88.3 and 97.0%, respectively. 3. The rates of aggregated pairs which obtained from 2-, 4- and 8-cell embryos developed to morula were 96.7, 95.6 and 96.9%, respectively, and embryos developed to blastocysts were 88.5, 89.7 and 90.8%, respectively. 4. Conspicuous differences in size of volume and inner cell masses between single and double blastocysts were observed. Although a single blastocolic cavity was formed in most double blastocysts, several formed two distinct cavities from the very beginning.

• Journal of Microbiology and Biotechnology
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• 제15권1호
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• pp.1-6
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• 2005

Optimal criterion for the scale-up production of schizophyllan, a fungal polysaccharide secreted by Schizophyllum commune, was investigated. For the production of the polysaccharide in a 150-l bioreactor, the culture conditions optimized in a 15-l bioreactor were applied to a 150-l bioreactor with scale-up process, by changing impeller speed and airflow rate. The optimized impeller speed in the 15-l bioreactor was 50 rpm in a technical medium based on barley. For establishment of the scale-up process, 3 kinds of criteria were used while the gas throughput number was kept constant, as follows; constant volume-related power input, constant tip speed of stirrer, and constant Reynolds number. In the 150-l bioreactor, the highest values for the maximum specific growth rate (1.17/day) and productivity (0.63 g/L${\cdot}$day) were achieved in the culture condition from constant volumerelated power input criterion.

• 한국미생물·생명공학회지
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• 제25권6호
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• pp.580-585
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• 1997

We selected two mutants namely strain D5 and Nu23 by mutagenesis of anthracycline producing Streptomyces: the former is an $\varepsilon$-rhodomycinone overproducing mutant selected from Streptomyces sp. C5, a baumycin producer and the latter, a blocked mutant of early pathway for doxorubicin biosynthesis obtained from Streptomyces peucetius ATCC 27952, a doxorubicin producer. The mutant strain Nu23 does not produce anthracycline metabolites but retains the most of enzyme activities converting aklavinone to doxorubicin and the mutant strain D5 produced $\varepsilon$-rhodomycinone at a level of 150 $\mu$g/ml. These strains were grown separately in NDYE medium and each was mixed at day 3 by equal volume of culture broth but the quantity of doxorubicin produced was far below an estimation based on the level of $\varepsilon$-rhodomycinone normaly produced by the strain D5. On the other hand doxorubicin was reached at maximum level after 4 days in the mixed culture condition which was composed of culture broth of strain D5 grown for 6 day and that of strain Nu23 grown for 3 day. It was turned out that the growth of mutant strain D5 was inhibited by the accumulation of daunorubicin and doxorubicin in mixed culture broth, which cause the limitation of $\varepsilon$-rhodomycinone.

• KSBB Journal
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• 제13권5호
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• pp.491-496
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• 1998

The production of fibrinolytic enzyme from Bacillus subtilis BK-17 was studied in the shake flask cultures. The important medium components studied include nitrogen source, carbon source and inorganic salts. The environmental conditions include initial pH, temperature, shaking speed and working volume. Among various N-sources, C-sources and inorganic salts tested, soybean flour, D-glucose and Na2HPO4 gave the best results, and their optimal concentrations were 1.5%, 0.5% and 0.05%, respectively. The optimal pH and temperature were 9.0 and 37$^{\circ}C$. With decreasing working volume in the range of 25∼100ml in the 250ml flask or increasing shaking speed in the range of 100∼300rpm, the enzyme production was greatly enhanced. The enzyme activity under the optimal conditions was about 1400I.U./ml with urokinase as a standard.

• Mycobiology
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• 제29권3호
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• pp.160-163
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• 2001

Sikhae is a Korean traditional beverage of saccharified rice. Its factory waste(SFW) is usually thrown away instead of being used. We developed a cheap substrate of SFW for use in liquid spawn that is known for its higher fruit body yields than grain spawn in sawdust cultivation. Mycelia of Lentinula edodes ASI 3046, which is regarded as the most suitable strain for sawdust cultivation, were cultured on six kinds of previous known media and SFW. As the seven kinds of media were applied, a Sikhae Factory Waste(SFW) was most excellent in growth. The dried mycelial weight in SFW was almost four times as much as that in the other media. In the flask culture, optimum culture conditions for the mycelial growth were obtained after 13 days of cultivation at media volume of 100 ml, 100 rpm, initial pH 4.5, and $25^{\circ}C$. The best mycelial growth was observed when $MgSO_4{\cdot}7H_2O$ and D-sucrose were added as a supplement in SFW. SWM must be a remarkable medium for L. edodes because of its simple preparation and low cost.