• Journal of Microbiology and Biotechnology
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• 제13권2호
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• pp.175-181
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• 2003

The production of a carotenoid astaxanthin, a growth-associated principal pigment, is limited in a batch cultivation, because a high glucose concentration severely inhibits the cell growth and also influences the carotenoid production. Therefore, a fermentation strategy including effective chemicals for the high-level production of cells and astaxanthin by a mutant strain Phaffia rhodozyma AJ-6-1 was developed in a fed-batch culture. First, a production medium for maximizing the cell and astaxanthin yields was formulated and optimized. Using this optimized medium, the highest cell and astaxanthin concentrations obtained were about 38.25 g/1 and 34.77 mg/1, respectively. In addition, an attempt was made to increase the amount of astaxanthin using effective chemicals such as ethanol and acetic acid, which are known at an inducer and/or precursor of carotenoid synthesis. When either 10g/1 ethanol or 5 g/1 acetic acid was added to investigate the resulting astaxanthin content, a relatively high astaxanthin concentration or 45.62 mg/l and 43.87 mg/1, respectively, was obtained, and the cell concentrations also increased slightly under these conditions. Therefore, these results imply that a fed-batch culture of the mutant strain P. rhodozyma AJ-6-1 could be effectively employed in the commercial production of astaxanthin, although the factors affecting the productivity remain to be elucidated.

• Journal of Microbiology and Biotechnology
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• 제25권3호
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• pp.358-365
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• 2015

ε-Poly-L-lysine (ε-PL) is a homopolymer of L-lysine molecules connected between the epsilon amino and alpha carboxyl groups. This polymer is currently used as a natural preservative in food. Insufficient biomass is a major problem in ε-PL fermentation. Here, to improve cell growth and ε-PL productivity, various nitrogen-rich nutrients were supplemented into flask cultures after 16 h cultivation, marking the onset of ε-PL biosynthesis. Yeast extract, soybean powder, corn powder, and beef extract significantly improved cell growth. In terms of ε-PL productivity, yeast extract at 0.5% (w/v) gave the maximum yield (2.24 g/l), 115.4% higher than the control (1.04 g/l), followed by soybean powder (1.86 g/l) at 1% (w/v) and corn powder (1.72 g/l) at 1% (w/v). However, supplementation with beef extract inhibited ε-PL production. The optimal time for supplementation for all nutrients examined was at 16 h cultivation. The kinetics of yeast-extract-supplemented cultures showed enhanced cell growth and production duration. Thus, the most commonly used two-stage pH control fed-batch fermentation method was modified by omitting the pH 5.0-controlled period, and coupling the procedure with nutrient feeding in the pH 3.9-controlled phase. Using this process, by continuously feeding 0.5 g/h of yeast extract, soybean powder, or corn powder into cultures in a 30 L fermenter, the final ε-PL titer reached 28.2 g/l, 23.7 g/l, and 21.4 g/l, respectively, 91.8%, 61.2%, and 45.6% higher than that of the control (14.7 g/l). This describes a promising option for the mass production of ε-PL.

• KSBB Journal
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• 제15권5호
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• pp.458-463
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• 2000

Production of acetic acid from cellulosic biomass by Simultaneous Saccharification and Extractive Fermentation (SSEF) was investigated. The homoacetate organism used in this study was a strain of Clostridium thermoaceticum, ATCC # 49707. A batch operation of Simultaneous Saccharification and Fermentation(SSF) using ${\alpha}$-cellulose at pH 5.5 and 55$^{\circ}C$ yielded 40% conversion of cellulose to acetic acid, while a fed-batch SSF operation produced a maximum acetic acid concentration of 25 g/L, with 50% overall yield. In-situ extractive fermentation to reduce the end-product inhibition on both bacteria and enzyme was carried out. in a batch SSEF using 200 g/L IRA-400 resin, acetic acid concentration reached to 23.9 g/L and acetic acid yield and productivity were observed to be 48% and 0.20 g/L-hr, respectively.

• Journal of Microbiology
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• 제44권2호
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• pp.233-242
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• 2006

The objectives of this study were to optimize submerged culture conditions of a new fungal isolate, Ganorderma resinaceum, and to enhance the production of bioactive mycelial biomass and exopolysaccharides (EPS) by fed-batch culture. The maximum mycelial growth and EPS production in batch culture were achieved in a medium containing 10 g/l glucose, 8 g/l soy peptone, and 5 mM $MnCl_2$ at an initial pH 6.0 and temperature $31^{\circ}C$. After optimization of culture medium and environmental conditions in batch cultures, a fed-batch culture strategy was employed to enhance production of mycelial biomass and EPS. Five different EPS with molecular weights ranging from 53,000 to 5,257,000 g/mole were obtained from either top or bottom fractions of ethanol precipitate of culture filtrate. A fed-batch culture of G. resinaceum led to enhanced production of both mycelial biomass and EPS. The maximum concentrations of mycelial biomass (42.2 g/l) and EPS (4.6 g/l) were obtained when 50 g/l of glucose was fed at day 6 into an initial 10 g/l of glucose medium. It may be worth attempting with other mushroom fermentation processes for enhanced production of mushroom polysaccharides, particularly those with industrial potential.

• KSBB Journal
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• 제14권4호
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• pp.495-502
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• 1999

The optimization of fed-batch yeast fermentation process has been performed using genetic algorithm(GA). Three strategies were designed and applied to obtain the optimal feed rate profiles. Genes in the chromosome (input variables for optimization) included feed rates on fixed time intervals (strategy I), or swiching times $t_s1\;and\;t_s2$, and feed rates on singular arc (strategy II), or feed rates and the length of time interval (strategy III). Strategy III showed the best results for all initial conditions due to efficient utilization of genetic information. Simulation results using GA showed similar or better performance compared with previous results by variational caculus and singular control approach.

• 한국식품과학회지
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• 제29권2호
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• pp.343-347
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• 1997

Lactobacillus brevis, L. fermentum과 L. plantarum의 혼합젖산균을 사용하여 배양조건이 밀가루 용액의 발효에 미치는 영향을 살펴보았다. 온도변화에 따른 pH와 총적정산도의 변화를 살펴본 결과 pH의 감소와 총적정산도의 증가가 가장 크게 나타난 $35^{\circ}C$를 밀가루 발효의 최적온도로 선정하였다. 5L발효조에서 질소가스를 1.0 vvm으로 첨가한 혐기적 조건보다 공기를 첨가한 호기적 조건에서 총적정산도의 증가와 pH의 감소가 더 크게 나타났다. 최적 산소공급 조건을 찾기 위하여 통기량을 1.0 vvm으로 고정하고 교반속도를 달리하여 밀가루 용액의 발효한 결과 총적정산도가 가장 크게 나타난 교반속도 250 rpm에 해당되는 산소 전달속도상수 $60\;hr^{-1}$에서 최적이었다. 선정된 최적 배양조건에서 밀가루 발효액의 생산성을 높이기 위하여 반복비율을 변화시키면서 pH-stat를 이용한 반복 유가식 배양을 수행하였다. 반복비율이 증가할수록 반복 간격시간은 증가하였으나 밀가루 발효의 최대 작동시간은 감소하였다. 최적 반복비율을 결정하기 위하여 단위시간 당 배양부피 당 생산된 밀가루 발효액의 부피와 최대가동시간 동안 배양부피 당 생산된 총 밀가루 발효액의 부피를 살펴본 결과 20%의 반복비율에서 최대값을 나타내었다. 혼합젖산균을 이용한 밀가루용액의 유가식 배양에서 최적조건은 배양온도 $35^{\circ}C$, 통기량 1.0 vvm, 산소전달속도 $60\;hr^{-1}$, 반복비율 20%로 나타났다.

• 한국미생물·생명공학회지
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• 제35권3호
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• pp.226-230
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• 2007

산소를 많이 소비하는 발효공정일수록 배양액중의 용존산소의 농도가 목적생산물의 생산성에 많은 영향을 주는 경우가 많다. 때문에 고농도 발효에 앞서, 발효조의 sparging hole로부터 임펠러 높이에 따른 산소전달 능력을 알아본 결과 공기공급이 1 vvm, 교반속도가 600 rpm에서 산소전달계수($K_La$)는 2.67($min^{-1}$)으로 가장 높았다. 배양 시 용존산소 농도를 20% 이상 유지시켰을 때 온도에 따른 k6ub/IFN-${\alpha}1$ 생성은 $30^{\circ}C$에서 세포증식을 하고 $25^{\circ}C$에서 IPTG로 Induction 하였을 때 발현율이 6.43mg/ml로 total protein의 37%로 가장 많은 양이 발현되는 것을 알 수 있었다. 용존산소 농도에 따른 k6ub/IFN-${\alpha}1$의 발현양은 용존산소 농도가 35%일 때 가장 높은 수율을 나타냈다. 용존산소량은 산소소비 속도를 측정함으로써 정확한 임계점을 찾을 수 있었는데 용존산소량이 35% 유지될 때 산소 전달 속도와 비교하여 가장 적당한 산소공급량임을 확인할 수 있었다.

• 한국미생물·생명공학회지
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• 제21권4호
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• pp.354-365
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• 1993

A novel control method with automatic tuning of PID controller parameters has been developed for efficient regulation of dissolved oxygen concentration in fed-batch fermentations of Escherichia coli. Agitation speed and oxygen partial pressure in the inlet gas stream were chosen to be the manipulated variables. A heuristic reasoning allowed improved tuning decisions from the supervision of control performance indices and it coule obviate the needs for process assumptions or disturbance patterns. The control input consisted of feedback and feedforword parts. The feedback part was determined by PID control and the feedforward part is determined from the feed rate. The proportional gain was updated on-line by a set of heuristics rules based on the supervision of three performance indices. These indices were output error covariance, the average value of output error, and input covariance, which were calculated on-line using a moving window. The integral and derivative time constants were determined from the period of output response. The specific growth rate was maintained at a low level to avoid acetic acid accumulation and thus to achieve a high cell density. The specific growthe rate was estimated from the carbon dioxide evolution rate. In fed-batch fermentation, the simutaneous control of dissolved oxygen concentration (at 0.2; fraction of saturated value) and specific growth rate (at 0.25$hr^{-1}$) was satisfactory for the entire culture period in spite of the changes in the feed rate and the switching of control input.

• KSBB Journal
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• 제29권4호
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• pp.303-306
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• 2014

Antifreeze proteins (AFP) inhibit ice growth to permit the survival of polar organisms in the cold environments. The recombinant AFP from an Antarctic bacterium, Flavobacterium frigoris PS1, FfIBP (Flavobacterium frigoris ice-binding protein), was produced using Pichia pastoris expression system. The optimum fermentation temperature ($30^{\circ}C$) and pH (5) for FfIBP production were determined using a fed-batch culture system. The maximal cell density and purified FfIBP were 112 g/L and 70 mg/L, respectively. The thermal hysteresis (TH) activity (0.85) of FfIBP obtained using a glycerol-methanol fed-batch culture system was 2-fold higher than that of the LeIBP (Leucosporidium ice-binding protein). This work allows for large-scale production of FfIBP, which could be extended to further application studies using recombinant AFPs.

• 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.