• Biotechnology and Bioprocess Engineering:BBE
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• 제10권6호
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• pp.528-534
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• 2005

Hairy root cultures have demonstrated great promise in terms of their biosynthetic capability toward the production of secondary metabolites, but continue to constitute a major challenge with regard to large-scale cultures. In order to assess the possibility of conducting mass production of biomass, and the extraction of useful metabolites from Panax ginseng. P. ginseng hairy roots, transformed by Rhizobium rhizogenes KCTC 2744, were used in bioreactors of different types and sizes. The most effective mass production of hairy roots was achieved in several differently Sized air bubble bioreactors compared to all other bioreactor types. Hairy root growth was enhanced by aeration, and the production increased with increasing aeration rate in a 1 L bioreactor culture. It was determined that the hairy root growth rate could be substantially enhanced by increases in the aeration rate upto 0.5vvm, but at aeration rates above 0.5vvm, only slight promotions in growth rates were observed. In 20 L air bubble bioreactors, with a variety of inoculum sizes, the hairy roots exhibited the most robust growth rates with an inoculum size of 0.1% (w/v), within the range 0.1 to 0.7% (w/v). The specific growth rates of the hairy root decreased with increases in the inoculum size.

• 한국미생물·생명공학회지
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• 제22권4호
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• pp.436-442
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• 1994

The characteristics of activated sludge affecting the biodegradation of plastic materials under aerobic condition were studied using cellophane film as a model system. The activated sludges of site 3, which treat a mixture of domestic sewage and supernatant of septic tank, obtained from December 1993 to April 1994 showed similar biodegradation activities. Biodegradations for 28 days reached around 80%. Viable cell number of inoculums maintained at a level of 10$^{6}$~10$^{7}$ /ml. In this range, viable cell number showed no relationship with biodegradation activities. The activa- ted sludges of site 2, which treat a mixture of domestic sewage and anaerobic digest of nightsoil, obtained four times from April 1993 to April 1994 showed very different biodegradation activities ranged from 20% to 80% for 28 days. Inoculum size affects biodegradation significantly. One percent inoculum showed the best biodegradation among the inoculum sizes of 0.1, 1.0 and 10%. Ten percent inoculum revealed inhibitory effects on the biodegradation activity which can be greatly reduced by centrifugation and filtration. Filtration was better than centrifugation in reducing inhibitory effects.

• Journal of Microbiology and Biotechnology
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• 제18권5호
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• pp.913-917
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• 2008

The cyclic undecapeptide, cyclosporin A (CyA), is one of the most commonly prescribed immunosuppressive drugs. It is generated nonribosomally from a multifunctional cyclosporin synthetase enzyme complex by the filamentous fungus Tolypocladium niveum. In order to maximize the production of CyA by wild-type T. niveum (ATCC 34921), each of three culture stages (sporulation culture, growth culture, and production culture) were sequentially optimized. Among the three potential sporulation media, the SSMA medium generated the highest numbers of T. niveum spores. The SSM and SM media were then selected as the optimal growth and production culture media, respectively. The addition of valine and fructose to the SM production medium was also determined to be crucial for CyA biosynthesis. In this optimized three-stage culture system, 3% of the spore inoculum generated the highest level of CyA productivity in a 15-day T. niveum production culture, thereby implying that the determination of an appropriate size of T. niveum spore inoculum plays a critical role in the maximization of CyA production.

• 한국미생물·생명공학회지
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• 제31권4호
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• pp.408-412
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• 2003

오염토양에 유류분해능을 가진 Nocardia sp. H17-1의 접종시 고려되어야 할 인자중 하나인 초기 오염농도에 의한 탄화수소분해능과 초기 접종농도에 의한 분해능 및 균주의 생육을 조사하였다. H17-1은 실험 50일 동안 초기 오염농도 10, 50, 100 g Arabian light oil/kg of soil에 대해 각각 78.5%, 94.3%, 53.2%의 탄화수소를 제거하였으며, 오염농도가 높을수록 분해속도 상수(k) 낮아졌다. $CO_2$의 생성량 또한 오염농도가 높을수록 증가하였으나, 100 g/kg-soil의 오염농도에서는 균의 생육이 저해를 받는 것으로 나타났다. H17-1의 초기 접종농도에 의한 영향은 균의 접종량에 따라 최종 남은 TPH의 양은 큰 차이를 나타내지 않았으나, 분해속도상수(k)는 균의 접종량이 늘어남에 따라 크게 증가되었으며, $CO_2$의 생성량 또한 균의 접종농도에 따라 증가하였다.

• 한국토양비료학회지
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• 제23권2호
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• pp.146-151
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• 1990

Rhizobium균을 이용하여 근류균 접종제를 생산하기 위한 기초연구로 우수한 증량제(carrier material)를 선발하고, 근류균의 접종량, carrier의 숙성 및 보존온도, 멸균조건등 근류균 접종제 생산을 위한 각종 조건을 확립하기 위하여 실험한 결과는 다음과 같다. 1. 근류균 접종제 생산을 위한 carrier material source로는 퍼라이트 및 토탄이 우수한 것으로 나타났다. 2. 근류균의 접종량(inoculum size)에 따른 경시적 생존율은 접종초기 $10^4cells/g$ 이하의 균주를 접종하여도 1~2주후에는 $5{\times}10^8cells/g$로 증가하여 $10^8cells/g$ 접종구와 비슷한 균수를 나타내었다. 3. Carrier의 숙성 및 보존온도에 따른 근류균의 경시적 생존율은 특별한 보온이나 냉장시설없이 실온($20^{\circ}C$) 보존에서도 $10^9cells/g$의 높은 균수가 12주째까지 유지되었다. 4. Carrier의 멸균방법에 따른 근류균의 경시적 생존율은 가압습열멸균 처리구에서 생존율이 가장 양호하였으나 각 처리에 따른 큰 차이는 볼 수 없었고, 특히 퍼라이트의 경우에는 멸균처리없이도 12주째 까지 $10^9cells/g$ 이상의 높은 균수를 유지하였다.

• Journal of Microbiology and Biotechnology
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• 제24권4호
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• pp.453-464
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• 2014

The current research was focused on the extracellular biosynthesis of bactericidal silver nanoparticles (AgNPs) using cell-free supernatant of a local isolate previously identified as a novel Streptomyces aegyptia NEAE 102. The biosynthesis of silver nanoparticles by Streptomyces aegyptia NEAE 102 was quite fast and required far less time than previously published strains. The produced particles showed a single surface plasmon resonance peak at 400 nm by UV-Vis spectroscopy, which confirmed the presence of AgNPs. Response surface methodology was chosen to evaluate the effects of four process variables ($AgNO_3$ concentration, incubation period, pH levels, and inoculum size) on the biosynthesis of silver nanoparticles by Streptomyces aegyptia NEAE 102. Statistical analysis of the results showed that the linear and quadratic effects of incubation period, initial pH, and inoculum size had a significant effect (p < 0.05) on the biosynthesis of silver nanoparticles by Streptomyces aegyptia NEAE 102. The maximum silver nanoparticles biosynthesis (2.5 OD, at 400 nm ) was achieved in runs number 5 and 14 under the conditions of 1 mM $AgNO_3$ (1-1.5% (v/v)), incubation period (72-96 h), initial pH (9-10), and inoculum size (2-4% (v/v)). An overall 4-fold increase in AgNPs biosynthesis was obtained as compared with that of unoptimized conditions. The biosynthesized silver nanoparticles were characterized using UV-VIS spectrophotometer and Fourier transform infrared spectroscopy analysis, in addition to antimicrobial properties. The biosynthesized AgNPs significantly inhibited the growth of medically important pathogenic gram-positive (Staphylococcus aureus) and gram-negative bacteria (Pseudomonas aeruginosa) and yeast (Candida albicans).

• Journal of Microbiology and Biotechnology
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• 제15권3호
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• pp.568-572
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• 2005

Saccharomyces yeast spores are more resistant to drying and storage than vegetative cells. For the mass production of yeast spores, compressed yeast was directly inoculated into a sporulation medium (SM). The effects of inoculum size and the addition of rice wine cake (RWC) into SM on the sporulation were examined using flasks. With $1\%$ inoculum of compressed yeast, $1.45{\times}10^8/ml$ of asci was obtained. The addition of $0.5\%$ RWC into SM improved the cell growth and spore yield, and the number of asci formed was $2.31{\times}10^8/ml$. The effects of culture temperature, temperature-shift, and concentrations of inoculum, potassium acetate, and RWC on the sporulation were also evaluated using a jar fermentor. The optimum temperature for spore formation was $22^{\circ}C$ where the number of asci formed was $2.46{\times}10^8/ml$. The shift of culture temperature from initial $30^{\circ}C$ for 1 day to $22^{\circ}C$ for 3 days increased the number of asci formed to $2.96{\times}10^8/ml$. The use of $2\%$ (w/v) inoculum of compressed yeast, $2\%$ potassium acetate, and $1\%$ (w/v) RWC in SM with the shift of culture temperature of initial $30^{\circ}C\;to\;22^{\circ}C$ resulted in $90\%$ sporulation ratio and formation of $6.18{\times}10^8\;asci/ml$.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.529-532
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• 2001

Korean food waste, one of the abundantly available but environmentally problematic organic wastes in Korea, was utilized as solid-substrate by fungal strain Aspergillus niger ATcC 6275 for the production of enzymemixture containing amylase, cellulase and xylanase. The enzyme mixture can be used as high value-added animal feed. Solid-state fermentation method yielded a 84-fold enhancement in xylanase activity compared with submerged fermentation method. The effect of incubation period, incubation temperature, pH of medium, moisture content, inoculum size and enrichment of the medium with nitrogen and carbon sources were observed for optimal production of these enzymes The optimal amylase activity of 33.10 U/g, cellulase activity of 24.41 U/g, xylanase activity of 328.84 U/g were obtained at 8 days incubation with 50%(w/w) soy bean flake, with incubation temperature of $25^{\circ}C$, pH of 6.38, optimal moisture content of 55% and with inoculum size of $3.8{\times}10^6$spore/g. Enzyme activities were enhanced when ImM $CaSO_4$, 2% Malt extract and 2% galactose were added as mineral, nitrogen and carbon enrichment respectively.

• Journal of Microbiology and Biotechnology
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• 제19권11호
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• pp.1385-1392
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• 2009

This work evaluates the effect of different amino acids on production of Cyclosporin (CyA) production in solid-state fermentation that was previously optimized for different fermentation parameters by one factor at-a-time for the maximum production of CyA by Tolypocladium inflatum MTCC557. Based on the Plackett-Burman design, glycerol, ammonium sulfate, $FeCl_3$, and inoculum size were selected for further optimization by response surface methodology (RSM). After identifying effective nutrients, RSM was used to develop mathematical model equations, study responses, and establish the optimum concentrations of the key nutrients for higher CyA production. It was observed that supplementation of medium containing (% w/w) glycerol, 1.53; ammonium sulfate, 0.95; $FeCl_3$, 0.18; and inoculum size 6.4 ml/5g yielded a maximum of 7,106 mg/kg as compared with 6,480 mg CyA/kg substrate using one factor at-a-time. In the second step, the effect of amino acids on the production of CyA was studied. Addition of $_L$-valine and $_L$-leucine in combination after 20 h of fermentation resulted in maximum production of 8,166 mg/kg.

• Journal of Ginseng Research
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• 제24권4호
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• pp.157-161
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• 2000

인삼배양세포로부터 항산화 물질 고함유 세포주를 선발하기 위하여 인삼육성계통별로 배양기 내에서 callus를 유기하여 callus생장에 미치는 식물호르몬의 영향을 조사하였고, 페놀성 화합물 및 항산화 물질의 함량을 조사함으로써 생리활성물질 고함유 세포주를 선발하고자 하였다. callus 생장은 2mg/L의 CPA를 함유한 MS기본배지에서 가장 높았고, CPA혼합배지로부터 CPA혼합배지로 계대배양할 때 가장 \ulcorner효과적이었으며 생장률은 계대시 Ig/flask의 callus를 접종할 때 가장 효율적이었다. 생리활성물질인 페놀성 화합물의 함량은 공시계통 callus가 재배 인삼뿌리보다 높았다. 특히 계통번호20601은 페놀성 화합물의 함량과 항산화 활성에서 가장 높았으며 Callus의 생장도 매우 양호하였다. 또한 페놀성 화합물의 TLC pattern은 재배 인삼근과 비슷하였으나 함량은 재배 인삼근에 비해 표준품인 caffeic acid의 위치에 더 많은 페놀성 물질이 분포하였다.