Response Surface Optimization of the Critical Medium Components for Pullulan Production by <Emphasis Type="Italic">Aureobasidium pullulans</Emphasis> FB-1

Culture conditions for pullulan production by Aureobasidium pullulans were optimized using response surface methodology at shake flask level without pH control. In the present investigation, a five-level with five-factor central composite rotatable design of experiments was employed to optimize the levels of five factors significantly affecting the pullulan production, biomass production, and sugar utilization in submerged cultivation. The selected factors included concentration of sucrose, ammonium sulphate, yeast extract, dipotassium hydrogen phosphate, and sodium chloride. Using this methodology, the optimal values for concentration of sucrose, ammonium sulphate, yeast extract, dipotassium hydrogen phosphate, and sodium chloride were 5.31%, 0.11%, 0.07%, 0.05%, and 0.15% (w/v), respectively. This optimized medium has projected a theoretically production of pullulan of 4.44%, biomass yield of 1.03%, and sugar utilization of 97.12%. The multiple correlation coefficient ‘R’ was 0.9976, 0.9761 and 0.9919 for pullulan production, biomass production, and sugar utilization, respectively. The value of R being very close to one justifies an excellent correlation between the predicted and the experimental data.     

Production of ethanol from cellulosic biomass by Clostridium thermocellum SS19 in submerged fermentation

Carotenogenesis of the lactose-negative yeast Rhodotorula rubra GED5 was studied by cocultivation with Kluyveromyces lactis MP11 in whey ultrafiltrate (WU) (35, 50, and 70 g of lactose/L). Maximum yields of cell mass (24.3 g/L) and carotenoids (10.2 mg/L of culture fluid or 0.421 μ g/g of dry cells) were obtained by growing the microbial association in WU (50 g of lactose/L) in a fermentor with an airflow rate of 0.8 L/(L·min), agitation of 220 rpm, and temperature of 30°C. The identified carotenoid pigments—β-carotene, torulene, and torularhodin—reached maximum concentrations (133, 26.9, and 222.3 μg/g of dry cells, respectively) on d 5 for torulene and d 6 for β-carotene and torularhodin.     

Analyses of the temperature and pH effects on the complexation of magnesium and calcium in human blood plasma: an approach using artificial neural networks

The temperature and pH effects on the equilibrium of a blood plasma model have been studied on the basis of artificial neural networks. The proposed blood plasma was modeled considering two important metals, calcium and magnesium, and six ligands, namely, alanate, carbonate, citrate, glycinate, histidinate and succinate. A large data set has been used to simulate different concentrations of magnesium and calcium as a function of temperature and pH and these data were used for training the neural network. The proposed model allowed different types of analyses, such as the effects of pH on calcium and magnesium concentrations, the competition between calcium and magnesium for ligands and the effects of temperature on calcium and magnesium concentrations. The model developed was also used to predict how the variation of calcium concentration can affect magnesium concentrations. A comparison of neural network predictions against experimental data produced errors of about 3%. Moreover, in agreement with experimental measurements (Wang et al. in Arch. Pathol. 126:947–950, 2002; Heining et al. in Scand. J. Clin. Lab. Invest. 43:709–714, 1983), the artificial neural network predicted that calcium and magnesium concentrations decrease when pH increases. Similarly, the magnesium concentrations are less sensitive than calcium concentrations to pH changes. It is also found that both calcium and magnesium concentrations decrease when the temperature increases. Finally, the theoretical model also predicted that an increase of calcium concentrations will lead to an increase of magnesium concentration almost at the same rate. These results suggest that artificial neural networks can be efficiently applied as a complementary tool for studying metal ion complexation, with especial attention to the blood plasma analysis. Figure Artificial neural networks for predicting the behavior of calcium and magnesium as a function of pH and temperature in human blood plasma     

Response Surface Optimization of Medium Components for Naringinase Production from <Emphasis Type="Italic">Staphylococcus xylosus MAK2</Emphasis>

Response surface methodology was used to optimize the fermentation medium for enhancing naringinase production by Staphylococcus xylosus. The first step of this process involved the individual adjustment and optimization of various medium components at shake flask level. Sources of carbon (sucrose) and nitrogen (sodium nitrate), as well as an inducer (naringin) and pH levels were all found to be the important factors significantly affecting naringinase production. In the second step, a 22 full factorial central composite design was applied to determine the optimal levels of each of the significant variables. A second-order polynomial was derived by multiple regression analysis on the experimental data. Using this methodology, the optimum values for the critical components were obtained as follows: sucrose, 10.0%; sodium nitrate, 10.0%; pH 5.6; biomass concentration, 1.58%; and naringin, 0.50% (w/v), respectively. Under optimal conditions, the experimental naringinase production was 8.45 U/mL. The determination coefficients (R ²) were 0.9908 and 0.9950 for naringinase activity and biomass production, respectively, indicating an adequate degree of reliability in the model.     

Effect of Zeolite NaY and Ca-Montmorillonite on Ethanol Production Using Synthetic Molasses

The influence of Ca-Montmorillonite (Ca-MNT) and zeolite NaY addition on ethanol production from synthetic molasses by S. cerevisiae 251 TP(3-2) was studied by the measurement of biomass concentrations and metal ion concentration with respect to fermentation time. Addition of 5 g/L Ca-MNT and 10 g/L zeolite NaY resulted in an increase in both ethanol concentration and ethanol production rate. This increase was 24 and 40% for ethanol concentration and 65 and 87% for production rate, respectively. From the ion analyses, it was observed that the NaY added to the medium decreased the toxic concentration of zinc, manganese, and iron cations and acted also as a pH regulator. Ca-MNT added to the medium decreased the concentration of Na⁺ ions, which is known to have a toxic effect on glycolysis and cell concentration. These effects caused improvement in the ethanol production rate.     

Ethanol Production from Residual Wood Chips of Cellulose Industry: Acid Pretreatment Investigation,Hemicellulosic Hydrolysate Fermentation,and Remaining Solid Fraction Fermentation by SSF Process

Current research indicates the ethanol fuel production from lignocellulosic materials, such as residual wood chips from the cellulose industry, as new emerging technology. This work aimed at evaluating the ethanol production from hemicellulose of eucalyptus chips by diluted acid pretreatment and the subsequent fermentation of the generated hydrolysate by a flocculating strain of Pichia stipitis. The remaining solid fraction generated after pretreatment was subjected to enzymatic hydrolysis, which was carried out simultaneously with glucose fermentation [saccharification and fermentation (SSF) process] using a strain of Saccharomyces cerevisiae. The acid pretreatment was evaluated using a central composite design for sulfuric acid concentration (1.0–4.0 v/v) and solid to liquid ratio (1:2–1:4, grams to milliliter) as independent variables. A maximum xylose concentration of 50 g/L was obtained in the hemicellulosic hydrolysate. The fermentation of hemicellulosic hydrolysate and the SSF process were performed in bioreactors and the final ethanol concentrations of 15.3 g/L and 28.7 g/L were obtained, respectively.     

Optimization of Fermentation Parameters to Enhance the Production of Ethanol from Palmyra Jaggery Using <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> in a Batch Fermentor

Application of statistical experimental designs for optimization of fermentation parameters to enhance ethanol production, which is an economical and renewable energy source using Saccharomyces cerevisiae NCIM 3090 from palmyra jaggery, was studied in a batch fermentor. Using Plackett–Burman design, impeller speed, concentrations of CoCl₂ and KH₂PO₄ were identified as significant variables, which highly influenced ethanol production, and these variables were further optimized using a central composite design (CCD). The ethanol production was adequately approximated with a full quadratic equation obtained from three factors and five levels of CCD. Maximum ethanol concentration of 132.56 g/l (16.8% [v/v]) was obtained for an impeller speed of 247.179 (≈250) rev/min, CoCl₂ of 0.263 g/l and KH₂PO₄ of 2.39 g/l. A second-order polynomial regression model was fitted and was found adequate with R ² of 0.8952. This combined statistical approach enables rapid identification and investigation of significant parameters for improving the ethanol production and could be very useful in optimizing processes.     

Optimization of Ultrasound-Assisted Extraction of Yields and Total Methoxyflavone Contents from Kaempferia parviflora Rhizomes

The major bioactive components of Kaempferia parviflora (KP) rhizomes, 3,5,7,3′,4′-pentamethoxyflavone (PMF), 5,7-dimethoxyflavone (DMF), and 5,7,4′-trimethoxyflavone (TMF), were chosen as the quantitative and qualitative markers for this plant material. In order to extract bioactive components (total methoxyflavones) from KP rhizomes, ultrasound-assisted extraction (UAE) was proposed as part of this study. Plackett–Burman design (PBD) and Box–Behnken design (BBD) were utilized to optimize the effects of UAE on extraction yields and total methoxyflavone contents in KP rhizomes. First, PBD was utilized to determine the effect of five independent variables on total yields and total methoxyflavone contents. The results indicated that the concentration of the extracting solvent (ethanol), the extraction time, and the ratio of solvent to solid were significant independent terms. Subsequently, BBD with three-level factorial experiments was used to optimize the crucial variables. It was discovered that the concentration of ethanol was the most influential variable on yields and total methoxyflavone contents. Optimum conditions for extraction yield were ethanol concentration (54.24% v/v), extraction time (25.25 min), and solvent-to-solid ratio (49.63 mL/g), while optimum conditions for total methoxyflavone content were ethanol concentration (95.00% v/v), extraction time (15.99 min), and solvent-to-solid ratio (50.00 mL/g). The relationship between the experimental and theoretical values was perfect, which proved that the regression models used were correct and that PBD and BBD were used to optimize the conditions in the UAE to obtain the highest yield and total methoxyflavone content in the KP rhizomes.     

金属氯化物对硼氢化钠还原α-氯代苯乙酮的影响

用NaBH₄还原α-氯代苯乙酮制备相应的β-氯代-α-苯乙醇时, 在较高的反应温度下产生许多副产物, 钙、镧、镁、锰、锌等金属氯化物的存在能有效抑制副反应的发生, 高产率地得到产物.     

Alcoholic Fermentation with Flocculant Saccharomyces cerevisiae in Fed-Batch Process

Studies have been conducted on selecting yeast strains for use in fermentation for ethanol production to improve the performance of industrial plants and decrease production costs. In this paper, we study alcoholic fermentation in a fed-batch process using a Saccharomyces cerevisiae yeast strain with flocculant characteristics. Central composite design (CCD) was used to determine the optimal combination of the variables involved, with the sucrose concentration of 170 g/L, a cellular concentration in the inoculum of 40 % (v/v), and a filling time of 6 h, which resulted in a 92.20 % yield relative to the theoretical maximum yield, a productivity of 6.01 g/L h and a residual sucrose concentration of 44.33 g/L. With some changes in the process such as recirculation of medium during the fermentation process and increase in cellular concentration in the inoculum after use of the CCD was possible to reduce the residual sucrose concentration to 2.8 g/L in 9 h of fermentation and increase yield and productivity for 92.75 % and 9.26 g/L h, respectively. A model was developed to describe the inhibition of alcoholic fermentation kinetics by the substrate and the product. The maximum specific growth rate was 0.103 h^?1, with K _I and K _s values of 109.86 and 30.24 g/L, respectively. The experimental results from the fed-batch reactor show a good fit with the proposed model, resulting in a maximum growth rate of 0.080 h^?1.     

Characterization of the health of southern Appalachian red spruce (Piceae rubens) through determination of calcium, magnesium, and aluminum concentrations in foliage and soil

The red spruce (Picea rubens) is a conifer whose southern native range is limited to high elevation sites in the Appalachian Mountains. Red spruce decline has been reported to involve atmospheric deposition of acidic anions, which may affect cation mobility in soil, primarily calcium, magnesium, and aluminum. In this work, the concentrations of these elements were determined in red spruce foliage and surrounding soil at high elevation locations in the Great Smoky Mountains and on the Blue Ridge Parkway by inductively coupled plasma optical emission spectrometry (ICP-OES). Few differences in foliar or soil concentrations were observed at the sites, indicating that elevation and geography do not affect these values. A comparison of foliar concentrations in seedlings, saplings, and mature trees demonstrated few differences, suggesting that life stage does not affect these elemental concentrations. A previously-developed model to estimate forest health using soil calcium/aluminum ratios suggested that all sites had a significant risk of adverse health effects. However, comparisons of foliar calcium/aluminum ratios and calcium and magnesium concentrations to values obtained in the mid-1990s showed an improvement in these parameters, presumably due to decreased acidic deposition.     

Optimization of Microwave-Assisted Extraction (MAE) for the Isolation of Antioxidants from Basil (Ocimum basilicum L.) by Response Surface Methodology (RSM)

Abstract

The recovery of antioxidants from basil (Ocimum basilicum L.) was modeled with the aid of response surface methodology (RSM) using microwave-assisted extraction (MAE). Face-centered central design (FCCD) was employed to optimize the MAE operational parameters including the extraction time (1 to 7?min), extraction temperature (30 to 120?°C), solid-to-solvent ratio (0.1 to 0.4), and solvent concentration (20 to 80% ethanol, v/v), and to obtain the best possible combinations of these parameters for a high antioxidant yield from basil. The total antioxidant capacity (TAC) was expressed in trolox (TR) equivalents per gram of dried sample (DS). Three of the operational parameters (temperature, extraction time and solvent concentration) were shown to have significant effect on the extraction efficiency of antioxidants in basil extracts (p?<?0.05). The solvent concentration was shown to be the most significant factor on antioxidant yield obtained by MAE. There was a close relationship between experimental and predicted values using the proposed method. This optimized MAE method shows an application potential for the efficient extraction of antioxidants from basil in the food and pharmaceutical industries.     

Improvement of Ethanol Production Using <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> by Enhancement of Biomass and Nutrient Supplementation

Optimization of ethanol production through addition of substratum and protein-lipid additives was studied. Oilseed meal extract was used as protein lipid supplement, while rice husk was used as substratum. The effect of oil seed meal extract and rice husk was observed at varying concentration of medium sugar from 8% to 20%. Of the three oil seed meal extracts used, viz. groundnut, safflower, and sunflower, safflower was found to be most efficient. The use of oilseed meal extract at 4% was found to enhance ethanol production by almost 50% and enhanced sugar tolerance from 8% to 16%. A further increase of almost 48% ethanol was observed on addition of 2 g of rice husk per 100 ml of medium. An increase in cell mass with better sugar attenuation was observed. Further optimization was sought through use of sugarcane juice as the sugar source. While 8.9% ethanol yield with 75% sugar attenuation was observed at 20% sucrose concentration, it was found to increase to 12% (v/v) with almost complete utilization of medium sugar when sugarcane juice was used. Cell weight was also observed to increase by 26%.     

Extrapolation studies on adsorption of thorium and uranium at different solution compositions on soil sediments

Study on adsorption of thorium and uranium radionuclides by a soil sediment as a function of ionic composition of Ca, Mg and Na has been carried out. Experimentally determined slopes represents an average of adsorption on soil sediments having different relative affinities for thorium, uranium, calcium and magnesium. Both thorium and uranium were found to be adsorbed to ion-exchange sites together with calcium and magnesium cations as effective competitors An extrapolated equation for the distribution coefficientK _d was formed for both radionuclides thorium and uranium at the specified site where the soil sediments were sampled. The combined cation concentration of both calcium and magnesium in solution correlates linearly with the measuredK _d ^Th,U values.     

Production of Lactic Acid from Raw Sweet Potato Powders by Rhizopus Oryzae Immobilized in Sodium Alginate Capsules

Rhizopus oryzae immobilized in calcium alginate was applied in lactic acid fermentation with unhydrolyzed raw sweet potato powders as the sole carbon source. The effects of sodium alginate concentration, calcium chloride concentration, and the immobilized bead diameter on lactic acid production were investigated. Increase in sodium alginate concentration during the gelation process would harden the immobilized capsule, which led to a decrease in lactic acid production. The increase in calcium chloride would increase the thickness of the immobilized capsule, which would increase the mass transfer resistance. Nevertheless, while the calcium chloride was lower than 15%, it would not have obvious effects on lactic acid production. A larger bead could have more space for cell growth, which led to the maximum lactic acid production observed at the 5-mm bead diameter. Moreover, results of repeated-batch operation suggested that immobilized cells could have higher stability in lactic acid production than free cells. The total cumulative lactic acid in immobilized-cell operation could increase by 55% as compared with free-cell operation after 216 h (seven repeated-batches), and no loss of amylolytic activity was observed. The results indicated that immobilized R. oryzae by Ca-alginate could be suitable for lactic acid production from unhydrolyzed raw potato powders.     

Shear flocculation of celestite with sodium oleate and tallow amine acetate: effects of cations

Sodium oleate and tallow amine acetate (TAA) were used as surfactants for the shear flocculation of celestite. The shear-flocculation power values obtained with sodium oleate were higher than those obtained with TAA in terms of the concentrations used in the shear-flocculation experiments. In addition, sodium oleate and TAA were more effective on the celestite suspension in the pH ranges of 7-11 and 6-10, respectively. For the shear-flocculation experiments with sodium oleate at pH 11, with preaddition of calcium or magnesium ions at 5 x 10(-5) M and lower concentrations into the suspension, the shear flocculation of the celestite suspension was promoted by the coagulation process due to the calcium and magnesium cations added. However, the shear-flocculation power values decreased due to the interaction between surfactant and cations at concentration values higher than 5 x 10(-5) M for magnesium ions and 10(-3) M for calcium ion. Particularly, magnesium ions significantly reduced the shear-flocculation power values by slime coating of Mg(OH)2 precipitates.     

Determination of true selectivity coefficients of neutral carrier calcium selective electrode

Literature data concerning selectivity coefficients of neutral carrier calcium selective electrode show large discrepancies up to several orders of magnitude for sensors containing the same active component. It can be shown theoretically that determination of selectivity coefficients in non-complexing media gives too high values because of additional sources of the main ion in the membrane vicinity. Determination of selectivity coefficients in solutions containing a complexing agent gives the possibility to evaluate true selectivity coefficients not influenced by additional factors. Measurements performed in complexing as well as in non-complexing media and calculation of the detection limit indicate a good agreement between expectations and experimental results. This has been confirmed using the calcium selective electrode with the neutral carrier ETH 1001 witho-nitrophenyl octyl ether or dioctylsebacate as mediators. As interferents sodium, potassium and magnesium ions were tested.     

Fermentation kinetics of ethanol production from glucose and xylose by recombinant Saccharomyces 1400(pLNH33)

Fermentation kinetics of ethanol production from glucose, xylose, and their mixtures using a recombinant Saccharomyces 1400 (pLNH33) are reported. Single-substrate kinetics indicate that the specific growth rate of the yeast and the specific ethanol productivity on glucose as the substrate was greater than on xylose as a substrate. Ethanol yields from glucose and xylose fermentation were typically 95 and 80% of the theoretical yield, respectively. The effect of ethanol inhibition is more pronounced for xylose fermentation than for glucose fermentation. Studies on glucose-xylose mixtures indicate that the recombinant yeast co-ferments glucose and xylose. Fermentation of a 52.8 g/L glucose and 56.3 g/L xylose mixture gave an ethanol concentration of 47.9 g/L after 36 h. Based on a theoretical yield of 0.51 g ethanol/g sugars, the ethanol yield from this experiment (for data up to 24 h) was calculated to be 0.46 g ethanol/g sugar or 90% of the theoretical yield. The specific growth rate of the yeast on glucose-xylose mixtures was found to lie between the specific growth rate on glucose and the specific growth rate on xylose. Kinetic studies were used to develop a fermentation model incorporating the effects of substrate inhibition, product inhibition, and inoculum size. Good agreements were obtained between model predictions and experimental data from batch fermentation of glucose, xylose, and their mixtures.     

Optimisation of the extraction of olive (Olea europaea) leaf phenolics using water/ethanol-based solvent systems and response surface methodology

An experimental setup based on a 2³ full-factorial, central-composite design was implemented with the aim of optimising the recovery of polyphenols from olive leaves by employing reusable and nontoxic solutions composed of water/ethanol/citric acid as extracting media. The factors considered were (i) the pH of the medium, (ii) the extraction time and (iii) the ethanol concentration. The model obtained produced a satisfactory fit to the data with regard to total polyphenol extraction (R ² = 0.91, p = 0.0139), but not for the antiradical activity of the extracts (R ² = 0.67, p = 0.3734). The second-order polynomial equation obtained after analysing the experimental data indicated that ethanol concentration and time mostly affected the extraction yield, but that increased pH values were unfavourable in this regard. The maximum theoretical yield was calculated to be 250.2 ± 76.8 mg gallic acid equivalent per g of dry, chlorophyll-free tissue under optimal conditions (60% EtOH, pH 2 and 5 h). Liquid chromatography–electrospray ionisation mass spectrometry of the optimally obtained extract revealed that the principal phytochemicals recovered were luteolin 7-O-glucoside, apigenin 7-O-rutinoside and oleuropein, accompanied by smaller amounts of luteolin 3′,7-O-diglucoside, quercetin 3-O-rutinoside (rutin), luteolin 7-O-rutinoside and luteolin 3′-O-glucoside. Simple linear regression analysis between the total polyphenol and antiradical activity values gave a low and statistically insignificant correlation (R ² = 0.273, p > 0.05), suggesting that it is not the sheer amount of polyphenols that provides high antioxidant potency; instead, this potency is probably achieved through interactions among the various phenolic constituents.