Plackett-Burman Research Articles

Optimization of Independent Variables for the Production of Extracellular Alpha Amylase by Bacillus subtilis IMD34 Using Plackett-Burman Design

Bacillus species are widely used for thermostable α-amylase production for various applications such as in the detergent and food industries. This research aimed to study the production of α-amylase by B. subtilis IMD34 in submerged fermentation conditions using Plackett-Burman (PB) design, along with the optimization of certain environmental factors for optimal growth of amylolytic bacteria. Plackett-Burman design was used to evaluate the effect of ten independent variables on α-amylase production. Maximum enzyme activity was recorded in a medium containing (g/l); starch (6.0), peptone (12.0), K2HPO4 (1.0), Na2HPO4 (2.0), NaCl (0.6), Na2SO4 (0.1), MgSO4.7H20 (1.0), FeCl3.6H2O (0.02), CaCl2.2H2O (0.02) and with an agitation speed (200 rpm). From the pareto chart it was observed that the most important independent variables for amylase production were starch, peptone, NaCl, Mg2SO4, CaCl2.2H2O and agitation. Optimization of environmental parameters to assess their effect on the growth of bacteria showed that optimal pH, incubation time and temperature were recorded at 6, 48h and 37˚C. Evaluating independent variables statistically using PB design revealed that some of the medium’s components had a positive impact on α-amylase production by B. subtilis IMD34.

Determination of Discrete Element Modelling Parameters of a Paddy Soil with a High Moisture Content (>40%)

Discrete element modelling (DEM) parameters are of great importance for the accurate prediction of soil properties and disturbance. This study aimed to provide an efficient method for accurately determining the DEM parameters of a paddy soil with a high moisture content (45.66%). The DEM parameters of the paddy soil modelled using the Hertz–Mindlin with JKR contact model were determined by using the Plackett–Burman (PB), steepest ascent, and central composite tests. The accuracies of the developed DEM models were evaluated using actual slumping tests. Based on the PB test, the surface energy of soil, coefficients of soil–soil rolling friction, and coefficients of soil–steel static friction exerted larger influences on the overall relative error (δZH). The optimization results showed that to achieve a minimum δZH (5.96%), the surface energy of soil, the coefficients of soil–soil rolling friction, and the coefficients of soil–steel static friction should be 0.808 J m−2, 0.11, and 0.6, respectively. The optimized DEM model had an overall relative error of 7.27% with a coefficient of variation of 1.32%, indicating that the DEM parameters of the calibrated paddy soil had good accuracy.

Prospecting and engineering yeasts for ethanol production under inhibitory conditions: an experimental design analysis.

The recently discovered wild yeast Wickerhamomyces sp. UFFS-CE-3.1.2 was analyzed through a high-throughput experimental design to improve ethanol yields in synthetic media with glucose, xylose, and cellobiose as carbon sources and acetic acid, furfural, formic acid, and NaCl as fermentation inhibitors. After Plackett-Burman (PB) and central composite design (CCD), the optimized condition was used in a fermentation kinetic analysis to compare this yeast's performance with an industrial Saccharomyces cerevisiae strain (JDY-01) genetically engineered to achieve a higher xylose fermentation capacity and fermentation inhibitors tolerance by overexpressing the genes XYL1, XYL2, XKS1, and TAL1. Our results show that furfural and NaCl had no significant effect on sugar consumption by UFFS-CE-3.1.2. Surprisingly, acetic acid negatively affected glucose but not xylose and cellobiose consumption. In contrast, the pH positively affected all the analyzed responses, indicating a cell's preference for alkaline environments. In the CCD, sugar concentration negatively affected the yields of ethanol, xylitol, and cellular biomass. Therefore, fermentation kinetics were carried out with the average concentrations of sugars and fermentation inhibitors and the highest tested pH value (8.0). Although UFFS-CE-3.1.2 fermented glucose efficiently, xylose and cellobiose were mainly used for cellular growth. Interestingly, the genetically engineered strain JDY-01 consumed ~ 30% more xylose and produced ~ 20% more ethanol. Also, while UFFS-CE-3.1.2 only consumed 32% of the acetic acid of the medium, JDY-01 consumed > 60% of it, reducing its toxic effects. Thus, the overexpressed genes played an essential role in the inhibitors' tolerance, and the applied engineering strategy may help improve 2G ethanol production.

Effective visible-light-driven photocatalytic degradation of fenitrothion by s-gC3N4/Ag-Au bimetallic nanocomposite

ABSTRACT This paper reports on the optimization of fenitrothion photocatalytic degradation in visible light based on Plackett Burman (PB) design and central composite design (CCD) in response surface methodology (RSM). A herbicide routinely used with a negative impact on the environment is fenitrothion, which must be degraded to minimize the impact on the environment. For fenitrothion degradation, Ag-Au bimetallic nanoparticles on the semiconducting s-doped gC3N4 surface were synthesized using the galvanic exchange. The properties of s-gC3N4/Ag-Au bimetallic nanocomposite were confirmed by various methods. Significant factors responsible for fenitrothion photocatalytic degradation were determined using Plackett-Burman (PB) design and were catalyst dosage, initial fenitrothion concentration, H2O2 concentration, pH, and rotational speed. Central composite design (CCD) design was used for further optimization. The optimum conditions for the maximum degradation of fenitrothion (100%) constraints were found to be 100% an amount of H2O2 concentration 60 mM, pH 10, rotational speed 700 rpm. These results showed that s-gC3N4/Ag-Au bimetallic nanocomposite could act as a suitable photocatalyst under visible light in the degradation of fenitrothion. By removing fenitrothion from real water samples, as well as by maintaining its stability and reusability in five successive cycles, the practicality of this nanocomposite was demonstrated.

Production and statistical optimization of cholesterol-oxidase generated by Streptomyces sp. AN strain

BackgroundCholesterol oxidases (CHOs) have attracted enormous attention because of their wide biotechnological potential. The present study explores the production of CHOs by Streptomyces sp. AN. Evaluation of culture conditions affecting enzyme production, medium optimization and released metabolite characteristics were also investigated. ResultsThe current work reports the isolation of 37 colonies (bacteria/actinobacteria) with different morphotypes from different soil/water samples. The isolate-coded AN was selected for its high potency for CHO production. Morphological characteristics and the obtained partial sequence of 16srRNA of AN showed 99.38% identity to Streptomyces sp. strain P12–37. Factors affecting CHO production were evaluated using Plackett-Burman (PB) and Box-Behnken (BB) statistical designs to find out the optimum level of the most effective variables, namely, pH, starch, NH4NO3 and FeSO4.7H2O with a predicted activity of 6.56 U/mL. According to this optimization, the following medium composition was considered to be optimum (g/L): cholesterol 1, starch 6, MgSO4.7H2O 0.1, CaCl2 0.01, FeSO4.7H2O 0.1, NH4NO3 23.97, yeast extract (YE) 0.2, K2HPO4 0.01, KH2PO4 0.1, NaCl 0.01, Tween 20 0.01, pH 6.36 and incubation temperature (30 °C) for 9 days. Spectophotometric analysis for released metabolites against cholesterol (standard) via Fourier-transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC) was carried out. FTIR spectrum showed the appearance of new absorption peaks at 1644 and 1725cm−1; this confirmed the presence of the Keto group (C=O) stretch bond. Besides, fermentation caused changes in thermal properties such as melting temperature peak (99.26; 148.77 °C), heat flow (− 8; − 3.6 Mw/mg), capacity (− 924.69; − 209.77 mJ) and heat enthalpy (− 385.29; 69.83 J/g) by comparison to the standard cholesterol as recognized through DSC thermogram. These changes are attributed to the action of the CHO enzyme and the release of keto derivatives of cholesterol with different properties. ConclusionStreptomyces sp. AN was endowed with the capability to produce CHO. Enzyme maximization was followed using a statistical experimental approach, leading to a 2.6-fold increase in the overall activity compared to the basal condition. CHO catalyzed the oxidation of cholesterol; this was verified by the appearance of a new keto group (C=O) peak at 1644 and 1725 cm−1 observed by FTIR spectroscopic analysis. Also, DSC thermogram demonstrates the alteration of cholesterol triggered by CHO.

Bioprocess development for biosurfactant production by Natrialba sp. M6 with effective direct virucidal and anti-replicative potential against HCV and HSV

Halophilic archaea is considered an promising natural source of many important metabolites. This study focused on one of the surface-active biomolecules named biosurfactants produced by haloarchaeon Natrialba sp. M6. The production trend was optimized and the product was partially purified and identified using GC–Mass spectrometry. Sequential optimization approaches, Plackett–Burman (PB) and Box–Behnken Designs (BBD) were applied to maximize the biosurfactants production from M6 strain by using 14 factors; pH, NaCl, agitation and glycerol; the most significant factors that influenced the biosurfactant production were used for Response Surface Methodology (RSM). The final optimal production conditions were agitation (150 rpm), glycerol (3%), NaCl (20.8%), pH (12) and cultivation temperature (37°C). GC–Mass spectrometry for the recovered extract revealed the presence of a diverse group of bipolar nature, hydrophobic hydrocarbon chain and charged function group. The majority of these compounds are fatty acids. Based on results of GC–MS, compositional analysis content and Zetasizer, it was proposed that the extracted biosurfactant produced by haloarchaeon Natrialba sp. M6 could be a cationic lipoprotein. The antiviral activity of such biosurfactant was investigated against hepatitis C (HCV) and herpes simplex (HSV1) viruses at its maximum safe doses (20 μg/mL and 8 μg/mL, respectively). Its mode of antiviral action was declared to be primarily via deactivating viral envelopes thus preventing viral entry. Moreover, this biosurfactant inhibited RNA polymerase- and DNA polymerase-mediated viral replication at IC50 of 2.28 and 4.39 μg/mL, respectively also. Molecular docking studies showed that surfactin resided well and was bound to the specified motif with low and accepted binding energies (ΔG = − 5.629, − 6.997 kcal/mol) respectively. Therefore, such biosurfactant could be presented as a natural safe and effective novel antiviral agent.

Enhancing the ability of two kinds of oil-degrading bacteria to treat oily sludge by optimising the growth conditions using a response surface methodology.

Biosurfactant are Surfactants produced by certain microorganisms. These biosurfactants increase the biodegradability of insoluble pollutants. In this study, the fermentation products of Pseudomonas stutzeri Lh-42 (PS) and Rhodococcus sp. PR-1 (RD) were studied by Oil spreading method, emulsifying activity and infrared spectrum analysis. It was proved that these fermentation products were biosurfactant. And then the fermentation conditions of PS, RD were optimised by Placket-Burman (PB) design, hill-climbing experiment and response surface methodology (RSM). N source and liquid loading were significant factors in the fermentation of PS, while C source and speed were significant factors in the fermentation of RD. The surface tension was found to be as low as 39.53 ± 0.25 mN/m for the fermentation conditions of PS with an N source of 4.62 ± 0.41 g and a liquid loading of 28.4 ± 0.3%. The surface tension was 40.70 ± 0.47 mN/m for the incubation conditions of RD with a C source of 26.94 ± 0.62 g and a rotational speed of 210 r/min. Finally, the experimental results for the degradation of oily sludge showed that the degradation rate of oily sludge was improved when the fermentation conditions were optimised. The results of the infrared spectroscopy analysis showed that the organic matter content of the oily sludge treated with PS bacteria was significantly reduced after the optimised fermentation. This study provides a theoretical reference for further use of these bacteria to produce biosurfactants to treat organic matter.

Media Optimization by Response Surface Methodology for the Enhanced Production of Acidic Extracellular Pectinase by the Indigenously Isolated Novel Strain Aspergillus cervinus ARS2 Using Solid-State Fermentation

Pectinolytic enzymes are related enzymes that hydrolyze pectic substances. Pectinolytic enzymes are of great interest in industrial applications for softening fruits, extracting and clarifying juices, extracting olive oil, retting textile fibers, preparing gel, and isolating protoplasts. The current work presents acidic extracellular pectinase production using low-cost agro-industrial waste with the indigenously isolated novel strain Aspergillus cervinus. Two fungal isolates, ARS2 and ARS8, with maximum pectinase activity, 41.88 ± 1.57 IU/mL and 39.27 ± 1.14 IU/mL, respectively, were screened out of 27 isolates from decayed fruit peels (orange, banana, and lemon) and soil containing decomposed vegetables. The isolate ARS2, identified as Aspergillus cervinus by molecular characterization, showed the highest pectinase activity of 43.05 ± 1.38IU/mL during screening and was further used for media component screening and optimization studies. To understand their effect on pectinase activity, one-factor-at-a-time (OFAT) studies were conducted on carbon sources, nitrogen sources, and mineral salts. The OFAT results showed the highest pectinase activity for orange peel (carbon source) at 44.51 ± 1.33 IU/mL, peptone (nitrogen source) at 45.05 ± 1.04 IU/mL, and NaH2PO4 (mineral salts) at 43.21 ± 1.12 IU/mL. The most significant media components screened by the Plackett–Burman (PB) design based on the p-value, Pareto chart, and main effect plot, were orange peel (p < 0.001), peptone (p < 0.001), NaH2PO4 (p < 0.001), and KH2PO4 (p < 0.001), which were further optimized using Response Surface Methodology (RSM) and Central Composite Design (CCD). The optimization results for the media components showed a maximum pectinase activity of 105.65 ± 0.31 IU/mL for 10.63 g orange peel, 3.96 g/L peptone, 2.07 g/L KH2PO4, and 2.10 g/L NaH2PO4. Thus, it was discovered that the indigenously isolated novel strain Aspergillus cervinus ARS2 was able to successfully produce a significant amount of pectinase using agro-industrial waste. Therefore, it can be considered for the large-scale optimized production of pectinase to meet industrial demands.

Oil-degrading bacterial consortium from Gulf of Mexico designed by a factorial method, reveals stable population dynamics

We describe an assembled marine bacterial consortium designed for bioremediation of oil-contaminated seawater, based on a statistical method using a Plackett-Burman (PB) experimental approach. The final consortium consists of four bacteria isolated from the Gulf of Mexico, from four genera: Pseudomonas, Halopseudomonas, Paenarthrobacter, and Alcanivorax. Individually, bacterial oil removal by these microorganisms was evaluated by gravimetry, reaching 39% at maximum after 75 days, whereas in consortium it was ~62%. We also measured biodegradation levels by Gas Chromatography/Mass Spectrometry (GC-MS) observing 12 polyaromatic hydrocarbons (PAHs) degradation analyzed and n-alkanes degradation with a preference for specific chain length. Consortium population analysis using the V3-V4 region of 16S rRNA showed a stable community, suggesting that the metabolic load was distributed among bacteria and that stable dynamic interactions were achieved. In this work, we show that the use of a factorial method for synthetic consortium design offers the possibility of improving oil degradation efficiency with stable bacterial populations.

Quality by Design: Optimization of Letrozole Solid Lipid Nanoparticle for Breast Cancer

Abstract: Aim: Hormone responsive breast cancer is the most prevalent cancer worldwide. Letrozole is a third-generation aromatase inhibitor widely used for the treatment of advanced breast cancer. The primary objective of the present work is to develop and optimize an injectable solid lipid nanoparticle incorporating letrozole to circumvent the side-effects of a marketed conventional formulation and thereby improve patient compliance. Materials and Methods: Emulsification solvent evaporation and melt dispersion techniques were used for formulating the said solid lipid nanoparticles. Quality by design concept was used for the development and optimization of formulation and process variables. Results: The optimum level selected is 60 mg lipid, 30 mg surfactant, and co-surfactant, 15000 psi HPH pressure, and 15 HPH passes. Conclusion: Glyceryl dibehenate was selected as suitable lipid based on solubility and partition coefficient. With an increase in lipid content there is increase in particle size and PDI and decrease in entrapment efficiency. Higher surfactant and co-surfactant concentrations result in lower particle size, PDI, higher zeta potential, and lower entrapment efficiency. An increase in HPH pressure reduced particle size and PDI up to a certain level, however, the increase in HPH pressure from 15000 to 20000 psi increased particle size. An increase in the number of HPH passes reduces particle size and PDI. The drug release mechanism for LTR-SLN was found to follow the first order and higuichi model. Keywords: Solid lipid nanoparticle, Letrozole, Ishikawa diagram, Plackett burman design, Central composite design, Mathematical modeling.

Characterisation of catechins and their oxidised derivatives in Ceylon tea using multi-dimensional liquid chromatography and high-resolution mass spectrometry

Tea is a complex food matrix comprising of many structurally diverse compounds, of which catechins and their oxidised derivatives are of particular interest due to their nutritional functionality. However, these catechins and derivatives exist in various isomeric forms with few or no pure standards available, rendering their analysis challenging. A method combining multi-dimensional liquid chromatography (MDLC) and high-resolution mass spectrometry (HRMS) was developed for the characterisation of these compounds using Ceylon tea as a model. Based on a Plackett–Burman (PB) design, flow rate and initial methanol percentage were identified as the most significant factors (p < 0.05) affecting chromatogram coverage and resolution (Rs) for comprehensive two-dimensional LC (LCxLC) and heart-cutting two-dimensional LC (LC-LC) respectively. Central composite design (CCD) was then applied using these parameters for method optimisation and to identify second-order relationships between screened parameters. The optimised LCxLC (flow rate: 2.18 mL/min and initial methanol percentage: 28.0%) and LC-LC (flow rate: 0.86 mL/min and initial methanol percentage for different cuts: A- 10.0%; B- 15.8%; and C- 18.7%) methods were applied to the analysis of Ceylon tea samples from seven regions of Sri Lanka and demonstrated an improved separation of co-eluting isomeric compounds. Finally, with the mass spectral information from HRMS, a total of 31 compounds (eight monomers, 17 dimers, five trimers and one tetramer) were detected and putatively identified in Ceylon tea.

A DEM-Based Modeling Method and Simulation Parameter Selection for Cyperus esculentus Seeds

To build a DEM model of Cyperus esculentus seed particles, the shape and size of the Cyperus esculentus seed particles were measured and analyzed. The results showed that the dispersity in size had a normal distribution. Additionally, a certain functional relationship between the primary dimension and secondary dimensions was determined. The width of the seed was the primary dimension, and the other secondary dimensions (length and thickness) were calculated based on their relationships with the primary dimension. On this basis, an approach for modeling Cyperus esculentus seed particles based on the multi-sphere (MS) method was proposed. The discrete element analysis models of three varieties of Cyperus esculentus seeds were established with different numbers of filing spheres. Moreover, to obtain more accurate simulation parameters, first, a range of values of the simulation parameters was obtained by the experimental method. Second, the Plackett–Burman (PB) test and the path of steepest ascent method were both adopted to correct and calibrate the simulation parameters, which were difficult to obtain through experiments, and simulation of the direct shear test was used for calibration. All of the methods guaranteed that the selected parameters were reasonable. The test results showed that the static friction coefficient of seed–seed had a significant effect on the simulation results. Finally, piling tests and the bulk density test were used for modeling verification. By comparing the simulated results and experimental results in the piling tests and bulk density test, when the number of filing spheres increased, the simulated results were close to those obtained experimentally. Therefore, the feasibility and validity of the modeling method for Cyperus esculentus seed particles that we proposed and the simulation parameters that were obtained were verified.

On the identification of active factors in nonregular two‐level designs with a small number of runs

AbstractNonregular two‐level designs are attractive screening designs due to their good projection properties and flexible run sizes. In particular, the 12‐run Plackett–Burman (PB) design has become quite popular. However, existing methods struggle with the identification of active factors when the number of active factors exceeds the projectivity of the designs. This is especially the case when interactions are present, the variance is high and the number of runs is small. In this paper, we propose a method for analysing nonregular two‐level designs that particularly addresses the issues above. It exploits the projection properties of designs and is here applied on the 12‐run PB design and the 16‐run no‐confounding (NC) designs. In the construction of the method, the use of test‐ and penalty‐based procedures are avoided. Instead, the number of allowed terms in a model is restricted. The effectiveness of the method and comparison between designs are evaluated by simulations for different scenarios. Ways to evaluate the reliability of the screening procedure are pointed out. An example with real data is given to demonstrate how one might perform the analysis in practice.

Successive approach of medium optimization using one-factor-at-a-time and response surface methodology for improved β-mannanase production from Streptomyces sp.

β-Mannanase is an enzyme catalyzing the β-1,4-mannosidic linkages of the hemi-cellulolytic structure. Optimization of medium components was studied for β-mannanase production from Streptomyces sp. RDA1496 using One-Factor-At-a-Time and resulting activity was found to be 244.54 IU/mL. Subsequently, Plackett-Burman (PB) design was adopted to evaluate the effect of guar gum, coprameal, peptone, yeast extract, fructose, inoculum size, NaCl, K2HPO4, KH2PO4 and CaCl2. Analysis of variance revealed the correlation coefficient (R2) of 0.9996 for the performed model at a significant level (p < 0.05). Four significant variables were optimized using central composite design of response surface methodology (RSM) at five different coded levels. The optimum concentration of significant factors namely fructose, guar gum, peptone, and K2HPO4 were found to be 4.81 g/L, 7.09 g/L, 4.91 g/L, and 2.05 g/L respectively. PB and RSM-guided experimental design, resulted in maximum β-mannanase production of 415.96 IU/mL, which is 5.55-fold increase from un-optimized condition.

Isolation and optimization of extracellular PHB depolymerase producer Aeromonas caviae Kuk1-(34) for sustainable solid waste management of biodegradable polymers

Bioplastics, synthesized by several microbes, accumulates inside cells under stress conditions as a storage material. Several microbial enzymes play a crucial role in their degradation. This research was carried to test the biodegradability of poly-β-hydroxybutyrate (PHB) utilizing PHB depolymerase, produced by bacteria isolated from sewage waste soil samples. Potent PHB degrader was screened based on the highest zone of hydrolysis followed by PHB depolymerase activity. Soil burial method was employed to check their degradation ability at different incubation periods of 15, 30, and 45 days at 37±2°C, pH 7.0 at 60% moisture with 1% microbial inoculum of Aeromonas caviae Kuk1-(34) (MN414252). Without optimized conditions, 85.76% of the total weight of the PHB film was degraded after 45 days. This degradation was confirmed with Fourier-transform infrared spectroscopy (FTIR) and Scanning electron microscope (SEM) analysis. The presence of bacterial colonies on the surface of the degraded film, along with crest, holes, surface erosion, and roughness, were visible. Media optimization was carried out in statistical mode using Plackett Burman (PB) and Central Composite Design (CCD) of Response Surface Methodology (RSM) by considering ten different factors. Analysis of Variance (ANOVA), Pareto chart, response surface plots, and F-value of 3.82 implies that the above statistical model was significant. The best production of PHB depolymerase enzyme (14.98 U/mL) was observed when strain Kuk1-(34) was grown in a media containing 0.1% PHB, K2HPO4 (1.6 gm/L) at 27 ℃ for seven days. Exploiting these statistically optimized conditions, the culture was found to be a suitable candidate for the management of solid waste, where 94.4% of the total weight of the PHB film was degraded after 45 days of incubation.

Synergistic antimicrobial activity of ε-polylysine, chestnut extract, and cinnamon extract against Staphylococcus aureus.

A mixed natural preservative composed of ε-polylysine (ε-PL), chestnut 70% ethanol extract (NE), and cinnamon hydrothermal extract (CW), was investigated for the reduction of Staphylococcus aureus. The minimum inhibitory concentration (MIC) and minimum bacterial concentration (MBC) of seven natural extracts were investigated against a cocktail of three strains of S. aureus (ATCC 25923, ATCC 33591, and ATCC 33594). Three important factors (ε-PL, NE, and CW) were selected by using the Plackett-Burman (PB) design for the response surface model (P < 0.001). Following a central composite design, S. aureus were treated with mixtures of natural preservatives that included ε-PL, NE, and CW. The MIC and MBC of ε-PL and the natural extracts and ranged from 1 to 16mg/mL (R2 = 0.9857). The mixed natural preservative presented a synergistic antibacterial effect, at the optimum point. These results suggest that mixed natural preservatives of ε-PL, NE, and CW can lower the economic cost of food processing.

Measurement and Calibration of the Discrete Element Parameters of Coated Delinted Cotton Seeds

To simulate the interactions between a pneumatic cotton precision seed-metering device and coated delinted cotton seeds accurately, physical and simulation experiments based on a rotating drum apparatus were combined to calibrate the discrete element simulation parameters of E`kangmian-10 cotton seeds. Firstly, the contact parameters and the dynamic repose angle of the cotton seeds were measured through physical tests. Based on the particle size requirement of the Discrete Element Method (DEM) and Computational Fluid Dynamics (CFD) coupling simulation and the reverse engineering technology, the cotton seed discrete element bonded-particle model (BPM) was established. Secondly, taking the contact parameters as calibration objects and the simulated dynamic repose angle as the evaluation index, a Plackett–Burman (PB) test was designed for significance screening. The results of the screening test showed that the static friction coefficient of cotton seed–tough photosensitive resin, the impact recovery coefficient of cotton seed–cotton seed, and the static friction coefficient of cotton seed–cotton seed had a highly significant effect on the simulated dynamic repose angle. Next, a Box–Behnken Design (BBD) test was adopted to establish the quadratic regression model between significant parameters and the simulated dynamic repose angle, and then the multi-factor optimization solution was carried out to obtain the optimal combination of parameters: the static friction coefficient of cotton seed–tough photosensitive resin and the impact recovery coefficient and static friction coefficient of cotton seed–cotton seed were 0.33, 0.06 and 0.10, respectively. Lastly, verification tests on the rotating drum apparatus and the seed-metering device were performed, and their relative errors were less than 2%, which indicated that the discrete element models and the contact parameters of the coated delinted cotton seeds were reliable. This study provides a reference for the selection of the discrete element parameters of coated delinted cotton seeds for DEM-CFD coupling simulation and the optimal design of precision seed-metering device for cotton.

Optimization of Carotenoids Production from Rhodotorula sp. Strain ATL72 for Enhancing Its Biotechnological Applications.

Rhodotorula yeasts which are known as carotenogenic yeasts have a great industrial value due to their ability to produce carotenoids. In particular, the isolated yeast Rhodotorula sp. (strain ATL72) has been reported to be a promising producer of high concentrations of carotenoids. A combination of central composite design (CCD) and Plackett–Burman (PB) design was used to optimize carotenoids produced by this yeast. The optimum production of carotenoids was completed when the yeast was grown in a production medium composed of 3.7 g/L malt extract, 7.7 g/L fructose, 9 g/L urea, 35 g/L NaCl, and 1 g/L yeast extract at 27.5 °C, pH 6.7, and 180 rpm. Two batch runs in 1 L and 7 L bioreactors were conducted which increased the productivity of carotenoid concentration from 21.5 mg/L after 98 h of incubation at the level of the shake flask to 229.9 mg/L after 47 h of incubation at the level of 7 L bioreactor. The carotenoid pigment was extracted in dimethylsulfoxide (DMSO), acetone, petroleum ether, and sodium chloride, and subsequently identified and characterized using UV-visible scanning, thin layer chromatography, and gas chromatography/mass spectrometry.

Extracting oil from grape seed using a combined wet enzymatic process and pressing

The novel method of wet enzymatic extraction and pressing (WEEP) was used to extract grape seed oil. Low-field nuclear magnetic resonance (LF-NMR) was applied to elucidate the water mobility and distribution of grape seeds during the process. The extraction variables were selected in accordance with a Plackett-Burman (PB) design, and the statistical model was constructed using response surface methodology (RSM). The results revealed that water mobility variations of the samples could be discriminated using the signal amplitude and peak position of the T2 inversion spectrum. The increase in water content was affected by the time and temperature of the process. The water migration of the samples was monitored clearly and intuitively using magnetic resonance imaging (MRI). Three significant factors out of the six variables (enzyme additive amount, hydrolysis temperature, hydrolysis time, comminution degree, screw speed, squeezed water) were determined. The ideal values of the significant factors were confirmed to be 1% of enzyme additive amount, 60 mesh of comminution degree, and 8% of squeezed water. The residual oil proportion to the initial oil content decreased significantly with parameter optimization (33.24% ± 5.98%) compared with that of the unoptimized parameters (57.79% ± 4.13%).

Statistical sequential optimization of process parameters for inulinase production by Kluyveromyces marxianus ATCC 36907 in solid-state fermentation using beer residue

The biotechnological production of inulinase , a hydrolytic enzyme with important applications in the food and pharmaceutical industry, is presented as a possible route for the valorization of beer residues. The current study employs a sequential optimization strategy, based on statistical experimental designs, was employed to optimize inulinase production by Kluyveromyces marxianus ATCC 36907 in solid-state fermentation (SSF), including the Plackett-Burman (PB) design and the Central Composite Rotatable Design (CCRD). The most important variables selected of SSF using dried beer residue were the concentration of sugarcane molasses , concentration of yeast extract (additional source of nitrogen) and the incubation temperature . The CCRD, which through the response surface methodology (RSM), determined the optimal fermentation conditions for these three variables, which were 47.1% (w/w), 0.032 g gds −1 and 37.5 °C, respectively. It was found that the RSM model has a predicted R 2 value of 0.9512 and adjusted R 2 of 0.8884, and can significantly predict (p < 0.05) the response variables of enzyme activity . Under these optimal conditions, it was possible to obtain a maximum inulinase activity of 239.38 U gds −1 of dried beer residue and maximum productivity at 48 h of 3.69 U gds.h −1 . While there is ample literature on inulinase production by solid-state fermentation using K. marxianus , this is the first report on the use of beer residue as a potential source for the production of this enzyme with significant high yield that is on par with the enzymatic activities reported so far using different substrates and different lineages of K. marxianus. • Two statistical designs were used sequentially to optimize inulinase production. • The dried beer residue proved to be effective as a new substrate and support in solid-state fermentations. • Pre-treated sugarcane molasses as an enzyme inducer, contributed to increase inulinase activity. • The high incubation temperature proved to be effective for increased inulinase activity. • The application of central composite design increased the inulinase activity 3.57-folds.