• Journal of Microbiology and Biotechnology
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• v.23 no.8
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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.

• Microbiology and Biotechnology Letters
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• v.45 no.4
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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.

• Food Science and Biotechnology
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• v.15 no.6
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• pp.825-832
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• 2006

The effect of alginate coating in combination with an anti-browning agent on increasing the post-cutting shelf life and improving the quality of minimally processed apple cubes was studied during storage at room temperature for 5 days. A simple coating technique involving the chemical cross-linkage of alginate by calcium was used. Statistical-based experimental designs were applied to improve the quality of the alginate-coated apple cubes (ACAC). Plackett-Burman design was first used to determine the main factors influencing the preservation of the original weight, color, and texture of ACAC. Among these variables, alginate concentration ($X_1$), dipping time ($X_2$), and dipping temperature ($X_3$) significantly influenced the ACAC weight and color (confidence levels above 90%). Subsequently, the effects of the 3 main factors were further investigated by a central composite design. The polynomial models developed by response surface methodology were adequate to describe the relationships between the studied factors and the responses. Overall optimization conducted by superimposing the curves of the responses enabled the determination of an optimal range of the independent variables in which the five responses were simultaneously optimized. The point chosen as representative of this optimal area corresponded to $X_1=2.98%$, $X_2=0.85\;min$, and $X_3=55^{\circ}C$ and under these conditions the model predicted weight loss=0.522%, relative hardness=1.517, ${\Delta}E=1.423$, browning inhibition=93.403%, and ${\Delta}L=0.158$.

• Journal of Microbiology and Biotechnology
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• v.23 no.12
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• pp.1757-1764
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• 2013

Trichoderma harzianum ZF-2 producing laccase was isolated from decaying samples from Shandong, China, and showed dye decolorization activities. The objective of this study was to optimize its culture conditions using a statistical analysis of its laccase production. The interactions between different fermentation parameters for laccase production were characterized using a Plackett-Burman design and the response surface methodology. The different media components were initially optimized using the conventional one-factor-at-a-time method and an orthogonal test design, and a Plackett-Burman experiment was then performed to evaluate the effects on laccase production. Wheat straw powder, soybean meal, and $CuSO_4$ were all found to have a significant influence on laccase production, and the optimal concentrations of these three factors were then sequentially investigated using the response surface methodology with a central composite design. The resulting optimal medium components for laccase production were determined as follows: wheat straw powder 7.63 g/l, soybean meal 23.07 g/l, $(NH_4)_2SO_4$ 1 g/l, $CuSO_4$ 0.51 g/l, Tween-20 1 g/l, $MgSO_4$ 1 g/l, and $KH_2PO_4$ 0.6 g/l. Using this optimized fermentation method, the yield of laccase was increased 59.68 times to 67.258 U/ml compared with the laccase production with an unoptimized medium. This is the first report on the statistical optimization of laccase production by Trichoderma harzianum ZF-2.

• Journal of Microbiology and Biotechnology
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• v.21 no.9
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• pp.960-967
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• 2011

Tannin acyl hydrolase, also known as tannase, is an enzyme with important applications in the food, feed, pharmaceutical, and chemical industries. However, despite a growing interest in the catalytic properties of tannase, its practical use is very limited owing to high production costs. Several studies have already demonstrated the advantages of solid-state fermentation (SSF) for the production of fungal tannase, yet the optimal conditions for enzyme production strongly depend on the microbial strain utilized. Therefore, the aim of this study was to improve the tannase production by a locally isolated A. niger strain in an SSF system. The SSF was carried out in packed-bed bioreactors using polyurethane foam as an inert support impregnated with defined culture media. The process parameters influencing the enzyme production were identified using a Plackett-Burman design, where the substrate concentration, initial pH, and incubation temperature were determined as the most significant. These parameters were then further optimized using a Box-Behnken design. The maximum tannase production was obtained with a high tannic acid concentration (50 g/l), relatively low incubation temperature ($30^{\circ}C$), and unique low initial pH (4.0). The statistical strategy aided in increasing the enzyme activity nearly 1.97-fold, from 4,030 to 7,955 U/l. Consequently, these findings can lead to the development of a fermentation system that is able to produce large amounts of tannase in economical, compact, and scalable reactors.

• Journal of Microbiology and Biotechnology
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• v.31 no.5
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• pp.717-725
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• 2021

This study aimed to optimize medium composition and culture conditions for enhancing the biomass of Lactobacillus plantarum 200655 using statistical methods. The one-factor-at-a-time (OFAT) method was used to screen the six carbon sources (glucose, sucrose, maltose, fructose, lactose, and galactose) and six nitrogen sources (peptone, tryptone, soytone, yeast extract, beef extract, and malt extract). Based on the OFAT results, six factors were selected for the Plackett-Burman design (PBD) to evaluate whether the variables had significant effects on the biomass. Maltose, yeast extract, and soytone were assessed as critical factors and therefore applied to response surface methodology (RSM). The optimal medium composition by RSM was composed of 31.29 g/l maltose, 30.27 g/l yeast extract, 39.43 g/l soytone, 5 g/l sodium acetate, 2 g/l K₂HPO₄, 1 g/l Tween 80, 0.1 g/l MgSO₄·7H₂O, and 0.05 g/l MnSO₄·H₂O, and the maximum biomass was predicted to be 3.951 g/l. Under the optimized medium, the biomass of L. plantarum 200655 was 3.845 g/l, which was similar to the predicted value and 1.58-fold higher than that of the unoptimized medium (2.429 g/l). Furthermore, the biomass increased to 4.505 g/l under optimized cultivation conditions. For lab-scale bioreactor validation, batch fermentation was conducted with a 5-L bioreactor containing 3.5 L of optimized medium. As a result, the highest yield of biomass (5.866 g/l) was obtained after 18 h of incubation at 30℃, pH 6.5, and 200 rpm. In conclusion, mass production by L. plantarum 200655 could be enhanced to obtain higher yields than that in MRS medium

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.153-156
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• 2003

The MRS medium is widely used as an optimized medium for the growth of Lactobacillus spp. and also used for the growth of Leuconostoc spp. Leuconostoc mesenteroides shows quite different physicochemical properties compared to Lactobacilli spp. and it is one of the major strain of kimchi fermenting microorganisms with its usefulness in our traditional foods and availability in biotechnology in the future, specifically tailor-made medium is necessary for the growth of Leuconostoc mesenteroides. Sequential experimental designs (Plackett-Burman, fractional factorial, steepest ascent, central composite design and response surface methodology) were introduced to optimize and improve the Leuconostoc medium. Fifteen medium ingredients were investigated and fructose, sodium acetate and ammonium citrate were determined to give a critical and positive effect for cell-growth. The yield of biomass using the optimal medium was improved more than that of the MRS medium and the result of fed-batch culture showed the capability of the improvement in cell mass similar to the E.coli system.

• Proceedings of the Korean Society for Quality Management Conference
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• 2004.04a
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• pp.513-516
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• 2004

Evolutionary operation is useful to improve on-line full-scale manufacturing processes by systematically changing the levels of the process variables while meeting production schedule. Evolutionary operation was developed using two or three process variables for process operators who are not good at statistics. Recently, when a product is developed, it is very important for the engineers to make the production line stable as soon as possible. And there are many causes which have influences to the product performance. This paper presents an evolutionary operation procedure with many process variables using saturated two level fractional factorial designs including Plackett-Burman design.

• Journal of Microbiology and Biotechnology
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• v.16 no.9
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• pp.1410-1415
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• 2006

Media components, including date juice, sodium acetate, peptone, and $K_{2}HPO_4$, which were screened by Plackett-Burman fractional factorial design, were optimized for lactic acid production from date juice using the response surface method (RSM). Sodium acetate, peptone (p<0.0001), and $K_{2}HPO_4$ (p=0.0029) were highly significant in influencing the lactic acid production. Close correlationship between predicted and experimental values was observed. When the optimum values of the parameters obtained through RSM (25.0 g/l date sugar, 15.0 g/l sodium acetate, 19.1 g/l peptone, and 4.7 g/l $K_{2}HPO_4$) were applied, lactic acid production (22.7 g/l) increased by 50.33%, compared with unoptimized media (15.1 g/l). The subsequent validation experiments confirmed the validity of the statistical model.

• 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.