• Microbiology and Biotechnology Letters
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• v.49 no.2
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• pp.181-191
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• 2021

The present study addresses isolation, optimization, partial purification, and characterization of a haloalkaline serine protease from a newly isolated haloarchaeal strain isolated from Wadi El Natrun in Egypt. We expected that a two-step sequential statistical approach (one variable at a time, followed by response surface methodology) might maximize the production of the haloalkaline serine protease. The enzyme was partially purified using Hiprep 16/60 sephacryl S-100 HR gel filtration column. Molecular identification revealed the newly isolated haloarchaeon to be Natrialba hulunbeirensis strain WNHS14. Among several tested physicochemical determinants, casamino acids, KCl, and NaCl showed the most significant effects on enzyme production as determined from results of the One-Variable-At-A-time (OVAT) study. The BoxBehnken design localized the optimal levels of the three key determinants; casamino acids, KCl, and NaCl to be 0.5% (w/v), 0.02% (w/v), and 15% (w/v), respectively, obtaining 62.9 U/ml as the maximal amount of protease produced after treatment at 40℃, and pH 9 for 9 days with 6-fold enhancement in yield. The enzyme was partially purified after size exclusion chromatography with specific activity, purification fold, and yield of 1282.63 U/mg, 8.9, and 23%, respectively. The enzyme showed its maximal activity at pH, temperature, and NaCl concentration optima of 10, 75℃, and 2 M, respectively. Phenylmethylsulfonyl fluoride (PMSF, 5 mM) completely inhibited enzyme activity.

• Journal of Microbiology and Biotechnology
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• v.32 no.7
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• pp.892-901
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• 2022

The rising demand for carotenoids can be met by microbial biosynthesis as a promising alternative to chemical synthesis and plant extraction. Several species of lactic acid bacteria (LAB) specifically produce C₃₀ carotenoids and offer the added probiotic benefit of improved gut health and protection against chronic conditions. In this study, the recently characterized Lactiplantibacillus plantarum subsp. plantarum KCCP11226^T produced the rare C₃₀ carotenoid, 4,4'-diaponeurosporene, and its yield was optimized for industrial production. The one-factor-at-a-time (OFAT) method was used to screen carbon and nitrogen sources, while the abiotic stresses of temperature, pH, and salinity, were evaluated for their effects on 4,4'-diaponeurosporene production. Lactose and beef extract were ideal for optimal carotenoid production at 25℃ incubation in pH 7.0 medium with no salt. The main factors influencing 4,4'-diaponeurosporene yields, namely lactose level, beef extract concentration and initial pH, were enhanced using the Box-Behnken design under response surface methodology (RSM). Compared to commercial MRS medium, there was a 3.3-fold increase in carotenoid production in the optimized conditions of 15% lactose, 8.3% beef extract and initial pH of 6.9, producing a 4,4'-diaponeurosporene concentration of 0.033 A₄₇₀/ml. To substantiate upscaling for industrial application, the optimal aeration rate in a 5 L fermentor was 0.3 vvm. This resulted in a further 3.8-fold increase in 4,4'-diaponeurosporene production, with a concentration of 0.042 A₄₇₀/ml, compared to the flask-scale cultivation in commercial MRS medium. The present work confirms the optimization and scale-up feasibility of enhanced 4,4'-diaponeurosporene production by L. plantarum subsp. plantarum KCCP11226^T.

• Journal of Electrochemical Science and Technology
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• v.15 no.3
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• pp.388-404
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• 2024

In the present work, the operating parameters were optimized using Box Behnken Design (BBD) in response surface methodology (RSM) to maximize the hydrogen production rate (R₁) and hydrogen production rate per unit watt consumed (R₂) of a proton exchange membrane electrolysis cell (PEMEC), a third response (R₃) which was the sum of the scaled values of R₁ and R₂ were selected to be maximized so that both hydrogen production rate and hydrogen production rate per unit watt consumed could be maximized. The major parameters which were influencing the experiment for enhancing the output responses were oxygen electrode/anode electrocatalyst loading (A), current supplied (B) and water inlet temperature (C). The commercial proton exchange membrane Nafion^® was used as the electrolyte. The acetylene black carbon (C_AB) supported IrO₂ was used as the electrocatalyst for preparing oxygen electrode/anode whereas commercial Pt (40 wt%)/C_HSA was used as the H₂ electrode/cathode electrocatalyst. The quadratic model was developed to predict the output/ responses and their proximity to the experimental output values. The developed model was found to be significant as the P values for both the responses were < 0.0001 and F values were greater than 1. The optimum condition for both the responses were O₂ electrode/anode electrocatalyst loading of 1.78 mg/cm², supplied current of 0.33 A and water inlet temperature of 54℃. The predicted values for hydrogen production rate (R₁) and hydrogen production rate per unit watt consumed (R₂) were 2.921 mL/min and 2.562 mL/(min·W), respectively obtained from the quadratic model. The error % between the predicted response values and experimental values were 1.47% and 3.08% for R₁ and R₂, respectively. This model predicted the optimum conditions reasonably in good agreement with the experimental conditions for the enhancement of the output responses of the developed PEM based electrolyser.

• Journal of Microbiology and Biotechnology
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• v.23 no.2
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• pp.211-217
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• 2013

Corynebacterium glutamicum is one of the well-studied industrial strain that is used for the production of nucleotides and amino acids. Recently, it has also been studied as a possible producer of organic acids such as succinic acid, based on its ability to produce organic acids under an oxygen deprivation condition. In this study, we conducted the optimization of medium components for improved succinate production from C. glutamicum under an oxygen deprivation condition by Plackett-Burman design and applied a response surface methodology. A Plackett-Burman design for ten factors such as glucose, ammonium sulfate, magnesium sulfate, potassium phosphate ($K_2HPO_4$ and $KH_2PO_4$), iron sulfate, manganese sulfate, biotin, thiamine, and sodium bicarbonate was applied to evaluate the effects on succinate production. Glucose, ammonium sulfate, magnesium sulfate, and dipotassium phosphate were found to have significant influence on succinate production, and the optimal concentrations of these four factors were sequentially investigated by the response surface methodology using a Box-Behnken design. The optimal medium components obtained for achieving maximum concentration of succinic acid were as follows: glucose 10 g/l, magnesium sulfate 0.5 g/l, dipotassium phosphate ($K_2HPO_4$) 0.75 g/l, potassium dihydrogen phosphate ($KH_2PO_4$) 0.5 g/l, iron sulfate 6 mg/l, manganese sulfate 4.2 mg/l, biotin 0.2 mg/l, thiamine 0.2 mg/l, and sodium bicarbonate 100 mM. The parameters that differed from a normal BT medium were glucose changed from 40 g/l to 10 g/l, dipotassium phosphate ($K_2HPO_4$) 0.5 g/l changed to 0.75 g/l, and ammonium sulfate ($(NH_4)_2SO_4$) 7 g/l changed to 0 g/l. Under these conditions, the final succinic acid concentration was 16.3 mM, which is about 1.46 fold higher than the original medium (11.1 mM) at 24 h. This work showed the improvement of succinate production by a simple change of media components deduced from sequential optimization.

• Journal of The Korean Society of Agricultural Engineers
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• v.62 no.2
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• pp.17-29
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• 2020

The objective of this study was to find an optimum methane yield condition in anaerobic digestion of sewage sludge with microwave pretreatment. The pretreatment process was carried out using a lab scale industrial microwave unit (2,450 MHz frequency). The digestion efficiency of pretreated sludge was evaluated by biochemical methane potential (BMP) test. Box-Behnken design and Response Surface Analysis (RSA) were applied to determine the optimal combination of sludge mixing ratio (0 to 100%), power (400 to 1600 W), holding time (0 to 10 min) and pretreatment temperature (60 to 100℃). BMP test results showed that Volatile Solid (VS) removal efficiency was up to 48% at a condition of 0% for mixing ratio, 1600 W for power, 5 min for holding time, and 80℃ for pretreatment temperature. Methane production was up to 832.3 mL/g VS_removed at a condition of 50% for mixing ratio, 1000 W for power, 5 min for holding time, and 80℃ for pretreatment temperature. The results of the variance analysis (ANOVA) showed that the p-value of the power and pretreatment temperature among the independent variables were significant (p<0.05), and in particular, the pretreatment temperature significantly affected on the solubilization and methane production. The optimum condition for the maximum methane yield (847 mL/g VS_removed) was consist of 38.4% of mixing ratio, 909.1 W of power, 4.1 min of holding time, and 80℃ of temperature within the design boundaries.

• Korean Journal of Food Science and Technology
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• v.53 no.2
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• pp.187-194
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• 2021

This study aimed to optimize the preparation method of citron (Citrus junos Sieb.) beverages with hibiscus using response surface methodology (RSM). The experimental conditions were established using a central composite design with three independent variables as follows: ratios of citron (40~60%), citric acid (0.34~0.94%), and hibiscus (0.3~0.7%). The results indicate that an increase in the citron ratio contributed to increased sweetness and as the concentration of citron and hibiscus increased, the brightness of the citron beverage decreased and the yellowness increased. The citron ratio showed a significant correlation with the ABTS radical scavenging capacity. Among the 15 experimental groups, 4 representative samples showing statistical significance were selected, and sensory tests were performed, in comparison with commercially available products. As a result of the sensory test, four beverages prepared with the selected recipes showed higher preference than commercial beverages, and optimal recipe conditions were 40% citron, 0.34% citric acid, and 0.5% hibiscus.

• Journal of Life Science
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• v.18 no.10
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• pp.1377-1383
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• 2008

A bacterium producing non- or partially digestible dextran was isolated from kimchi broth by enrichment culture technique. The bacterium was identified tentatively as Leuconostoc sp. strain SKY. We established the response surface methodology (Box-Behnken design) to optimize the principle parameters such as culture pH, temperature, and yeast extract concentration for maximizing production of dextran. The ranges of parameters were determined based on prior screening works done at our laboratory and accordingly chosen as 5.5, 6.5, and 7.5 for pH, 25, 30, and $35^{\circ}C$ for temperature, and 1, 5, and 9 g/l yeast extract. Initial concentration of sucrose was 100 g/l. The mineral medium consisted of 3.0 g $KH_2PO_4$, 0.01 g $FeSO_4{\cdot}H_2O$, 0.01 g $MnSO_4{\cdot}4H_2O$, 0.2 g $MgSO_4{\cdot}7H_2O$, 0.01 g NaCl, and 0.05 g $CaCO_3$ per 1 liter deionized water. The optimum values of pH and temperature, and yeast extract concentration were obtained at pH (around 7.0), temperature (27 to $28^{\circ}C$), and yeast extract (6 to 7 g/l). The best dextran yield was 60% (dextran/g sucrose). The best dextran productivity was 0.8 g/h-l.

• Journal of Life Science
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• v.28 no.4
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• pp.483-487
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• 2018

Cypermethrin, a commonly used domestic and agricultural pyrethroid pesticide, is widely considered detrimental to the environment and to many organisms because of its residual property and toxicity. Cellulophaga lytica DAU203, isolated from coastal sediment, was chosen because it degrade cypermethrin. Cellulophaga lytica DAU203 effectively degraded cypermethrin, as the utilized carbon source and substrate, in a mineral salt medium. Effective factors, such as carbon source, nitrogen source, initial pH, and temperature, for cypermethtin biological degradation by Cellulophaga lytica DAU203 were analyzed by one factor at a time method. Temperature ($22{\sim}42^{\circ}C$), initial pH (5~9), and yeast extract concentration (0.1~2.5%[w/v]) were selected as the three most important factors. There were optimized at $33.4^{\circ}C$, pH 7.7, and 2.4%(w/v) by response surface methodology, respectively. The Box- Behnken design consisting of 46 experimental runs with three replicates was used to optimize the independent variables which significantly influenced the cypermethrin biological degradation. This model for cypermethrin degradation by Cellulophaga lytica DAU203 is highly significant (p<0.05). Under the optimized condition, Cellulophaga lytica DAU203 degraded approximately 83.7 % of the cypermethrin within 5 days. These results suggest that Cellulophaga lytica DAU203 may be useful for the biological degradation of cypermethrin in cypermethrin-contaminated environments.

• Journal of The Korean Society of Grassland and Forage Science
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• v.33 no.1
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• pp.58-66
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• 2013

This study was conducted in order to develop slaughterhouse rumen content (SRC) as a potential feed additive. The moisture content of SRC can reach 80%, and therefore an appropriate dewatering process is required before it can be used. In this study, the effects of heating temperature, heating time, and filler content during the dewatering process on the activity of various enzymes in SRC were investigated. The Box-Behnken experimental design was employed, involving a total of 45 experimental runs, consisting of three variables (heating time, heating temperature, and filler content) with three levels per variable (12, 30 and 48 hr; 60, 75 and $90^{\circ}C$; 12, 22.5 and 33% for heating time, heating temperature, and filler content, respectively). For enzyme activities, xylanase, cellulase, and amylase were examined, and the results were subjected to an analysis of variance. Heating time, heating temperature and filler content had significant effects on the activity of each enzyme (p<0.05). Cellulase and amylase activities decreased (p<0.05) at elevated heating temperatures, whereas xylanase was reasonably stable around $90^{\circ}C$. The activities of all enzymes decreased (p<0.05) with increased heating time. Optimum filler contents for xylanase, cellulase, and amylase activities were 22.5, 12 and 33%, respectively. However, optimum conditions for all variables that simultaneously maximize the activity of all three enzymes could not be ascertained in this study. Nevertheless, the results from the current study can be useful as basic information for the development of SRC as a feed additive enriched with improved major enzymes for livestock feed digestion.

• Steel and Composite Structures
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• v.47 no.1
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• pp.91-102
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• 2023

To study the evaluation standard and control limit of mortar filling layer void length, in this paper, the train sub-model was developed by MATLAB and the track-bridge sub-model considering the mortar filling layer void was established by ANSYS. The two sub-models were assembled into a train-track-bridge coupling dynamic model through the wheel-rail contact relationship, and the validity was corroborated by the coupling dynamic model with the literature model. Considering the randomness of fastening stiffness, mortar elastic modulus, length of mortar filling layer void, and pier settlement, the test points were designed by the Box-Behnken method based on Design-Expert software. The coupled dynamic model was calculated, and the support vector regression (SVR) nonlinear mapping model of the wheel-rail system was established. The learning, prediction, and verification were carried out. Finally, the reliable probability of the amplification coefficient distribution of the response index of the train and structure in different ranges was obtained based on the SVR nonlinear mapping model and Latin hypercube sampling method. The limit of the length of the mortar filling layer void was, thus, obtained. The results show that the SVR nonlinear mapping model developed in this paper has a high fitting accuracy of 0.993, and the computational efficiency is significantly improved by 99.86%. It can be used to calculate the dynamic response of the wheel-rail system. The length of the mortar filling layer void significantly affects the wheel-rail vertical force, wheel weight load reduction ratio, rail vertical displacement, and track plate vertical displacement. The dynamic response of the track structure has a more significant effect on the limit value of the length of the mortar filling layer void than the dynamic response of the vehicle, and the rail vertical displacement is the most obvious. At 250 km/h - 350 km/h train running speed, the limit values of grade I, II, and III of the lengths of the mortar filling layer void are 3.932 m, 4.337 m, and 4.766 m, respectively. The results can provide some reference for the long-term service performance reliability of the ballastless track-bridge system of HRS.