• 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).

• Journal of Life Science
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• v.26 no.5
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• pp.571-583
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• 2016

Biogenic amines are produced primarily by microorganisms found in fermented foods and are often implicated in poisoning incidents in humans. In this study, 620 strains of microorganisms were isolated from traditional Korean fermented food in Sunchang in order to screen for non-biogenicamine-producing microorganisms present in these foods. One strain was identified and named Bacillus subtilis SCJ1, by using 16S rRNA sequencing and biochemical characterization. We investigated the cell growth of this organism in order to understand its potential for industrial application. To this end, we optimized the culture medium constituents by using the response surface methodology. The Plackett-Burman experimental design was used for screening of the medium constituents, such as molasses, yeast extract and peptone, for improving cell growth. In order to determine the optimal concentration of each constituent, we used a central composite design. Consequently, the optimized concentrations of molasses, yeast extract and peptone were predicted to be 27.5 g/l, 7.5 g/l and 17.5 g/l, respectively. By model verification, we confirmed that a 1.49-fold increase in dry cell weight compared to the basal medium-from 1.32 g/l, to 1.9722 g/l-was achieved.

• Korean Journal of Microbiology
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• v.51 no.1
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• pp.48-60
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• 2015

612 strains isolated from Korean traditional fermented food in Sunchang and their investigated biochemical characterization and ability of biogenic amines non-producing. We selected the SCJ4 having various activity by measurement of extracellular enzyme, antioxidant and antimicrobial activities. Selected strain SCJ4 by 16S rRNA sequencing and biochemical characterization was named Bacillus subtilis SCJ4. And then, we investigated cell growth of SCJ4, and optimized of culture medium constituents using response surface methodology as statistically method. Response surface methodology used Plackett-Burman experimental design for screening of medium constituent. Tryptone, peptone and $MgSO_4$ as medium constituent improving cell growth selected. In order to find out optimal concentration on each constituent, we carried out central composite design. Consequently, optimized concentrations of tryptone, peptone and $MgSO_4$ were predicted to be 15.35 g/L, 12.235 g/L, and 3.5 g/L respectively. Through the model verification, we confirmed about 1.28-fold improvement of the dried cell weight from 0.8767 g/L to 1.1222 g/L when compared to basal medium.

• KSBB Journal
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• v.23 no.6
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• pp.463-469
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• 2008

During the last decade, monacolin-K biosynthesized by fermentation of red yeast rice (Monascus strains) was proved to have an efficient cholesterol lowering capability, leading to rapid increase in the market demand for the functional red yeast rice. In this study, the production medium composition and components were optimized on a shake flask scale for monacolin-K production by Monascus pilosus (KCCM 60160). The effect of three different soybean flours on the monacolin-K production were studied in order to replace the nitrogen sources of basic production medium (yeast extract, malt extract and beef extract). Among the several experiments, the production medium with dietary soybean flour to replace a half of yeast extract was very good for monacolin-K production. Plackett-Burman experimental design was used to determine the key factors which are critical to produce the biological products in the fermentation. According to the result of Plackett-Burman experimental design, a second order response surface design was applied using yeast extract, beef extract and $(NH_4)_2SO_4$ as factors. Applying this model, the optimum concentration of the three variables was obtained. The maximum monacolin-K production (369.6 mg/L) predicted by model agrees well with the experimental value (418 mg/L) obtained from the experimental verification at the optimal medium. The yield of monacolin-K was increased by 67% as compared to that obtained with basic production medium in shake flasks.

• KSBB Journal
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• v.22 no.5
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• pp.305-312
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• 2007

It is crucial to develop a miniaturized cultivation method for large and rapid screening of high-yielding mutants of monacolin-K, a powerful anti-hypercholesterolemic secondary metabolite biosynthesized by the fungal cells of Monascus ruber. In order to investigate as many strains as possible in a short time, a miniaturized fermentation method especially suitable for the cultivation of the filamentous Monascus mutants was developed using $50m{\ell}$ culture-tube ($7m{\ell}$ of working volume) instead of the traditional $250m{\ell}$ flask ($50m{\ell}$ of working volume). Generally, in filamentous fungal cell fermentations, morphologies in growth and production cultures should be maintained as thick filamentous and compact-pelleted (usually less than 1 mm in diameter) forms, respectively, for enhanced production of secondary metabolites in final production cultures. In this study, we intended to induce the respective optimal morphologies in the miniaturized culture system for the purpose of rapid screening of overproducers. Miniaturized growth culture system was successfully developed due to the mass production of spores in the statistically optimized solid medium. When large amounts of spores were inoculated into the growth cultures, and brown rice flour (20 g/L) was also supplemented to the growth medium, dense filamentous morphologies were successfully induced in the growth cultures performed with the 50 ml culture tubes. It was implied that the amounts of spores inoculated into the growth tube-cultures and the growth medium components should be the key factors for the induction of the filamentous forms in the growth fermentations. Furthermore, in order to statistically optimize production medium, multiple experiments based on Plackett-Burman design and response surface method (RSM) were carried out, resulting in more than 2 fold enhanced production of monacolin-K in the final production cultures with the optimized production medium. Notably, under the production culture conditions with the statistically optimized medium, optimal pellet sizes below 1 mm in diameter were reproducibly induced, in contrast to the thick and viscous filamentous morphologies observed in the previous production cultures.

• Journal of Life Science
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• v.22 no.5
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• pp.575-581
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• 2012

This study was conducted to develop an economical optimum medium composition for the mass production of $Lactobacillus$ $plantarum$ and $Lactobacillus$ $reuteri$, livestock probiotics. Medium ingredient factors were selected on the basis of MRS broth composition, and the 15 ingredient variables were as follows: sucrose, glucose, molasses, yeast extract, corn steep liquor, soy peptone, dipotassium phosphate, manganese chloride, magnesium chloride, tween 80, sodium chloride, sodium acetate, ammonium citrate, sodium sulphate, and ferrous sulphate. The Plackett Burman design, consisting of 20 runs, was employed for the analysis of ingredient effects on cell growth of $L.$ $plantarum$ and $L.$ $reuteri$. As a result, sucrose, glucose, molasses, yeast extract, corn steep liquor, soy peptone, sodium acetate, and ammonium citrate positively influenced the growth of $L.$ $plantarum$. Additionally, yeast extract, soy peptone, $K_2PHO_4$, and tween 80 positively influenced the growth of $L.$ $reuteri$. Positive effects were found from sucrose, yeast extract, and soy peptone in the integrated analysis of the effects of both $L.$ $plantarum$ and $L.$ $reuteri$. Finally, effective medium components for both strains were found as follows: sucrose (20.0 g/l), glucose (5.0 g/l), soy peptone (11.0 g/l), yeast extract (5.0 g/l), $K_2PHO_4$ (0.2 g/l), $CH_3COONa$ (2 g/l), and $MgCl_2$ (0.02 g/l).

• Journal of Microbiology and Biotechnology
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• v.18 no.12
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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 of Animal Resources
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• v.30 no.5
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• pp.764-772
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• 2010

Staphylococcus aureus lipase is regarded as a virulence factor. The response of lipase activity to various factors can provide important insights concerning the prevention of S. aureus during meat fermentation. This study was conducted to evaluate the main effects of nutrients used in culture media, and their combined effects on the inhibition of lipase activity and cell growth of pathogenic S. aureus SK1593 isolated from fermented pork meat. A Plackett-Burman design was used to evaluate the main effects of variables, including olive oil, soybean oil, grapeseed oil, sesame oil, $CuSO_4$, $MgCl_2$, $KNO_3$, $CaCl_2$, and KCl. Significant negative effects on lipase activity were detected with soybean oil, grapeseed oil, $KNO_3$, and $CaCl_2$. Additionally, these nutrients were further selected as variables for the investigation of their combined effect on lipase activity, via response surface methodology. In order to confirm the regression model, a situation that only inhibits lipase activity was simulated. The predicted lipase activity and cell growth of the simulated situation were 14.0 U/mL and $9.6\;{\log}_{10}$ (CFU/mL), respectively, and the estimated value of those in the same medium showed 15.14 U/mL and $9.4\;{\log}_{10}$(CFU/mL) respectively. The lipase activity of the simulated medium was inhibited approximately 5-fold as compared to the basal medium, but no significant differences in cell counts were noted to exist between the basal and simulated media. These results suggest that soybean oil, grapeseed oil, $KNO_3$, and $CaCl_2$ can be used to inhibit the growth of pathogenic S. aureus during the process of meat fermentation.

• Journal of Microbiology and Biotechnology
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• v.19 no.4
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• pp.378-386
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• 2009

A sequential optimization strategy, based on statistical experimental designs, is employed to enhance the production of alkaline protease by a Bacillus pseudofirmus local isolate. To screen the bioprocess parameters significantly influencing the alkaline protease activity, a 2-level Plackett-Burman design was applied. Among 15 variables tested, the pH, peptone, and incubation time were selected based on their high positive significant effect on the protease activity. A near-optimum medium formulation was then obtained that increased the protease yield by more than 5-fold. Thereafter, the response surface methodology(RSM) was adopted to acquire the best process conditions among the selected variables, where a 3-level Box-Behnken design was utilized to create a polynomial quadratic model correlating the relationship between the three variables and the protease activity. The optimal combination of the major medium constituents for alkaline protease production, evaluated using the nonlinear optimization algorithm of EXCEL-Solver, was as follows: pH of 9.5, 2% peptone, and incubation time of 60 h. The predicted optimum alkaline protease activity was 3,213 U/ml/min, which was 6.4 times the activity with the basal medium.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.4
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• pp.288-292
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• 2006

Through statistically designed experiments, lysis agents were optimized to effectively disrupt bacterial cells in a microfluidic device. Most surfactants caused the efficient lysis of Gram-positive microbes, but not of Gram-negative bacteria. A Plackett-Burman design was used to select the components that increase the efficiency of the lysis of the Gram-negative bacteria Escherichia coli. Using this experimental design, both lysozyme and benzalkonium chloride were shown to significantly increase the cell lysis efficiency, and ATP was extracted in proportion to the lysis efficiency. Benzalkonium chloride affected the cell membrane physically, while lysozyme destroyed the cell wall, and the amount of ATP extracted increased through the synergistic interaction of these two components. The two-factor response-surface design method was used to determine the optimum concentrations of lysozyme and benzalkonium chloride, which were found to be 202 and 99 ppm, respectively. The lysis effect was further verified by microscopic observations in the microchannels. These results indicate that Gram-negative cells can be lysed efficiently in a microfluidic device, thereby allowing the rapid detection of bacterial cells using a bioluminescence-based assay of the released ATP.