• KSBB Journal
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• v.22 no.5
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• pp.297-304
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• 2007

For large and rapid screening of high-yielding mutants of lovastatin produced by filamentous fungal cells of Aspergillus terreus, one of the most important stage is to test as large amounts of mutated strains as possible. For this purpose, we intended to develop a miniaturized cultivation method using $7m{\ell}$ culture tube instead of traditional $250m{\ell}$ flask (working volume $50m{\ell}$). For obtaining large amounts of conidiospores to be used as inoculums for miniaturized cultures, 4 components i.e., glucose, sucrose, yeast extract and $KH_2PO_4$ were intensively investigated, which had been observed to show positive effect on enhancement of spore production through Plackett-Burman design experimet. When optimum concentrations of these components that were determined through application of response surface method (RSM) based on central composite design (CCD) were used, maximum spore numbers amounting to $1.9\times10^{10}$ spores/plate were obtained, resulting in approximately 190 fold increase as compared to the commonly used PDA sporulation medium. Using the miniaturized cultures, intensive strain development programs were carried out for screening of lovastatin high-yielding as well as highly reproducible mutants. It was observed that, for maximum production of lovastatin, the producers should be activated through 'PaB' adaptation process during the early solid culture stage. In addition, they should be proliferated in condensed filamentous forms in miniaturized growth cultures, so that optimum amounts of highly active cells could be transferred to the production culture-tube as reproducible inoculums. Under these highly controlled fermentation conditions, compact-pelleted morphology of optimum size (less than 1 mm in diameter) was successfully induced in the miniaturized production cultures, which proved essential for maximal utilization of the producers' physiology leading to significantly enhanced production of lovastatin. As a result of continuous screening in the miniaturized cultures, lovastatin production levels of the 81% of the daughter cells derived from the high-yielding producers turned out to be in the range of 80%$\sim$120% of the lovastatin production level of the parallel flask cultures. These results demonstrate that the miniaturized cultivation method developed in this study is efficient high throughput system for large and rapid screening of highly stable and productive strains.

• 한국신재생에너지학회:학술대회논문집
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• 2010.06a
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• pp.249.2-249.2
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• 2010

최근의 고유가와 환경오염에 대한 대응 수단으로 수송용 바이오연료의 보급에 대한 관심이 세계적으로 높아지고 있다. 이 중 바이오디젤은 동식물성 기름으로부터 메탄올과의 전이에스테르화 반응에 의해 생산되는 경유대체 연료로서 환경 친화성과 지속가능성이 인정됨에 따라 그 생산량이 급격히 증가하고 있다. 바이오디젤의 생산량이 증가함에 따라 대두유, 유채유, 팜유 등의 원료유 가격 상승 및 수급 불안정 문제가 대두되고 있으며 식량자원과의 충돌 문제도 발생되고 있다. 이를 해결하기 위한 방안으로 유리지방산 함량이 높은 저가유지 자원(폐식용유, 폐돈지, 폐우지, soapstock, trapped grease)을 이용한 공정 개발 연구가 활발히 진행되고 있다. 본 연구에서는 비활용되고 있는 해외 열대작물 열매씨앗에서 착유한 식물성 오일의 바이오디젤 원료유로서의 사용 가능성을 검토하였다. 열대작물 오일의 물성 분석 결과 고형물, 수분, 인, 유리지방산 함량이 대두원유보다 매우 높게 나타났다. 오일 중의 인지질은 바이오디젤 제조 반응후 에스테르와 글리세린의 층분리를 방해하여 공정 효율을 감소시키고 유리지방산은 염기촉매와 결합하여 지방산염을 생성해 생산수율을 감소시키는 문제를 일으킨다. 고형물과 수분은 여과와 감압증발에 의해 쉽게 제거가 가능하였다. 15~20%의 유리지방산 함유 열대작물 오일의 전처리를 위해 균질계 산촉매와 비균질 고체 산촉매를 이용해 에스테르화 반응 효율을 조사한 결과 황산이 가장 높은 효율을 보였다. 반응표면분석법(Response Surface Method, RSM)을 적용해 메탄올과 촉매량의 2변수 에스테르화반응 최적화를 수행한 결과 메탄올 26%, 촉매 0.98%로 최적 조건이 도출되었으며 초기 산가 33mgKOH/g에서 0.98mgKOH/g으로 감소됨을 확인하였다. 전처리 정제한 오일의 물성분석 결과 고형물 0.1%, 수분 0.10%, 산가 1.0mgKOH/g, 인함량 20ppm 이하로 바람직한 원료유가 생산됨을 알 수 있었다. 제조된 원료유를 이용해 전이에스테르화 반응 최적화 실험을 RSM에 근거하여 진행한 결과 KOH 0.8%, 메탄올:오일 몰비 6.2:1, 반응온도 $60^{\circ}C$, 교반속도 200rpm, 반응시간 30분으로 나타났으며 증류 정제전 97.3%, 증류후 100.0%의 바이오디젤을 생산 할 수 있었다. 열대작물 오일의 전처리 공정은 메탄올을 과잉양으로 사용함으로 효과적인 알콜 회수 공정이 중요하다. 전처리 후 층분리를 통해 회수되는 메탄올 중의 수분함량은 2%~7%로서 이를 전처리 반응에 재사용하기 위해서는 0.3%이하의 수분함량으로 정제가 필요하다. 본 연구에서는 고가의 증류탑 형태가 아닌 단증류방식으로 2단계 내지 3단계로 0.3% 수분의 메탄올 회수 조건을 도출하였으며 파일롯 공정 설계를 진행하고 있다. 이로서 본 연구의 열대작물 오일은 저가로 충분한 물량의 확보가 가능하다면 바이오디젤 원료 자원으로서 큰 활용가치가 있는 것으로 판단된다.

• Journal of the Korean GEO-environmental Society
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• v.12 no.4
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• pp.25-31
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• 2011

In this study, the experimental design methodology was applied to optimize 1,4-dioxane treatment in E-beam process. Main factor was mathematically described as a function of parameters 1,4-dioxane removal efficiencies(%), TOC removal efficiencies(%) modeled by the use of the central composite design(CCD) method among the response surface methodology(RSM). Concentration of 1,4-dioxane is designated as "$x_1$" and Irradiation intensity is designated as "$x_2$". The regression equation in coded unit between the 1,4-dioxane concentration and removal efficiencies(%) was $y=71.00-10.85x_1+20.67x_2+{1.53x_1}^2-{7.92x_2}^2-1.23x_1x_2$. The regression equation in coded unit between the 1,4-dioxane concentration and TOC removal efficiencies(%) was $y=44.48-13.25x_1+9.54x_2+{5.43x_1}^2-{1.35x_2}^2+4.45x_1x_2$. The model predictions agreed well with the experimentally observed results $R^2$(Adj) over 90%. Toxicity test using algae Pseudokirchneriella Subcapitata showed that the inhibition was reduced according to increasing an E-beam irradiation.

• Journal of Biosystems Engineering
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• v.30 no.1 s.108
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• pp.8-16
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• 2005

The extrusion-cooking condition on Cordyceps pruinosa was designed using twin-screw extruder. Response surface methodology (RSM) was used to investigate extrusion-cooking using a central composition design with varying die temperature $(114-146^{\circ}C)$, feed moisture $(22-38\%)$, feed rate (4-14 ka/h) and screw speed (120-280 rpm). System parameters (die pressure and specific mechanical energy (SME)) and extrudate parameters (density and water solubility index (WSI)) were statically analyzed using RSH. Die pressure was significantly affected by temperature, moisture contents and feed rate. SM was affected by screw speed and feed rate. When die temperature is $130^{\circ}C$ and moisture content $25\%$, the optimum pressure is shown. SME is about 20 Wh/kg, when feed rate is $10\~12kg/min$ and screw speed $200\~250rpm$. WSI was affected by temperature and moisture contents. Density was not affected by any factor. WSI increases by $7\%$ from about $23\%$ to about $30\%$, as temperature is raised from $120^{\circ}C\;to\;140^{\circ}C$. The WSI of Cordyceps pruinosa pulverized after extruding (PE) is about $26.97\%$ higher than that of raw material and $10\%$ higher than that of pulverized after drying (PD). The content of unsaturated fatty acid were not significantly different in PD and PE. Anti-oxidative activity of PE was 1.67-2.2 times higher than that of PD in Cordyceps pruinosa using 1- dipheny1-2-picrylhydrazyl method (DPPH).

• Transactions of the Korean Society of Mechanical Engineers A
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• v.36 no.3
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• pp.331-337
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• 2012

In this study, the front side member is optimized using a topography optimization technique. Optimization of a simple beam is conducted before optimization of the front side member. The objective function is set to minimize the first buckling factor in the longitudinal direction. The design variable corresponds to the perturbation of nodes normal to the shell's mid-plane space. The crash analysis is conducted on a simple beam, which is optimized by Response Surface Method and the topography optimization technique. In order to verify the topography optimization technique, the results of the RSM and topography optimization model are compared. Consequently, we confirm the satisfactory performance of the topography optimization technique, and apply this topography optimization to the front side member. Thus, the front side member is optimized and its crashworthiness is increased.

• Food Science of Animal Resources
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• v.40 no.1
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• pp.118-131
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• 2020

It is necessary to add protective agents for protecting the probiotic viability in the preparation process of probiotics starter. In this study, we used whey protein concentrate (WPC), pullulan, trehalose, and sodium glutamate as the protective agent and optimized the proportion of protective agent and spray-drying parameters to achieve the best protective effect on Lactobacillus plantarum. Moreover, the viable counts of L. plantarum in starter stored at different temperatures (-20℃, 4℃, and 25℃) for 360 days were determined. According to response surface method (RSM), the optimal proportion of protective agent was 24.6 g/L WPC, 18.8 g/L pullulan, 16.7 g/L trehalose and 39.3 g/L sodium glutamate. The optimum spray-drying parameters were the ratio of bacteria to protective agents 3:1 (v: v), the feed flow rate 240 mL/h, and the inlet air temperature 115℃ through orthogonal test. Based on the above results, the viable counts of L. plantarum was 12.22±0.27 Log CFU/g and the survival rate arrived at 85.12%. The viable counts of L. plantarum stored at -20℃ was more than 10¹⁰ CFU/g after 200 days.

• Korean Chemical Engineering Research
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• v.54 no.6
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• pp.822-829
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• 2016

Our aim was to optimize the production of cellulase-free thermoactive xylanase by Aureobasidium pullulans CBS 135684 with statistical methodology based on experimental designs. Among eleven variables, the nutrient sources that had significant effect on xylanase production were corncob, $(NH_4)_2SO_4$, xylose, $KH_2PO_4$ and tween 80, identified by the initial screening method of Plackett-Burman. The optimum concentrations of these five components were subsequently investigated using response surface methodology. The optimal concentrations ($g{\cdot}l^{-1}$) for maximum production of xylanase were corncob, 39.0; $(NH_4)_2SO_4$, 3.0; xylose, 1.8; $KH_2PO_4$ 1.4; and tween 80, 1.4, respectively. An improved xylanase yield of $8.74{\pm}0.84U{\cdot}ml^{-1}$ was obtained with optimized medium which is 2.1-fold higher production than previously obtained results ($4.10{\pm}0.10U{\cdot}ml^{-1}$) after 48 h of cultivation. In addition, the xylanase production under optimal condition reached $10.09{\pm}0.27U{\cdot}ml^{-1}$ after 72 h of cultivation.

• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.381-384
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• 2002

Response surface methodology (RSM) was successfully applied to optimize for the production of Ganoderma lucidum in batch fermentations using the whey (40,000 mg latose/L) as substrate. This study was performed according to the central composite design (CCD) with respect to pH and temperature, where the designed intervals were 3.3$22.9^{\circ}C$$37.1^{\circ}C$, respectively. A second-order factorial design of the experiments was used to build empirical models providing a quantitative interpretation of the relationships between the two variables. The optimum conditions to maximize the production of G. lucidum were pH 4.2 and $28.3^{\circ}C$. At optimum conditions, the mycelial dry weight (MDW) and residual soluble COD (SCOD) were simultaneously used to evaluate the biokinetic coefficients assocoated with substrate inhibition model by nonlinear least squares method with 95% confidence interval. The. maximum microbial growth rates (${\mu}m$), half saturation coefficient ($K_s$), and the inhibition substrate concentration ($K_{is}$) were determined to be 0.095 l/hr, 128,000 mg SCOD/L and 49,000 mg SCOD/L, respectively. And the microbial yield coefficient (Y), biomass decay rate coefficient ($K_d$), and the maintenance energy coefficient ($m_s$) were determined to be 0.37 mg MDW/mg SCOD, 0.001 1/hr, and 0.0015 1/hr, respectively.

• Journal of Microbiology and Biotechnology
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• v.22 no.1
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• pp.92-99
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• 2012

The optimal physical factors affecting enzyme production in an airlift fermenter have not been studied so far. Therefore, the physical parameters such as aeration rate, pH, and temperature affecting PLA-degrading enzyme production by Actinomadura keratinilytica strain T16-1 in a 3 l airlift fermenter were investigated. The response surface methodology (RSM) was used to optimize PLA-degrading enzyme production by implementing the central composite design. The optimal conditions for higher production of PLA-degrading enzyme were aeration rate of 0.43 vvm, pH of 6.85, and temperature at $46^{\circ}C$. Under these conditions, the model predicted a PLA-degrading activity of 254 U/ml. Verification of the optimization showed that PLA-degrading enzyme production of 257 U/ml was observed after 3 days cultivation under the optimal conditions in a 3 l airlift fermenter. The production under the optimized condition in the airlift fermenter was higher than un-optimized condition by 1.7 folds and 12 folds with un-optimized medium or condition in shake flasks. This is the first report on the optimization of environmental conditions for improvement of PLA-degrading enzyme production in a 3 l airlift fermenter by using a statistical analysis method. Moreover, the crude PLA-degrading enzyme could be adsorbed to the substrate and degraded PLA powder to produce lactic acid as degradation products. Therefore, this incident indicates that PLA-degrading enzyme produced by Actinomadura keratinilytica NBRC 104111 strain T16-1 has a potential to degrade PLA to lactic acid as a monomer and can be used for the recycle of PLA polymer.

• KSBB Journal
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• v.20 no.3
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• pp.215-219
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• 2005

Production of alkaline pretense by Bacillus clausii I-52 was optimized by experimental design methods. Among 7 medium components, three (wheat flour, sodium citrate, sodium carbonate) were selected as components affecting the pretense activity significantly by Plackett-Burman methods. Furthermore the ranges of effective concentrations were determined by Box-Behnken methods. The objective function describing the alkaline pretense production was obtained and optimum concentration of 3 components was determined by using response-surface methods (RSM). Theoretical maximum production was 74000 U/mL (Wheat flour: 0 g/L, Sodium citrate: 5 g/L, Sodium carbonate: 10 g/L). With the optimized medium composition, 92000 U/mL alkaline protease was produced experimentally, resulting in $90\%$ increase compared to before-optimization production (49000 U/mL).