• 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.

• Computers and Concrete
• /
• v.30 no.6
• /
• pp.393-407
• /
• 2022

Due to the enormous cement content, pozzolanic materials, and the use of different aggregates, sustainable controlled low-strength material (CLSM) has a higher material cost than conventional concrete and sustainable construction issues. However, by selecting appropriate materials and formulations, as well as cement and aggregate content, whitethorn costs can be reduced while having a positive environmental impact. This research explores the desire to optimize plastic properties and 28-day unconfined compressive strength (UCS) of CLSM containing powder content from unprocessed-fly ash (u-FA) and recycled fine aggregate (RFA). The mixtures' input parameters consist of water-to-cementitious material ratio (W/CM), fly ash-to-cementitious materials (FA/CM), and paste volume percentage (PV%), while flowability, bleeding, segregation index, and 28-day UCS were the desired responses. The central composite design (CCD) notion was used to produce twenty CLSM mixes and was experimentally validated using MATLAB by an Artificial Neural Network (ANN). Variance analysis (ANOVA) was used for the determination of statistical models. Results revealed that the plastic properties of CLSM improve with the FA/CM rise when the strength declines for 28 days-with an increase in FA/CM, the diameter of the flowability and bleeding decreased. Meanwhile, the u-FA's rise strengthens the CLSM's segregation resistance and raises its strength over 28 days. Using calcareous powder as a substitute for cement has a detrimental effect on bleeding, and 28-day UCS increases segregation resistance. The response surface method (RSM) can establish high correlations between responses and the constituent materials of sustainable CLSM, and the optimal values of variables can be measured to achieve the desired response properties.

• Journal of Microbiology and Biotechnology
• /
• v.18 no.1
• /
• pp.110-117
• /
• 2008

Three kinds of prenylated flavonols, icariside I, icariside II, and icaritin, were isolated from an icariin hydrolysate and their effects on melanogenesis evaluated based on mushroom tyrosinase inhibition and quantifying the melanin contents in melanocytes. Although none of the compounds had an effect on tyrosinase activity, icariside II and icaritin both effectively inhibited the melanin contents with an $IC_{50}$ of 10.53 and $11.13{\mu}M$, respectively. Whereas icariside II was obtained from a reaction with ${\beta}$-glucosidase and cellulase, the icariin was not completely converted into icariside II. Thus, for the high-purity production of icariside II, the reaction was optimized using the response surface methodology, where an enzyme concentration of 5.0mg/ml, pH 7, $37.5^{\circ}C$, and 8 h reaction time were selected as the central conditions for the central composite design (CCD) for the enzymatic hydrolysis of icariin into icariside II using cellulase. Empirical models were developed to describe the relationships between the operating factors and the response (icariside II yield). A statistical analysis indicated that all four factors had a significant effect (p<0.01) on the icariside II production. The coefficient of determination $(R^2)$ was good for the model (0.9853), and the optimum production conditions for icariside II was an enzyme concentration of 7.5mg/ml, pH 5, $50^{\circ}C$, and 12 h reaction time. A good agreement between the predicted and experimental data under the designed optimal conditions confirmed the usefulness of the model. A laboratory pilot scale was also successful.

• 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 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.

• Microbiology and Biotechnology Letters
• /
• v.41 no.1
• /
• pp.52-59
• /
• 2013

The nutritional requirements for the maximum production of lipopeptides by Bacillus subtilis N7 (B. subtilis N7) were investigated and optimized using response surface methodology (RSM) under shake flask fermentation. A one-factor-at-a-time experimental setup was used to screen carbon and nitrogen sources. A Plackett-Burman design (PBD) was employed to screen the most critical variables for lipopeptides production amongst ten nutritional elements. The central composite experimental design (CCD) was finally adopted to elucidate the composition of the fermentation medium. Statistical analyses (analysis of variance, ANOVA) of the results showed that KCl, $MnSO_4$ and $FeSO_4{\cdot}6H_2O$ were important components and that their interactions were strong. Lipopeptide production was predicted to reach 709.87 mg/L after a 60 h incubation using an optimum fermentation medium composed of glucose 7.5 g/L, peanut oil 1.25 g/L, $MgSO_4$ 0.37 g/L, $KH_2PO_4$ 0.75 g/L, monosodium glutamate 6.75 g/L, yeast extract and $NH_4Cl$ (5:3 w/w) 10 g/L, KCl 0.16 g/L, $FeSO_4{\cdot}6H_2O$ 0.24 mg/L, $MnSO_4$ 0.76 mg/L, and an initial pH of 7.0. Lipopeptide production ($706.57{\pm}3.70$ mg/L) in the optimized medium confirmed the validity of the predicted model.

• 한국신재생에너지학회:학술대회논문집
• /
• 2011.11a
• /
• pp.114.1-114.1
• /
• 2011

Bioenergy production from lignocellulosic biomass and agriculture wastes have been attracted because of its sustainable and non-edible source. Especially, canola is considered as one of the best feedstock for renewable fuel production. Oil extracted canola and its agriculture residues are reuseable for bioethanol production. However, a pretreatment step is required before enzymatic hydrolysis to disrupt recalcitrant lignocellulosic matrix. To increase the sugar conversion, more efficient pretreatment process was necessary for removal of saccharification barriers such as lignin. Alkaline pretreatment makes the lignocellulose swollen through solvation and induces more porous structure for enzyme access. In our previous work, aqueous ammonia (1~20%) was utilized for alkaline reagent to increase the crystallinity of canola residues pretreatment. In this study, significant factors for efficient soaking in aqueous ammonia pretreatment on canola residues was optimized by using the response surface method (RSM). Based on the fundamental experiments, the real values of factors at the center (0) were determined as follows; $70^{\circ}C$ of temperature, 17.5% of ammonia concentration and 18 h of reaction time in the experiment design using central composition design (CCD). A statistical model predicted that the highest removal yield of lignin was 54% at the following optimized reaction conditions: $72.68^{\circ}C$ of temperature, 18.30% of ammonia concentration and 18.30 h of reaction time. Finally, maximum theoretical yields of soaking in aqueous ammonia pretreatment were 42.23% of glucose and 22.68% of xylose.

• Transactions of the Korean Society of Mechanical Engineers B
• /
• v.39 no.5
• /
• pp.435-441
• /
• 2015

In this study, a method for optimal design of impeller and diffuser blades in the centrifugal compressor using response surface method (RSM) and multi-objective genetic algorithm (MOGA) was evaluated. A numerical simulation was conducted using ANSYS CFX with various values of impeller and diffuser parameters, which consist of leading edge (LE) angle, trailing edge (TE) angle, and blade thickness. Each of the parameters was divided into three levels. A total of 45 design points were planned using central composite design (CCD), which is one of the design of experiment (DOE) techniques. Response surfaces that were generated on the basis of the results of DOE were used to determine the optimal shape of impeller and diffuser blade. The entire process of optimization was conducted using ANSYS Design Xplorer (DX). Through the optimization, isentropic efficiency and pressure recovery coefficient, which are the main performance parameters of the centrifugal compressor, were increased by 0.3 and 5, respectively.

• Earthquakes and Structures
• /
• v.17 no.5
• /
• pp.511-519
• /
• 2019

The concrete gravity dam is one of the most important parts of the nation's infrastructure. Besides the benefits, the dam also has some potentially catastrophic disasters related to the life of citizens directly. During the lifetime of service, some degradations in a dam may occur as consequences of operating conditions, environmental aspects and deterioration in materials from natural causes, especially from dynamic loads. Cumulative Absolute Velocity (CAV) plays a key role to assess the operational condition of a structure under seismic hazard. In previous researches, CAV is normally used in Nuclear Power Plant (NPP) fields, but there are no particular criteria or studies that have been made on dam structure. This paper presents a method to calculate the limitation of CAV for the Bohyeonsan Dam in Korea, where the critical Peak Ground Acceleration (PGA) is estimated from twelve sets of selected earthquakes based on High Confidence of Low Probability of Failure (HCLPF). HCLPF point denotes 5% damage probability with 95% confidence level in the fragility curve, and the corresponding PGA expresses the crucial acceleration of this dam. For determining the status of the dam, a 2D finite element model is simulated by ABAQUS. At first, the dam's parameters are optimized by the Minitab tool using the method of Central Composite Design (CCD) for increasing model reliability. Then the Response Surface Methodology (RSM) is used for updating the model and the optimization is implemented from the selected model parameters. Finally, the recorded response of the concrete gravity dam is compared against the results obtained from solving the numerical model for identifying the physical condition of the structure.

• 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.