• Analytical Science and Technology
• /
• v.32 no.2
• /
• pp.48-55
• /
• 2019

The simple and effective analytical method for the quality control of a novel scalp tonic formulation has been developed and optimized in terms of HPLC conditions and sample preparation method, meanwhile, the optimization of preparation condition was using response surface methodology (RSM) based on central composite design (CCD). Oleanolic acid was selected as marker compound because of its bioactivities for alopecia therapy. The developed analytical method and extraction condition were successfully qualified. Coefficient of determination ($r^2$) for the calibration was 0.9997 with a line passing through the origin point in the range of 0.1-100 mg/mL. The limit of detection (LOD) and the limit of quantitation (LOQ) were 17.5 ng/mL and 55.0 ng/mL, respectively. The intra-day and inter-day precision of the method were 0.5-1.4 % and 0.7-1.8 % in relative standard deviation, respectively, while those accuracy were 99.5-100.9 % and 100.0-102.2 %, respectively. The repeatability of oleanolic acid in samples ranged of 0.3-1.9 % based on peak area and 0.3-0.7 % for retention time. Recoveries from samples were 95.0-99.4 % with lower than 1.8 % in relative standard deviation. Overall, the developed analytical method will be used for quality control of this commercial scalp tonic products successfully.

• Microbiology and Biotechnology Letters
• /
• v.49 no.1
• /
• pp.45-56
• /
• 2021

Improved amylases were developed from protoplast fusants of two amylase-producing Aspergillus species. Twenty regenerated fusants were screened for amylase production using Remazol Brilliant Blue agar. Crude enzyme extracts produced by solid state fermentation of rice bran were assayed for activity. Three variable factors (temperature, pH and enzyme type) were optimized to increase the amylase activity of the parents and selected fusants using rice bran medium and solid state fermentation. Analysis of this optimization was completed using the Central Composite Design (CCD) of the Response Surface Methodology (RSM). Amylase activity assays conducted at room temperature and 80℃ demonstrated that Aspergillus designates, T5 (920.21 U/ml, 966.67 U/ml), T13 (430 U/ml, 1011.11 U/ml) and T14 (500.63 U/ml, 1012.00 U/ml) all exhibited improved function making them the preferred fusants. Amylases produced from these fusants were observed to be active over the entire pH range evaluated in this study. Fusants T5 and T14 demonstrated optimal activity under acidic and alkaline conditions, respectively. Fusants T13 and T14 produced the most amylase at 72 h while parents TA, TC and fusant T5 produced the most amylase after 96 h of incubation. Response surface methodology examinations revealed that the enzyme from fusant T5 was the optimal enzyme demonstrating the highest activity (1055.17 U/ml) at pH 4 and a temperature of 40℃. This enzyme lost activity with further increases in temperature. Starch hydrolysis using fusant T5 gave the highest yield of glucose (1.6158 g/100 ml). The significant activities of the selected fusants at 28 ± 2℃ and 80℃ and the higher sugar yields from cassava starch hydrolysis over their parental strains indicate that it is possible to improve amylase activity using the protoplast fusion technique.

• Korean Chemical Engineering Research
• /
• v.60 no.1
• /
• pp.70-79
• /
• 2022

Biodiesel production has attracted attention as a sustainable source of fuel and is a competitive alternate to diesel engines. The glycerol that is produced as a by-product is generally discarded as waste and can be converted to green chemicals such as acetins to increase bio-diesel profitability. Acetins find application in fuel, food, pharmaceutical and leather industries. Batch experiments and analysis have been previously conducted for synthesis of acetins using glycerol esterification reaction aided by sulfated metal oxide catalysts (SO₄^2-/CeO₂-ZrO₂). The aim of this study was to optimize process parameters: effects of mole ratio of reactants (glycerol and acetic acid), catalyst concentration and reaction temperature to maximize glycerol conversion/acetin selectivity. The optimum conditions for this reaction were determined using response surface methodology (RSM) designed as per a five-level-three-factor central composite design (CCD). Statistica software 10 was used to analyze the experimental data obtained. The optimized conditions obtained were molar ratio - 1:12, catalyst concentration - 6 wt.% and temperature -90 ℃. A packed bed reactor was fabricated and column studies were performed using the optimized conditions. The breakthrough curve was analyzed.

• Journal of the Korean Society of Food Culture
• /
• v.23 no.2
• /
• pp.243-251
• /
• 2008

This purpose of this study was to develop a functional muffin by adding yam powder in the shape of a muffin as a partial surrogate for wheat flour. The yam has been found to be effective for liver and kidney function, as well as the digestion of protein, since it produces glucuronic acid in the body. Therefore, the purpose of this study was to determine the optimal mixing conditions of yam muffins by adjusting the amounts yam powder, butter, and sugar. The mixing conditions for the yam muffins included 3 categories: yam powder $(X_1)$, sugar $(X_2)$, and butter $(X_3)$ by Central Composite Design (CCD) which was optimized by Response Surface Methodology (RSM). The effects of the three variable additions on muffin quality were examined via physical and chemical experiments, such as the analysis of texture (hardness, cohesiveness, springiness, gumminess), coloration (lightness, redness, yellowness), and height. Lastly, we performed a sensory test, which revealed significant findings for gumminess, color, appearance, flavor, softness (p<0.05), redness, and overall quality (p<0.01). Consequently, the optimal mixing rate which best satisfied the sensory items were 34.35g of yam powder, 80.15 g of sugar, and 80.55 g of butter.

• Journal of Microbiology and Biotechnology
• /
• v.33 no.11
• /
• pp.1531-1541
• /
• 2023

Lipase is a well-known and highly in-demand enzyme. During the last decade, several lipase optimization studies have been reported. However, production costs have always been a bottleneck for commercial-scale microbial enzyme production. This research aimed to optimize the conditions for lipase production by Limtongozyma siamensis DMKU-WBL1-3 via a One-Factor-At-a-Time (OFAT) approach combined with statistical methods while using a low-cost substrate. Results suggest that low-cost substrates can be substituted for all media components. An optimal medium was found, using response surface methodology (RSM) and central composite design (CCD), to consist of 0.50% (w/v) sweet whey, 0.40% (w/v) yeast extract (food grade), and 2.50% (v/v) palm oil with the medium pH adjusted to 4 under shaking flask cultivation. From an economic point of view, this work was successful in reducing production costs while increasing lipase productivity. The medium costs were reduced by 87.5% of the original cost while lipase activity was increased by nearly 6-fold. Moreover, lipase production was further studied in a 2-L stirred-tank fermentor. Its activity was 1,055.6 ± 0.0 U/ml when aeration and agitation rates were adjusted to 1 vvm and 170 rpm, respectively. Interestingly, under this optimal lipase production, the yeast showed accumulated lipids inside the cells. The primary fatty acid is a monounsaturated fatty acid (MUFA) that is typically linked to health benefits. This study hence reveals promising lipase production and lipid accumulation by L. siamensis DMKU-WBL1-3 that are worthy of further study.

• Journal of Life Science
• /
• v.25 no.6
• /
• pp.680-685
• /
• 2015

Previously, cellulase and xylanase producing microorganism, Bacillus subtilis NC1, was isolated from soil. Based on the 16S rRNA gene sequence and API 50 CHL test the strain was identified as Bacillus subtilis, and named as B. subtilis NC1. We cloned and sequenced the genes for cellulase and xylanase. Plus, the deduced amino acid sequences from the genes of cellulase and xylanase were determined and were also identified as glycosyl hydrolases family (GH) 5 and 30, respectively. In this study to optimize the medium parameters for cellulase production by B. subtilis NC1 the RSM (response surface methodology) based on CCD (central composite design) model was performed. Three factors, tryptone, yeast extract, and NaCl, for N or C source were investigated. The cellulase activity was measured with a carboxylmethyl cellulose (CMC) plate and the 3,5-dinitrosalicylic acid (DNS) methods. The coefficient of determination (R²) for the model was 0.960, and the probability value (p=0.0001) of the regression model was highly significant. Based on the RSM, the optimum conditions for cellulase production by B. subtilis NC1 were predicted to be tryptone of 2.5%, yeast extract of 0.5%, and NaCl of 1.0%. Through the model verification, cellulase activity of Bacillus subtilis NC1 increased from 0.5 to 0.62 U/ml (24%) compared to the original medium.

• Korean Journal of Microbiology
• /
• v.54 no.4
• /
• pp.384-397
• /
• 2018

The aim of this study is to screen the strains of Bacillus spp. possessing safety, probiotic activity, and so on, which can be utilized as probiotic resource for using the feed and supplement food of companion animal. About 300 isolates were isolated from traditional Korean sauces, four isolates that did not have or produce the six kinds of B. cereus type vomiting and diarrhea toxin genes, ${\beta}$-hemolytic, and three kinds of carcinogenic enzymes were selected. Antibiotic gene retention, cell surface hydrophobicity, antibiotic sensitivity, and glucose utilization were analyzed for four isolates, and finally SRCM 100731 was selected. SRCM 100731 was named as Bacillus amyloliquefaciens SRCM 100731 16S rRNA sequencing analysis, and carried out optimization of cell growth for industrial applications such as pet food and feed. The effects of 14 different components on cell growth were investigated and three significant positive factors, molasses, sodium chloride, and potassium chloride were selected as the main factors based on a Plackett-Burman design. In order to find out optimal concentration on each constituent, we carried out central composite design. The predicted optimized concentrations were 7% molasses, 1.1% sodium chloride, 0.5% potassium chloride. Finally, an overall about 7-fold increase in dry cell weight yield ($12.6625{\pm}0.0658g/L$) was achieved using the optimized medium compared with the non-optimized medium ($1.8273{\pm}0.0214g/L$). This research is expected to be highly utilized in the growing pet industry by establishing optimal cultivation conditions for industrial application as well as screening Bacillus amyloliquefaciens SRCM 100731 as probiotic resource for companion animal.

• Journal of Life Science
• /
• v.21 no.8
• /
• pp.1083-1093
• /
• 2011

A microorganism utilizing rice hulls as a substrate for the production of carboxymethylcellulase (CMCase) was isolated from seawater and identified as Bacillus lincheniformis by analyses of its 16S rDNA sequences. The optimal carbon and nitrogen sources for production of CMCase were found to be rice hulls and ammonium nitrate. The optimal conditions for cell growth and the production of CMCase by B. lincheniformis LBH-52 were investigated using the response surface method (RSM). The analysis of variance (ANOVA) of results from central composite design (CCD) indicated that a highly significant factor ("probe>F" less than 0.0001) for cell growth was rice hulls, whereas those for production of CMCase were rice hulls and initial pH of the medium. The optimal conditions of rice hulls, ammonium nitrate, initial pH, and temperature for cell growth extracted by Design Expert Software were 48.7 g/l, 1.8 g/l, 6.6, and 35.7$^{\circ}C$, respectively, whereas those for the production of CMCase were 43.2 g/l, 1.1 g/l, 6.8, and 35.7$^{\circ}C$. The maximal production of CMCase by B. lincheniformis LBH-52 from rice hulls under optimized conditions was 79.6 U/ml in a 7 l bioreactor. In this study, rice hulls and ammonium nitrate were developed to be substrates for the production of CMCase by a newly isolated marine microorganism, and the time for production of CMCase was reduced to 3 days using a bacterial strain with submerged fermentation.

• Journal of the Korean Wood Science and Technology
• /
• v.38 no.3
• /
• pp.191-198
• /
• 2010

This research was performed to investigate the effect of hydrogen peroxide on the stain removal in japanese hackberry. Response surface method (RSM) was used to optimize the bleaching conditions such as reaction temperature, reaction time and the concentration of hydrogen peroxide. Fifteen different bleaching conditions were selected according to $2^3$ factorial central composite design (CCD). The bleaching effect were evaluated by lightness differences of wood surface before and after the bleaching. The RSM model was determined and its $R^2$ values were 0.93, showing it well represented the bleaching effect. The most affecting factor on the stain removal was the concentration of hydrogen peroxide, followed by reaction time and reaction temperature. Second degree of concentration was proved to have an effect on the bleaching. Bleaching rates above 3% concentrations of hydrogen peroxide were tended to be slightly decreased, and low bleaching effect was found at $20^{\circ}C$. The determined RSM model may offer very practical ways to obtain the desired levels of bleaching because it offers multiple solutions.

• Korean Journal of Food Science and Technology
• /
• v.41 no.3
• /
• pp.245-250
• /
• 2009

This study examined the optimization of alcohol extraction conditions for maximizing the total polyphenols derived from apple pomace, by response surface methodology (RSM). The effects of four independent variables, including $X_1$ (ratio of solvent to sample content), $X_2$ (dipping time), $X_3$ (extraction time), and $X_4$(extraction temperature), were investigated at five levels using central composite design (CCD). $Y_1$ (yield) and $Y_2$ (total polyphenols) were chosen as dependent variables. The coefficients of determination, $R^2$, were greater than 0.900 (0.9042 and 0.9555). The results showed that the model fit was very significant (p<0.001). The optimum extraction conditions were as follows: 13.00 mL/g for the ratio of solvent to sample content, 89.02 min for dipping time, 180 min for extraction time, and $70^{\circ}C$ for extraction temperature. At these conditions, the predicted total polyphenol content was 29.68 mg catechin equiv./g.