• Journal of Microbiology and Biotechnology
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• 제18권12호
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• pp.1903-1907
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• 2008

Prodigiosin is a natural red pigment with algicidal activity against Cochlodinium polykrikoides, a major harmful red-tide microalga. To increase the yield of prodigiosin production by Hahella chejuensis KCTC 2396, significant medium components were determined using a two-level Plackett-Burman statistical design technique. Among 12 components included in basal medium, $NaHCO_3$, ${Na}_{2}{SiO}_{3}$, ${NH_4}{NO_3}$, ${Na}_{2}{SO}_{4}$ and $CaCl_2$ were determined to be important for prodigiosin production. The medium formulation was finally optimized using a Box-Behnken design as follows: 1% sucrose; 0.4% peptone; 0.1 % yeast extract; and (g/l): NaCl, 20.0; ${Na}_{2}{SO}_{4}$, 9.0; $CaCl_2$, 1.71; KCl, 0.4; and (mg/l): ${H_3}{BO_3}$, 10.0; KBr, 50.0; NaF, 2.0; $NaHCO_3$, 45.0; ${Na}_{2}{SiO}_{3}$, 4.5; ${NH_4}{NO_3}$, 4.5. The predicted maximum yield of prodigiosin in the optimized medium was 1.198 g/l by the Box-Behnken design, whereas the practical production was 1.495 g/l, which was three times higher than the basal medium (0.492 g/l).

• Food Science and Biotechnology
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• 제18권2호
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• pp.442-448
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• 2009

In this study Bacillus subtilis PTCC 1023 was used for the production of protopectinase using soybean based media. The use of isolated soybean protein (ISP) and soybean flour resulted in similar protopectinase production and growth rates. The effect of medium composition on protopectinase production was studied using central composite design (CCD) methodology. The change in the concentration of ISP (1-7%), glucose (0-10%), and phosphate (0.1-0.3 M) was found to affect the protopectinase activity (response variable) after 24 hr of cultivation. In the range studied, ISP and glucose had a negative effect on the response variable, whereas phosphate had a positive effect. A statistically significant interaction was identified between phosphate and ISP, suggesting that correct optimization of medium formulation in this case can only be obtained using factorial design of experiments. Protopectinase activity exceeding 215 U/mL was obtained in a medium containing 4% ISP, 0.3M phosphate, and no added sugar.

• Journal of Microbiology and Biotechnology
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• 제23권8호
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• pp.1084-1091
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• 2013

In this paper, two statistical methods were applied to optimize medium components to improve the production of the milk-clotting enzyme by Bacillus amyloliquefaciens D4. First, wheat bran juice, skim milk powder, and $Na_2HPO_4$ were shown to have significant effects on D4 enzyme production using the Plackett-Burman experimental design. Subsequently, an optimal medium was obtained using the Box-Behnken method, which consisted of 3.31 g/l of skim milk powder, 5.0 g/l of sucrose, 0.1 g/l of $FeSO_4{\cdot}7H_2O$, 0.1 g/l of $MgSO_4{\cdot}7H_2O$, 0.1 g/l of $MnSO_4{\cdot}2H_2O$, 0.1 g/l of $ZnSO_4{\cdot}7H_2O$, 1.52 g/l of $Na_2HPO_4$, and 172.45 g/l of wheat bran juice. With this optimal medium, the milk-clotting enzyme production was remarkably enhanced. The milk-clotting enzyme activity reached 3,326.7 SU/ml after incubation of 48 h, which was 1.76-fold higher than that of the basic medium, showing that the Plackett-Burman design and Box-Behnken response surface method are effective to optimize medium components, and B. amyloliquefaciens D4 possessed a high rennet-producing capacity in the optimal medium.

• Journal of Microbiology and Biotechnology
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• 제24권2호
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• pp.188-196
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• 2014

Extracellular production of recombinant human bone morphogenetic protein-7 (rhBMP-7) was carried out through the fermentation of Bacillus subtilis. Three significant fermentation conditions and medium components were selected and optimized to enhance the rhBMP-7 production by using the response surface methodology (RSM). The optimum values of the three variables for the maximum extracellular production of rhBMP-7 were found to be 2.93 g/l starch, 5.18 g/l lactose, and a fermentation time of 34.57 h. The statistical optimization model was validated with a few fermentations of B. subtilis in shake flasks under optimized and unoptimized conditions. A 3-L jar fermenter using the shake-flask optimized conditions resulted in a higher production (413 pg/ml of culture medium) of rhBMP-7 than in a shake flask (289.1 pg/ml), which could be attributed to the pH being controlled at 6.0 and constant agitation of 400 rpm with aeration of 1 vvm.

• Food Science and Biotechnology
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• 제18권4호
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• pp.959-964
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• 2009

Production of the antioxidant substances by Bacillus polyfermenticus SCD was investigated using shake-flask fermentation. The one-factor-at-a-time method was first employed to determine the key ingredients for optimal medium composition, then further investigation of the medium composition was performed using response surface methodology (RSM). The antioxidant activity was measured using 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) assays. After screening various elements, fructose, tryptone, and $MgSO_4\;7H_2O$ were chosen as the main factors for study in the statistical experimental design. Central composite design (CCD) was then used to determine the optimal concentrations of these 3 components. Under the proposed optimized medium containing 2.8% fructose, 1.34% tryptone, 0.015% $MgSO_4\;7H_2O$), 0.5% NaCl, and 0.25% $K_2HPO_4$, the model predicted an antioxidant activity of 80.5% ($R^2=0.9421$. The actual experimental results were in agreement with the prediction.

• Journal of Microbiology and Biotechnology
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• 제15권5호
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• pp.930-937
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• 2005

The Plackett-Burman design and the response surface method (RSM) with a central composite design (CCD) were employed to develop a culture medium for ansamitocin P-3 production using Actinosynnema pretiosum ATCC 31565. Among the 11 nutrients tested using the Plackett-Burman design, two carbon sources, sucrose and dextrin, and two nitrogen sources, polypeptone and yeast extract, were selected. Optimization of the concentrations of the selected nutrients was then performed using RSM with CCD. After two rounds of RSM, the optimum concentrations ($\%w/v$) of sucrose, dextrin, polypeptone, and yeast extract were identified as 4.5, 4.5, 0.16, and 0.89, respectively. The maximum ansamitocin P-3 titer was 45.2 mg/l with the optimized medium, which was about 6 times higher than that (7.315 mg/l) obtained with an $R_{2}YE$ medium before optimization.

• Journal of Microbiology and Biotechnology
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• 제16권9호
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• pp.1338-1346
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• 2006

Ansamitocin P-3 is a potent antitumor agent produced by A. pretiosum. A deletion mutant of A. pretiosum was constructed by deleting the asm2 gene, a putative transcriptional repressor. The deletion mutant showed a 9-fold enhanced ansamitocin P-3 productivity. The response surface method with central composite design was employed to further optimize the culture medium composition for ansamitocin P-3 production by the deletion mutant. The concentrations of four medium ingredients, dextrin, maltose, cotton seed flour, and yeast extract, which have been reported as major components for ansamitocin production, were optimized through a series of flask culture experiments. The optimum concentrations of the selected factors were found to be dextrin 6.0%; maltose 3.0%; cotton seed flour 0.53%; and yeast extract 0.45%. The maximum titer of ansamitocin P-3 was 78.3 mg/l with the optimized composition, about 15-folds higher than the unoptimized titer of 5.0 mg/l obtained with YMG medium.

• ALGAE
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• 제37권2호
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• pp.175-183
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• 2022

Polar microorganisms produce physiologically active substances to adapt to harsh environments, and these substances can be used as biomedical compounds. The green microalga Micractinium variabile KSF0031, which was isolated from Antarctica, produced phytol, a natural antimicrobial agent. Furthermore, several polyunsaturated fatty acids (PUFAs), including omega-3, exhibit antioxidant properties. Here statistical methods (Plackett-Burman design and Box-Behnken design) were used to optimize the culture medium of KSF0031 to improve biomass production, and K₂HPO₄, MgSO₄·7H ₂O, and ammonium ferric citrate green (AFCg) were selected as significant components of the culture medium. Changes in the concentration of K₂HPO₄ and MgSO₄·7H ₂O as positive factors and AFCg as a negative factor affected cell growth to a remarkable degree. The biomass production in a 100 L culture using the optimized medium for 24 d at 18℃ was improved by 37.5% compared to that obtained using the original BG-11 medium. The quantities of PUFAs and phytol obtained were 13 mg g^-1 dry cell weight (DCW) and 10.98 mg g^-1 DCW, which represent improved yields of 11.70% and 48.78%, respectively. The results of this study could contribute to an improved production of phytol and fatty acids from Antarctic microalgae in the biomedical industry.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2005년도 생물공학의 동향(XVI)
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• pp.157-157
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• 2005

• Biotechnology and Bioprocess Engineering:BBE
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• 제9권6호
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• pp.459-464
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• 2004

The objective of this study is to improve cephalosporin C (CPC) production byoptimization of medium and culture conditions. A statistical method was introduced to optimize the main culture medium. The main medium for CPC production was optimized using a statistical method. Glucose and corn steep liquor (CSL) were found to be the most effective factors for CPC production. Glucose and CSL were optimized to 2.84 and $6.68\%$, respectively. CPC produc­tion was improved $50\%$ by feeding of $5\%$ rice oil at day 3rd and 5th day during the shake flask culture of C acremonium M25. The effect of agitation speeds on CPC production in a 2.5-L bio­reactor was also investigated with fed-batch mode. The maximum cell mass (54.5 g/L) was obtained at 600 rpm. However, the maximum CPC production (0.98 g/L) was obtained at 500 rpm. At this condition, the maximum CPC production was improved about $132\%$ compared to the re­sult with batch flask culture.