Plackett-Burman Design Research Articles

Preparation of Au-MoS2 photocatalyst with a green synthesis method and its application in water contaminant degradation

In the current research, a successful and fast synthesis of gold nanoparticles (AuNPs) by utilizing the leaf extract of Eryngium planum is reported. In order to analyze the influential synthesis parameters, the Plackett-Burman (PB) design was utilized, while the response surface methodology (RSM) was employed for optimization purposes. Furthermore, the achieved AuNPs were applied in the production of the Au-MoS2 nanocomposite. The efficiency of the visible-light photocatalyst was examined in the photocatalytic decolorization of the refractory azo dye, Acid Blue 113 (AB113). A high dye removal efficiency (94%) was observed under the optimal conditions of pH 7.6 using 15.5 mg of Au-MoS2, and 14 min. These results proved that the fabricated Au-MoS2 photocatalyst using biogenic AuNPs was more efficient than pure MoS2 and could be used as a promising catalyst for the degradation of dye-containing effluents.

Agro-industrial Waste Utilization, Medium Optimization, and Immobilization of Economically Feasible Halo-Alkaline Protease Produced by Nocardiopsis dassonvillei Strain VCS-4.

The oceanic actinobacteria have strong potential to secrete novel enzymes with unique properties useful for biotechnological applications. The Nocardiopsis dassonvillei strain VCS-4, associated with seaweed Caulerpa scalpeliformis, was a halo-alkaline protease producer. Further investigation focuses on medium optimization and the use of agro-industrial waste for economically feasible, high-yield protease production. A total of 12 experimental runs were designed using Minitab-20 software and Placket-Burman design. Among the 7 physicochemical parameters analyzed, incubation time and gelatin were detected as significant factors responsible for higher protease production. Incubation time and gelatin were further analyzed using OVATs. Optimal protease production was achieved with 2% gelatin, 0.1% yeast extract, 0.1% bacteriological peptone, 7% NaCl, pH 8, 5% inoculum, and a 7-day incubation period, resulting in a maximum protease activity (Pmax) of 363.97 U/mL, generation time of 11.9h, specific growth rate of 0.161g/mL/h, and protease productivity (Qp) of 61.65 U/mL/h. Moreover, utilizing groundnut cake as an agro-industrial waste led to enhanced production parameters: Pmax of 408.42 U/mL, generation time of 9.74h, specific growth rate of 0.361g/mL/h, and Qp of 68.07 U/mL/h. The immobilization of crude protease was achieved using Seralite SRC 120 as a support matrix resulting in 470.38 U/g immobilization, 88.20% immobilization yield, and 28.90% recovery activity. Characterization of both crude and immobilized proteases revealed optimal activity at pH 10 and 70°C. Immobilization enhanced the shelf-life, reusability, and stability of VCS-4 protease under extreme conditions.

Extraction of pesticides from soil using direct-immersion SPME LC-Tips followed by GC–MS/MS: Evaluation and proof-of-concept

A new method was evaluated and developed for the analysis of pesticides in sandy-loam soil by direct-immersion solid phase microextraction (DI-SPME) followed by gas chromatography tandem-mass spectrometry (GC–MS/MS) determination. Ten pesticides were selected based on a literature survey of the compounds reported to be present in EU soils. The extraction was performed using SPME LC-Tips, a new SPME configuration with the coated fibers attached to a disposable and easy-to-handle micropipette tip, which was immersed into a soil slurry made by the addition of an aqueous solution to the soil sample. Ten experimental parameters were evaluated with a Plackett-Burman design, after which the extraction time and percentage of organic solvent in the aqueous extraction were optimized separately. The two fiber chemistries available (PDMS/DVB and C18) were evaluated in parallel for the entire work. In the final method, slurry samples were made by adding an aqueous solution (6 % methanol v/v) to 2 g of soil. The fiber was conditioned and then inserted, for extraction, into the samples, stirred by a magnetic bar. Afterwards, the analytes were desorbed onto 100 µL of methanol. After the addition of analyte protectants (ethylglycerol, gulonolactone, and sorbitol) the extract was injected into the GC–MS/MS system. Isotopically labelled penconazole was used as internal standard. A calibration was performed by extracting spiked soil with analyte concentrations of 0.1–50 µg/kg. Coefficients of determination of the linear calibration were between 0.94–0.98 for the PDMS/DVB and 0.92–0.99 for the C18. Limits of detection range between 0.01–10 µg/kg for the PDMS/DVB and 0.1–10 µg/kg for the C18. Overall, the C18 analytically outperformed the PDMS/DVB but required a longer extraction time (120 min vs 75 min for the PDMS/DVB). This method allows automation and generates low residual toxic waste, having the potential to be introduced as a greener and simpler alternative to currently used sample preparation methodologies.

Optimized analytical strategy based on high-resolution mass spectrometry for unveiling associations between long-term chemical exposome in hair and Alzheimer’s disease

Exposure to environmental pollutants or contaminants is correlated with detrimental effects on human health, such as neurodegenerative diseases. Adopting hair as a biological matrix for biomonitoring is a significant innovation, since it can reflect the long-term chemical exposome, spanning months to years. However, only a limited number of studies have developed analytical strategies for profiling the chemical exposome in this heterogeneous biological matrix. In this study, a systematic investigation of the chemical extraction procedure from human hair was conducted, using a design of experiments and a high-resolution mass spectrometry (HRMS)-based suspect screening approach. The PlackettBurman (PB) design was applied to identify the significant variables influencing the number of detected features. Then, a central composite design was implemented to optimize the levels of each identified significant variable. Under the optimal conditions—15-minute pulverization, 25 mg of hair weight, 40 min of sonication, and a sonication temperature of 35 °C—approximately 32,000 and 15,000 aligned features were detected in positive and negative ion modes, respectively. This optimized analytical procedure was applied to hair samples from patients with Alzheimer's disease (AD) and individuals with normal cognitive function. Overall, 307 chemicals were identified using the suspect screening approach, with 37 chemicals differentiating patients with AD from controls. This study not only optimized an analytical procedure for characterizing the long-term chemical exposome in human hair but also explored the associations between AD and environmental factors.

Restaurant food waste valorization by microwave-assisted hydrolysis: Optimization, typological and biochemical analysis

Annually, a substantial volume of food waste is being released into the environment. Restaurant food waste (RFW) valorization using microwave-assisted hydrolysis (MAH) is a sustainable approach to produce fermentable sugars. However, RFW is composed of different foodstuffs with different physicochemical, nutritional, and degradation rates. This study explored the typological, chemical, and elemental analysis of RFW. Results revealed that the four main types of RFW were vegetable (33.2 %), meat (19.3 %), rice (15.2 %), and bread waste (11.0 %). The key parameters impacting the MAH of typologically sorted RFW were identified using the Plackett–Burman design (PBD). Then the central composite design (CCD) with 30 runs was used to increase reducing sugar content (RSC). The optimized condition was as follows: temperature 96.0 °C, microwave power 340 W, HCl concentration of 1.45 %, and microwave heating time 11.1 min. The derived hydrolysates were characterized for their biochemical and monosaccharide composition.

Extraction, purification, chemical characterization, and in vitro hypoglycemic activity of polysaccharides derived from Rosa laevigata Michx.

This study aimed to optimize the extraction process of polysaccharides from Rosa laevigata Michx. (RLMP) using an ultrasound-microwave-assisted method and investigate its in vitro hypoglycemic activity. Key factors affecting RLMP yield were identified using a Plackett–Burman design, followed by a Box–Behnken design and response–surface methodology, to determine the optimal extraction conditions. RLMP was purified using DEAE-52 cellulose, yielding two homogeneous fractions: RLMP-1 and RLMP-2. Monosaccharide composition was analyzed by gas chromatography, and structural characterization of RLMP, RLMP-1, and RLMP-2 was performed using FT-IR, SEM, and TEM. Methylation analysis and NMR were used to elucidate the sugar-chain structure of RLMP-1. In vitro hypoglycemic activity analysis showed that RLMP improved the glucose consumption and glycogen synthesis and enhanced the activities of pyruvate kinase and hexokinase in IR-HepG2 cells. Moreover, RLMP significantly increased the activities of antioxidant enzymes, such as CAT, SOD, and GSH-Px and decreased those of ROS and MDA. Western blotting analysis confirmed that RLMP enhances glucose and lipid metabolism and reduces oxidative stress by activating the PI3K/Akt/GLUT-4 signaling pathway, thereby exerting its hypoglycemic effect. These findings suggest that RLMP is a promising candidate for developing novel antioxidant agents or hypoglycemic drugs.

Fungal carbonatogenesis process mediates zinc and chromium removal via statistically optimized carbonic anhydrase enzyme

IntroductionWith rapid elevation in population, urbanization and industrialization, the environment is exposed to uncontrolled discharge of effluents filled with broad-spectrum toxicity, persistence and long-distance transmission anthropogenic compounds, among them heavy metals. That put our ecosystem on the verge or at a stake of drastic ecological deterioration, which eventually adversely influence on public health. Therefore, this study employed marine fungal strain Rhodotorula sp. MZ312369 for Zn2+ and Cr6+ remediation using the promising calcium carbonate (CaCO3) bioprecipitation technique, for the first time.ResultsInitially, Plackett–Burman design followed by central composite design were applied to optimize carbonic anhydrase enzyme (CA), which succeeded in enhancing its activity to 154 U/mL with 1.8-fold increase comparing to the basal conditions. The potentiality of our biofactory in remediating Zn2+ (50 ppm) and Cr6+ (400 ppm) was monitored through dynamic study of several parameters including microbial count, CA activity, CaCO3 weight, pH fluctuation, changing the soluble concentrations of Ca2+ along with Zn2+ and Cr6+. The results revealed that 9.23 × 107 ± 2.1 × 106 CFU/mL and 10.88 × 107 ± 2.5 × 106 CFU/mL of cells exhibited their maximum CA activity by 124.84 ± 1.24 and 140 ± 2.5 U/mL at 132 h for Zn2+ and Cr6+, respectively. Simultaneously, with pH increase to 9.5 ± 0.2, a complete removal for both metals was observed at 168 h; Ca2+ removal percentages recorded 78.99% and 85.06% for Zn2+ and Cr6+ remediating experiments, respectively. Further, the identity, elemental composition, functional structure and morphology of bioremediated precipitates were also examined via mineralogical analysis. EDX pattern showed the typical signals of C, O and Ca accompanying with Zn2+ and Cr6+ peaks. SEM micrographs depicted spindle, spherical and cubic shape bioliths with size range of 1.3 ± 0.5–23.7 ± 3.1 µm. Meanwhile, XRD difractigrams unveiled the prevalence of vaterite phase in remediated samples. Besides, FTIR profiles emphasized the presence of vaterite spectral peaks along with metals wavenumbers.ConclusionCA enzyme mediated Zn2+ and Cr6+ immobilization and encapsulation inside potent vaterite trap through microbial biomineralization process, which deemed as surrogate ecofriendly solution to mitigate heavy metals toxicity and restrict their mobility in soil and wastewater.

Eco-friendly biotransformation of penicillin G by free and immobilized marine halophilic Bacillus pseudomycoides AH1

BackgroundSeveral antibiotics are partially metabolized by patients after administration and end up in municipal sewage systems. The fate of biodegradation in aquatic environments and the role of biodegradation in the development of bacterial resistance are poorly understood. Thus, as a crucial step in an environmental risk assessment, the biodegradability of many therapeutically significant antibiotics was investigated.ResultsA marine halophilic bacteria that degrades penicillin G (PEN-G) was isolated and identified based on morphology, physio-biochemical characteristics, and 16S rDNA sequences as Bacillus pseudomycoides AH1 (accession no. MF037698). The effects of various concentrations of PEN-G and carbon and nitrogen sources on the biotransformation ability at 30°C and pH 7.0 were evaluated. Cells grown in medium supplemented with glucose as an additional carbon source and yeast extract as a nitrogen source exhibited maximal PEN-G biotransformation efficiency and rate (71.678% ±1.28 and 2.99 mg/h, respectively). The culture conditions for B. pseudomycoides AH1 cells were optimized using a Plackett–Burman design (PBD). Six key determinants (p < 0.05) significantly affected the process outcome, as deduced by regression analysis of the PBD data, and modified MSM broth achieved PEN-G biotransformation efficiency (100%) under aerobic shaking conditions at 35°C, irrespective of HPLC analysis. Additionally, the present investigation could strongly support the application of immobilization approaches for the removal of PEN-G-contaminated environmental sites.ConclusionTo the best of the authors’ knowledge, this is the first detailed study on the efficient biotransformation of PEN-G by an alginate-bacteria system as a simple, green, and inexpensive process, as well as a promising method.

Polysaccharides from Hericium erinaceus and its immunomodulatory effects on RAW 264.7 macrophages

This study aimed to optimize the extraction of Hericium erinaceus polysaccharides (HEP) using ultrasound-assisted enzymatic extraction combined with Plackett-Burman design (PBD) and response surface methodology (RSM). The optimal extraction conditions were identified as: 33 min extraction time, 30:1 liquid to material ratio, 38 °C extraction temperature, 9 g/kg cellulase amount, pH 4, and 20 % ethanol concentration. Under these conditions, the extraction yield of HEP was 5.87 ± 0.16 %, consistent with the predicted results. Additionally, the potential immunomodulatory activity of HEP on RAW 264.7 macrophage was evaluated. The results revealed that HEP improved the immunostimulatory activity of RAW264.7 cells, evident from increased production of IL-6 and TNF-α. These findings suggest that HEP is capable of enhancing the immune activity of RAW 264.7 macrophage.

Combining the natural deep eutectic solvents and high-speed-shearing-enzymatic-assisted extraction to recover flavonoids and terpenoids from perilla leaves

This research was conducted for the purpose of extracting flavonoids and terpenoids from perilla leaves (Perilla frutescens var. crispa) using high-speed-shearing-enzymatic-assisted extraction (HSSEAE) in combination with natural deep eutectic solvents (NADES). Various solvents were examined to select the most suitable for further extraction experiments. The one-factor experiment was employed to investigate the influence of different parameters of HSSEAE on the efficacy of flavonoid and terpenoid extraction from perilla leaves. The Plackett–Burman Design Model was used to evaluate the significance of independent variables on the dependent responses in the optimal condition ranges. The Box-Behnken Design Model was employed to determine the optimal extracting conditions for obtaining bioactive compounds from perilla leaves. The highest yield of total flavonoid contents (TFC) and total terpenoid contents (TTC) using HSSEAE was 40.37 ± 1.02 mg rutin acid equivalent per gram of dry weight (mg RE/g dw) and 37.93 ± 1.59 mg ursolic acid equivalent per gram of dry weight (mg UE/g dw), respectively. Meanwhile, the traditional extraction (TrE) process using acidified ethanol as solvent yielded lower results: TFC: 8.7 ± 0.17 mg RE/g dw; and TTC: 8.04 ± 0.55 mg UE/g dw. The second-order extraction kinetics model was used to figure out the principle of the HSSEAE process. Scanning electron microscope and X-ray diffraction were employed to indicate the surface morphology crystallinity of perilla leaves before and after HSSEAE. This extraction approach can be considered highly effective and sustainable for the recovery process of bioactive compounds originating from plants.

Harnessing the power of Neobacillus niacini AUMC-B524 for silver oxide nanoparticle synthesis: optimization, characterization, and bioactivity exploration

BackgroundBiotechnology provides a cost-effective way to produce nanomaterials such as silver oxide nanoparticles (Ag2ONPs), which have emerged as versatile entities with diverse applications. This study investigated the ability of endophytic bacteria to biosynthesize Ag2ONPs.ResultsA novel endophytic bacterial strain, Neobacillus niacini AUMC-B524, was isolated from Lycium shawii Roem. & Schult leaves and used to synthesize Ag2ONPS extracellularly. Plackett–Burman design and response surface approach was carried out to optimize the biosynthesis of Ag2ONPs (Bio-Ag2ONPs). Comprehensive characterization techniques, including UV–vis spectral analysis, Fourier transform infrared spectroscopy, transmission electron microscopy, X-ray diffraction, dynamic light scattering analysis, Raman microscopy, and energy dispersive X-ray analysis, confirmed the precise composition of the Ag2ONPS. Bio-Ag2ONPs were effective against multidrug-resistant wound pathogens, with minimum inhibitory concentrations (1–25 µg mL−1). Notably, Bio-Ag2ONPs demonstrated no cytotoxic effects on human skin fibroblasts (HSF) in vitro, while effectively suppressing the proliferation of human epidermoid skin carcinoma (A-431) cells, inducing apoptosis and modulating the key apoptotic genes including Bcl-2 associated X protein (Bax), B-cell lymphoma 2 (Bcl-2), Caspase-3 (Cas-3), and guardian of the genome (P53).ConclusionsThese findings highlight the therapeutic potential of Bio-Ag2ONPs synthesized by endophytic N. niacini AUMC-B524, underscoring their antibacterial efficacy, anticancer activity, and biocompatibility, paving the way for novel therapeutic strategies.

Analysis of polycyclic aromatic hydrocarbons in Brazilian human milk: A simple and effective approach

Human milk is the best source of nutrition for infants due to its essential biologically active components. However, it can contain potentially harmful chemicals, such as polycyclic aromatic hydrocarbons, originating from contaminated air and food, which can cause adverse effects to infants. Using a Plackett-Burman design, a QuEChERS protocol was optimized to determine 13 polycyclic aromatic hydrocarbons in 79 Brazilian human milk samples by gas chromatography high-resolution mass spectrometry. Limits of quantification below 1 μg/kg were achieved for all compounds, with recoveries ranging between 61 % and 126 %. Six PAHs, including benzo(g,h,i)perylene, dibenzo(a,h)anthracene, fluoranthene, indeno(1,2,3-cd)pyrene, phenanthrene, and pyrene were detected in 40 samples. Mature milk exhibited the highest incidence of PAHs, with approximately 44 % of the positive samples, compared to 27 % for both colostrum and transitional milk. The concentrations detected varied between < LOQ and 1.14 μg/kg, with pyrene being the most frequently found PAH in the human milk samples. The estimated daily intake and margin of exposure of PAHs showed the highest values for babies aged 0–15 days for all PAHs evaluated. Despite the low levels of PAHs found in the analyzed samples, discontinuing breastfeeding is not recommended. This represents the first study on the incidence of PAHs in Brazilian human milk.

Optimized phenol degradation and lipid production by Rhodosporidium toruloides using response surface methodology and genetic algorithm-optimized artificial neural network

Oleaginous yeast can produce lipids while degrading phenol in wastewater treatment. In this study, a Plackett-Burman Design (PBD) was adopted to identify key factors of phenol degradation and lipid production using R toruloides 9564T. While temperature, inoculum size, and agitation were significant for both the processes (p < 0.05), pH and incubation were significant for lipid production, and phenol removal, respectively. Results from four factors (pH, temperature, inoculum size, and incubation period) central composite design (CCD) experiment were used to formulate quadratic and genetic algorithm-optimized ANN models. The reduced quadratic model for phenol degradation (R2: 0.993) and lipid production (R2: 0.958) were marginally inferior to ANN models (R2: 0.999, 0.982, respectively) on training sets. Multi-objective optimization with equal importance suggests phenol degradation between 106.4 and 108.76%, and lipid production of 0.864–0.903 g/L, by polynomial and ANN models. Complete phenol degradation (100%) and 3.35-fold increment (0.918 g/L) in lipid production were obtained at pH 6.07, inoculum size 14.68% v/v, at 29.5 °C in 92.17 h experimentally.

Development and validation of stability indicating HPLC-DAD assay of adapalene, methylparaben and propylparaben in a cosmeceutical gel matrix with blueness, greenness and whiteness assessment via relative ranking: An ab initio-sub finem juxtaposed non-traditional approaches

Adapalene (ADA) is primarily used as a cosmeceutical gel in the treatment of mild-moderate acne and is also used off-label to treat keratosis pilaris as well as other skin conditions. A stability-indicating fit-for-purpose (FFP) method has been developed for co-determination of ADA, methylparaben (MPN), and propylparaben (PPN) combined in a cosmeceutical gel dosage form. This method was based on a reversed-phase RP-HPLC separation of the cited analytes. It avoided the use of tetrahydrofuran (THF), which is commonly used by both compendial and published procedures for determination of ADA, thus improving method blueness (applicability), greenness and whiteness. The method development and validation followed non-traditional eco-friendly approaches including chemical simulations and empirical modelling. The HPLC separation was performed on an RP Zorbax Eclipse XDB-C8 analytical column [150 × 4.6 mm, 5 µm (80 Å)] with gradient elution system of HPLC grade acetonitrile and 50 mM aqueous sodium dihydrogen phosphate buffer containing 5 mM ethylenediaminetetraacetic acid (EDTA) adjusted to pH 8.0 as the mobile phase at a flow rate of 2.5 mL min−1 and column oven was set at 25 °C. EDTA played an important role for reproducibility of retention time of the three analytes especially for ADA. It controlled deprotonation of the three acidic analytes under study and hence reproduce their ionization, hydrophobicity, and hence retention time. Quantitation was achieved with the photodiode array detection (DAD) at wavelengths of 256 nm for both MPN and PPN and 312 nm for ADA. Target Measurement Uncertainty (TMU) for the three analytes was determined via two approaches; misclassification rates approach employing 100,000 Monte Carlo simulations (MCS) and in-specification probability (ISP) to ascertain fitness-for-purpose of the proposed method. Microsoft Excel workbooks were developed for both TMU calculation approaches in addition to calculation of procedure capability index (Ppk) and sigma level for each analyte and attached in the Supplementary material. Moreover, linearity of the method underwent additional assessment using the previously mentioned TMU and procedure capability determination techniques. The robustness of the method was checked using Plackett-Burman experimental design (PBD) as an empirical modelling technique which was examined thoroughly to detect some interactions. The calibration graphs for each drug were rectilinear in the range of 0.34–7.08, 0.38–7.00 and 0.08–1.75 µg mL−1 for ADA, MPN, and PPN, respectively. The stress testing revealed photostability of the three components and variable degrees of degradation in the other conditions. The proposed HPLC-DAD method was successfully validated and applied for the determination of the investigated analytes in a gel matrix. In addition, method blueness and greenness were comparatively evaluated via very recent approaches of Blue Applicability Grade Index (BAGI), Green Solvents Selection Tool (GSST), Spider Diagram for Assessment of the Green Analytical Index (SDAGI), Analytical Method Greenness Score (AMGS), Carbon Footprint (CF), Modified Complementary Green Analytical Procedure Index (ComplexMoGAPI), and Analytical GREEnness Metric Approach (AGREE). Furthermore, the method whiteness was assessed via Red Green Blue Twelve (RGB 12) model following a relative ranking approach.

Optimizing Polyhydroxyalkanoate production using a novel Bacillus paranthracis isolate: A response surface methodology approach

Microorganisms have emerged as promising resources for producing economical and sustainable bioproducts like Polyhydroxyalkanoate (PHA), a biodegradable polymer that can replace synthetic plastics. In this study, we screened a novel isolate, Bacillus paranthracis RSKS-3 strain, to produce PHA from sewage water, identifying it using Whole Genome Sequence. This study represents the first report on optimizing PHA production using B. paranthracis RSKS-3, employing Design Expert 12.0 software. Our findings reveal that four factors (temperature, inoculum size, potassium dihydrogen phosphate, and magnesium sulfate) significantly affect PHA production in the Plackett-Burman design experiment. Through Response Surface Methodology, we optimized PHA production to 0.647 g/L with specific values for potassium dihydrogen phosphate (0.55 %), inoculum size (3 %), magnesium sulfate (0.055 %), and a temperature of 35 °C, in agreement with the predicted value of 0.630 g/L. This optimization resulted in a substantial 13.29-fold increase in PHA production from 0.34 g/L to 4.52 g/L, underscoring the promising role of B. paranthracis RSKS-3 in eco-friendly PHA production and advancing sustainable bioproduct development.

3D-printed microfluidics under ultrasonic cooling crystallization for nano-precipitation of ezetimibe: Effect of process parameters and PBD approach

The current research focuses on the antisolvent precipitation of Ezetimibe (EZB) via microfluidics under continuous ultrasonic cooling crystallization. One factor at a time was applied to study the effect of parameters: organic-aqueous flow rate ratio (1:2–1:5), EZB concentration (2.5–12.5, mg/mL), polymer concentration (0.025–0.3, % w/v), ultrasonic frequency (22–42, kHz), ultrasonic bath temperature (10–25, °C), ultrasonic amplitude (20–100, %), confluence angle of micromixer inlet (90°–180°) on the particle size (PS, nm), % yield, % encapsulation efficiency (% EE), and % dissolution rate (% DR). A PBD (Plackett–Burman design) was also employed for the screening studies of influencing factors for aforementioned responses. PS, Zeta potential (ZP, mv), Field Emission Gun Transmission Electron microscope (FEG-TEM), and X-ray diffraction (XRD) were analyzed for raw EZB and nano-precipitated EZB. An average EZB PS of 101 ± 1.1 nm, PDI of 0.027 ± 0.01, 80 ± 1.7 % yield, 78.83 ± 0.5 % DR, 90.67 ± 0.1 % EE, and −20 mV ZP were obtained under best parametric condition. FEG-TEM revealed the final nano EZB morphology was likely to be elongated cuboidal or rod-shaped. An approximately 5-fold increase in %DR was observed for the confluence angle of 90° (├ shaped) micromixer as compared to raw EZB. However, 94.14 % decreament in avg EZB PS was achieved for the confluence angle of 90° (├ shaped) micromixer and utilizing ultrasonic microfluidic system (USMS) under cooling crystallization as a process intesification approach.

Enrichment of Fruit Peels' Nutritional Value by Solid-State Fermentation with Aspergillus ibericus and Rhizopus oryzae.

The fruit processing industry is responsible for disposing of huge amounts of byproducts, especially fruit peels (FPs), which are often discarded in landfills. Using FPs in biotechnological processes contributes to a circular economy, reducing the environmental burden of FPs and increasing the revenue of the fruit processing industry. This study was focused on upgrading the nutritional value of orange (OPs) and banana (BPs) peels by solid-state fermentation (SSF) with filamentous fungi. SSF factors (moisture, fermentation time, inoculum size, ammonium sulfate (AS), and corn steep liquor (CSL)) and fungi species (Aspergillus ibericus and Rhizopus oryzae) were studied by a variable screening Plackett-Burman design. Both fungi grew on untreated FPs, increasing their protein content and antioxidant activity. Moisture, AS, and CSL were further studied by a Box-Behnken design with A. ibericus. Fermented OPs at 70% moisture and 0.005 g/g AS increased their protein content by 200%, whereas BPs at 70% moisture and 0.005 g/g CSL increased by 123%. Fermented peels were enriched in protein, fiber, and minerals, with a low content of carbohydrates and soluble sugars. Fermented OPs and BPs showed higher antioxidant activity than unfermented peels. The SSF of these FPs is an innovative approach that contributes to obtaining rich nutrient-fermented peels for food.

Optimization of 1,3-propanediol production from fermentation of crude glycerol by immobilized Bacillus pumilus

This study investigates the application of crude glycerol to the production of 1,3-propanediol by immobilized cells of Bacillus pumilus. This is a novel application of a naturally occurring producer obtained from a wastewater storage pond in Thailand. Crude glycerol was obtained through the methanolysis of palm oil, which was catalyzed using rice bran lipase. Ten components of the fermentation medium were screened using a Plackett-Burman design. The statistical significance of the results was determined using multiple linear regression with a backward elimination approach. The significance level was set to 5 % (p < 0.05). Only crude glycerol, (NH4)2SO4, MgSO4, and CaCl2 significantly affected 1,3-propanediol production by immobilized B. pumilus. Furthermore, preliminary screenings of environmental conditions used for 1,3-propanediol production were conducted using a Plackett-Burman design. The results showed that the temperature, time, and quantity of immobilized cells were factors that significantly affected 1,3-propanediol yield. Therefore, the quantities of crude glycerol, (NH4)2SO4, MgSO4, and CaCl2 and the temperature, time, and quantity of immobilized cells were optimized using response surface methodology based on a Box-Behnken design. The model predicted a maximum 1,3-propanediol yield of 45.68 g/L with the following conditions: 60 g/L crude glycerol, 5 g/L (NH4)2SO4, 0.55 g/L MgSO4, 0.05 g/L CaCl2, a fermentation duration of 101 h, and a temperature of 25 °C, with 250 g of immobilized cells. The validation trials confirmed a production level of 44.12 ± 1.81 g/L, indicating a 2.86-fold production increase relative to the control group. Overall, this study demonstrates the potential of using crude glycerol as a substrate to improve the yields of 1,3-propanediol produced by B. pumilus.

Optimization of the chromium (Cr+6) reduction from waterways using chemically and bacterially treated agro-waste.

Heavy metals, a major source of pollution in the environment, pose a substantial threat due to their non-biodegradability and ability to accumulate in living organisms, causing health problems. Recently, researchers have been searching for cost-effective and safe ways to remove heavy metals from polluted waterways using agricultural waste substitutes. The present study focused on the low-cost treatments for the reduction of chromium Cr+6 metal from the effluent, wherein it has been found that chemically and bacterially treated agro-waste had increased heavy metal ion adsorption capabilities. A sequential optimization of the process parameters was attempted using Plackett-Burman design (PBD) and central composite design of response surface methodology (CCD-RSM) for the maximum reduction of the chromium metal from the effluent. A total of eight parameters were screened out using a 12-run PBD experiment. Out of the eight parameters, time, HCl, NaOH, and bacterial treatments were found to be significantly affecting the maximum reduction of Cr+6 from the effluent. To investigate the interactions' effects of the chosen parameters, they were evaluated using CCD-RSM. Maximum 74% Cr+6 reduction was achieved under the optimum treatment to rice husk of HCl 4.52 N, NaOH 3.53 N, bacterial suspension 7.41%, and with an interaction time 14.32 min using 30 run CCD-RSM experiment. A scanning electron microscope was used to confirm the effects of selected variables on the agro-waste for the Cr+6 reductions, as well as a Fourier transform infrared spectrometer.

Recovering bioactive compounds from yacon (Smallanthus sonchifolius) using the ultrasonic-microwave-assisted extraction technique

The aim of this study was to recover bioactive compounds from Smallanthus sonchifolius roots, including flavonoids, terpenoids, phenolics, vitamin C, and reducing sugar contents, as well as to examine antioxidant activities using ultrasonic-assisted and microwave-assisted extraction technique. Single-factor experiments were employed to explore the impact of various independent variables, including concentration, extraction method, power, duration, temperature, and pH, on the dependent variables of yacon extraction using the ultrasonic-microwave-assisted extraction method. The Plackett-Burman Design Model was used to screen the parameters that significantly influenced the extraction efficiency. Box-Behnken Design Model was utilized to optimize the ultrasonic-microwave-assisted extracting process to acquire bioactive compounds. The highest yield of total flavonoid contents was obtained with 4.00 of pH, 743.33 W of microwave power, and 39.08°C of ultrasonic temperature. According to reducing sugar contents yield optimization, the optimum conditions were 3.98 of pH, 36.17 seconds of microwave time and 40°C of ultrasonic temperature. To optimize vitamin C concentration, the optimized conditions were 1:40 g/mL of sample-to-solvent ratio, 3.99 of pH, 45.33 seconds of microwave time, and 451.09 W of ultrasonic power. Regarding total terpenoids yield, optimized conditions for extracting were 1:40 g/mL of sample-to-solvent ratio, 2.48 of pH, 30.49 seconds of microwave time, and 49.48 °C of ultrasonic temperature. Under optimized conditions, the recovery yield of flavonoid, terpenoid, vitamin C, and reducing sugar contents were 7.95 mg rutin equivalents/g, 47.93 mg ursolic acid equivalents/g, 107.78 mg ascorbic acid/g, and 230.39 mg D-glucose equivalents/g, respectively. Ultrasonic-microwave-assisted extraction showed significantly higher performance than singular extracting methods and is regarded as a sustainable and effective approach to recover target compounds from yacon roots.