Plackett-Burman Design Research Articles

Improvement of Bacillus subtilis PI agarase production, hydrolysate scavenging capability assessment, and saccharification of algal biomass for green ethanol generation

The goal of the current work was to optimize the growth parameters needed to manufacture agarase enzyme from a non-marine PI strain of Bacillus subtilis on an agar-based medium. Using Plackett–Burman design (PBD), nine process parameters were evaluated, and agar, peptone, and yeast-extract were identified as the most significant independent factors influencing agarase production with confidence levels more than 90%. To evaluate the optimal concentrations of the indicated process parameters on agarase production, the Box–Behnken design (BBD) was applied. After optimization, B. subtilis strain PI produced 119.8 U/ml of agarase, representing a 1.36-fold increase. In addition the agar hydrolysate fermented products contain the liberated oligosaccharide acts as strong antioxidant which has 62.4% scavenging activity. Also, the agarase yields increased (1141.12, 1350.253, 1684.854 and 1921.863 U/ml) after substitution the agar with algal biomass of Carolina officinalis at different concentrations (2, 5, 10 and 15%), respectively. After completing the saccharification process, the resulted hydrolysate was used to produce ethanol through fermentation with Pichia pastoris yeast strain as an economical method giving yields (6.68317, 7.09748, 7.75648 and 8.22332 mg/ml), that are higher than using yeast extract peptone dextrose (YPD) medium (4.461 mg/ml).

Optimization of Four Different Rosemary Extraction Techniques Using Plackett-Burman Design and Comparison of Their Antioxidant Compounds.

Rosemary has many medicinal and therapeutic properties and therefore it is important to study how to maximize the recovery of its bioactive compounds. In the present study, four different extraction techniques were used, namely stirring extraction (STE), pulsed electric field-assisted extraction (PEF), ultrasound probe-assisted extraction (UPAE), and ultrasound bath-assisted extraction (UBAE). First, some primary experiments were carried out in order to optimize each technique individually through the Plackett-Burman design. Then, each technique was applied under optimal conditions and the results were compared with each other. The optimal total polyphenol content (TPC) of STE is ~19 mg gallic acid equivalents per gram of dry weight (dw), while the antioxidant activity of the extract is 162 μmol ascorbic acid equivalents (AAEs) per gram of dw via FRAP and ~110 μmol AAE per gram of dw via DPPH. As for PEF, the optimal TPC is ~12 mg GAE/g dw, and the FRAP and DPPH values are ~102 and ~70 μmol AAE per gram of dw, respectively. When it comes to UPAE, the optimal TPC is ~16 mg GAE/g dw and the antioxidant capacity of the extract is ~128 μmol AAE/g dw through FRAP and ~98 μmol AAE/g dw through DPPH. UBAE optimal extract yielded ~17 mg GAE/g dw TPC, ~146 μmol AAE/g dw for FRAP, and ~143 μmol AAE/g dw for DPPH. The highest flavonoid content (~6.5 mg rutin equivalent/g dw) and DPPH (~143 μmol ascorbic acid equivalent/g dw) is obtained through UBAE. UPAE has been shown to be more efficient in recovering ascorbic acid (~20 mg/g dw). Additionally, the chlorophyll-to-carotenoid ratios of UPAE and UBAE were 2.98 and 2.96, respectively, indicating that the extracts had a generally positive impact on health. Considering the environmental impact of each extraction technique but also which antioxidant factor needs to be maximized, the most suitable extraction technique will be chosen.

Combinatorial Delivery of Docetaxel- and Erlotinib-Loaded Functionalized Nanostructured Lipid Carriers for the Treatment of Triple-Negative Breast Cancer Using Quality-by-Design Approach.

This study explored the combined administration of docetaxel (DOC) and erlotinib (ERL) using nanostructured lipid carriers (NLCs), with folic acid (FA) conjugation to enhance their synergistic anticancer efficacy against triple-negative breast cancer. NLCs were developed through hot melt homogenization-ultrasound dispersion, and optimized by a quality-by-design (QbD) approach using Plackett-Burman design and Box-Behnken design. Plots were generated based on maximum desirability. Spherical, nanosized dispersions (<200 nm) with zeta potential ranging from -16.4 to -14.15 mV were observed. These nanoformulations demonstrated ~95% entrapment efficiency with around 5% drug loading. Stability tests revealed that the NLCs remained stable for 6 months under storage conditions at 4 °C. In vitro release studies indicated sustained release over 24 h, following Higuchi and Korsmeyer-Peppas models for NLCs and FA NLCs, respectively. Additionally, an in vitro pH-stat lipolysis model exhibited a nearly fivefold increase in bioaccessibility compared to drug-loaded suspensions. The DOC-ERL-loaded formulations exhibited dose- and time-dependent cytotoxicity, revealing synergism at a 1:3 molar ratio in MDA-MB-231 and 4T1 cells, with combination indices of 0.35 and 0.37, respectively. Co-treatment with DOC-ERL-loaded FA NLCs demonstrated synergistic anticancer effects in various in vitro assays.

Optimization of biotransformation processes of Camarosporium laburnicola to improve production yields of potent telomerase activators

BackgroundTelomerase activators are promising agents for the healthy aging process and the treatment/prevention of short telomere-related and age-related diseases. The discovery of new telomerase activators and later optimizing their activities through chemical and biological transformations are crucial for the pharmaceutical sector. In our previous studies, several potent telomerase activators were discovered via fungal biotransformation, which in turn necessitated optimization of their production. It is practical to improve the production processes by implementing the design of experiment (DoE) strategy, leading to increased yield and productivity. In this study, we focused on optimizing biotransformation conditions utilizing Camarosporium laburnicola, a recently discovered filamentous fungus, to afford the target telomerase activators (E-CG-01, E-AG-01, and E-AG-02).ResultsDoE approaches were used to optimize the microbial biotransformation processes of C. laburnicola. Nine parameters were screened by Plackett-Burman Design, and three significant parameters (biotransformation time, temperature, shaking speed) were optimized using Central Composite Design. After conducting validation experiments, we were able to further enhance the production yield of target metabolites through scale-up studies in shake flasks (55.3-fold for E-AG-01, 13-fold for E-AG-02, and 1.96-fold for E-CG-01).ConclusionFollowing a process optimization study using C. laburnicola, a significant increase was achieved in the production yields. Thus, the present study demonstrates a promising methodology to increase the production yield of potent telomerase activators. Furthermore, C. laburnicola is identified as a potential biocatalyst for further industrial utilization.

A green liquid phase microextraction using phthalic acid as switchable hydrophilicity solvent for the HPLC determination of sildenafil (Viagra®) in human urine

Herein, a green liquid phase microextraction protocol using phthalic acid as switchable hydrophilicity solvent (SHS) is reported for the quantification of sildenafil in authentic human urine. The analyte was extracted onto phthalic acid solid particles which were produced through acidification of the sample. Its solidification was accomplished at ambient conditions without sample cooling. The determination of the sildenafil was carried out using high performance liquid chromatography-ultraviolet detection (HPLC-UV). The microextraction parameters that affect the extraction efficiency of the drug (i.e. SHS type and its concentration, acid type and concentration, extraction time, filter type) have been studied. The optimized analytical protocol involved the mixing of 300 μL of phthalate solution (0.75 M) with 600 μL of sample, followed by the addition of 50 μL of concentrated H3PO4. The produced solid was collected using membrane syringe filter (0.45 μm) and was finally dissolved in 500 μL of CH3OH. Method validation data showed determination coefficient ≥ 0.99 for the linear range of 50 – 2000 ng/mL. The limit of detection (LOD) and the lower limit of quantitation (LLOQ) were 30 and 100 ng/mL, respectively. The accuracy (expressed as % recovery) of the method ranged between 88.0 – 108.9 % while the precision (expressed as % RSD) was less than 17.8 % in all cases. The robustness of the microextraction procedure and the instrumental method were investigated using Plackett-Burman experimental designs. The applicability of the method was demonstrated by analyzing both spiked and authentic human urine samples after oral administration of drug-containing pharmaceutical formulation. The developed protocol offers cost-efficiency, handling simplicity, and high throughput. Its green character was evaluated using Green Analytical Procedure Index and Blue Applicability Grade Index.

Improvement in Natural Antioxidant Recovery from Sea Buckthorn Berries Using Predictive Model-Based Optimization

Sea buckthorn berries (SBB) are well known for being rich in natural bioactive compounds with high pharmacological activity. In this study, the optimization of extraction parameters was performed to recover phenolic compounds with high antioxidant activity from SBB. This study involved a systematic optimization approach, including screening for a variety of parameters, including temperature, time, ethanol concentration, agitation, and solid loading. On the basis of Plackett–Burman design (PBD) model, the two most significant parameters (agitation and solid loading) were selected, and the correlation model between those parameters and multiple responses was derived via response surface methodology (RSM). As a result, the optimal extraction condition for maximizing phenolic content and antioxidant activity was determined to be agitation at 109.54 rpm and a solid loading of 172.67 g/L. Under optimal conditions, SBB extract showed a total phenolic content of 0.21 mg/mL and ABTS and DPPH activities of 27.27% and 58.16%, respectively. The SBB extract prepared under optimal conditions was found to contain caffeic acid, vanillic acid, rutin, and vitamin B1 (thiamine). This work is the first challenge to design an optimization model for the efficient recovery of antioxidants from SBB and is significant in that the model can be applied simply and economically to conventional extraction processes.

Ammonia recovery via direct contact membrane distillation: Modeling and performance optimization

Ammonia recovery from wastewater has positive environmental benefits, avoiding eutrophication and reducing production energy consumption, which is one of the most effective ways to manage nutrients in wastewater. Specifically, ammonia recovery by membrane distillation has been gradually adopted due to its excellent separation properties for volatile substances. However, the global optimization of direct contact membrane distillation (DCMD) operating parameters to maximize ammonia recovery efficiency (ARE) has not been attempted. In this work, three key operating factors affecting ammonia recovery, i.e., feed ammonia concentration, feed pH, and DCMD running time, were identified from eight factors, by a two-level Plackett-Burman Design (PBD). Subsequently, Box-Behnken design (BBD) under the response surface methodology (RSM) was used to model and optimize the significant operating parameters affecting the recovery of ammonia though DCMD identified by PBD and statistically verified by analysis of variance (ANOVA). Results showed that the model had a high coefficient of determination value (R2 = 0.99), and the interaction between NH4Cl concentration and feed pH had a significant effect on ARE. The optimal operating parameters of DCMD as follows: NH4Cl concentration of 0.46 g/L, feed pH of 10.6, DCMD running time of 11.3 h, and the maximum value of ARE was 98.46%. Under the optimized conditions, ARE reached up to 98.72%, which matched the predicted value and verified the validity and reliability of the model for the optimization of ammonia recovery by DCMD process.

Optimization, gene cloning, expression, and molecular docking insights for enhanced cellulase enzyme production by Bacillus amyloliquefaciens strain elh1

BackgroundIn this study, we isolated a cellulase-producing bacterium, Bacillus amyloliquefaciens strain elh, from rice peel. We employed two optimization methods to enhance the yield of cellulase. Firstly, we utilized a one-variable-at-a-time (OVAT) approach to evaluate the impact of individual physical and chemical parameters. Subsequently, we employed response surface methodology (RSM) to investigate the interactions among these factors. We heterologously expressed the cellulase encoding gene using a cloning vectorin E. coli DH5α. Moreover, we conducted in silico molecular docking analysis to analyze the interaction between cellulase and carboxymethyl cellulose as a substrate.ResultsThe bacterial isolate eh1 exhibited an initial cellulase activity of 0.141 ± 0.077 U/ml when cultured in a specific medium, namely Basic Liquid Media (BLM), with rice peel as a substrate. This strain was identified as Bacillus amyloliquefaciens strain elh1 through 16S rRNA sequencing, assigned the accession number OR920278 in GenBank. The optimal incubation time was found to be 72 h of fermentation. Urea was identified as the most suitable nitrogen source, and dextrose as the optimal sugar, resulting in a production increase to 5.04 ± 0.120 U/ml. The peak activity of cellulase reached 14.04 ± 0.42 U/ml utilizing statistical optimization using Response Surface Methodology (RSM). This process comprised an initial screening utilizing the Plackett–Burman design and further refinement employing the BOX -Behnken Design. The gene responsible for cellulase production, egl, was effectively cloned and expressed in E. coli DH5α. The transformed cells exhibited a cellulase activity of 22.3 ± 0.24 U/ml. The egl gene sequence was deposited in GenBank with the accession number PP194445. In silico molecular docking revealed that the two hydroxyl groups of carboxymethyl cellulose bind to the residues of Glu169 inside the binding pocket of the CMCase. This interaction forms two hydrogen bonds, with an affinity score of −5.71.ConclusionsOptimization of cultural conditions significantly enhances the yield of cellulase enzyme when compared to unoptimized culturing conditions. Additionally, heterologous expression of egl gene showed that the recombinant form of the cellulase is active and that a valid expression system can contribute to a better yield of the enzyme.

Functionalized solid lipid nanoparticles combining docetaxel and erlotinib synergize the anticancer efficacy against triple-negative breast cancer

The goal of the study was to fabricate folic acid functionalized docetaxel (DOC)/erlotinib (ERL)-loaded solid lipid nanoparticles (SLNs) to synergistically increase the anticancer activity against triple-negative breast cancer. DOC/ERL-SLNs were prepared by the high shear homogenization – ultrasound dispersion method (0.1 % w/v for DOC, and 0.3 %w/v for ERL) and optimized using Plackett Burman Design (PBD) followed by Box Behnken Design (BBD). The optimized SLNs demonstrated particle size < 200 nm, PDI < 0.35, and negative zeta potential with entrapment and loading efficiency of ∼80 and ∼4 %, respectively. The SLNs and folic acid functionalized SLNs (FA-SLNs) showed sustained release for both drugs, followed by Higuchi and Korsemeyer-Peppas drug release models, respectively. Further, the in vitro pH-stat lipolysis model demonstrated an approximately 3-fold increase in the bioaccessibility of drugs from SLNs compared to suspension. The TEM images revealed the spherical morphology of the SLNs. DOC/ERL loaded SLNs showed dose- and time-dependent cytotoxicity and exhibited a synergism at a molar ratio of 1:3 in TNBC with a combination index of 0.35 and 0.37, respectively. FA-DOC/ERL-SLNs showed enhanced anticancer activity as evidenced by MMP and ROS assay and further inhibited the colony-forming ability and the migration capacity of TNBC cells. Conclusively, the study has shown that SLNs are encouraging systems to improve the pharmaceutical attributes of poorly bioavailable drugs.

In-vivo pharmacokinetic study of ibrutinib-loaded nanostructured lipid carriers in rat plasma by sensitive spectrofluorimetric method using harmonized approach of quality by design and white analytical chemistry

Ibrutinib, an antineoplastic agent tackling chronic lymphocytic leukemia, mantle cell lymphoma, and Waldenstrom’s Macroglobulinemia, falls under the category of BCS class II drugs, characterized by a puzzling combination of low solubility and high permeability. Its oral bioavailability remains a perplexing challenge, merely reaching 2.9 % due to formidable first-pass metabolism hurdles. In a bid to surmount this obstacle, researchers embarked on a journey to develop ibrutinib-loaded NLCs (Nanostructured Lipid Carriers) using a methodology steeped in complexity: a Design of Experiments (DoE)-based hot melted ultrasonication approach. Despite a plethora of methods for analyzing ibrutinib in various matrices, the absence of a spectrofluorimetric method for assessing it in rat plasma added to the enigma. Thus emerged a spectrofluorimetric method, embodying principles of white analytical chemistry and analytical quality by design, employing a Placket-Burman design for initial method exploration and a central composite design for subsequent refinement. This method underwent rigorous validation in accordance with ICH guidelines, paving the way for its application in scrutinizing the in-vivo pharmacokinetics of ibrutinib-loaded NLCs, juxtaposed against commercially available formulations. Surprisingly, the optimized NLCs exhibited a striking 1.82-fold boost in oral bioavailability, shedding light on their potential efficacy. The environmental impact of this method was scrutinized using analytical greenness tools, affirming its eco-friendly attributes. In essence, the culmination of these efforts has not only propelled advancements in drug bioavailability but also heralded the dawn of a streamlined and environmentally conscious analytical paradigm.

Optimization of polysaccharide conditions and analysis of antioxidant capacity in the co-culture of Sanghuangporus vaninii and Pleurotus sapidus

Fungal polysaccharides are commonly utilized in the food industry and biomedical fields as a natural and safe immune modulator. Co-culturing is a valuable method for enhancing the production of secondary metabolites. This study used intracellular polysaccharide (IPS) content as a screening index, co-culturing seven different fungi with Sanghuangporus vaninii. The seed pre-culture liquid culture time was selected through screening, and conditions were assessed using single factor experimentation, a Plackett-Burman (PB) design, and response surface methodology (RSM) optimization. RSM optimization was conducted, leading to the measurement of antioxidant capacity. Results indicated that the co-culture of S. vaninii and Pleurotus sapidus exhibited the most effective outcome. Specifically, pre-culturing S. vaninii and P. sapidus seed cultures for 2 days and 0 days, respectively, followed by co-culturing, significantly increased IPS content compared to single-strain culturing. Further optimization of co-culture conditions revealed that yeast extract concentration, liquid volume, and S. vaninii inoculum ratio notably influenced IPS content in the order of yeast extract concentration &gt; liquid volume &gt; S. vaninii inoculum ratio. Under the optimal conditions, IPS content reached 69.9626 mg/g, a 17.04% increase from pre-optimization co-culture conditions. Antioxidant capacity testing demonstrated that co-cultured IPS exhibited greater scavenging abilities for DPPH and ABTS free radicals compared to single strain cultures. These findings highlight the potential of co-culturing S. vaninii and P. sapidus to enhance IPS content and improve antioxidant capacity, presenting an effective strategy for increasing fungal polysaccharide production.

Hyper β-glucosidase producer Beauveria bassiana SAN01—optimization of fermentation conditions and evaluation of saccharification potential

AbstractThe hyper-production of β-glucosidase by a local strain of Beauveria bassiana under submerged conditions is reported in this study. The initial screening of seven agricultural residues showed that the haulm of Bambara—an underutilized African legume—supported the highest β-glucosidase production; hence, statistical optimization of enzyme production was done using this biomass as the sole carbon source. Plackett–Burman design identified the concentrations of Bambara haulm, KCl, and NaCl as well as agitation speed and incubation time as the most significant factors affecting enzyme production. Subsequently, the central composite design predicted the optimal conditions (Bambara 57 g/L, KCl 302 mg/L, NaCl 154 mg/L, agitation speed 150 rpm, and incubation 223 h) for B. bassiana β-glucosidase production, which were further validated. The generated quadratic model was deemed significant judging from its F-value (201.63), adequate precision ratio (45.74), as well as the R2 (0.9988), adjusted R2 (0.9938), and predicted R2 (0.9195) values. The optimization resulted in a ~5.36-fold increase in enzyme levels from the unoptimized production of ~133 to 711 U/mL. The enzyme was also demonstrated to efficiently hydrolyze cellobiose, converting more than 90% of the substrate to glucose. These results further establish the resourcefulness of the B. bassiana strain for the production of β-glucosidase enzyme, having immense potential, especially in the food and energy industries.

Parameter Calibration of Discrete Element Model of Wine Lees Particles

In order to investigate the contact characteristics of the mechanical parts of the brewing robot with wine lees particles, it is essential to calibrate the parameters of the discrete elemental model of wine lees particles. This paper proposes a method based on tests of the angle of repose. The simulation test is conducted to establish a regression model and combined with physical tests to find optimization. The contact model used in simulation modeling is Hertz-Mindlin with Johnson-Kendall-Roberts. Not all discrete element model parameters of wine lees particles have a significant impact on the angle of repose, so screening through Plackett-Burman Design is performed. The results indicate that the restitution coefficient between wine lees particles and restitution coefficient between wine lees particle and steel plate have a significant impact on angle of repose. Additionally, another parameter that is difficult to obtain, namely surface energy (JKR), also plays a crucial role. The optimal value interval for these three parameters is determined by the steepest ascent test, and a linear regression model for angle of repose is built through Box-Behnken Design. The optimal values obtained are as follows: restitution coefficient between wine lees particles—0.603; restitution coefficient between wine lees particle and steel plate—0.595; JKR—0.083. Finally, in order to verify the accuracy of calibrated parameters, simulation verification tests are carried out which show that there is only a relative error rate at 0.18% between simulated angle of repose and actual angle of repose, indicating that accurate calibration parameters were achieved. This study can provide reference for selecting discrete element model parameters for wine lees particles in future research endeavors.

Bio-inspired synthesis of silver nanoparticles using Salsola imbricata and its application as antibacterial additive in glass ionomer cement

Nanotechnology has gained immense popularity and observed rapid development due to the remarkable physio-chemical properties of nanoparticles (NPs) and related nanomaterials. The green production of NPs has many benefits over traditional techniques because the current procedures are expensive, time-consuming, and involve harmful substances that limit their applicability. This study aimed to use a novel green source, the Salsola imbricata (SI) plant, which is commonly found in Central Asia and known for its medicinal properties as a reducing and stabilizing agent for the synthesis of AgNPs. The current study also utilized efficient statistical design, the Plackett–Burman Design (PBD) of Experiment method to synthesize the NPs. The characterization of NPs was carried out using UV-Vis spectroscopy, Fourier transform infrared spectroscopy, and scanning electron microscopy (SEM). The PBD results showed that only two out of four factors i.e. AgNO3 concentration and incubation time, were significant for the synthesis of SI-AgNPs. While remaining factors, incubation temperature and plant extract: AgNO3 ratio were non-significant. The SEM analysis result showed that SI-AgNPs had a size of 20–50 nm. The SI-AgNPs demonstrated strong antibacterial activity against oral pathogens such as S. mutans and Lactobacillus acidophilus, with the highest efficacy observed at a concentration of 2 mg ml−1. The addition of SI-AgNPs in glass ionomer cement significantly increased the antibacterial activity of GIC against S. mutans. Based on the results of the current study, the plant based AgNPs can be further evaluated in detail as alternate antimicrobial agent either alone or in combination with other antimicrobial agents for different dental applications.

Analytical quality by design-based development of a capillary electrophoresis method for Omeprazole impurity profiling

Omeprazole (OME) is a proton pump inhibitor used to treat gastroesophageal reflux disease associated conditions. The current study presents an Analytical Quality by Design-based approach for the development of a CE method for OME impurity profiling. The scouting experiments suggested the selection of solvent modified Micellar ElectroKinetic Chromatography operative mode using a pseudostationary phase composed of sodium dodecyl sulfate (SDS) micelles and n-butanol as organic modifier in borate buffer. A symmetric three-level screening matrix 37//16 was used to evaluate the effect of Critical Method Parameters, including Background Electrolyte composition and instrumental settings, on Critical Method Attributes (critical resolution values, OME peak width and analysis time). The analytical procedure was optimized using Response Surface Methodology through a Central Composite Orthogonal Design. Risk of failure maps made it possible to define the Method Operable Design Region, within which the following optimized conditions were selected: 72 mM borate buffer pH 10.0, 96 mM SDS, 1.45 %v/v n-butanol, capillary temperature 21 °C, applied voltage 25 kV. The method was validated according to ICH guidelines and robustness was evaluated using a Plackett-Burman design. The developed procedure enables the simultaneous determination of OME and seven related impurities, and has been successfully applied to the analysis of pharmaceutical formulations.

Pomegranate seed as a novel source of plant protein: Optimization of protein extraction and evaluation of invitro digestibility, functional, and thermal properties.

This research was carried out to optimize the extraction process of proteins from pomegranate seeds and characterize their invitro digestibility as well as their thermal and functional properties. For this purpose, the study screened five parameters (liquid/solid ratio, pH, temperature, NaCl concentration, and time) that could potentially influence the extraction process. This screening was conducted using a two-level Placket-Burman design (PBD). The significant parameters (pH and NaCl concentration) were subsequently optimized using a three-level face-centered central composite design (FCCD) to determine the optimum extraction conditions. A maximum protein recovery of 83.8% was obtained at pH 11.0 and NaCl concentration of 0.0 M. Pomegranate seed protein isolate (PSPI) with a protein content of 92.4% (w/w) was obtained through the isoelectric precipitation of pomegranate seed protein extracted under the optimized conditions. An emulsifying activity index of 14.1 m2 g-1 was observed at the isoelectric pH, where the emulsion stability index was at 8.2%. PSPI also showed high water- and oil-holding capacities (3.7 and 4.3 g g-1, respectively). The essential amino acid levels in PSPI (except for valine and isoleucine) exceeded the recommended amounts set by WHO/FAO/UNU for adults, highlighting its high nutritional value. Based on thermal analysis data, denaturation of PSPI could occur at 89.5°C. The invitro digestibility of PSPI was found to be 74.3%. PSPI shows a potential as a novel ingredient for substituting animal-based proteins in various food applications.

Application of Response Surface Methodology in the Formulation of an Eco-friendly Degreaser using Rhamnolipid Biosurfactant

Formulating a bio-based degreaser emulsion can be challenging because it requires maximizing the performance to be as good as a commercial chemical-based product without compromising the cost and effect on the environment. This study aimed to develop an eco-friendly degreaser using natural Rhamnolipid biosurfactant through response surface methodology (RSM), which was then validated and assessed compared to chemical-based commercial products. The Plackett–Burman design (PBD) and Box–Behnken design (BBD) were used to screen and optimize factors for the formulation. From the PBD's result, the amount of Rhamnolipid, with other environmentally friendly ingredients of D-limonene, sodium citrate, and calcium carbonate, were shortlisted as significant factors in the formulation. Next, the BBD optimization study revealed that only rhamnolipid and sodium citrate significantly affected the two-way interaction of the factor on the result of the surface tension and oil displacement test (ODT). In the validation study, the experimental value was very close to the rate predicted by the model for the response surface tension and ODT (24.14 mN/m and 241 mm, respectively). A comparison of the performance with commercial products proved that this formulation was comparable to that commercial product. Overall, this study revealed that the effective eco-friendly degreaser formulation was successfully produced using a minimal amount of rhamnolipid biosurfactant with a response surface methodology as a tool.

Statistical optimization of bioprocess parameters for enhanced production of bacterial cellulose from K. saccharivorans BC-G1.

Bacterial Cellulose (BC) offers a wide range of applications across various industries, including food, biomedical, and textiles, owing to its distinctive properties. Its unique 3D reticulated network of cellulose nanofibers, imparts excellent mechanical qualities, a high water-holding capacity, and thermal stability. Additionally, it possesses remarkable biocompatibility, biodegradability, high crystallinity, and purity. These attributes have offered significant interest in BC within both academic and industrial sectors. However, BC production is associated with high costs due to the use of expensive growth media and low yields. The study reports the potential of our indigenous isolate, Komagataeibacter saccharivorans BC-G1, as BC producer. Statistical optimization of BC production was carried out using Placket-Burman design and Central composite design, by selecting different parameters. Eight significant factors such as temperature, pH, glucose, yeast, peptone, acetic acid, incubation time and % inoculum were studies using ANOVA-based response surface methodology. Results showed that BC yield (8.5g/L) with 1.8-fold after optimization of parameters. Maximum cellulose production (8.5 ± 1.8g/L) was obtained using 2% glucose, 0.3% yeast extract, 0.3% peptone, 0.75% (v/v) acetic acid at pH 7.0 for 10 days of incubation with 4% inoculum at 25°C under static culture. Main effect graph showed incubation time and acetic acid concentration as the most significant parameters affecting BC production in our study. The physicochemical characterization of produced BC was done using FTIR, XRD and SEM techniques.

Maximization of uricase production in a column bioreactor through response surface methodology-based optimization

Uricase (EC 1.7.3.3) is an oxidoreductase enzyme that is widely exploited for diagnostic and treatment purposes in medicine. This study focuses on producing recombinant uricase from E. coli BL21 in a bubble column bioreactor (BCB) and finding the optimal conditions for maximum uricase activity. The three most effective variables on uricase activity were selected through the Plackett–Burman design from eight different variables and were further optimized by the central composite design of the response surface methodology (RSM). The selected variables included the inoculum size (%v/v), isopropyl β-d-1-thiogalactopyranoside (IPTG) concentration (mM) and the initial pH of the culture medium. The activity of uricase, the final optical density at 600 nm wavelength (OD600) and the final pH were considered as the responses of this optimization and were modeled. As a result, activity of 5.84 U·ml−1 and a final OD600 of 3.42 were obtained at optimum conditions of 3% v/v inoculum size, an IPTG concentration of 0.54 mM and a pH of 6.0. By purifying the obtained enzyme using a Ni-NTA agarose affinity chromatography column, 165 ± 1.5 mg uricase was obtained from a 600 ml cell culture. The results of this study show that BCBs can be a highly effective option for large-scale uricase production.

Preparation of Bacterial Ghosts from Campylobacter jejuni Strain PP3894181

In the industrialized world, campylobacter jejuni is a prevalent foodborne bacterial infection. And is zoontic disease and causing gastroenteritis and others disease in the study using this bacteria as vaccine by preparing of bacterial ghost from C. jejuni This is the aim of the study for vaccine was prepared by determining the MIC, MGC of the bacteria that were grown on N.B medium after that a group of chemicals compound SDS, NaOH, CaCO3 and H2O2 were added at specific concentration and according to the Sponglike Protocol and using Plackett-Burman Experiment design and conducting a viability test for the bacteria, the concentration of MIC and MGC were determined.