• Journal of Microbiology and Biotechnology
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• 제9권6호
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• pp.751-756
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• 1999

For mass production of poly(3-hydroxybutyrate) (PHB), high cell density cultures of Ralstonia eutropha were carried out in 2.5-1 and 60-1 fermentors by two fed-batch culture techniques of nitrogen and phosphate limitation. When the nitrogen limitation technique was employed using both an on-line glucose monitoring and control system, a high concentration level of PHB (121g/l) was obtained in the small-scale fermentor of 2.5 1. However, the PHB concentration obtained in a large-scale fermentor of 60 1 only turned out to be 60g/l. In contrast, when another fed-batch culture technique of the phosphate-limitation employing dissolved oxygen (DO) stat glucose feeding was used, a large amount of PHB was successfully produced in both 60-1 and 2.5-1 fermentors. In a 2.5-1 fermentor, concentrations of PHB and cells obtained in 58 h were 175 and 210 g/l, respectively, which corresponded to the PHB productivity level of 3.02 g/l/h. In a 60-1 fermentor, a final cell concentration of 221 g/l and a PHB concentration of 180 g/l with PHB productivity level of 3.75 g/l/h were obtained in 48h. PHB content and yield from glucose were 81% and 0.38g PHB/g glucose, respectively. These data suggest that the phosphate limitation technique is more effective compared to nitrogen limitation in the mass production of PHB by R. eutropha of a large scale.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2005년도 생물공학의 동향(XVII)
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• pp.347-347
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• 2005

• Journal of Microbiology and Biotechnology
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• 제20권4호
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• pp.844-851
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• 2010

Controlling the dissolved oxygen (DO) in the fed-batch culture of the medicinal mushroom Ganoderma lucidum led to a 2-fold increase of the maximum biomass productivity compared with uncontrolled DO conditions. By contrast, extracellular polysaccharide (EPS) production was two times higher under oxygen limitation (uncontrolled DO) than under increased oxygen availability (controlled DO). Morphologically, dispersed mycelium was predominant under controlled DO conditions, with highly branched hyphae, consistent with the enhanced culture growth noted under these conditions, whereas in the uncontrolled DO process mycelial clumps were the most common morphology throughout the culture. However, in both cultures, clamp connections were found. This is an exciting new finding, which widens the applicability of this basidiomycete in submerged fermentation. In rheological terms, broths demonstrated shear-thinning behavior with a yield stress under both DO conditions. The flow curves were best described by the Herschel-Bulkley model: flow index down to 0.6 and consistency coefficient up to 0.2 and 0.6 Pa $s^n$ in uncontrolled and controlled cultures DO, respectively. The pseudoplastic behavior was entirely due to the fungal biomass, and not to the presence of EPS (rheological analysis of the filtered broth showed Newtonian behavior). It is clear from this study that dissolved oxygen tension is a critical process parameter that distinctly influences G. lucidum morphology and rheology, affecting the overall performance of the process. This study contributes to an improved understanding of the process physiology of submerged fermentation of G. lucidum.

• Journal of Microbiology and Biotechnology
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• 제14권2호
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• pp.225-230
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• 2004

A plasminogen kringle domain 1 to 3, rKl-3, was expressed in Escherichia coli under the control of T7 promoter. For the cost-effective production of rKl-3, the induction process was analyzed and optimized. Induction characteristics with lactose were analyzed in terms of induction time and inducer concentration in various culture conditions including batch and high-cell-density fed-batch cultures. In the fed-batch culture, the induction around 6 h after initiation of the DO-stat fed-batch culture resulted in the highest expression level of rKI-3 among the induction points examined. The highest demand of oxygen at this point was crucial for the maximum expression level of rKI-3. As the lactose concentration increased, the expression level also increased, though the expression level showed a plateau above a concentration of 14 mM of lactose. Lactose acted less specifically than IPTG since most of it was hydrolyzed to glucose and galactose. However, using lactose, the cell growth and the maximum expression level of rKl-3 increased by 20% and 24%, respectively, compared with those using IPTG in the fed-batch culture. The lactose seemed to be hydrolyzed by intracellular and extracellular $\beta$-galactosidase liberated by cell lysis at the same time. Residual concentration of glucose was maintained to a a limit of detection by high performance liquid chromatography, and galactose was not consumed by the host strain Escherichia coli BL2l(DE3).

• Biotechnology and Bioprocess Engineering:BBE
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• 제1권1호
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• pp.36-40
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• 1996

For the production of cis, cis-muconic acid via biocatalytic conversion reactions from a toxic cosubstrate, benzoic acid, a fed-batch process using computer-controlled DO-stat feeding was developed. The mutant strain of Pseudomonas putida BM014 produced cis, cis-muconic acid from benzoic acid with high conversion yield. More than 32 g/L of cis, cis-muconic acid was accumulated in 42h and a productivity of 1.4g/(L.h)was achieved.

• KSBB Journal
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• 제6권1호
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• pp.15-25
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• 1991

The optimal glucose concentration for the high-density culture of recombinant yeasts was obtained using dynamic simulation. An adaptive and predictive algoritilm complimented by the rule base was proposed for the control of the fed-batch fermentation process. The measurement of process variables has relatively long sampling period and relatively long time delay characteristics. As one of the solution on these problems, prediction techniques and rule bases were added to a classical recursive identification and control algorithm. Rule bases were used in the determination of control input considering the difference between the predicted value and the measured value. A mathelnatical model was used in the estimation and interpretation of the changes of state variables and parameters. Better performances were obtained by employing the control algorithm proposed in the present study compared to the conventional adaptive control method.

• 미생물학회지
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• 제16권1호
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• pp.21-29
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• 1978

In the fed-batch fermentation of penicillin specific uptake rates of carbon source and ammonia nitrogen, and specific production rate of penicillin as the most important process variables were evaluated over the fermentation course and their effects on the productivity studied. As the results, glucose and lactose each as a major carbon source showed the following values, respectively ; the specific uptake rates of 47-93 mg hexose per gm-DCW per he and 37-44 mg hexose per gm-DCW per hr, the specific uptake rates of 4.6-6.8 mg $NH_3-N$ per gm-DCW per hr, and 1.2 mg $NH_3-N$ per gm-DCW per he and the specific production rates of 32-42 arbitrary unit per gm-DCW per hr and 46-50 arbitrary unit per gm-DCW per hr. The productivity of penicillin could be improved by controlling the feed rates of glucose and ammonia nitrogen to meet the uptake rates.

• 한국미생물·생명공학회지
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• 제20권6호
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• pp.668-676
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• 1992

Alcaligenes eutrophus에 의한 poly-Beta-hydroxybuty-rate(PHB) 생산을 최적화하기 위해서 1단계에서는 성장을 최대화하고, 2단계에서는 생산을 최대화하는 이단 유가식 배양을 채택하였다. 성장단계에서 탄소원과 질소원의 최적농도는 회분식 배양 모델로부터 결정되었는데, 각기 16.64g/l와 0.54g/l이었다. 생산단계에서 질소원의 경우, 유가식 배양실험을 통해 0.07g/l가 최적인 것으로 나타났다. 탄소원의 최적농도는 모사로부터 결정되었고 이때 최적 전환시간도 함께 결정되었다. 생산공정의 제어를 위하여 시간지연을 고려한 적응비례 되먹임제어를 이용한 결과, 탄소원 농도 및 질소원농도를 최적의 상태로 유지시킬 수 있었다.

• Journal of Microbiology and Biotechnology
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• 제31권9호
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• pp.1305-1310
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• 2021

Shikimate is a key high-demand metabolite for synthesizing valuable antiviral drugs, such as the anti-influenza drug, oseltamivir (Tamiflu). Microbial-based strategies for shikimate production have been developed to overcome the unstable and expensive supply of shikimate derived from traditional plant extraction processes. In this study, a microbial cell factory using Corynebacterium glutamicum was designed to overproduce shikimate in a fed-batch culture system. First, the shikimate kinase gene (aroK) responsible for converting shikimate to the next step was disrupted to facilitate the accumulation of shikimate. Several genes encoding the shikimate bypass route, such as dehydroshikimate dehydratase (QsuB), pyruvate kinase (Pyk1), and quinate/shikimate dehydrogenase (QsuD), were disrupted sequentially. An artificial operon containing several shikimate pathway genes, including aroE, aroB, aroF, and aroG were overexpressed to maximize the glucose uptake and intermediate flux. The rationally designed shikimate-overproducing C. glutamicum strain grown in an optimized medium produced approximately 37.3 g/l of shikimate in 7-L fed-batch fermentation. Overall, rational cell factory design and culture process optimization for the microbial-based production of shikimate will play a key role in complementing traditional plant-derived shikimate production processes.

• 한국미생물·생명공학회지
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• 제32권1호
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• pp.52-59
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• 2004

Fructose와 galactose를 배지에 동시에 사용하였을 때 catabolite repression에 의한 PAFS생산성 저해 현상을 극복하기 위해서 본 연구에서는 두 가지 탄소원의 공급량과 유속을 달리하는 유가식 배양으로 PAFS 생산성을 향상시키고자 하였다. 또한 통계적으로 성공률이 매우 높다고 밝혀진 실험방법에 의해 모균주로부터 고생산변이주를 선별하여 생산성이 가장 우수한 것으로 판명된 AP-20 균주를 생물반응기를 이용한 회분식 배양 및 유가식 배양 실험에 이용하였다. 생물반응기에서의 유가식 배양이 회분식 배양에서보다 PAFS생산성이 약 4배 이상 향상되었음을 확인할 수 있었다. 또한 생산균주의 배양생리학적 특성으로서 Pseudomonas aeruginosa는 galactose를 이용해서 세포성장과 이차대사산물 생합성과 관련된 유전자의 발현을 하고 fructose를 이용하여 PAFS를 생합성하는 것으로 추론되며, 너무 느린 탄소원의 공급은 세포성장에 제한요소로 작용하여 이차대사산물의 생합성을 저해하는 것으로 판단되었다. 한편 유가식 배양일지라도 배양 조건에 따라 그 생산성이 뚜렷이 차이가 나는 것으로 관찰되었다 즉 galactos의 초기량이 20g/L이고 fructose 30g/L를 0.032 mL/min의 속도로 공급했을 경우의 PAFS생산량을 100%로 정의했을 때, 초기부터 40 g/L의 galactose가 존재하고 20 g/L의 fructose를 0.032 mL/min의 속도로 공급한 경우에 PAFS생산성이 약 580%향상된 것으로 나타났다.