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.

Enhancement of Streptomyces thinghirensis WAE1 for production of bioactive metabolites under different optimization strategies.

Isolation of novel bioactive metabolites from Streptomyces strains is a promising source for drug discovery. However, conventional screening approaches have limitations in identifying new leads due to redundant discoveries. Optimization of culture conditions is important but traditionally optimized one factor at a time, failing to consider interactions. This study addressed these gaps by enhancing metabolite production from Streptomyces thinghirensis WAE1 through statistical optimization. Various chemical and physical factors impacting metabolite production were identified. Response surface methodology with a central composite design was applied to optimize significant factors like carbon source, nitrogen source, inoculum size, pH, temperature and incubation period. This optimized production against Streptococcus pneumoniae, increasing antibacterial activity by 74.92%. Gas chromatography-mass spectrometry revealed 19 bioactive compounds, including 1,25-dihydroxyvitamin D3 inhibiting cell wall development. This highlights S. thinghirensis WAE1's potential as a bioresource and emphasizes studying metabolite production from novel Streptomyces strains to discover new antibacterial drugs.

Characterization and optimization of exopolysaccharide extracted from a newly isolated halotolerant cyanobacterium, Acaryochloris Al-Azhar MNE ON864448.1 with antiviral activity.

Several antiviral agents lost their efficacy due to their severe side effects and virus mutations. This study aimed to identify and optimize the conditions for exopolysaccharide (EPS) production from a newly isolated cyanobacterium, Acaryochloris Al-Azhar MNE ON864448.1, besides exploring its antiviral activity. The cyanobacterial EPS was purified through DEAE-52 cellulose column with a final yield of 83.75%. Different analysis instruments were applied for EPS identification, including Fourier-transform infrared (FT-IR) spectroscopy, thermogravimetric analysis (TGA), and gas chromatographic-mass spectrometry (GC-MS). Plackett-Burman's design demonstrated that working volume (X1), EDTA (X2), inoculum size (X3), CaCl2 (X4), and NaCl (X5) are the most important variables influencing EPS production. Central composite design (CCD) exhibited maximum EPS yield (9.27 mg/mL) at a working volume of 300 mL in a 1 L volumetric flask, EDTA 0.002 g/L, inoculum size 7%, CaCl2 0.046 g/L, and NaCl 20 g/L were applied. EPS showed potent antiviral activities at different stages of herpes simplex virus type-1 and 2 (HSV-1, HSV-2), adenovirus (ADV) and coxsackievirus (A16) infections. The highest half-maximal inhibitory concentration (IC50) (6.477 µg/mL) was recorded during HSV-1 internalization mechanism, while the lowest IC50 (0.005669 µg/mL) was recorded during coxsackievirus neutralization mechanism.

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 of media components for enhanced carotenoid production by Paracoccus marcusii RSPO1 and assessment of their cytotoxicity against A549 and vero cells.

In this study, we tried to explore the influence of various tricarboxylic acid (TCA) cycle intermediates on carotenoid production and with a focus on enhancing pigment biosynthesis, we conducted two statistical analysis. In case of TCA intermediates influence on pigment production by Paracoccus marcusii RSPO1; fumaric acid, and malic acid were observed as potent enhancers of pigment biosynthesis. Further, to optimize key media components for enhanced carotenoid production, the Plackett-Burman design was employed encompassing carbon, nitrogen sources, TCA cycle intermediates, and metal salts. The selected factors after Plackett Burman were fine-tuned through Response Surface Methodology and the optimal concentrations that have remarkably elevated carotenoid production were starch-2.24 g/l, MgSO4-0.416 g/l, ZnSO4-0.0157 g/l, and fumaric Acid-16 mM. Further, evaluation of pigment cytotoxicity against normal (Vero) and Non-Small Cell Carcinoma (A549) cells was performed. The resultant IC50 values were quantified as 161.3 µg/ml and 7.623 µg/ml for Vero and A549 cells, respectively. Moreover, a reactive oxygen species (ROS) determination study in A549 cells was done which have shown a noteworthy threefold ROS production in A549 cells through fluorescence spectroscopic observation. This implies that the bacterial carotenoids can act as potent pro-oxidants against cancerous cells while being nontoxic toward normal cells.

Application of sequential design for enhanced L-asparaginase synthesis from Ganoderma australe GPC191.

With an increasing demand for L-asparaginase in pharmaceutical and food sectors for its cytostatic and acrylamide-reducing qualities, there's a need to discover novel, highly productive enzyme sources with improved pharmacokinetic profiles. Keeping this in mind, the present study aimed at maximizing the potential of Ganoderma australe GPC191 to produce L-asparaginase by fermentation medium optimization using statistical validation. Of the 11 physicochemical parameters evaluated under submerged fermentation conditions through one-factor-at-a-time approach and Plackett-Burman design, only four parameters (inoculum load, L-asparagine, soybean meal, and initial pH) influenced L-asparaginase production, significantly (p < 0.001). The optimal levels and interaction effects of these on the overall production were further evaluated by the central composite rotatable design of response surface methodology. Post-optimization, 27.34 U/mL was predicted as the maximum activity at pH 7 with 5n inoculum load and 15 g/L each of L-asparagine and soybean meal. Experimental validation yielded an activity of 28.52 U/mL, indicating an overall 18.17-fold increase from the unoptimized stage. To our knowledge, this is the first report signifying the L-asparaginase production aptitude of G. australe with sequential statistical validation using agricultural waste, which can serve as a model to enhance its yields, offering a sustainable and cost-effective solution for industrial application.

Enhancement of α-galactosidase production using novel Actinoplanes utahensis B1 strain: sequential optimization and purification of enzyme.

α-Galactosidase is an important exoglycosidase belonging to the hydrolase class of enzymes, which has therapeutic and industrial potential. It plays a crucial role in hydrolyzing α-1,6 linked terminal galacto-oligosaccharide residues such as melibiose, raffinose, and branched polysaccharides such as galacto-glucomannans and galactomannans. In this study, Actinoplanes utahensis B1 was explored for α-galactosidase production, yield improvement, and activity enhancement by purification. Initially, nine media components were screened using the Plackett-Burman design (PBD). Among these components, sucrose, soya bean flour, and sodium glutamate were identified as the best-supporting nutrients for the highest enzyme secretion by A. Utahensis B1. Later, the Central Composite Design (CCD) was implemented to fine-tune the optimization of these components. Based on sequential statistical optimization methodologies, a significant, 3.64-fold increase in α-galactosidase production, from 16 to 58.37 U/mL was achieved. The enzyme was purified by ultrafiltration-I followed by multimode chromatography and ultrafiltration-II. The purity of the enzyme was confirmed by Sodium Dodecyl Sulphate-Polyacrylamide Agarose Gel Electrophoresis (SDS-PAGE) which revealed a single distinctive band with a molecular weight of approximately 72 kDa. Additionally, it was determined that this process resulted in a 2.03-fold increase in purity. The purified α-galactosidase showed an activity of 2304 U/mL with a specific activity of 288 U/mg. This study demonstrates the isolation of Actinoplanes utahensis B1 and optimization of the process for the α-galactosidase production as well as single-step purification.

Effective remediation of mercury (II) from aqueous solutions using CS-MnFe2O4-CoS-MWCNTs: Box-Behnken design optimization and adsorption isotherm, kinetics, and thermodynamics evaluation.

Mercury (Hg) is a hazardous heavy metal, non-biodegradable and toxic, posing a serious threat to aquatic life and human health. Therefore, the removal of Hg ions from contaminated water using effective and eco-friendly adsorbents is necessary. In the present study, three magnetic chitosan-based organic-inorganic nanocomposites, such as CS-MnFe2O4, CS-MnFe2O4-CoS, and CS-MnFe2O4-CoS-MWCNTs, were designed and constructed to investigate their capacity for adsorbing Hg ions from aqueous solutions. The physicochemical properties of prepared composites were characterized by various analyses. The BET analyses indicated their high surface area and porous structure, and the N2 adsorption-desorption showed that the modification of CS in three stages by MnFe2O4 and crosslinking reaction, CoS preparation, and MWCNT incorporation resulted in increased N2 adsorption. The XRD confirms the synthesis of MnFe2O4 and CoS in the CS matrix and also the distinct peaks of MWCNTs. The CS-MnFe2O4-CoS-MWCNTs showed acceptable thermal stability with 45% char yields and superparamagnetic properties with magnetic saturation (Ms) of 16 emu g-1. The interactive impacts of independent variables (pH, contact time, and adsorbent dosage) on the removal percentage of Hg(II) onto three prepared adsorbents, as well as the process optimization, were assessed by the Box-Behnken design. The optimum conditions were identified, and the data from the analysis of variance showed that the three independent factors (pH, contact time, and adsorbent dosage) significantly influenced the adsorption of Hg(II). The adsorption isotherm and thermodynamics analysis investigation showed that at low concentrations of Hg(II), the adsorption process was both endothermic and spontaneous for the studied adsorbents.

Statistical Modelling of Thermostable Cellulase Production Conditions of Thermophilic Geobacillus sp. TP-1 Isolated from Tapovan Hot Springs of the Garhwal Himalayan Mountain Ranges, India.

A thermo-alkali stable cellulase from Geobacillus sp. TP-1 was isolated from Tapovan hot spring soil sample. The BLASTn sequence analysis of 16S rRNA sequence revealed that the isolate belonged to the Geobacillus genus and shared the highest degree of sequence similarity (99.43%) with the different strains of Geobacillus subterraneus. The neighbour joining method of multiple sequence alignment revealed that the 16S rRNA sequence of Geobacillus sp. TP-1 shows maximum similarity with Geobacillus stearothermophilus strain S_YE6-1017-022. One-Factor-At-a-Time analysis was used to optimize the carbon source, nitrogen source, pH, temperature, inoculum size and growth profile with respect to cellulase production. When compared to un-optimized basal media, optimised medium increased cellulase production by around 3.6 times. The Plackett Burman factorial design was employed to identify the critical medium components influencing cellulase activity and temperature was determined to have a significant effect on overall cellulase production. The current strain was capable of utilising lignocellulosic waste as an alternative carbon source. The use of sugarcane molasses and wheat bran as carbon sources resulted in a significant increase (~ 7.2 fold) in cellulase production in the current study, indicating the bacterium's potential for valorising lignocellulosic biomass into value-added products, which encourages its use in lignocellulosic-based bio refineries. Supplementary Information: The online version contains supplementary material available at 10.1007/s12088-024-01258-x.

Development and validation of HPLC method for simultaneous estimation of erlotinib and niclosamide from liposomes optimized by screening design.

The emerging drug resistance to the approved first-line drug therapy leads to clinical failure in cancer. Drug repurposing studies lead to the identification of many old drugs to be used for cancer treatment. Combining the repurposed drugs (niclosamide) with first-line therapy agents like erlotinib HCl showed improved efficacy by inhibiting erlotinib HCl acquired resistance. But there is a need to develop a sensitive, accurate, and excellent analytical method and drug delivery system for successfully delivering drug combinations. In the current study, an HPLC method was developed and validated for the simultaneous estimation of niclosamide and erlotinib HCl. The retention time of niclosamide and erlotinib hydrochloride was 6.48 and 7.65 min at 333 nm. The developed method was rapid and sensitive to separating the two drugs with reasonable accuracy, precision, robustness, and ruggedness. A Plackett-Burman (PBD) screening design was used to identify the critical parameters affecting liposomal formulation development using particle size, size distribution, zeta potential, and entrapment efficiency as the response. Lipid concentration, drug concentration, hydration temperature, and media volume were critical parameters affecting the particle size, polydispersity index (PDI), ZP, and %EE of the liposomes. The optimized NCM-ERL liposomes showed the particle size (126.05 ± 2.1), PDI (0.498 ± 0.1), ZP (-16.2 ± 0.3), and %EE of NCM and ERL (50.04 ± 2.8 and 05.42 ± 1.3). In vitro release studies indicated the controlled release of the drugs loaded liposomes (87.06 ± 9.93% and 42.33 ± 0.89% in 24 h).

Optimisation of ß-Glucosidase Production in a Crude Aspergillus japonicus VIT-SB1 Cellulase Cocktail Using One Variable at a Time and Statistical Methods and its Application in Cellulose Hydrolysis.

Pulp and paper mill sludge (PPMS) is currently disposed of into landfills which are reaching their maximum capacity. Valorisation of PPMS by enzymatic hydrolysis using cellulases is an alternative strategy. Existing commercial cellulases are expensive and contain low titres of ß-glucosidases. In this study, ß-glucosidase production was optimised by Aspergillus japonicus VIT-SB1 to obtain higher ß-glucosidase titres using the One Variable at a Time (OVAT), Plackett Burman (PBD), and Box Behnken design (BBD)of experiments and the efficiency of the optimised cellulase cocktail to hydrolyse cellulose was tested. ß-Glucosidase production was enhanced from 0.4 to 10.13 U/mL, representing a 25.3-fold increase in production levels after optimisation. The optimal BBD production conditions were 6 days of fermentation at 20 °C, 125 rpm, 1.75% soy peptone, and 1.25% wheat bran in (pH 6.0) buffer. The optimal pH for ß-glucosidase activity in the crude cellulase cocktail was (pH 5.0) at 50 °C. Optimal cellulose hydrolysis using the crude cellulase cocktail occurred at longer incubation times, and higher substrate loads and enzyme doses. Cellulose hydrolysis with the A. japonicus VIT-SB1 cellulase cocktail and commercial cellulase cocktails resulted in glucose yields of 15.12 and 12.33 µmol/mL glucose, respectively. Supplementation of the commercial cellulase cocktail with 0.25 U/mg of ß-glucosidase resulted in a 19.8% increase in glucose yield.

Optimization of medium composition of Lactobacillus plantarum Y44 using Plackett-Burman and Box-Behnken designs.

The biomass of Lactobacillus strains depends on the culture media and culture conditions. The purpose of this study was to optimize the culture medium composition and culture conditions of Lactobacillus plantarum Y44 to improve its biomass. The utilization of different carbon sources and nitrogen sources by L. plantarum Y44 was assessed by single factor experiment to screen out the economical carbon and nitrogen sources for L. plantarum Y44 growth. Through optimization experiments, the optimized culture medium for L. plantarum Y44 growth consists of soybean peptone 44.1 g/L, yeast extract 22.1 g/L, sucrose 35.6 g/L, hydrogen diamine citrate 2 g/L, anhydrous sodium acetate 8.5 g/L, dipotassium hydrogen phosphate 4 g/L, Tween-80 2 mL/L, manganese sulfate 0.25 g/L, and magnesium sulfate 0.58 g/L, and the initial pH 6.7. The concentration of viable bacteria cells of L. plantarum Y44 culturing in the optimized medium at 37 °C for 16 h was up to 3.363 × 1010 CFU/mL, as 6.11 times higher than that in the MRS medium.

Enhancing docosahexaenoic acid production of Schizochytrium sp. by optimizing fermentation using central composite design.

Docosahexaenoic acid (DHA) can improve human and animal health, particularly including anti-inflammatory, antioxidant, anticancer, neurological, and visual functions. Schizochytrium sp. is a marine heterotrophic protist producing oil with high DHA content, which is widely used in animal and food production. However, different fermentation conditions have intensive impacts on the growth and DHA content of Schizochytrium sp. Thus, this study aimed to enhance the DHA yield and concentration of Schizochytrium sp. I-F-9 by optimizing the fermentation medium. First, a single-factor design was conducted to select a target carbon and nitrogen source from several generic sources (glucose, sucrose, glycerol, maltose, corn syrup, yeast extract, urea, peptone, and ammonium sulfate). The Plackett-Burman design and the central composite design (CCD) were utilized to optimize the fermentation mediums. Schizochytrium sp. in 50-mL fermentation broth was cultured in a 250 mL shake flask at 28 °C and 200 rpm for 120 h before collecting the cell pellet. Subsequently, the cell walls were destroyed with hydrochloric acid to extract the fatty acid using n-hexane. The DHA content was detected by gas chromatography. The single-factor test indicated that glucose and peptone, respectively, significantly improved the DHA content of Schizochytrium sp. compared to the other carbon and nitrogen sources. Glucose, sodium glutamate, and sea crystal were the key factors affecting DHA production in the Plackett-Burman test (P = 0.0247). The CCD result showed that DHA production was elevated by 34.73% compared with the initial yield (from 6.18 ± 0.063 to 8.33 ± 0.052 g/L). Therefore, the results of this study demonstrated an efficient strategy to increase the yield and content of DHA of Schizochytrium sp.

Optimization of melanin pigment production from the halotolerant black yeast Hortaea werneckii AS1 isolated from solar salter in Alexandria.

BACKGROUND: Melanins are one of the magnificent natural pigments synthesized by a wide range of microorganisms including different species of fungi and bacteria. Marine black yeasts appear to be potential prospects for the synthesis of natural melanin pigment. As a result, the goal of this research was to isolate a marine black yeast melanin-producing strain and improve the culturing conditions in order to maximize the yield of such a valuable pigment. RESULTS: Among five locally isolated black yeast strains, the only one that demonstrated a potent remarkable melanin pigment production was identified using ITS rDNA as Hortaea werneckii AS1. The extracted pigment's physiochemical characterization and analytical investigation with Ultraviolet-Visible (UV) spectrophotometry, Fourier Transform-Infrared spectroscopy (FTIR), and Scanning Electron Microscope (SEM) confirmed its nature as a melanin pigment. The data obtained from the polynomial model's maximum point suggested that CaCl2, 1.125 g/L; trace element, 0.25 ml/L; and a culture volume 225 mL/500 mL at their optimal values were the critical three elements impacting melanin production. In comparison with the baseline settings, the response surface methodology (RSM) optimization approach resulted in a 2.0 - fold improvement in melanin output. CONCLUSIONS: A maximum melanin yield of 0.938 g/L proved the halotolerant H. werneckii AS1 potentiality as a source for natural melanin pigment synthesis 'when compared to some relevant black yeast strains' and hence, facilitating its incorporation in a variety of pharmaceutical and environmental applications.

Statistical optimization of chromium (VI) reduction using response surface methodology (RSM) by newly isolated Stenotrophomonas sp. (a novel strain).

Isolation of Microorganisms capable of reducing toxic chromium (VI) into less toxic one (Cr (III)) has been given attention due to their significance in bioremediation of the contaminated sites. In the present study, Stenotrophomonas sp. Crt94-4A an isolated strain from tannery wastewater and identified genetically by 16s rRNA gene sequencing was able to grow at concentrations up to 354 mg/L of Cr (VI). The results revealed 1% (w/v) NaCl, 2% (v/v) (2 × 106 CFU) inoculum size, and PH 7 in culture containing glucose and peptone as carbon and nitrogen sources respectively were the best conditions for Cr (VI) reduction. Statistical optimization was performed using Plackett-Burman design where peptone, inoculum size, and NaCl had significant effects on Cr (VI) reduction which were tested by three factors Box-Behnken design (BBD) to determine their correlation. The reduction capacity of Cr (VI) by Stenotrophomonas Sp. Crt94-4A was increased from 82, 55, and 23 to 96, 76, and 45% at 88.5, 177 and 354 mg/L of Cr (VI) respectively, which make this strain a good candidate for bioremediation of Cr (VI).

Medium optimization for submerged fermentative production of ß-cyclodextrin glucosyltransferase by isolated novel alkalihalophilic Bacillus sp. NCIM 5799 using statistical approach.

ß-cyclodextrin glucosyltransferase (ß-CGTase) is an essential enzyme to catalyse the biotransformation of starch into ß-cyclodextrins (ß-CD). ß-CD has widespread applications in the biomedical, pharmaceutical and food industries. The present study focused on ß-CGTase production using an efficient natural microbial strain and statistical production optimization for enhanced production. The isolated organism Bacillus sp. NCIM 5799 was found to be 5 µm short bacilli under FE-SEM and alkalihalophilic in nature. The ß-CGTase production was optimized using a combination of Plackett-Burman design (PBD) and Central Composite Design-Response Surface Methodology (CCD-RSM). On PBD screening Na2 CO3 , peptone and MgSO4 .7H2 O were found to be significant for optimal ß-CGTase production, whereas the soluble starch and K2 HPO4 concentrations were found to be nonsignificant for ß-CGTase production. The significant factors obtained after PBD were further optimized using CCD-RSM design. Peptone was found to have a significant interaction effect with Na2 CO3 , and MgSO4 ·7H2 O and Na2 CO3 exhibited a significant effect on the production of CGTase. The production of ß-CGTase was enhanced in the presence of peptone (3%) and Na2 CO3 (0·8%). CGTase production obtained was 156·76 U/ml when optimized using CCD-RSM. The final optimized medium (RSM) shows 7·7- and 5·4-fold high productions as compared to un-optimized and one factor at a time production media.

Optimization and kinetic modeling of media composition for hyaluronic acid production from carob extract with Streptococcus zooepidemicus.

Hyaluronic acid (HA), a mucopolysaccharide belonging to the glycosaminoglycan family, consists of repeating disaccharide units and has been used directly or indirectly in numerous human health practices. This study focused on evaluating carob pods for microbial HA production and kinetic modeling of HA fermentation. Therefore, the optimal medium composition was determined using Plackett-Burman Design (PBD) for HA production from carob extract with Streptococcus zooepidemicus. Maximum HA production of shake flask fermentation was 2.6 g/L (1.25 × 106) in the optimum medium, comprising 10°Bx of carob pods extract, 0.5 g/L of MgSO4.7H2O, 10.0 g/L of casein, 2.5 g/L of KH2PO4, 2.0 g/L of NaCl, 1.5 g/L of K2HPO4, 0.002 g/L of FeSO4 and 10.0 g/L of beef extract. In the continuation of the study, the fermentation performed with the optimal medium composition was modeled using three different models including the logistic model for biomass production, the Luedeking-Piret model for HA production, and the modified Luedeking-Piret model for substrate consumption. Based on the results, the experimental HA production data agreed with the Luedeking-Piret model with an R2 of 0.989. Since the α value was 63-fold higher than the value of ß, the HA production is growth-associated. Consequently, carob extract can be evaluated as a promising carbon source for producing HA.

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

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

Statistical optimization of medium components for biosurfactant production by Pseudomonas guguanensis D30.

Biosurfactant production by Pseudomonas guguanensis D30 was reported using mineral oil in submerged condition. Twelve medium components were tested at two levels by Plackett-Burman design, among them, mineral oil, yeast extract, peptone, MgSO4, and CaCl2 found significant on the basis of emulsification index. These five significant components were further optimized through central composite design (CCD). The experimental design was successfully used for regression analysis and the significant model suggested the solution of 10% (v/v) mineral oil, 3.0 g/L (w/v) yeast extract and 0.2 g/L (w/v) peptone for 13.14 g/L predicted biosurfactant production. We kept the suggested concentrations of medium components and got 13.34 ± 0.08 g/L biosurfactant production, which is almost double the conventional one-factor-at-a-time production (7.126 ± 0.12 g/L). It reduced the surface tension of the medium up to 28 ± 1.2 mN/m. We found ethyl acetate a suitable solvent for biosurfactant extraction amongst methanol, chloroform, and methanol:chloroform. The partially purified biosurfactant was chemically characterized as lipopeptide by Fourier transform infrared spectroscopy (FT-IR).

Applicability of Saccharomyces cerevisiae Strains for the Production of Fruit Wines Using Cocoa Honey Complemented with Cocoa Pulp.

Research background: Cocoa honey and cocoa pulp are both highly appreciated fruit pulp, but until now, cocoa honey has been less processed than cocoa pulp. In this work, we investigate the applicability of Saccharomyces cerevisiae strains to ferment cocoa honey complemented with cocoa pulp to obtain fruit wines and improve cocoa honey commercialization. Experimental approach: The strain, previously isolated from cachaçaria distilleries in Brazil, was selected based on its fermentation performance. The following conditions for fermentation with S. cerevisiae L63 were then studied: volume fraction of cocoa honey (φ CH) complemented with cocoa pulp, sucrose addition (γ suc), temperature (t) and inoculum size (N o). The best conditions were applied in order to obtain fermentation profiles. Results and conclusions: S. cerevisiae L63 (N o=107-108 cell/mL) is capable of fermenting φ CH=90 and 80% for 24 or 48 h with γ suc=50 and 100 g/L at t=28-30 °C resulting in wines with ethanol volume fractions from 8 to 14%. Additionally, the wine produced from φ CH=90% had lower residual sugar concentration (<35 g/L) than the wine produced from φ CH=80% (~79 g/L) which could be classified as a sweet wine. In general, S. cerevisiae L63 resulted in a similar fermentation performance as a commercial strain tested, indicating its potential for fruit pulp fermentation. Novelty and scientific contribution: Saccharomyces cerevisiae L63 can ferment cocoa honey complemented with cocoa pulp to produce fruit wines with good commercial potential, which may also benefit small cocoa producers by presenting a product with greater added value.