Recovery and separation of glycyrrhizic acid from Natural Deep Eutectic Solvent (NADES) extract by macroporous resin: adsorption kinetics and isotherm studies.

Natural Deep Eutectic Solvents (NADESs) have emerged as a green and sustainable alternative to conventional organic solvents to extract bioactive compounds. However, the recovery of bioactive compounds from the NADES extracts is challenging, restricting their large-scale applications. The present work investigated the recovery of glycyrrhizic acid (GA) from choline-chloride/lactic acid NADES extract using macroporous resins. GA possesses a wide spectrum of biological activities, and it is extracted from the well-known herb Glycyrrhiza glabra. During resin screening, DIAIONTM SP700 showed high adsorption and desorption capacities. The adsorption kinetics study demonstrated that the adsorption of GA on SP700 followed Pseudo First-order kinetic model. Moreover, the adsorption behaviors were elucidated by the Freundlich isotherm using a correlation coefficient based on a static adsorption study at different temperatures and pH. Furthermore, the thermodynamic parameters, for instance, the change of Gibbs free energy (ΔG*), entropy (ΔS*), and enthalpy (ΔH*), showed that the adsorption process was spontaneous, favorable and exothermic. In addition, the sample after macroporous resin treatment, which is enriched with GA exhibited good anticancer potential analyzed by SRB assay. The regenerated NADES solvent was recycled twice, keeping more than 90% extraction efficiency, indicating good reusability of NADES in the GA extraction process by using macroporous resin.

Process Intensification of Enzymatic Synthesis of Flavor Esters: A Review.

The conventional flavor synthesis method suffers from low yields, time inefficiency, and extreme reaction conditions. Therefore, there is a necessity for the green and novel synthesis approach to overcome these limitations. The current review presents a holistic insight into different aspects associated with the synthesis of flavor esters using the immobilized enzyme. The application of process intensification tools such as ultrasound and microwave irradiation can enhance the reaction efficiency because of higher product recovery, less formation of by-products, and decreased energy consumption. This review presents the process intensification of value-added flavor esters synthesis and the mechanism of ultrasound and microwave action on the enzyme to enhance the enzyme activity and increase the reaction rate. It also summarizes the role of process intensification in enzymatic flavor ester synthesis, followed by specific examples as reported in the literature.

Extraction of total antioxidants from Azadirachta indica (neem) using three phase partitioning and its process intensification using ultrasound.

The majority of the naturally occurring antioxidants are obtained from plant sources. The antioxidant activity is mostly exhibited by polyphenols present in the plant cells. Azadirachta Indica (Neem) leaves are renowned for their medicinal applications due to their anti-inflammatory, antimalarial, antifungal, antibacterial, antiviral, antioxidant, and anticarcinogenic properties. This work aims to optimize the extraction of Azadirachta Indica (Neem) leaf antioxidants using three-phase partitioning (TPP). The optimized conditions are operating time 15 minutes, slurry ratio 1:30 (g/mL), salt concentration 30% (w/v), aqueous to solvent ratio 1:1.5 (v/v), and stirring speed 400 rpm that infer 74.66% extraction yield. Additionally, ultrasonic pretreatment was also employed to increase the extraction yield up to 86.61%. Sonication pretreatment for 4 min operated at 30 W power, and 75% duty cycle was observed to offer maximum antioxidant extraction about 3.3 mg/g.

Evaluation of nutritional and medicinal potential of defatted Sapindus mukorossi seed kernel.

This work deals with the evaluation of nutritional and medicinal potential of a defatted kernel of Sapindus mukorossis seed. Defatted sapindus seed kernel is a rich source of proteins (33.4 ± 2.12%), which show balanced amino acid composition for the requirement of adults as per the World Health Organization. Protein isolate possesses 29 kDa molecular weight peptide, which shows trypsin inhibitor activity. It also showed around 31.2% reduction in amylase activity while aqueous Ethanol and ethanol extracts showed 55% and 72.83%, respectively. Aqueous ethanol and ethanol extracts were found to contain polyphenols and saponins. Polyphenol content in aqueous ethanol and ethanol extract was 4.50 ± 0.15 mg/g and 5.7 ± 0.34 mg/g ferulic acid equivalent, respectively, while 0.72 ± 0.68% and 1.2 ± 0.23% Oleonolic acid equivalent saponins, respectively. Both these extracts showed potent antioxidant activity, and the rate of DPPH scavenging activity was higher than the ferulic acid standard. The deffated seed also contains dietary fibers in which 3.8 ± 0.01% are soluble, and 2.2 ± 0.03% are insoluble fibers.

Valorization of waste Syzygium cumini seed kernels by three-phase partitioning extraction and evaluation of in vitro antioxidant and hypoglycemic potential.

Diabetes is one of the leading causes of morbidity and mortality worldwide. Replacement of the chemically synthesized medicines with phytomedicines is of great importance. Syzygium cumini fruit has around 30% hard seed kernel part that is considered solid waste and can be a good source of bioactive phenolic compounds. Thus, this work was performed to valorize Syzygium cumini kernels powder (SCKP) using a three-phase partitioning technique (TPP) and evaluate in vitro antioxidant and hypoglycemic activities. The extraction yield of catechin, gallic acid, TFC, and TPC obtained through TPP was 0.52, 33.24 mg/g, 7.78 mg CE/g, and 82.66 mg GAE/g, respectively, in 45 min, 40% ammonium sulfate, 1:1 (V/V) extraction slurry to t-butanol, 30 ± 2 °C temperature, pH 4, 1:20 (g/mL) SCKP to water and SCKP size 106 µm. IC50 values 12.15, 9.33, and 7.55 in µg/mL for TPP extract were obtained from DPPH, α-amylase, and α-glucosidase in vitro assays, respectively. TPP for extraction of bioactive compounds was found superior over traditional extraction techniques (devoid of t-butanol and ammonium sulfate). The extract obtained shows enormous antioxidant and hypoglycemic potential for use in nutraceutical and pharmaceutical industries.HighlightsFirst report on three-phase partitioning (TPP) extraction of catechin and gallic acid from SCKP.Effective extraction of total phenolic and flavonoid compounds.Proved the enormous antioxidant and hypoglycemic potential of SCKP's TPP extract.TPP extraction, the selective, efficient, and economically viable option.

Rapid extraction of watermelon seed proteins using microwave and its functional properties.

Solid food industry waste like watermelon seed is an excellent source of value-added components such as proteins, oil, and carbohydrate. In the present study, protein extraction was carried out using microwave energy from defatted watermelon seeds (DWS), containing 50% of proteins. Microwave-assisted extraction (MAE) was optimized with different parameters, namely, solid to solvent ratio (1:10-1:40), pH (7-10), microwave power (30 W, 50 W, 70 W), extraction time (30 s-8 min) and moisture content or pre-leaching effect. Maximum protein recovery was achieved with 50 W microwave power, solid to solvent ration of 1:30, and pH 10 in 2 minutes of microwave irradiation time. MAE gave higher yield in less time compared to conventional extraction. SDS-PAGE confirmed the molecular weight of watermelon seed proteins (WSP) in the range of 25-250 kDa. A comparative study showed 90% protein recovery with MAE in 2 min with 1:30 (w/v) solid to solvent ratio, whereas ultrasound gave 87% in 9 min with 1:50 (w/v) ratio and batch 72% in 25 min with 1:70 (w/v) ratio. Watermelon seed proteins obtained from MAE method possess excellent functional properties with reference to conventional extraction method indicating its application in food products.

Antidiabetic potential evaluation of aqueous extract of waste Syzygium cumini seed kernel's by in vitro α-amylase and α-glucosidase inhibition.

Syzygium cumini, owing to higher bioactive constituents, its parts principally kernels are used for the antidiabetic purpose since the olden days. The current manuscript illustrated batch extraction of phenolic compounds from S. cumini using a stirred extractor. The yields 0.61 mg/g, 35.9 mg/g, 79.89 mg GAE/g, and 7.29 mg CE/g of catechin, gallic acid, TPC and TFC, respectively, were obtained in 105 min. at 1:20 SCKP to water, 50 ± 2 °C temperature, 4 pH, at 250 rpm and 106 µm particle size of SCKP. In vitro evaluation of the antioxidant and antidiabetic potential of the obtained aqueous extract was carried out by DPPH, α-amylase, and α-glucosidase inhibitory assays. The IC50 values of SCKP aqueous extract obtained were 12.97, 9.03, and 7.13 µg/mL for DPPH scavenging, inhibition of α-amylase, and α-glucosidase, respectively. The cost required to extract 1 kg of catechin, gallic acid, TPC, and TFC was Rs 6691.6, 113.7, 51.1, and 559.93/-, respectively. Stirred batch extraction technique manifests traditional but simple, ecofriendly, and efficient compared to other traditional techniques. The output of this research bestows support to utilize the SCKP stirred batch extract as a promising source of antioxidant and antidiabetic compounds in ayurvedic formulations.

Lipase-catalyzed synthesis of propyl-phenyl acetate: a kinetic and thermodynamic study.

This study focuses on the synthesis of propyl-phenyl acetate via esterification reaction in the presence of immobilized Candida antartica lipase-B (CAL-B). In this work, the effect of relevant factors (kinetics and thermodynamic) on total percent conversion and process optimization was studied. The reaction was performed in heptane medium with 1:2 molar ratio of benzoic acid: n-propanol with 0.6% (w/v) biocatalyst loading at 40 °C to attain a maximum conversion of 96.1% within 40 min of reaction time. Effect of increase in temperature on ∆G values indicates that lipase is more promising at moderate temperature (40 °C). A second-order kinetic model was proposed to evaluate apparent kinetic constants that indicate a good agreement between the experimental and theoretical data (0.94 ≤ R2 ≤ 0.99) with high initial reaction rate (113.5 mM/min). Finally, the catalyst CAL-B was successfully reused eight times without any significant decrease in relative activity.

Response surface methodology for the extraction of wedelolactone from Eclipta alba using aqueous two-phase extraction.

Aqueous two-phase extraction of wedelolactone from Eclipta alba was studied using the polymer-salt system. The system consisted of polyethylene glycol (PEG) as a top phase (polymer) and sodium citrate as a bottom phase (salt). Process parameters such as PEG concentration, PEG molecular weight, salt concentration, and pH have been optimized using response surface methodology (RSM) with the help of central composite design (CCD). The optimized conditions for aqueous two-phase system (ATPS), in the case of one factor at a time approach, were found as PEG 6000, PEG concentration 18% (w/v), salt concentration 16% (w/v), and pH 7; with maximum extraction yield of 6.52 mg/g. While, RSM studies showed maximum extraction yield of 6.73 mg/g with the optimized parameters as PEG 6000, PEG concentration 18% (w/v), salt concentration 17.96% (w/v), and pH 7. ATPS was found to give a 1.3 fold increase in the extraction yield of wedelolactone as compared to other conventional extraction methods.

Extraction of watermelon seed proteins with enhanced functional properties using ultrasound.

Watermelon seed is the potential source of value-added proteins, oils, and carbohydrates. The present study evaluates the extraction, and functional properties of watermelon seed protein (WMSP) obtained by ultrasound-assisted extraction (UAE) method from watermelon seed (WMS). The optimization of various operating parameters, such as pH (9), WMS powder to solvent ratio (1:50 w/v), temperature (30 ± 2 °C), ultrasound power (90 W), frequency (25 kHz), and duty cycle (75%) has been carried out. The extraction yield obtained was 87% and the extraction time was lowered down to 9 min from 120 min of conventional batch extraction. It contains all essential amino acids in an adequate amount required for adults as per FAO/WHO guidelines while for 2-5 years old children, the content of valine and isoleucine are above the required range. Methionine and lysine contents are adequate for both children and adults. Functional properties of ultrasonic extracted proteins were found superior to conventionally extracted proteins.highlightsThe UAE method is more efficient for watermelon seed protein extraction.Impact of extraction parameters on the extraction yield was studied.Protein isolate with enhanced functional properties was obtained.Essential amino acid content was determined.

Role of ultrasound in assisted fermentation technologies for process enhancements.

Biological molecules are widely produced by fermentation technology using bacteria, fungi or yeast. Fermentation is a biochemical process wherein the rate of bioconversion is governed by the organisms involved. The growth of the organism is mainly limited by mass transfer rates of nutrients and gases that directly affect the product formation in fermentation. Attempts have been made to enhance the growth rate and yield using mutational, recombinant strain development approach at microbial level as well as fed batch and continuous processing approach at bioprocess level in the past. The growth rate of microbes can be accelerated by increased mass transfer rates and cell wall permeability with the use of controlled low frequency ultrasound irradiation. The present review provides insights into the application of acoustic cavitation in process intensification of fermentation approaches and the role of various factors involved are highlighted with typical examples.

Kinetics and thermodynamics of lipase catalysed synthesis of propyl caprate.

OBJECTIVE: To investigate kinetics and thermodynamics of lipase-catalyzed esterification of capric acid with 1-propyl alcohol in a solvent-free system for synthesis of propyl caprate. RESULTS: The capric acid conversion of 83.82% is achieved at temperature 60 °C, speed of agitation 300 rpm, molar ratio acid:alcohol 1:3, enzyme loading 2% (w/w) and molecular sieves loading 5% (w/w). The activation energy (Ea) for the reaction was determined as 37.79 kJ mol-1. Furthermore, enthalpy (ΔH), entropy (ΔS) and Gibbs free energy (ΔG) values were found out to be + 90.45 kJ mol-1, + 278.99 J mol-1 K-1 and - 2.35 kJ mol-1 respectively. CONCLUSIONS: The results showed that the lipase-catalyzed esterification exhibits an ordered bi-bi mechanism with capric acid inhibiting the reaction and forming the dead-end complex with the lipase. Under the given set of reaction conditions, the lipase catalysed esterification reaction was anticipated to be spontaneous, referring to the value of the Gibbs free energy change (ΔG). Moreover, the esterification process was found to be endothermic, based on the values of enthalpy (ΔH) and entropy (ΔS).

Extraction of curcuminoids from Curcuma longa: comparative study between batch extraction and novel three phase partitioning.

Curcuminoids, the active components of dried rhizome of Curcuma longa have been extracted using batch extraction and three-phase partitioning (TPP) process. The effect of different processing parameters, namely different solvents, extraction time, ammonium sulfate concentration, slurry to tert-butanol ratio, and solute to aqueous ratio on extraction efficiency of TPP, was studied to attain maximum extraction yield. The highest yield of 58.38 mg/g was achieved at 40±2 °C in 150 min, with saturated ammonium sulfate 30% (w/v), slurry to tert-butanol ratio 1:1 (v/v), and turmeric powder to water ratio 1:40 (w/v) in TPP. However, batch extraction using ethanol as a solvent yielded 52.77 mg/g in 180 min extraction time at 40±2 °C with 1:40 (w/v) turmeric powder to water ratio and 400 rpm agitation speed. In view of reference method, i.e., Soxhlet extraction (100%), TPP showed 65.63% yield in 150 min and batch exhibited 59.92% in 180 min. The turmeric extracts obtained by different methods exhibited excellent antioxidant and anti-inflammatory activities equivalent to their respective reference standards. Hence, TPP extraction process assures a rapid and improved recovery of curcuminoids with excellent therapeutic properties.

Enzyme-catalysed production of n-butyl palmitate using ultrasound-assisted esterification of palmitic acid in a solvent-free system.

This work exhibits the implementation of ultrasound technology in solvent-free synthesis study of n-butyl palmitate using Fermase CALB™10000. Sequential experimental design study was instrumental in determining the most significant process variables. The effect of acid-alcohol molar ratio, enzyme dose, temperature, power and duty cycle on the reaction kinetics was studied. Highest conversion of ~ 96.6% was observed in 50 min at 1:1 molar ratio of palmitic acid to n-butanol, 70 °C temperature, 4% w/w enzyme loading, 40 W power, 70% duty cycle, 25 kHz frequency and 100 rpm speed of impeller. Ping-pong bisubstrate model showed the best fit with kinetic parameters, Vmax = 21.88 M/min/g catalyst, KA = 0.011 M, KB = 8.74M, KiA = 0.014M, KiB = 0.00036M and SSE = 0.0000193. Ultrasound reduced the reaction time by over 70%. The enzyme was reusable for four successive cycles after which it showed decline in conversion.

Extraction of total phenolic content from Azadirachta indica or (neem) leaves: Kinetics study.

The objective of this work is to put forth the optimization and kinetics of total phenolic compounds extraction from Azadirachta indica leaves in a stirred batch extraction. Various experiential extraction parameters have been studied for maximum extraction of the total phenolic compounds. The maximum yield of total phenolic compounds was found to be 10.80 mg g-1 of dried neem powder under the optimized conditions. The extraction kinetics behavior followed first-order kinetics with diffusion coefficient ranged from 1.8 × 10-12 to 3.2 × 10-12 m2 s-1 for all sets. Activation energy (Ea) value for the extraction of the total phenolic compounds was found to be 22.87 kJ mol-1. The kinetic expression model developed by Spiro and Siddique showed a good agreement with the experimental outcomes. The obtained results can be used to scale up the operations for industrial purposes.

Optimization of novel and greener approach for the coproduction of uricase and alkaline protease in Bacillus licheniformis by Box-Behnken model.

This study explores a novel concept of coproduction of uricase and alkaline protease by Bacillus licheniformis using single substrate in single step. Seven local bacterial strains were screened for uricase production, amongst which B. licheniformis is found to produce highest uricase along with alkaline protease. Optimization of various factors influencing maximum enzyme coproduction by B. licheniformis is performed. Maximum enzyme productivity of 0.386 U/mL uricase and 0.507 U/mL alkaline protease is obtained at 8 hr of incubation period, 1% (v/v) inoculum, and at 0.2% (w/v) uric acid when the organism is cultivated at 25°C, 180 rpm, in a media containing xylose as a carbon source, urea as a nitrogen source, and initial pH of 9.5. The statistical experimental design method of Box-Behnken was further applied to obtain optimal concentration of significant parameters such as pH (9.5), uric acid concentration (0.1%), and urea concentration (0.05%). The maximum uricase and alkaline protease production by B. licheniformis using Box-Behnken design was 0.616 and 0.582 U/mL, respectively, with 1.6- and 1.13-fold increase as compared to one factor at a time optimized media. This study will be useful to develop an economic, commercially viable, and scalable process for simultaneous production of uricase and protease enzymes.

Adsorption kinetics, isotherm, and thermodynamics studies of acetyl-11-keto-ß-boswellic acids (AKBA) from Boswellia serrata extract using macroporous resin.

An acetyl-11-keto-ß-boswellic acid (AKBA) is potent anti-inflammatory agent found in Boswellia serrata oleogum resin. Adsorption characteristics of AKBA from B. serrata were studied using macroporous adsorbent resin to understand separation and adsorption mechanism of targeted molecules. Different macroporous resins were screened for adsorption and desorption of AKBA and Indion 830 was screened as it showed higher adsorption capacity. The kinetic equations were studied and results showed that the adsorption of AKBA on Indion 830 was well fitted to the pseudo first-order kinetic model. The influence of two parameters such as temperature (298, 303, and 308 K) and pH (5-8) on the adsorption process was also studied. The experimental data was further investigated using Langmuir, Freundlich, and Temkin isotherm models. It was observed that Langmuir isotherm model was found to be the best fit for AKBA adsorption by Indion 830 and highest adsorption capacity (50.34 mg/g) was obtained at temperature of 303 K. The values of thermodynamic parameters such as the change of Gibbs free energy (ΔG*), entropy (ΔS*), and enthalpy (ΔH*), indicated that the process of adsorption was spontaneous, favourable, and exothermic.

Extraction of rapamycin (sirolimus) from Streptomyces rapamycinicus using ultrasound.

The study was designed to investigate the use of ultrasound-assisted extraction (UAE) of rapamycin (sirolimus) from bacterial strain of Streptomyces rapamycinicus NRRL 5491. To achieve the maximum extraction yield, various parameters were optimized which include S. rapamycinicus (10 g) of biomass in toluene (50 mL), temperature (20°C), acoustic intensity (35.67 W/cm2), and duty cycle (40%) for 4 min extraction time with probe tip length of 0.5 cm dipped into extraction solvent from the surface. The maximum extraction yield 60.15 ± 0.01 mg/L was attained under the mentioned optimum parameters. The use of ultrasound for the extraction of rapamycin shows about twofold increase in the yield as compared to the conventional solid-liquid extraction (29.7 ± 0.2 mg/L). The study provides the effective UAE technique to produce potential value-added products.

Ultrasound assisted intensification of enzyme activity and its properties: a mini-review.

Over the last decade, ultrasound technique has emerged as the potential technology which shows large applications in food and biotechnology processes. Earlier, ultrasound has been employed as a method of enzyme inactivation but recently, it has been found that ultrasound does not inactivate all enzymes, particularly, under mild conditions. It has been shown that the use of ultrasonic treatment at appropriate frequencies and intensity levels can lead to enhanced enzyme activity due to favourable conformational changes in protein molecules without altering its structural integrity. The present review article gives an overview of influence of ultrasound irradiation parameters (intensity, duty cycle and frequency) and enzyme related factors (enzyme concentration, temperature and pH) on the catalytic activity of enzyme during ultrasound treatment. Also, it includes the effect of ultrasound on thermal kinetic parameters and Michaelis-Menten kinetic parameters (km and Vmax) of enzymes. Further, in this review, the physical and chemical effects of ultrasound on enzyme have been correlated with thermodynamic parameters (enthalpy and entropy). Various techniques used for investigating the conformation changes in enzyme after sonication have been highlighted. At the end, different techniques of immobilization for ultrasound treated enzyme have been summarized.

Application of mixed modal resin for purification of a fibrinolytic enzyme.

Recent advances in purification technologies for therapeutic molecules have stirred the research consortium. Mixed mode chromatography, having multiple interactions with the solute molecule, has drawn significant attention due to its overall advantage over traditional ion-exchange and reverse-phase chromatography. Capto adhere, a mixed mode chromatography resin with strong anion-exchange and reverse-phase interaction with solutes, was explored for purification of fibrinolytic enzyme from Bacillus sphaericus MTCC 3672. Static and dynamic resin binding study revealed that 30°C temperature, pH 8, and 0.5 mL/min flow rate were optimum for maximum binding of fibrinolytic enzyme. Maximum static dynamic binding and breakthrough capacities for Capto adhere were 249 and 196 U/mL of resin, respectively. Final purification with Sephadex G 100 gel chromatography resulted in 38-fold purity of fibrinolytic enzyme with 39% enzyme recovery. Purified enzyme was further characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis to homogeneity, and molecular mass was found to be around 55-70 kD. Like most of the serine alkaline proteases, purified fibrinolytic enzyme was stable in a temperature range of 25-40°C and pH range of 7-9. Offshoots of our research findings have revealed a broad application area of mixed mode chromatography.