Optimization of Lead Bioremediation by Marine Halomonas sp. ES015 Using Statistical Experimental Methods

Bioremediation technology is now used for treatment instead of traditional metal removal methods. A strain was isolated from Marsa Alam, Red sea, Egypt showed high resistance to high lead concentration and was identified by the 16S rRNA gene sequencing technique as Halomonas sp. ES015. Medium optimization was carried out using Plackett-Burman design, and the most significant factors were yeast extract, casamino acid and inoculums size. The optimized media obtained by the statistical design raised the removal efficiency from 84% to 99% from initial concentration 250 ppm of lead. Moreover, Box-Behnken experimental design was applied to study the relationship between yeast extract concentration, casamino acid concentration and inoculums size. The optimized medium increased removal efficiency to 97% from initial concentration 500 ppm of lead. Immobilized Halomonas sp. ES015 cells on sponge cubes, using optimized medium in loop bioremediation column, showed relatively constant lead removal efficiency when reused six successive cycles over the range of time interval. Also metal removal efficiency was not affected by flow rate changes. Finally, the results of this research refer to the possibility of lead bioremediation by free or immobilized cells of Halomonas sp. ES015. Also, bioremediation can be done in batch cultures and semicontinuous cultures using column technology.

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.

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.

Determination of Effective Variables on Arachidonic Acid Production by Mortierella alpina CBS 754.68in Solid-State Fermentation using Plackett-Burman Screening Design

In the present study, the oleaginous fungus Mortierella alpina CBS 754.68 was screened for arachidonic acidproduction using inexpensive agricultural by-products as substrate. Four oilcakes were analysed to choose the best substrate among them. Sunflower oilcake was the most effective substrate for ARA production followed by soybean, colza and olive oilcakes. In the next step, seven variables including substrate particle size, moisture content, time, temperature, yeast extract supply, glucose supply and glutamate supply were surveyed and effective variables for ARA production were determined using a Plackett-Burman screening design. Analysis results showed that time (12 days), substrate particle size (1-1.4 mm) and temperature (20ºC) were the most effective variables for the highest level of ARA production respectively.

Screening of Process Variables for the Production of Extracellular Lipase from Palm Oil by Trichoderma Viride using Plackett-Burman Design

Plackett-Burman statistical screening of media constituents and operational conditions for extracellular lipase production from isolate Trichoderma viride has been carried out in submerged fermentation. This statistical design is used in the early stages of experimentation to screen out unimportant factors from a large number of possible factors. This design involves screening of up to 'n-1' variables in just 'n' number of experiments. Regression coefficients and t-values were calculated by subjecting the experimental data to statistical analysis using Minitab version 15. The effects of nine process variables were studied in twelve experimental trials. Maximum lipase activity of 7.83 μmol /ml /min was obtained in the 6th trail. Pareto chart illustrates the order of significance of the variables affecting the lipase production. The present study concludes that the most significant variables affecting lipase production were found to be palm oil, yeast extract, K2HPO4, MgSO4 and CaCl2.