• Journal of Microbiology and Biotechnology
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• 제10권3호
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• pp.321-326
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• 2000

To examine the feasibility of the long-term production of the human granulocyte colony stimulating factor (hG-CSF) using the $_{L}-arabinose$ promoter system of Escherichia coli, flask relay culture and cyclic fed-batch culture were performed. In the flask relay culture, it was found that the pismid was maintained stably up to about 170 generations in an uninduced condition, whereby the cells could also maintain the capability of expressing hG-CSF expression were maintained stably up to at least 100 generations. In contrast, in the cyclid fed-batch culture, segregational plasmid instability was observed within about 4 generations after induction, even though the cell growth and hG-CSF production reached their maximum balues, 78.0 g/l of dry cell weight and 7.0 g/l of hG-CSF, respectively. It would appear that, when compared to the flask relay culture, the high-cell density and high-level expression of hG-CSF in the cyclic fed-batch cultrure led to the segregational plasmid instability; in other words, a severe metabolic burden existe on the cells due to the high-level expression of hG-CSF. Accordingly, based on these long-term cultures, the segregational and structural plasmid instability was observed and a strategy to overcome such problems could be designed.

• Journal of Genetic Medicine
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• 제2권2호
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• pp.79-81
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• 1998

Genetic amniocenteses were performed in a series of 127 patients as a routine study. Samples from the patients were cultured by in situ method, flask method or both according to the state of amniotic fluid. The overall success rate of culture was 97.6% and no culture failure was observed in the flask method. It took 5 days first of all and 8.15 days average from set-up to harvest and there were 7.2 colonies per dish in in situ method. Therefore, it is suggested that in situ method which decreased the mean culture days and made clonal analyses possible, is a clinically available and even more reliable method in parallel with flask method in prenatal diagnosis.

• The International Journal of Costume Culture
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• 제2권1호
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• pp.54-61
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• 1999

The purpose of this research was first to analyse and compare the types of microbes inherent among the unwashed and washed Kupori textiles, and second to investigate whether there is a difference between unwashed and washed Kupori textiles on the susceptibility of contamination by microorganism when exposed to the same microbial environment. Microbial identification procedure and the Shake Flask Test for investigating the effect of exposure to microbial environment were carried out separately. The result of microbial identification procedure indicated that a variety of bacteria and fungi were inherent in both unwashed and washed textiles and that the population of contaminated microorganism became more diverse after washing. The result of Shake Flask Test indicated that given the same exposure condition, the unwashed textiles tend to be more susceptible to bacterial contamination than the washed textiles. The results of the present study supported the current conservation procedures adopted in Korean museums which include washing and humigation procedures before long-term storage or display of exhumed textiles.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XII)
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• pp.398-401
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• 2003

Bacillus species predominantly outgrown in a night soil treatment system were isolated and characterized. Cell interactions took place among them and cell population changed under various culture conditions. Maximum removal of $NH_4\;^+-N$ and cell production occurred under the conditions of 30% DO and C/N ratio of 8. Additions of 0.8% peptone and 0.3% yeast extract to a basal medium influenced the growth of isolates and the removal of $NH_4\;^+-N$ in flask culture, and metal ions such as Ca, Fe and Mg also did. During the flask experiment of nitrogen removal under an aerobic condition, active nitrification by the isolates occurred largely in 1 h with the decrease of COD and alkalinity destructed was only 74.6% of theoretical value. From the nitrogen balance, the percentage of nitrogen lost in the flask culture was estimated to be 29.0%. This conversion of ammonia to $N_2$ under an aerobic condition was confirmed by GC analysis. The B3 process using the Bacillus species seemed to have some economic advantage.

• 생명과학회지
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• 제14권1호
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• pp.126-130
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• 2004

곰보버섯(Morchella esculenta)은 맛과 영양이 풍부한 버섯으로 전 세계적으로 분포하는 버섯이다. 현재까지 우리나라에서 곰보버섯에 관한 체계적인 연구가 이루어지지 않은 상태에서 본 연구는 곰보버섯(자실체) 인공대량 생산방법 개발 및 심부배양을 통한 균사체 및 다당체 대량생산 공정개발을 위한 기초자료를 얻고자 shaking flask culture를 이용한 곰보버섯 균사체 배양특성에 관하여 조사하였다. 곰보버섯 균사체의 최적 배양온도는 $25^{\circ}C$이며, 최적 초기 pH는 6.5, 최적배지는 BG 이었다. BG 배지를 기본배지로 하여 영양 요구성 실험을 한 결과 최적탄소원은 fructose이고 최적농도는 5.0% (w/v)이며 질소원 선발 및 최적농도에서는 peptone과 NH$_4$Cl를 혼합하여 사용하였을 때 최대 균사체생육을 보였고, 상기의 두 질소원을 1:1의 무게비로 혼합하여 4.0% (w/v) 농도로 첨가하였을 때 최대 균사생육을 보였다.

• 산업기술연구
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• 제21권A호
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• pp.337-341
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• 2001

This study was carried out to screen the effective polymeric additives preventing wall growth during mycelial submerged cultivation of Ganoderma. lucidum. Effects of additives on mycelial growth and exo-polysaccharide (EPS) production in flask culture and jar fermenter system under 3 different pH processes were investigated, and changes of mycelial morphology were also examined. From flask culture of G. lucidum with additives of different concentrations, 0.1%(w/v) polyacrylic acid was effective for EPS production. As the polyacrylic acid of 0.1%(w/v) was added in medium, wall growth of G. lucidum mycelium grown in jar fermenter system could be protected. The addition of 0.1%(w/v) polyacrylic acid to medium was also improved the mycelial growth and EPS production in the later of submerged culture G. lucidum and no changes of mycelial morphology were observed.

• 한국미생물·생명공학회지
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• 제11권1호
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• pp.59-66
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• 1983

유포자 유산균인 Lactobacillus sporogenes 가 특이하게 균체외 $\beta$-galactosidase를 다량 생산하는 것을 발견하여 효소생산을 위한 영양요구성과 배양조건을 조사하였다. 영양요구성으로는 탄소원으로서 lactose 1%, 유기질소원으로서 peptone 1.5%, 무기질소원으로서 ammonium sulfate 0.2%, phosphorus 원으로 ammonium phosphate, dibasic을 0.8%. mineral은 potassium chloride 0.05%, ferric chloride 0.001% 첨가했을 때 최대의 효소생산을 나타내었다. 배지의 최적 initial pH는 7.0, 최적배양온도는 37$^{\circ}C$, aeration효과는 500$m\ell$용 진탕 flask에 배지량을 50~200$m\ell$ 주입, 140 strokes/min (진폭 7cm)으로 진탕 배양하였을 때 최고의 효소생산을 나타냈다. 상기의 최적조건에서 균체증식은 24시간 배양에 최고에 달하는 반면 $\beta$-galactosidase 생산은 균체내 효소생산의 경우 균체증식 속도와 비례적으로 증가하여 배양 24시간 전후에서 30U/$m\ell$로 최고치를 나타내고 균체외 효소는 배양 40시간 전후 즉, 균증식의 후기 정지기에서 38U/$m\ell$로 가장 높은 효소생산량을 보였다. 또한 fermentor 실험 역시 flask배양과 거의 비슷한 배양 양상을 보였으며 균체외 효소역가는 45U/$m\ell$로 진탕 flask 배양 결과보다 다소 양호하였다.

• KSBB Journal
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• 제5권3호
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• pp.207-214
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• 1990

Growth properties of carrot hairy roots transformed by Agrobacterium rhizogenes were compared in flask and bioreactor. Oxygen transfer coefficient KLa was measured during the cultivation in bioreactor. In flask cultures, initially sucrose 30g/l was nearly exhausted after 20days. pH was dropped from initially 5.8 to 4.79 after 4 days, but it is stable after that time. Finally, after 28 days, hairy roots were grown about twelve times. In view of the results studied optimum conditions, hairy roots were maintained high growth rates in sucrose 50g/l, pH 5.8, total nitrogen 60mM. Also in bioreactor cultures, fixed stainless sieve in bottom and aerated 0.31 vvm, the results of cultivation by the use of sucrose 50g/l had grown about twenty-eight times and pH variations were liked in flask. As a results, growth rate of 1.756g fresh weight/day/g inoculum in bioreactor were higher about three times than 0.57g fresh weight/day/g inoculum in flask culture. KLa values were showed a tendency to decrease from 0.209 min-1 to 0.068 min-1.

• Korea-Australia Rheology Journal
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• 제14권1호
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• pp.11-16
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• 2002

Cephalosporium acremonium is a filamentous microorganism producing cephalosporin C. The morphological differentiation of C. acremonium in submerged culture is closely related with the rheological properties of culture broth and production of cephalosporin C. In this study, the rheological and morphological properties of culture broth of C. acremonium were investigated. In the seed broths of shake-flask and fermenter culture, the Herschel-Berkley equation was in excellent agreement with experimental results in the whole range of shear rate. In the seed broths of shake-flask culture, morphological differentiation into arthrospores affected to changes of apparent viscosity. But results in the fermenter culture, morphological factors such as mean hyphal thickness and the number of tips gave more effect on changes of apparent vitacosity than differentiation into arthrospores. Overall, it suggested that the morphological parameters measured by image analysis can be used as a good parameter to indicate the rheological properties of culture broth of C. acremonium M25.

• 생명과학회지
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• 제14권3호
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• pp.391-395
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• 2004

B. stearothermophilus NO2의 CGTase 유전자 (cgtS)를 구성적 $P_{JH}$ promoter 하류에 subcloning 하여 재조합 plasmid pIH-CGT1 (8.14 kb)을 구축하고 B. subtilis DB431에 형질 전환하였다. B. subtilis DB431/pJH-CGT1를 5가지 배지(LB, 2${\times}$LB, 5% molasses+2% CSL, CS, LBG)로 flask 배양하여 균체증식과 CGTase발현량 및 분비국재성을 조사하여 최적 배지를 결정하였다. 그 중 〔5% molasses+2% CSL〕 배지에서 9시간에 1.8 unit/$m\ell$의 CGTase가 발현$.$생산되었다. 이 결과를 토대로 3. subtilis DB431/pJH-CGT1를 〔10% molasses + 5% corn steep liquor〕 배지에서 발효조 회분 배양한 결과, 30시간 배양시 CGTase의 최대 발현량은 4.2 unit/$m\ell$, 90%의 분비 효율, 90% 이상의 plasmid 안정성을 나타내었다. 저렴한 산업용 molasses 배지로 발효조 회분배양시 플라스크 배양보다 균체증식과 CGTase 발현량이 2배 이상의 증가된 값을 얻었다.