Box-Behnken Technique Research Articles

Preparation and Optimization of Beta-Sitosterol Nanosuspension-Loaded In situ Gel by Using Box-Behnken Model for the Treatment of Prostate Cancer

Introduction: Prostate cancer is the second most often occurring cancer in males and the fourth most common cancer overall. Beta-sitosterol (β-Sit), the most prevalent plant phytosterol found in several plant species, has been reported to have inhibitory effects against several malignancies. Even though β-Sit has considerable potential, its therapeutic uses are limited due to its poor aqueous solubility (<0.1mg/ml), low bioavailability (0.41%), and poor absorption from GIT. Nanosuspension is one of the most innovative approaches to address problems linked to low solubility and poor absorption. Method: In the present research work, β-Sit nanosuspension has been fabricated by nanoprecipitation- ultrasonication, followed by high-pressure homogenization (Panda plus 2000) employing HPMC E5 and poloxamer (188 and 407) as stabilizers, optimized using a Box-Behnken technique. Subsequently, in situ nano gel was prepared by dispersion method using gellan gum as an ionsensitive polymer by incorporating optimized nanosuspension. Results: The optimized nanosuspension was evaluated for various parameters and has been found to have an average particle size (137 ± 5.07 nm), zeta potential (-24±4.99), PDI (0.207), and improved solubility up to 5 folds, being suitable for systemic absorption through the nasal route. The optimized in situ gel was characterized and showed the desired viscosity, good spreadability, acceptable gelation property, and sufficient mucoadhesive strength to adhere to nasal mucosa after ionic interaction. The in vitro release of pure drug, nanosuspension, and the optimized gel was compared, and optimized in situ gel showed a maximum release of 91.41 ± 1.32% up to 8 hours. Conclusion: It was concluded that the in situ nasal nanogel could be the best possible approach to delivering β-Sit into systemic circulation for the treatment of prostate cancer.

Optimization of Alkali Treatment Conditions of Ramie Fabrics Using Box–Behnken Method

Ramie is a widely used plant fiber for making textiles and reinforcement in biodegradable composites. Pretreating cellulosic fibers with alkali before producing composites is increasingly used to enhance adhesion with polymeric resin. In this work, response surface methodology (RSM) based on the Box-Behnken technique was utilized to investigate the impact of independent variables on ramie fabric characteristics and determine the optimal treatment condition. The impact of alkali concentration, treatment time, and temperature on the breaking load and elongation at break of woven ramie fabrics were evaluated using Design-Expert software, which established the design matrix and analyzed the experimental data employing numerical and graphical optimization methods. Moreover, the impact of alkali treatment conditions on the surface morphology, structural change of ramie fabrics, and thermal properties was investigated. Based on the analysis of variance (ANOVA) results, the suggested quadratic models can adequately predict the breaking load and elongation at break of the ramie woven fabrics within the range of conditions applied in this investigation. The RSM revealed that an alkali concentration of 6.12%, a treatment time of 30 min, and a temperature of 39.13°C resulted in an optimum treatment condition with a breaking load of 518.28 N and elongation at break of 23.36%.

Optimization of the cutting process on machining time of ankle foot as transtibial prosthesis components using response surface methodology

The purpose of this research is to determine the best milling process parameters for minimizing machining time of the ankle-foot as a component of transtibial prostheses. The ankle foot prosthesis is of the energy storing type with Al 6061 material. The experimental design employed the Box Behnken technique, with four factors and three levels for each factor, and machining time as a response. The machining parameters assessed in this study are spindle speed, feed rate, step-over, and toolpath technique. The physical trials were carried out using a three-axis CNC milling machine with a flat endmill cutting tool (diameter 5.5 mm, 2 flute with HSS material). The experiment outcomes were assessed using variance analysis, graphs, and numerical approaches. The response surface method (RSM) calculates the mathematical model of the ideal cutting combination parameters based on machining time. Based on the findings, the ideal cutting parameters in the ankle-foot prosthesis production process on a CNC milling machine were spindle speed of 6500 rpm, feed rate of 800 m/min, step over of 0.2 mm, and flowline toolpath strategy. To establish the machining time value of the ankle-foot prosthesis, the ideal circumstances were identified. The findings of this study revealed that CNC milling gave the quickest time for machining transtibial prosthesis components, which can be used to guide the development of machining methods for ankle-foot prostheses constructed of Al 6061 material. The optimal machining time is 424.4601 min.

Development of Asbestos-free Disc Brake Pad Using Periwinkle Shell Powder and Coconut Shell Ash as Base Materials

Brake pads are integral part of an automobile braking system that contribute to the full control of the vehicle. The composite materials for their production must therefore possess adequate chemical, mechanical, physical, and thermal properties. The development of asbestos-free disc brake pad using Periwinkle Shell (PS) powder and Coconut Shell Ash (CSA) as reinforcement and frictional filler material respectively is presented. This was done with a view of establishing suitability of PS powder and CSA as replacements for asbestos which has been found to be carcinogenic and capable of causing Asbestosis and Mesothelioma [1]. Other ingredients used are graphite as lubricant, epoxy resin as binder, copper, zinc, and aluminum powders as abrasives and also to impact mechanical strength. The optimum percentage formulation of these materials for the pad was arrived at by using the ANOVA tool of the Box-Behnken technique of Design Expert Software, DX-13. The CSA and PS powders were sieved into grades of 106 µm, 150 µm, 212 µm and 300 µm and then combined with other constituents to produce the brake pad. The pads were characterized in terms of their physical, mechanical and tribological properties. It was observed that increase in particle size distributions leads to decrease in densification and carbon crosslinking of the produced composite brake pads. Therefore, the 106 µm particle size sample has better properties than others. Comparison analysis shows that the performance parameters of the 106 μm size brake pad compares well, and in some cases better, with typical after-market replacement pads, an asbestos-based brake pad and brake pads developed from past research works.

RETRACTED: Multi-attribute optimization of sustainable aviation fuel production-process from microalgae source

The aviation sector has been one of the most significant contributors to greenhouse gas (GHG) emissions; there is an urgent need to transition from traditional fossil-based jet fuel to sustainable aviation fuel to meet net-zero targets by 2030. The use of renewable aviation fuel may be regarded as the most effective option for reducing GHG emissions in the aviation sector while allowing for long-term growth. In this study, the microalgae oil was transformed to develop hydrocarbons with the boiling point range of jet fuel. Following the transesterification of algal oil fatty acids to algal oil methyl ester, fractional distillation was carried out. The Box-Behnken technique was used to design experiments for efficient resource utilization by minimizing the number of tests during transesterification. The quantitative relationship function between input (molar ratio of methanol to oil, catalyst concentration, and temperature) and % yield of methyl ester as an output of the process was developed using analysis of variance (ANOVA). The ANFIS (adaptive neuro-fuzzy inference system) was utilized to develop a prognostic model with good prediction effectiveness. MORSM (multi-objective response surface methodology) was also used to build a prediction model using correlations. A set of statistical indicators and Theil's U2 were used to examine the prediction efficacy and model uncertainty of ANFIS and MORSM. On both statistical indices and Theil's U2, the ANFIS-based model outperformed. To get the best results, the desirability approach was used to improve operating parameters. According to the desirability approach, the best operating parameters were catalyst concentration at 2.09%, methanol to oil ratio at 9.17%, and temperature at 60.49 °C, which resulted in the greatest yield of 91%.

EXPERIMENTAL INVESTIGATION OF ELECTRICAL DISCHARGE MACHINING OF INCONEL 718 USING A TiB2-Cu SINTERED COMPOSITE ELECTRODE

Electrical discharge machining (EDM) is much preferred in modern precision-manufacturing industries owing to its ability to machine any metal regardless of its hardness. However, its constraint is that the selected metal should be an electrically conductive material. For the present investigation, an Inconel 718 alloy was selected for EDM, using a TiB2-Cu electrode made with powder metallurgy. Input factors, namely, the pulse current (Ip), pulse-on time (Ton) and gap voltage (Gv) were selected and their output responses were the surface roughness (SR) and material-removal rate (MRR). For the response surface, the Box Behnken technique was preferred when designing the experiments (DoE). An ANOVA test was performed to understand the influence of the selected input factors on the SR and MRR. The RSM integrated with a grey relational analysis (GRA) revealed that the optimal input parameters for better machining characteristics were: Ip = 10 A, Ton = 40 µs and Gv = 50 V. Besides, the results also showed that the pulse current more significantly influenced the output responses than the other parameters. Moreover, an increase in the gap voltage caused surface irregularities on the machined surface. Surface morphology of the machined surfaces was analysed through SEM and EDAX. Moreover, a certain amount of tool-material transfer was noted with the EDAX analysis.

Quorum quenching attenuation of rhizobacteria Chromobacterium violaceum virulence by Desmodium gangeticum root extract

Quorum sensing signal molecules used amongst bacteria for communicating cell density can be interfered with through quenching of the signal molecule, a process known as quorum quenching. Several bacteria depend upon a mechanism generally called as quorum sensing to identify their population density and initiate its colonization. The goal of this research was to test and optimize an anti-quorum sensing chemical derived from Desmodium gangeticum root extract against the biosensor strain Chromobacterium violaceum, a rhizosphere bacterium with an opportunistic pathogenic ability. Desmodium gangeticum root extract contained a chemical known as 1,6-octadien, 3,5-dimethyl-, cis, which had a promising bacterial silencing action. The studies suggested that the extract have a downregulating effect upon bacterial virulence. Response surface modelling (RSM) and the Box-Behnken technique were used to investigate the optimization of dependent variables such as bacterial density, rate of administration, and extract concentration. ANOVA and Cook's distance table were used to verify the mathematical model that resulted. The chemical exhibited anti-quorum sensing activity that is dose dependent, peaking at 300 μg/ml extract three times per day. The findings imply that the extract has no bactericidal effect but does affect the quorum sensing system.

Study of a Multicriterion Decision-Making Approach to the MQL Turning of AISI 304 Steel Using Hybrid Nanocutting Fluid.

The enormous use of cutting fluid in machining leads to an increase in machining costs, along with different health hazards. Cutting fluid can be used efficiently using the MQL (minimum quantity lubrication) method, which aids in improving the machining performance. This paper contains multiple responses, namely, force, surface roughness, and temperature, so there arises a need for a multicriteria optimization technique. Therefore, in this paper, multiobjective optimization based on ratio analysis (MOORA), VIseKriterijumska Optimizacija I Kompromisno Resenje (VIKOR), and technique for order of preference by similarity to ideal solution (TOPSIS) are used to solve different multiobjective problems, and response surface methodology is also used for optimization and to validate the results obtained by multicriterion decision-making technique (MCDM) techniques. The design of the experiment is based on the Box–Behnken technique, which used four input parameters: feed rate, depth of cut, cutting speed, and nanofluid concentration, respectively. The experiments were performed on AISI 304 steel in turning with minimum quantity lubrication (MQL) and found that the use of hybrid nanofluid (Alumina–Graphene) reduces response parameters by approximately 13% in forces, 31% in surface roughness, and 14% in temperature, as compared to Alumina nanofluid. The response parameters are analyzed using analysis of variance (ANOVA), where the depth of cut and feed rate showed a major impact on response parameters. After using all three MCDM techniques, it was found that, at fixed weight factor with each MCDM technique, a similar process parameter was achieved (velocity of 90 m/min, feed of 0.08 mm/min, depth of cut of 0.6 mm, and nanoparticle concentration of 1.5%, respectively) for optimum response. The above stated multicriterion techniques employed in this work aid decision makers in selecting optimum parameters depending upon the desired targets. Thus, this work is a novel approach to studying the effectiveness of hybrid nanofluids in the machining of AISI 304 steel using MCDM techniques.

FSSW process parameter optimization for AA2024 and AA7075 alloy

ABSTRACT The novelty of the present work is the investigation of friction stir spot welding between AA2024 and AA7075 aluminum plates performed by Box-Behnken technique of response surface methodology. The chemical composition of the aluminum alloys is confirmed using XRD analysis. The process parameters of the FSSW process are analyzed to find the significant parameter that influences the output responses; it was revealed that the Tool Rotation Speed (TRS) was the influencing parameter. Furthermore, the most influenced tensile and hardness analysis of FSS Welded joint are identified by empirical analysis and it was found that the Tensile Shear force (Ts) was the improved property due to optimized FSSW process. Desirability approach was used to predict the optimal Ts value. The optimized process parameters are 2168.813 rpm of TRS, 3.32 mm depth of plunge and 46.98s dwell time. The computed Ts values for the present work investigation were 4.0116 kN and it was found that these values are consistent within the results obtained by the validation experiments. The microstructure of the weld joint was analyzed by optical microscope. The correlations between the predicted and experimental results were established.

Characterizing and using a new bi-functional catalyst to sustainably synthesize methyl levulinate from biomass carbohydrates

In recent years, significant attention has been dedicated to converting biomass carbohydrates into alkyl levulinates (biofuel additives) in alcoholic media, in search of various cheap and active solid catalysts through relatively easy pathways and from non-dangerous precursors. The synthesis of methyl levulinate (ML) using a new Brønsted/Lewis acid sites catalyst was investigated, emphasizing the influence of three parameters (catalyst loading, the molar concentration of fructose in methanol, and reaction temperature) on the process. The synthetized catalyst, SO42−/TiO2–La2O3 coating Fe3O4, was characterized by different techniques (FT-IR, XRD, TGA, TEM-EDX, ICP-OES). The design of the ML synthesis experiment was carried out using the Box-Behnken technique, while the response surface methodology was applied to obtain the optimized process conditions. A mathematical model of the catalytic synthesis was built, based upon first principles, and the experimental time variation of the reaction mixture concentrations was used to find, through regression, the values of the constants of a newly proposed kinetic model.

Modeling and optimization of energy consumption and performance characteristics of a solar assisted fluidized bed dryer

The major problem regarding conventional rice mills is the use of fixed-bed drying systems, which is accompanied by high energy consumption. In this study, a novel solar-assisted fluidized-bed system for drying paddy grains was used. The system included a solar water heater, an infrared lamp powered by photovoltaic panels, along with a gas water heater, which were used together to provide the required thermal energy. The working factors were drying air temperature (35, 45, and 55 °C), drying air velocity (7, 8, and 9 m/s), and the angle of the desiccant wheel regeneration gate (0, 45, and 90 degrees). The performance characteristics of the drying system were measured in terms of drying rate, total energy consumption, solar fraction and energy efficiency. Several mathematical models were also applied to fit the moisture ratio curves. In order to analyze the results and obtain the optimized working conditions of the drying system, Response Surface Methodology (RSM) based on the Box–Behnken technique was used. In all of the evaluated conditions, the model entitled “Approximation of diffusion” had the best results for fitting the moisture ratio curves with a correlation coefficient value of at least 0.9729. The selected optimum point included drying air velocity of 7 m/s, drying air temperature of 52.77 oC and regeneration gate angle of 0. Under the mentioned working conditions, the drying time of 75.99 min, the total energy consumption of 0.297 kWh, solar energy fraction of 0.540 and energy efficiency of 47.67% could be obtained with a desirability value of 0.970.

An Experimental Study on Modelling the Physical Properties of Composite Psyllium, Alginate and Chitosan Fibers Using Box-Behnken Technique

Polysaccharides from different sources have found their applications in the wound care for their inherent abilities of gelling, biocompatibility, biodegradability, and antibacterial activity. Considering the low cost, ease of processing, abundance in nature and therapeutic benefits of the polysaccharides we developed psyllium, alginate and chitosan (PAC) composite fibers and modelled their physical properties namely; tenacity, linear density and absorption in both distilled water and saline solution; using Box-Behnken (BB) technique of Response Surface Methodology (RSM). The PAC fibers were produced using an in house developed wet spinning module; briefly the dope solutions containing psyllium and alginate were extruded into a hydrolyzed chitosan and CaCl2. The produced fibers were acetone washed to remove the polar solvent; in this case water; and later dried in air. The present work aimed at the development of statistical models for physical properties: linear density, tenacity, antibacterial efficacy and liquid absorption; of the composite PAC fibers. The factors under consideration were the raw material concentration namely; alginate and psyllium in dope solution and calcium chloride (CaCl2) and hydrolysed chitosan in the coagulation bath. The experimental work was designed and analyzed using Minitab® 17 statistical software. Regression models using the under consideration factors were successfully developed.

Pulsed laser butt welding of AISI 1005 steel thin plates

Pulsed laser welding is a joining process, which takes advantage of the laser energy quality with precise control equipment. Its characteristics allow joining thinner materials with reduced thermal distortion without filler material, fulfilling the industrial interest in this process. In the present work, AISI 1005 steel sheets of 1.7 mm thickness were subjected to different pulse Nd:YAG laser treatments to obtain one side bead-on-plate weld samples and double sided butt welds. Since the quality of the joints is strongly influenced by the process parameters, an experimental design methodology was adopted to define a suitable combination of the laser beam power, pulse duration and spot diameter. The Box-Behnken technique and response surface methodology associated with analyses of defects allowed the determination of the appropriate welding parameters. The microstructure characterisation revealed the melted fusion zones originated from the thermal effect of the overlapped pulses. Additionally, microhardness measurements at fusion zone exhibited higher values than the base material and heat affected zone, due to the hard microconstituents developed. The tensile test performed at samples extracted from double sided laser welds showed good resistance results, fracturing at the base material of joints. The present work demonstrates that pulsed laser welding methodology is suitable to obtain high quality joints of steel thin plates.

Efficient removal of hexavalent chromium from electroplating wastewater using polypyrrole coated on cellulose sulfate fibers

In this study, cellulose sulfate was synthesized through sulfonation of cotton, and polypyrrole was coated on the surface of fibers. Then, the optimum ratio of pyrrole to cellulose sulfate was evaluated, and the physical, chemical, and morphological properties of the composite were assessed by using FESEM, EDS, FTIR, BET, and TGA analysis. Furthermore, adsorption of hexavalent chromium using the composite adsorbent was studied by the results of designed experiments with the Box-Behnken technique to assess the effect of pH, contact time, adsorbent dose and the initial concentration of hexavalent chromium and optimize the adsorption process. The removal percentage was 99.9% under the optimum conditions (adsorbent dose, 4 g L-1; initial concentration of Cr(VI), 200 mg L-1; pH value, 2; contact time, 200 min). The results of adsorption isotherms illustrated that the adsorption process followed Redlich-Peterson, Freundlich, Radke-Prausnitz, and UT models, and the calculated maximum adsorption capacity by the Langmuir model was 198 mg g-1. Based on the kinetic and thermodynamic studies, the adsorption process followed the intraparticle diffusion model and showed the endothermic and spontaneous adsorption with an increase in entropy on the adsorbent surface. The presence of copper, nickel and zinc cations had no adverse effect on the removal percentage of hexavalent chromium significantly. The adsorbent was reused successfully in four sequential treatments. Consequently, the synthesized adsorbent is efficient due to the high efficiency of hexavalent chromium removal percentage from electroplating effluent (99.87%).

Optimization of photocatalytic degradation of methylene blue dye using titanate nanotube

Titanate nanotubes were evaluated for photocatalytic depletion of methylene blue (MB) dye under UV radiation in a cylindrical reactor. Titanate has been prepared using the method of hydrothermal treatment where the precursors are TiO2 and NaOH. Transmitting electron microscopy (TEM) and x-ray diffraction (XRD) were used to characterize the prepared nanotubes. The effect of some parameters was examined. It was shown that removing the dye is effective: dye initial concentration 5 to 10 ppm, dosage of catalyst 0.25 to 0.50 g catalyst/l of the solution, and mixture pH 1.4 to 12.6. A total of 17 runs have been designed to take into account the effective ranges of the effective variables using the experimental Box–Behnken technique. Two polynomial models were tailored to the experimental outputs created by the response surface methodology, demonstrating a functional correlation between MB’s percent degradation and experimental parameters. Design Expert program V.6.0.7 was utilized to maximize the reaction rate of the experimental parameters. The optimum parameter values were initial concentration of MB 5.45 ppm, a dose of TiO2 0.50 g / l, and pH of the mixture 11.89. Under these conditions, the percentage of degradation of MB was 99.0995%.

Characteristics and process optimization of rice snack (khao‐tan) puffing by home microwave oven

Khao‐tan is a popular Thai traditional glutinous rice snack that involves a deep frying process. Nowadays, consumers are concerned about the safety of fried food and its potential to contribute to obesity. Puffing using a home microwave oven is an alternative that is more convenient, requires a shorter processing time, has a lower cost, is healthy and environment‐friendly. This research investigated the optimal processing conditions (initial moisture content [IMC], microwave power [MWP], and puffing time [PT]) to puff khao‐tan from three rice varieties (sticky rice [SR], rice berry [RB], and black rice [BR]) using the microwave. Response surface methodology (the Box–Behnken technique) was applied. The process responses comprised puffing yield, expansion ratio, hardness, and yellowness. The results indicated that the optimized conditions for khao‐tan were 10% IMC, 1,200 W MWP, and 25 s PT with overall desirability values of 0.733, 0.642, and 0.899 for SR, RB, and BR, respectively. PRACTICAL APPLICATIONS: Recently, microwaves have become popular use in the food industry. This study provides a good technique to introduce microwave technology in the snack industry to puff a Thai traditional rice snack, namely, khao‐tan using a home microwave oven. The optimized conditions can help with puffing khao‐tan made with three varieties of rice (sticky rice, rice berry, and black rice) to achieve consumer acceptance in terms of product quality and a healthy product. This work can be applied commercially and on a small scale.

Development of Multi-Objective Optimization Model for Electromagnetic Die Forming of Aa5083 Aluminum Alloy Sheet

Electromagnetic die forming is a high-speed formation method that can be quite effective in increasing the limits on the shape of the metal sheet of low weight and of the strong electrical conductivity content. A coil is an essential instrument for transmitting electrical power to plastic energy, and the geometry of the coil plays a key role in macrofield transmission. Experiments of the electrical composition of 0.5 mm, 0.7 mm and 0.9 mm AA5083 aluminum sheets are performed via the box-behnken technique with a single layer spiral and a flat square-cross section and spiral. Evaluating characteristics like total height, rib height internal diameter, and external diameter for various charge voltage and current was investigated for the forming behaviour. The results show that the efficiency of production is very dependent on the divide and thickness of work pieces.

Fabrication and optimization of hybrid AA-6082-T6 alloy/8%Al2O3(Alumina)/2%Grp metal matrix composites using novel Box-Behnken methodology processed by wire-sinking electric discharge machining

This work reports the fabrication and studies the influence of Electric discharge machining parameters during machining of the hybrid AA-6082-T6 alloy/8%Al2O3(Alumina)/2%Graphite (Grp) alloy-based metal matrix composite. Experiments were designed & optimized via Novel Box-Behnken technique in Response surface methodology. The pulse off-time, pulse on-time, peak current, and applied voltage used during this research investigation considered as process parameters. These process parameters measured in terms of MRR (Material removal rate) and SR (Surface roughness) using a copper rod of Ø12 mm as an electrode. A scientific model was additionally created to achieve a better performance in optimizing process parameters. From the experimental study, it has concluded that both MRR and SR are directly and mostly influenced using Pulse-on time & peak current. The affected response parameters were also evident by Scanning Electron Microscopy (SEM) & Energy Dispersive Analysis of X-Ray (EDAX) observations. This experimental research study believed that our Novel approach to Design & Optimization plays a vital role for researchers who tend to work in the field of process parameters optimization during machining.

Box-Behnken technique based multi-parametric optimization of electrostatic spray coating in the manufacturing of thermoplastic composites

ABSTRACTAn electrostatic spray coating process was designed and developed in-house for consistent deposition of polypropylene powder on continuously fed carbon tow to produce a matrix coated towpreg. The electrostatic spray coating process was optimized for fiber volume fraction using Box-Behnken experimental design. An empirical multi-parametric equation was obtained to define the relationship between the process parameters (electrostatic voltage at the gun, velocity of the carbon towpreg, and the distance between the tip of gun and the towpreg) through response surface methodology. Experimental verification of the model showed a validation of ~93%. The velocity of the carbon towpreg was found to have a more profound effect on the output response of fiber volume fraction when compared to other process parameters.

Polyextremotolerant Amylase Produced from Novel Enterococcus with Potpourri of Applications

Amylolytic toluene-tolerant Enterococcus faecalis mercadA7 isolated from asbestos contaminated black soil from the Tokat province of Turkey was used for statistically optimized liquid phase fermentation to spawn elevated levels of amylase. Box Behnken technique of Response surface Methodology is followed to optimize three significant factors influencing Amylase production. To purify the amylase, ammonium sulfate fractionation, dialysis and gel filtration on Sephadex G-100 column chromatography were used. Submerged state fermentation at statistically optimized conditions augmented amylase production levels (8467 U ml−1) 8.74-fold compared to the conventional optimization. Purified amylase was found to possess specific activity of 1022 U mg−1, optimal activity at pH 9 and 60 °C, 80% stable at 8.0–10.0 pH range and 90% stable at 45–75 °C for 1 h; with Km value of 8.06 mg ml−1 and a Vmax of 4.36 mmol h−1 ml−1, noteworthy stability up to 43% toluene, and upto 8% NaCl. High titers of Amylase produced from Enterococcus faecalis mercadA7 might find applications in copious industrial processes due to noteworthy stability of the cell-free purified enzyme at elevated temperatures, alkaline, and high toluene or salt concentrated environments.