• Journal of Microbiology and Biotechnology
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• 제23권2호
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• pp.211-217
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• 2013

Corynebacterium glutamicum is one of the well-studied industrial strain that is used for the production of nucleotides and amino acids. Recently, it has also been studied as a possible producer of organic acids such as succinic acid, based on its ability to produce organic acids under an oxygen deprivation condition. In this study, we conducted the optimization of medium components for improved succinate production from C. glutamicum under an oxygen deprivation condition by Plackett-Burman design and applied a response surface methodology. A Plackett-Burman design for ten factors such as glucose, ammonium sulfate, magnesium sulfate, potassium phosphate ($K_2HPO_4$ and $KH_2PO_4$), iron sulfate, manganese sulfate, biotin, thiamine, and sodium bicarbonate was applied to evaluate the effects on succinate production. Glucose, ammonium sulfate, magnesium sulfate, and dipotassium phosphate were found to have significant influence on succinate production, and the optimal concentrations of these four factors were sequentially investigated by the response surface methodology using a Box-Behnken design. The optimal medium components obtained for achieving maximum concentration of succinic acid were as follows: glucose 10 g/l, magnesium sulfate 0.5 g/l, dipotassium phosphate ($K_2HPO_4$) 0.75 g/l, potassium dihydrogen phosphate ($KH_2PO_4$) 0.5 g/l, iron sulfate 6 mg/l, manganese sulfate 4.2 mg/l, biotin 0.2 mg/l, thiamine 0.2 mg/l, and sodium bicarbonate 100 mM. The parameters that differed from a normal BT medium were glucose changed from 40 g/l to 10 g/l, dipotassium phosphate ($K_2HPO_4$) 0.5 g/l changed to 0.75 g/l, and ammonium sulfate ($(NH_4)_2SO_4$) 7 g/l changed to 0 g/l. Under these conditions, the final succinic acid concentration was 16.3 mM, which is about 1.46 fold higher than the original medium (11.1 mM) at 24 h. This work showed the improvement of succinate production by a simple change of media components deduced from sequential optimization.

• Journal of Microbiology and Biotechnology
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• 제31권4호
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• pp.570-583
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• 2021

Pyrococcus furiosus α-amylase can hydrolyze α-1,4 linkages in starch and related carbohydrates under hyperthermophilic condition (~ 100℃), showing great potential in a wide range of industrial applications, while its relatively low productivity from heterologous hosts has limited the industrial applications. Bacillus subtilis, a gram-positive bacterium, has been widely used in industrial production for its non-pathogenic and powerful secretory characteristics. This study was conducted to increase production of P. furiosus α-amylase in B. subtilis through three strategies. Initial experiments showed that co-expression of P. furiosus molecular chaperone peptidyl-prolyl cis-trans isomerase through genomic integration mode, using a CRISPR/Cas9 system, increased soluble amylase production. Therefore, considering that native P. furiosus α-amylase is produced within a hyperthermophilic environment and is highly thermostable, heat treatment of intact culture at 90℃ for 15 min was performed, thereby greatly increasing soluble amylase production. After optimization of the culture conditions (nitrogen source, carbon source, metal ion, temperature and pH), experiments in a 3-L fermenter yielded a soluble activity of 3,806.7 U/ml, which was 3.3- and 28.2-fold those of a control without heat treatment (1,155.1 U/ml) and an empty expression vector control (135.1 U/ml), respectively. This represents the highest P. furiosus α-amylase production reported to date and should promote innovation in the starch liquefaction process and related industrial productions. Meanwhile, heat treatment, which may promote folding of aggregated P. furiosus α-amylase into a soluble, active form through the transfer of kinetic energy, may be of general benefit when producing proteins from thermophilic archaea.

• 한국미생물·생명공학회지
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• 제46권4호
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• pp.377-389
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• 2018

In this study, a novel microalgal strain, Desmodesmus sp. KAERI-NJ5, was isolated, identified, and evaluated as a candidate for biodiesel feedstock. In a preliminary study, the effects of four general microalgal growth factors, including temperature, pH, light intensity, and concentration of nitrogen source ($KNO_3$), on the microalgal photoautotrophic growth were evaluated. With the exception of light intensity, the growth factors needed to be optimized for the microalgal biomass production. Optimization was done using response surface methodology. The optimal conditions for biomass production were pH 6.54, $27.66^{\circ}C$, and 0.52 g/l $KNO_3$. The biomass production at the optimal conditions was 1.55 g/l, which correlated well with the predicted value of 1.5 g/l. The total lipid and fatty acid methyl ester contents of the cells grown at the optimal conditions were 49% and 21.2% of cell dry weight, respectively. To increase the lipid content of the biomass, microalgae were challenged by nitrogen starvation. Enhancement of total lipid and fatty acid content up to 52.02% and 49%, respectively, were observed. Lipid analysis of the nitrogen-starved cells revealed that C16 and C18 species accounted for 95.9% of the total fatty acids. Among them, palmitic acid (46.17%) and oleic acid (39.43%) dominantly constituted the algal fatty acids. These results suggest Desmodesmus sp. KAERI-NJ5 as a promising feedstock for biodiesel production.

• 생명과학회지
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• 제31권8호
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• pp.761-770
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• 2021

본 연구에서는 반응표면분석법을 이용하여 전통발효식품인 막걸리로부터 분리한 Bacillus amyloliquefaciens SRCM115785 균주에 대하여 protease 생산량을 증가시키기 위한 배지의 최적 농도를 확립하고자 하였다. 선정한 11개의 배지 성분 중 각 성분이 protease 생산에 미치는 영향에 대한 분석을 위해 Plackett-Burman design (PBD)를 설계하여 통계분석한 결과 glucose, yeast extract, beef extract를 protease 생산 향상을 위한 요인으로 최종 선별하였다. 선별된 3개의 성분에 대해 protease 생산을 위한 각 성분별 최적 농도를 결정하기 위해 central composite design (CCD)분석을 설계하여 protease 최대 생산을 위한 각 배지조성별 농도는 glucose 6.75 g/l, yeast extract 12.42 g/l, beef extract 17.48 g/l로 예측되었다. ANOVA 분석을 통해 실험모델의 적합성을 증명하였고, 설계한 최적배지에서 반복실험을 진행하여 protease 생산량을 측정한 결과 예측값과 매우 유사한 값을 나타냄을 확인하였다. 최종적으로 일반 배지에 비해 137% 환이 증가하였으며, 추가로 정량 분석 결과 기존 25.72 U/ml 대비 59.28 U/ml로 230.47% 증가함을 확인하였다. 본 연구를 통해 protease 생산량 증가를 위한 배지 성분의 최적화를 확립하였고, 이를 바탕으로 산업용 효소로서 protease의 효율적인 활용방안에 대한 기초자료로서 활용될 수 있을 것으로 기대된다.

• 한국축산식품학회지
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• 제30권1호
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• pp.66-72
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• 2010

Pediococcus pentosaceus SA131 was isolated from jeotgal, is the bacteriocin producer against bovine mastitis pathogens, Streptococcus uberis E290, Enterococcus gallinarum E362, and Staphylococcus epidermidis ATCC 12228. The medium composition for pediocin SA131 production by P. pentosaceus SA131 was optimized using response surface methodology. Component of medium was studied as carbon source (glucose, fructose, lactose, glycerol, sucrose, maltose, and mannitol), nitrogen source (beef extract, yeast extract, peptone, malt extract, and tryptone), mineral and surfactant ($MgSO_4$, $KH_2PO_4$, $(NH_4)_2SO_4$, $MnSO_4$, NaCl, sodium acetate, and Tween 80). Through one factor-at-a-time experiment, glucose, fructose, yeast extract, malt extract, NaCl, $MgSO_4$, and Tween 80 were determined as the good ingredient. The effects of major factors for pediocin SA131 production were investigated by two-level fractional factorial designs (FFD). By a $2^4$ FFD, fructose, yeast extract, and $MnSO_4$ were found to be the important factors for the bacteriocin production. Subsequently, a $2^3$ central composite design (CCD) was adopted to derive a statistical model for optimizing the composition of the fermentation medium. The estimated optimum composition for the production of pediocin SA131 by P. pentosaceus SA131 was as follows; 0.13% fructose, 1% glucose, 1.8% yeast extract, 2.58% $MnSO_4$, 0.2% NaCl, and 0.2% Tween 80. The pediocin production under optimized medium was increased to 1,000 AU/mL, compared to the 400 AU/mL in MRS medium.

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권4호
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• pp.275-287
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• 2000

Fermentation strategies for recombinant protein production in Pichia pastoris have been investigated and are reviewed here. Characteristics of the expression system, such as phenotypes and carbon utilization, are summarized. Recently reported results such as growth model establishment, app58lication of a methanol sensor, optimization of substrate feeding strategy, DOstat controller design, mixed feed technology, and perfusion and continuous culture are discussed in detail.

• 한국미생물·생명공학회지
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• 제45권4호
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• pp.330-342
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• 2017

Statistical design of experiments was employed to optimize the media composition for the production of laccase from Bacillus sp. PK4. In order to find the key ingredients for the best yield of enzyme production from the selected eleven variables viz yeast extract, glucose, zinc sulphate, copper sulphate, potassium chloride, magnesium sulphate, calcium chloride, ferrous sulphate, sodium chloride, potassium dihydrogen phosphate ($KH_2PO_4$) and dipotassium hydrogen phosphate ($K_2HPO_4$), Plackett-Burman design was applied. The $MgSO_4$, $FeSO_4$, and $CuSO_4$ showed positive estimate, and their concentration optimized further. The steepest ascent method and Box-Behnken method revealed that 1.5 mM $MgSO_4$, 0.33 g/l $FeSO_4$ and 1.41 mM $CuSO_4$ were optimal for the laccase production by Bacillus sp. PK4. This optimization strategy leads to enhancement of laccase production from 2.13 U/ml to 40.79 U/ml. Agro-wastes residues replace the carbon source glucose in the optimized media namely sugarcane bagasse, wheat bran, rice husk, and groundnut shell, among these groundnut shells (117 U/ml) was found to enhance the laccase production significantly. The laccase produced by Bacillus sp. PK4 was found to have the potential to degrade persistent organic pollutant benzo[a]pyrene.

• Management Science and Financial Engineering
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• 제3권1호
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• pp.75-88
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• 1997

Tabu search (TS) has been successfully applied for solving many complex combinatorial optimization problems in the areas of operations research and production control. However, TS is for single-objective problems in its present form. In this article, a TS-based heuristic is developed to determine Pareto-efficient solutions to a multi-objective combinatorial optimization problem. The developed algorithm is then applied to the discrete optimal design problem in statistics to demonstrate its usefulness.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2005년도 생물공학의 동향(XVI)
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• pp.197-197
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• 2005

• Journal of Applied Biological Chemistry
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• 제57권4호
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• pp.313-320
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• 2014

Black liquor (BL) is a by-product of rice straw pulping process. It is a low costs raw material for production value-adding proteins and enzymes, which has been paid more and more attention to reduce its environmental pollution. Mixed cultures of micelial fungi, Trichoderma reesei Northern Regional Research Laboratory (NRRL)11236, Trichoderma reesei NRRL 6165 and Aspergillus niger strains NRC 5A, NRC 7A, and NRC 9A were evaluated for their ability to produce xylanase using crude hemicellulose (CHC) prepared from BL and peat moss as an inert support under solid state fermentation (SSF). The most potent strains, A. niger NRC 9A (818.26 U/g CHC) and T. reesei NRRL 6165 ($100.9{\pm}57.14$ U/g CHC), were used in a mixed culture to enhance xylanase production by co-culturing under SSF. In the mixed culture, xylanase production ($1070.52{\pm}12.57$ U/g CHC) was nearly1.3 and 10.6-fold increases over the activities attained in their monocultures, A. niger NRC 9A and T. reesei NRRL 6165, respectively. Optimization of the culture parameters of the mixed culture SSF process, concentration of ammonium sulfate and corn steep liquor, CHC/peat moss ratio, inoculum size and ratios of the two strains, initial pH value, initial moisture content and incubation time, exhibited a significant increase ($2414.98{\pm}84.02$ U/g CHC) in xylanase production than before optimization.