Application of Statistical Approach for Optimizing CMCase Production by Bacillus tequilensis S28 Strain via Submerged Fermentation Using Wheat Bran as Carbon Source

Biofuels production has come forth as a future technology to combat the problem of depleting fossil fuels. Bio-based ethanol production from enzymatic lignocellulosic biomass degradation serves an efficient method and catching the eye of scientific community. High cost of the enzyme is the major obstacle in preventing the commercialization of this process. Thus main objective of the present study was to optimize composition of medium components for enhancing cellulase production by newly isolated strain of Bacillus tequilensis. Nineteen factors were taken into account using statistical Plackett-Burman Design. The significant variables influencing the cellulose production were further employed in statistical Response Surface Methodology using Central Composite Design for maximizing cellulase production. The optimum medium composition for cellulase production was: peptone (4.94 g/L), ammonium chloride (4.99 g/L), yeast extract (2.00 g/L), Tween-20 (0.53 g/L), calcium chloride (0.20 g/L) and cobalt chloride (0.60 g/L) with pH 7, agitation speed 150 rpm and 72 h incubation at 37oC. Analysis of variance (ANOVA) revealed high coefficient of determination (R2) of 0.99. Maximum cellulase productivity of 11.5 IU/ml was observed against the model predicted value of 13 IU/ml. This was found to be optimally active at 60oC and pH 5.5.

A β-mannanase from Fusarium oxysporum SS-25 via Solid State Fermentation on Brewer’s Spent Grain: Medium Optimization by Statistical Tools, Kinetic Characterization and Its Applications

This study is concerned with the optimization of fermentation parameters for the hyper production of mannanase from Fusarium oxysporum SS-25 employing two step statistical strategy and kinetic characterization of crude enzyme preparation. The Plackett-Burman design used to screen out the important factors in the culture medium revealed 20% (w/w) wheat bran, 2% (w/w) each of potato peels, soyabean meal and malt extract, 1% tryptone, 0.14% NH4SO4, 0.2% KH2PO4, 0.0002% ZnSO4, 0.0005% FeSO4, 0.01% MnSO4, 0.012% SDS, 0.03% NH4Cl, 0.1% NaNO3 in brewer’s spent grain based medium with 50% moisture content, inoculated with 2.8×107 spores and incubated at 30oC for 6 days to be the main parameters influencing the enzyme production. Of these factors, four variables including soyabean meal, FeSO4, MnSO4 and NaNO3 were chosen to study the interactive effects and their optimum levels in central composite design of response surface methodology with the final mannanase yield of 193 IU/gds. The kinetic characterization revealed the crude enzyme to be active over broader temperature and pH range. This could result in 26.6% reduction in kappa number with 4.93% higher tear index and 1% increase in brightness when used to treat the wheat straw based kraft pulp. The hydrolytic potential of enzyme was also demonstrated on both locust bean gum and guar gum.

Microwave-Assisted Alginate Extraction from Portuguese Saccorhiza polyschides – Influence of Acid Pretreatment

Brown seaweeds are abundant in Portuguese coastline and represent an almost unexploited marine economic resource. One of the most common species, easily available for harvesting in the northwest coast, is Saccorhiza polyschides grows in the lowest shore and costal rocky reefs. It is almost exclusively used by local farmers as natural fertilizer, but contains a substantial amount of valuable compounds, particularly alginates, natural biopolymers of high interest for many industrial applications. Alginates are natural polysaccharides present in cell walls of brown seaweed, highly biocompatible, with particular properties that make them of high interest for the food, biotechnology, cosmetics and pharmaceutical industries. Conventional extraction processes are based on thermal treatment. They are lengthy and consume high amounts of energy and solvents. In recent years, microwave-assisted extraction (MAE) has shown enormous potential to overcome major drawbacks that outcome from conventional plant material extraction (thermal and/or solvent based) techniques, being also successfully applied to the extraction of agar, fucoidans and alginates. In the present study, acid pretreatment of brown seaweed Saccorhiza polyschides for subsequent microwave-assisted extraction (MAE) of alginate was optimized. Seaweeds were collected in Northwest Portuguese coastal waters of the Atlantic Ocean between May and August, 2014. Experimental design was used to assess the effect of temperature and acid pretreatment time in alginate extraction. Response surface methodology allowed the determination of the optimum MAE conditions: 40 mL of HCl 0.1 M per g of dried seaweed with constant stirring at 20ºC during 14h. Optimal acid pretreatment conditions have enhanced significantly MAE of alginates from Saccorhiza polyschides, thus contributing for the development of a viable, more environmental friendly alternative to conventional processes.

Statistical Optimization of Medium Components for Biomass Production of Chlorella pyrenoidosa under Autotrophic Conditions and Evaluation of Its Biochemical Composition under Stress Conditions

The aim of the present work was to statistically design an autotrophic medium for maximum biomass production by Chlorella pyrenoidosa using response surface methodology. After evaluating one factor at a time approach, K2HPO4, KNO3, MgSO4.7H2O and NaHCO3 were preferred over the other components of the fog’s medium as most critical autotrophic medium components. The study showed that the maximum biomass yield was achieved while the concentrations of MgSO4.7H2O, K2HPO4, KNO3 and NaHCO3 were 0.409 g/L, 0.24 g/L, 1.033 g/L, and 3.265 g/L, respectively. The study reported that the biomass productivity of C. pyrenoidosa improved from 0.14 g/L in defined fog’s medium to 1.40 g/L in modified fog’s medium resulting 10 fold increase. The biochemical composition biosynthesis of C. pyrenoidosa was altered using nitrogen limiting stress bringing about 5.23 fold increase in lipid content than control (cell without stress), as analyzed by FTIR integration method.

Numerical Optimization of Trapezoidal Microchannel Heat Sinks

This study presents the numerical simulation of three-dimensional incompressible steady and laminar fluid flow and conjugate heat transfer of a trapezoidal microchannel heat sink using water as a cooling fluid in a silicon substrate. Navier-Stokes equations with conjugate energy equation are discretized by finite-volume method. We perform numerical computations for a range of 50 ≦ Re ≦ 600, 0.05W ≦ P ≦ 0.8W, 20W/cm2 ≦q"≦ 40W/cm2. The present study demonstrates the numerical optimization of a trapezoidal microchannel heat sink design using the response surface methodology (RSM) and the genetic algorithm method (GA). The results show that the average Nusselt number increases with an increase in the Reynolds number or pumping power, and the thermal resistance decreases as the pumping power increases. The thermal resistance of a trapezoidal microchannel is minimized for a constant heat flux and constant pumping power.

Enzymatic Synthesis of Olive-Based Ferulate Esters: Optimization by Response Surface Methodology

Ferulic acid has widespread industrial potential by virtue of its antioxidant properties. However, it is partially soluble in aqueous media, limiting their usefulness in oil-based processes in food, cosmetic, pharmaceutical, and material industry. Therefore, modification of ferulic acid should be made by producing of more lipophilic derivatives. In this study, a preliminary investigation of lipase-catalyzed trans-esterification reaction of ethyl ferulate and olive oil was investigated. The reaction was catalyzed by immobilized lipase from Candida antarctica (Novozym 435), to produce ferulate ester, a sunscreen agent. A statistical approach of Response surface methodology (RSM) was used to evaluate the interactive effects of reaction temperature (40-80°C), reaction time (4-12 hours), and amount of enzyme (0.1-0.5 g). The optimum conditions derived via RSM were reaction temperature 60°C, reaction time 2.34 hours, and amount of enzyme 0.3 g. The actual experimental yield was 59.6% ferulate ester under optimum condition, which compared well to the maximum predicted value of 58.0%.

Immobilization of Lipase Enzyme by Low Cost Material: A Statistical Approach

Immobilization of lipase enzyme produced from palm oil mill effluent (POME) by the activated carbon (AC) among the low cost support materials was optimized. The results indicated that immobilization of 94% was achieved by AC as the most suitable support material. A sequential optimization strategy based on a statistical experimental design, including one-factor-at-a-time (OFAT) method was used to determine the equilibrium time. Three components influencing lipase immobilization were optimized by the response surface methodology (RSM) based on the face-centered central composite design (FCCCD). On the statistical analysis of the results, the optimum enzyme concentration loading, agitation rate and carbon active dosage were found to be 30 U/ml, 300 rpm and 8 g/L respectively, with a maximum immobilization activity of 3732.9 U/g-AC after 2 hrs of immobilization. Analysis of variance (ANOVA) showed a high regression coefficient (R2) of 0.999, which indicated a satisfactory fit of the model with the experimental data. The parameters were statistically significant at p

Statistical Modeling for Permeabilization of a Novel Yeast Isolate for β-Galactosidase Activity Using Organic Solvents

The hydrolysis of lactose using β-galactosidase is one of the most promising biotechnological applications, which has wide range of potential applications in food processing industries. However, due to intracellular location of the yeast enzyme, and expensive extraction methods, the industrial applications of enzymatic hydrolysis processes are being hampered. The use of permeabilization technique can help to overcome the problems associated with enzyme extraction and purification of yeast cells and to develop the economically viable process for the utilization of whole cell biocatalysts in food industries. In the present investigation, standardization of permeabilization process of novel yeast isolate was carried out using a statistical model approach known as Response Surface Methodology (RSM) to achieve maximal b-galactosidase activity. The optimum operating conditions for permeabilization process for optimal β-galactosidase activity obtained by RSM were 1:1 ratio of toluene (25%, v/v) and ethanol (50%, v/v), 25.0 oC temperature and treatment time of 12 min, which displayed enzyme activity of 1.71 IU /mg DW.

Total Lipid of Mutant Synechococcus sp. PCC 7002

Microalgae lipid is a promising feedstock for biodiesel production. The objective of this work was to study growth factors affecting marine mutant Synechococcus sp. (PCC 7002) for high lipid production. Four growth factors were investigated; nitrogen-phosporus-potassium (NPK) concentration, light intensity, temperature and NaNO3 concentration on mutant strain growth and lipid production were studied. Design Expert v8.0 was used to design the experimental and analyze the data. The experimental design selected was Min-Run Res IV which consists of 12 runs and the response surfaces measured were specific growth rate and lipid concentration. The extraction of lipid was conducted by chloroform/methanol solvents system. Based on the study, mutant Synechococcus sp. PCC 7002 gave the highest specific growth rate of 0.0014 h-1 at 0% NPK, 2500 lux, 40oC and 0% NaNO3. On the other hand, the highest lipid concentration was obtained at 0% NPK, 3500 lux, 30oC and 1% NaNO3.

Response Surface Methodology Approach to Defining Ultrafiltration of Steepwater from Corn Starch Industry

In this work the concentration of steepwater from corn starch industry is monitored using ultrafiltration membrane. The aim was to examine the conditions of ultrafiltration of steepwater by applying the membrane of 2.5nm. The parameters that vary during the course of ultrafiltration, were the transmembrane pressure, flow rate, while the permeate flux and the dry matter content of permeate and retentate were the dependent parameter constantly monitored during the process. Experiments of ultrafiltration are conducted on the samples of steepwater, which were obtained from the starch wet milling plant „Jabuka“ Pancevo. The procedure of ultrafiltration on a single-channel 250mm lenght, with inner diameter of 6.8mm and outer diameter of 10mm membrane were carried on. The membrane is made of a-Al2O3 with TiO2 layer obtained from GEA (Germany). The experiments are carried out at a flow rate ranging from 100 to 200lh-1 and transmembrane pressure of 1-3 bars. During the experiments of steepwater ultrafiltration, the change of permeate flux, dry matter content of permeate and retentate, as well as the absorbance changes of the permeate and retentate were monitored. The experimental results showed that the maximum flux reaches about 40lm-2h-1. For responses obtained after experiments, a polynomial model of the second degree is established to evaluate and quantify the influence of the variables. The quadratic equitation fits with the experimental values, where the coefficient of determination for flux is 0.96. The dry matter content of the retentate is increased for about 6%, while the dry matter content of permeate was reduced for about 35-40%, respectively. During steepwater ultrafiltration in permeate stays 40% less dry matter compared to the feed.

Influence of Fermentation Conditions on Humic Acids Production by Trichoderma viride Using an Oil Palm Empty Fruit Bunch as the Substrate

Humic acids (HA) were produced by a Trichoderma viride strain under submerged fermentation in a medium based on the oil palm empty fruit bunch (EFB) and the main variables of the process were optimized by using response surface methodology. A temperature of 40°C and concentrations of 50g/L EFB, 5.7g/L potato peptone and 0.11g/L (NH4)2SO4 were the optimum levels of the variables that maximize the HA production, within the physicochemical and biological limits of the process. The optimized conditions led to an experimental HA concentration of 428.4±17.5 mg/L, which validated the prediction from the statistical model of 412.0mg/L. This optimization increased about 7–fold the HA production previously reported in the literature. Additionally, the time profiles of HA production and fungal growth confirmed our previous findings that HA production preferably occurs during fungal sporulation. The present study demonstrated that T. viride successfully produced HA via the submerged fermentation of EFB and the process parameters were successfully optimized using a statistics-based response surface model. To the best of our knowledge, the present work is the first report on the optimization of HA production from EFB by a biotechnological process, whose feasibility was only pointed out in previous works.

Optimization of Process Parameters Affecting Biogas Production from Organic Fraction of Municipal Solid Waste via Anaerobic Digestion

The aim of this study was to obtain the optimal conditions for biogas production from anaerobic digestion of organic fraction of municipal solid waste (OFMSW) using response surface methodology (RSM). The parameters studied were initial pH, substrate concentration and total organic carbon (TOC). The experimental results showed that the linear model terms of initial pH and substrate concentration and the quadratic model terms of the substrate concentration and TOC had significant individual effect (p < 0.05) on biogas yield. However, there was no interactive effect between these variables (p > 0.05). The highest level of biogas produced was 53.4 L/Kg VS at optimum pH, substrate concentration and total organic carbon of 6.5, 99gTS/L and 20.32 g/L respectively.

Artificial Intelligent Approach for Machining Titanium Alloy in a Nonconventional Process

Artificial neural networks (ANN) are used in distinct researching fields and professions, and are prepared by cooperation of scientists in different fields such as computer engineering, electronic, structure, biology and so many different branches of science. Many models are built correlating the parameters and the outputs in electrical discharge machining (EDM) concern for different types of materials. Up till now model for Ti-5Al-2.5Sn alloy in the case of electrical discharge machining performance characteristics has not been developed. Therefore, in the present work, it is attempted to generate a model of material removal rate (MRR) for Ti-5Al-2.5Sn material by means of Artificial Neural Network. The experimentation is performed according to the design of experiment (DOE) of response surface methodology (RSM). To generate the DOE four parameters such as peak current, pulse on time, pulse off time and servo voltage and one output as MRR are considered. Ti-5Al-2.5Sn alloy is machined with positive polarity of copper electrode. Finally the developed model is tested with confirmation test. The confirmation test yields an error as within the agreeable limit. To investigate the effect of the parameters on performance sensitivity analysis is also carried out which reveals that the peak current having more effect on EDM performance.

Development of Regression Equation for Surface Finish and Analysis of Surface Integrity in EDM

Electrical discharge machining (EDM) is a relatively modern machining process having distinct advantages over other machining processes and can machine Ti-alloys effectively. The present study emphasizes the features of the development of regression equation based on response surface methodology (RSM) for correlating the interactive and higher-order influences of machining parameters on surface finish of Titanium alloy Ti-6Al-4V. The process parameters selected in this study are discharge current, pulse on time, pulse off time and servo voltage. Machining has been accomplished using negative polarity of Graphite electrode. Analysis of variance is employed to ascertain the adequacy of the developed regression model. Experiments based on central composite of response surface method are carried out. Scanning electron microscopy (SEM) analysis was performed to investigate the surface topography of the EDMed job. The results evidence that the proposed regression equation can predict the surface roughness effectively. The lower ampere and short pulse on time yield better surface finish.

Compliance Modelling and Optimization of Kerf during WEDM of Al7075/SiCP Metal Matrix Composite

This investigation presents the formulation of kerf (width of slit) and optimal control parameter settings of wire electrochemical discharge machining which results minimum possible kerf while machining Al7075/SiCp MMCs. WEDM is proved its efficiency and effectiveness to cut the hard ceramic reinforced MMCs within the permissible budget. Among the distinct performance measures of WEDM process, kerf is an important performance characteristic which determines the dimensional accuracy of the machined component while producing high precision components. The lack of available of the machinability information such advanced MMCs result the more experimentation in the manufacturing industries. Therefore, extensive experimental investigations are essential to provide the database of effect of various control parameters on the kerf while machining such advanced MMCs in WEDM. Literature reviled the significance some of the electrical parameters which are prominent on kerf for machining distinct conventional materials. However, the significance of reinforced particulate size and volume fraction on kerf is highlighted in this work while machining MMCs along with the machining parameters of pulse-on time, pulse-off time and wire tension. Usually, the dimensional tolerances of machined components are decided at the design stage and a machinist pay attention to produce the required dimensional tolerances by setting appropriate machining control variables. However, it is highly difficult to determine the optimal machining settings for such advanced materials on the shop floor. Therefore, in the view of precision of cut, kerf (cutting width) is considered as the measure of performance for the model. It was found from the literature that, the machining conditions of higher fractions of large size SiCp resulting less kerf where as high values of pulse-on time result in a high kerf. A response surface model is used to predict the relative significance of various control variables on kerf. Consequently, a powerful artificial intelligence called genetic algorithms (GA) is used to determine the best combination of the control variable settings. In the next step the conformation test was conducted for the optimal parameter settings and found good agreement between the GA kerf and measured kerf. Hence, it is clearly reveal that the effectiveness and accuracy of the developed model and program to analyze the kerf and to determine its optimal process parameters. The results obtained in this work states that, the resulted optimized parameters are capable of machining the Al7075/SiCp MMCs more efficiently and with better dimensional accuracy.

Statistical Screening of Medium Components on Ethanol Production from Cashew Apple Juice using Saccharomyces diasticus

In the present study, effect of critical medium components (a total of fifteen components) on ethanol production from waste cashew apple juice (CAJ) using yeast Saccharomyces diasticus was studied. A statistical response surface methodology (RSM) based Plackett-Burman Design (PBD) was used for the design of experiments. The design contains a total of 32 experimental trails. The effect of medium components on ethanol was studied at two different levels such as low concentration level (-) and high concentration levels (+). The dependent variables selected in this study were ethanol concentration (g/L) and cellmass concentration (g/L). Data obtained from RSM on ethanol production were subjected to analysis of variance (ANOVA). In general, initial substrate concentration significantly influenced the microbial growth and product formation. Of the medium components evaluated, CAJ concentration, yeast extract, (NH4)2SO4, and malt extract showed significant effect on ethanol fermentation. A second-order polynomial model was used to predict the experimental data and the model fitted the data with a high correlation coefficient (R2 > 0.98). Maximum ethanol (15.3 g/L) and biomass (6.4 g/L) concentrations were obtained at the optimum medium composition and at optimum condition (temperature-30°C; initial pH-6.8) after 72 h fermentation using S.diasticus.

Studies on Various Parameters Involved in Conjugation of Starch with Lysine for Excellent Emulsification Properties Using Response Surface Methodology

The process parameters, starch-water ratio (A, (w/v) %), pH of suspension (B), Temperature(C, °C) and Time (D, hrs.)., were optimized for the preparation of starch-lysine conjugate and studying their effect on stability of emulsions by calculating emulsion stability index using response surface methodology. The optimized conditions are pH 9.0, temperature 60oC, reaction time 6 hrs, starch:water ratio 1:2.5, having emulsion stability index was 0.72.

Investigation of the Emulsifying Properties of Bambara Groundnut Flour and Starch

The current desire in food and industrial emulsification is the use of natural emulsifiers. Bambara groundnut flour (BGNF) and its starch (BGNS) will serve both emulsifying and nutritional purposes if found suitable. This current study was aimed at investigating the emulsifying properties of BGNF/BGNS. BGNS was extracted from the BGNF. Emulsions were prepared using a wide range of flour-oil-water and starch-oil-water composition as generated through the application of Response Surface (D-optimal) design. Prepared emulsions were investigated for stability to creaming/sedimentation (using the kinetic information from turbiscan) and flocculation/coalescence (by monitoring the droplet diameter growth using optical microscope) over 5 days. The most stable emulsions (one BGNF-stabilized and the other BGNS-stabilized) were determined. The optimal emulsifier/oil composition was 9g/39g for BGNF and 5g/30g for BGNS. The two emulsions had only 30% and 50% growth in oil droplet diameter respectively by day 5, compared to over 3000% in the unstable ones. The BGNF-stabilized emulsions were more stable than the BGNS-stabilized ones. Emulsions were successfully stabilized with BGNF and BGNS.

Optimization of Lipase Production Using Bacillus subtilis by Response Surface Methodology

A total of 6 isolates of Bacillus subtilis were isolated from oil mill waste collected in Namakkal district, Tamilnadu, India. The isolated bacteria were screened using lipase screening medium containing Tween 80. BS-3 isolate exhibited a greater clear zone than the others, indicating higher lipase activity. Therefore, this isolate was selected for media optimization studies. Ten process variables were screened using Plackett–Burman design and were further optimized by central composite design of response surface methodology for lipase production in submerged fermentation. Maximum lipase production of 16.627 U/min/ml were predicted in medium containing yeast extract (9.3636g), CaCl2 (0.8986g) and incubation periods (1.813 days). A mean value of 16.98 ± 0.2286 U/min/ml of lipase was acquired from real experiments.

Statistical Optimization of Process Conditions for Disinfection of Water Using Defatted Moringa oleifera Seed Extract

In this study, statistical optimization design was used to study the optimum disinfection parameters using defatted crude Moringa oleifera seed extracts against Escherichia coli (E. coli) bacterial cells. The classical one-factor-at-a-time (OFAT) and response surface methodology (RSM) was used. The possible optimum range of dosage, contact time and mixing rate from the OFAT study were 25mg/l to 200mg/l, 30minutes to 240 minutes and 100rpm to 160rpm respectively. Analysis of variance (ANOVA) of the statistical optimization using faced centered central composite design showed that dosage, contact time and mixing rate were highly significant. The optimum disinfection range was 125mg/l, at contact time of 30 minutes with mixing rate of 120 rpm.