Physicochemical Properties, Functionalities, and Antioxidant Activity of Protein Extracts from New Zealand Wild Sea Cucumbers (Australostichopus mollis).

This study investigated the physicochemical properties, functionalities, and antioxidant capacities of protein extracts from wild sea cucumber Australostichopus mollis collected from four distinct locations in New Zealand. Protein was extracted from sea cucumber body walls using trypsin enzymatic extraction, followed by cold acetone precipitation. The amino acid analysis revealed high glycine (189.08 mg/g), glutamic acid (119.45 mg/g), and aspartic acid (91.91 mg/g) concentrations in all samples. The essential amino acid indexes of the protein extracts (62.96, average) were higher than the WHO/FAO standard references, indicating the excellent protein quality of A. mollis. Furthermore, protein extracts from A. mollis demonstrated superior emulsifying activity (202.3-349.5 m2/g average) compared to commercial soy and whey protein isolates under all tested pH conditions, and enhanced foaming capacity (109.9-126.4%) and stability (52.7-72%) in neutral and acidic conditions. The extracts also exhibited good solubility, exceeding 70% across pH 3-11. Antioxidant capacities (ABTS and DPPH free radical scavenging activity and ferric reducing antioxidant power) were identified in A. mollis protein extracts for the first time, with clear variations observed among different locations. These findings elucidate the advantageous functional properties of protein extracts from wild New Zealand A. mollis and highlight their potential application as high-quality antioxidant food ingredients.

Phytochemical and Functional Diversity of Enzyme-Assisted Extracts from Hippophae rhamnoides L., Aralia cordata Thunb., and Cannabis sativa L.

Plant leaves are a source of essential phenolic compounds, which have numerous health benefits and can be used in multiple applications. While various techniques are available for recovering bioactive compounds from by-products, more data are needed on enzyme-assisted extraction (EAE). The aim of this study was to compare EAE and solid-liquid extraction (SLE), to evaluate the impact on bioactive compounds' extraction yield, phytochemical composition, and the antioxidant, antimicrobial, and antidiabetic properties of Aralia cordata leaves and roots, sea buckthorn Hippophae rhamnoides, and hemp Cannabis sativa leaves. The results indicate that EAE with Viscozyme L enzyme (EAE_Visc) extracts of the tested plant leaves possess the highest yield, antioxidant activity, and total phenolic content. Moreover, the EAE_Visc extract increased by 40% the total sugar content compared to the control extract of A. cordata root. Interestingly, the sea buckthorn leaf extracts exhibited α-glucosidase inhibitory activity, which reached an almost 99% inhibition in all extracts. Furthermore, the sea buckthorn leaves SLE and EAE_Visc extracts possess antibacterial activity against Staphylococcus aureus. Additionally, scanning electron microscopy was used to examine changes in cell wall morphology after EAE. Overall, this study shows that EAE can be a promising method for increasing the yield and improving the functional properties of the resulting extracts in a fast and sustainable way compared to SLE.

Stabilized chlorophyll-based food colorants from spinach: Kinetics of a tailored enzymatic extraction.

An organic solvent-free method based on limited dosing options (biocatalyst and zinc chloride) for the quick and mild recovery of chlorophyll (Chl) from spinach has been proposed. This tailored, custom-made protocol has been designed to produce stable green natural colorants. The kinetics of pigment extraction turned out to be a very useful tool to identify the proper conditions, in terms of biocatalyst dose (0.10-50 U/g), extraction time (1-48h), and ZnCl2 amount (50-300ppm), both for enhancing the recovery yield and preserving the green color. Considering the extraction kinetics, the recovery yield, and the colorimetric data, the suitable conditions to produce stable green and food-grade colorants are 0.10 U/g of enzyme, 3h, and 150ppm of ZnCl2 at 25°C. The extraction yield of Chl (4863µg/U) was about 51% greater than control, with a higher extraction rate constant (5.43 × 10-4 g/(µg min)). Considering the impact of ZnCl2 amount on Chl, its protective action resulted to be more noticeable toward Chl a: at 150ppm, an increased amount of about 2.5 and 1.5 times was found for Chl a and Chl b, respectively, in comparison to the reference (0ppm ZnCl2). PRACTICAL APPLICATION: This research demonstrates how a suitable kinetic approach helps to provide a tailored protocol, customized for the vegetable matrix, to produce stable green natural colorants from spinach. Lowering enzyme dosage and ZnCl2 amount during the extraction of chlorophyll at low temperature is crucial for its potential use as a colorant in food industry, providing high economic values through saving time and environmental protection.

Chemical and Antioxidant Properties of Solvent and Enzyme-Assisted Extracts of Fucus vesiculosus and Porphyra dioica.

Extraction strategies impact the efficiency and nature of extracted compounds. This work assessed the chemical composition and antioxidant capacity of ethanolic, hydroethanolic, and aqueous versus enzyme-assisted extracts (isolated or with the sequential use of alcalase®, cellulase®, and viscozyme®) of the macroalgae Fucus vesiculosus (brown, Phaeophyceae) and Porphyra dioica (red, Rhodophyta. For both macroalgae, enzyme-assisted extraction (EAE) was the most efficient process compared to solvent-assisted extraction (SAE), independent of solvent. Fucus vesiculosus extraction yields were higher for EAE than for SAE (27.4% to 32.2% and 8.2% to 30.0%, respectively). Total phenolics content (TPC) was at least 10-fold higher in EAE extracts (229.2 to 311.3 GAE/gextract) than in SAE (4.34 to 19.6 GAE/gextract) counterparts and correlated well with antioxidant capacity (ABTS and ORAC methods), with EAE achieving values up to 8- and 2.6-fold higher than those achieved by SAE, respectively. Porphyra dioica followed F. vesiculosus's trend for extraction yields (37.5% to 51.6% for EAE and 5.7% to 35.1% for SAE), TPC, although of a lower magnitude, (0.77 to 8.95 GAE/gextract for SE and 9.37 to 14.73 GAE/gextract for EAE), and antioxidant capacity. Aqueous extracts registered the highest DPPH values for both macroalgae, with 2.3 µmol TE/gextract and 13.3 µmol TE/gextract for F. vesiculosus and P. dioica, respectively. EAE was a more efficient process in the extraction of soluble protein and reducing sugars in comparison to SAE. Furthermore, an improved effect of enzyme-assisted combinations was observed for almost all analyzed parameters. This study shows the promising application of enzyme-assisted extraction for the extraction of valuable compounds from F. vesiculosus and P.dioica, making them excellent functional ingredients for a wide range of health and food industrial applications.

Optimisation of enzyme-assisted extraction of camptothecin from Nothapodytes nimmoniana and its characterisation.

INTRODUCTION: Efficient extraction of camptothecin (CPT), an anticancer agent from the commercial source Nothapodytes nimmoniana (J. Graham) Mabb in India, is of paramount importance. CPT is present in the highest concentration in the stem portion, and the stem can be readily harvested without uprooting the plant. The fluorescence microscopy mapping of the bark matrix for CPT revealed its presence in a free form within both the outer (epidermal and cortical tissues) and inner (xylem and phloem tissues) sections. The bark matrix primarily consists of cellulose, hemicellulose, and lignin, rendering it woody, rigid, and resistant to efficient solvent penetration for CPT extraction. We proposed a hypothesis that subjecting it to disruption through treatment with hydrolytic enzymes like cellulase and xylanase could enhance solvent diffusion, thereby enabling a swift and effective extraction of CPT. OBJECTIVE: The present study was aimed at enzyme-assisted extraction, using cellulase and xylanase for hydrolytic disruption of the cells to readily access CPT from the stem of the plant N. nimmoniana (J. Graham) Mabb. METHODOLOGY: The hydrolytic cell disruption of ground powder from the stem bark was studied using cellulase and xylanase enzymes. The enzymatically pretreated stem bark powder was subsequently recovered by filtration, dried, and subjected to extraction with methanol to isolate CPT. This process was optimised through a Box-Behnken design, employing a one-factor-at-a-time approach to assess parameters such as enzyme concentration (2-10% w/w), pH (3-7), incubation time (6-24 h), and solid-to-solvent ratio (1:30-1:70 g/mL). CPT was characterised using proton nuclear magnetic resonance (1H-NMR) and Fourier transform infrared (FTIR) spectra, and a high-performance liquid chromatography (HPLC) method was developed for quantification. RESULTS: The cellulase and xylanase treatment resulted in the highest yields of 0.285% w/w and 0.343% w/w, with efficiencies of 67% and 81%, respectively, achieved in a significantly shorter time compared to the untreated material, which yielded 0.18% with an efficiency of only 42%. Extraction by utilising the predicted optimised process parameters, a nearly two-fold increase in the yield, was observed for xylanase, with incubation and solvent extraction times set at 16 and 2 h, respectively. Scanning electron microscopy (SEM) images of the spent material indicated perforations attributed to enzymatic action, suggesting that this could be a primary factor contributing to the enhanced extraction. CONCLUSION: Enzyme-mediated hydrolytic cell disruption could be a potential approach for efficient and rapid isolation of CPT from the bark of N. nimmoniana.

Determination of process parameters and precipitation methods for potential large-scale production of sugar beet leaf protein concentrate.

BACKGROUND: Sugar beet is one of the most produced industrial plants in the world, and during manufacturing it produces a large quantity of leaf waste. Because this waste is rich in protein, this study aimed to identify an efficient method for producing large-scale protein concentrate from sugar beet leaves. RESULTS: Results showed that protein extraction from fresh leaves was more effective than from dried leaves. Maximum protein extraction was achieved at pH 9, compared with pH 7 or 8. Blanching as a pretreatment reduced protein yield during isoelectric precipitation, with a yield of 2.31% compared to 20.20% without blanching. Consequently, blanching was excluded from the extraction process. After extraction, isoelectric precipitation, heat coagulation, and isoelectric-ammonium sulfate precipitation were compared. Although the latter resulted in the highest protein yield, Fourier transform infrared analysis revealed that excessive salt was not removed during dialysis, making it unsuitable for scale-up due to its additional cost and complexity. Therefore, isoelectric precipitation was selected as the appropriate method for protein precipitation from sugar beet leaves. To increase yield, extractions were assisted by ultrasound or enzyme addition. Ultrasound-assisted extraction resulted in an increased protein yield from 20.20% to 28.60%, while Pectinex Ultra SP-L-assisted extraction was the most effective, increasing protein yield from 20.20% to 38.09%. CONCLUSION: Proteins were extracted from fresh sugar beet leaves using optimum conditions (50 °C, 30 min, pH 9) and precipitated at isoelectric point, with enzymatic-assisted extraction yielding the maximum protein recovery. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Sequential and enzyme-assisted extraction of algal bioproducts from Ecklonia maxima.

Brown algae are gaining recognition as sources of bio-compounds with diverse properties and potential applications in the food, nutraceutical, and pharmaceutical industries. Compounds such as polyphenols, alginates and fucoidan possess multiple bioactivities, including antidiabetic, antioxidant, anticancer, anti-inflammatory, and antibacterial properties. Conventional extraction methods provide low yields, posing challenges for the industrial applications of biocompounds. However, innovations are rapidly emerging to address these challenges, and one such approach is enzyme-assisted extraction. Furthermore, extracting single compounds undervalues algal biomass as valuable compounds may remain in the waste. Therefore, the aim of our study was to develop a framework for the sequential and enzyme-assisted extraction of various bio-compounds using the same biomass in a biorefinery process. The Ecklonia maxima algal biomass was defatted, and polyphenols were extracted using solid-liquid extraction with aqueous ethanol. The remaining residue was treated with an enzyme combination (Cellic® Ctec 2 and Viscozyme L) to liberate carbohydrates into solution, where an alginate and fucoidan fraction were isolated. A second alginate fraction was harvested from the residue. The phenolic fraction yielded about 11% (dry weight of extract/dry weight of seaweed biomass), the alginate fraction 35% and the fucoidan fraction 18%. These were analysed using a variety of biochemical methods. Structural analyses, including FTIR, NMR and TGA, were performed to confirm the integrity of these compounds. This study demonstrated that a sequential extraction method for various algal bioproducts is possible, which can pave the way for a biorefinery approach. Furthermore, our study primarily employed environmentally and eco-friendly extraction technologies promoting an environmentally sustainable industrial approach. This approach enhances the feasibility and flexibility of biorefinery operations, contributing to the development of a circular bio-economy.

Enzyme-Assisted Supercritical Fluid Extraction of Flavonoids from Apple Pomace (Malus×domestica).

Herein an enzyme-assisted supercritical fluid extraction (EA-SFE) was developed using the enzyme mix snailase to obtain flavonols and dihydrochalcones, subgroups of flavonoids, from globally abundant waste product apple pomace. Snailase, a commercially available mix of 20-30 enzymes, was successfully used to remove the sugar moieties from quercetin glycosides, kaempferol glycosides, phloridzin and 3-hydroxyphloridzin. The resulting flavonoid aglycones quercetin, kaempferol, phloretin and 3-hydroxyphloretin were extracted using supercritical carbon dioxide (scCO2) and minimum amounts of polar cosolvents. A sequential process of enzymatic hydrolysis and supercritical fluid extraction was developed, and the influence of the amount of snailase, pre-treatment of apple pomace, the time for enzymatic hydrolysis, the amount and type of cosolvent and the time for extraction, was studied. This revealed that even small amounts of snailase (0.25 %) provide a successful cleavage of sugar moieties up to 96 % after 2 h of enzymatic hydrolysis followed by supercritical fluid extraction with small amounts of methanol as cosolvent, leading up to 90 % of the total extraction yields after 1 h extraction time. Ultimately, a simultaneous process of EA-SFE successfully demonstrates the potential of snailase in scalable scCO2 extraction processes for dry and wet apple pomace with satisfactory enzyme activity, even under pressurized conditions.

Optimization of fucoidan recovery by ultrasound-assisted enzymatic extraction from South African kelp, Ecklonia maxima.

Fucoidan, a sulphated polysaccharide, is found exclusively in brown seaweeds and has been reported to possess a wide range of biological functionalities. Fucoidans are found within the cell wall of brown seaweeds, which is composed of recalcitrant cellulose and hemicellulose. This hampers the recovery of fucoidans. In addition, fucoidans are found within a network of viscous hydrocolloids, alginates, further complicating their recovery. Traditionally, the hot water extraction method is used to recover fucoidans from brown seaweed, however, this is characterized by low yields and long extraction time. To combat these issues, several novel extraction technologies have been introduced, these include ultrasound-assisted extraction and enzyme-assisted extraction. Thus, the main aim of this study was to investigate and optimize fucoidan recovery from Ecklonia maxima based on ultrasound-assisted enzymatic extraction. The impact of temperature (40-65 °C), ultrasound intensity (0-118 W·cm-2), enzyme dosage (0-0.05 ml·g-1) and pH (4.5-6) on total dissolved, total carbohydrates and inorganic sulphates yields was studied. The application of ultrasound-assisted enzymatic extraction mainly improved the extraction of total carbohydrates. Ultrasound significantly improves the kinetics and extraction of fucoidan, but there was no merit when it was applied with enzymes. Results reveal that at optimized conditions, the fucoidan extracted 79.13 mg⋅g-1 (7.9 % w/w) of algal dry weight. The present study provides insight into the extraction potentials of enzyme-assisted extraction, ultrasound-assisted extraction, and ultrasound-assisted enzymatic extraction.

Phenolic compounds profile of optimised green and eco-friendly extracts of Eugenia pyriformis leaves: an alternative for antioxidant and antibacterial applications.

The Eugenia pyriformis Cambess (uvaia) is a well-known source of bioactive compounds. This study investigated the efficiency of Ultrasound-Assisted Extraction (UAE) and Enzyme-Assisted Extraction (EAE) in obtaining uvaia leaf extracts with high antioxidant and antibacterial activity. In a first step, different variables of the leaves were employed to define the best conditions for obtaining the extract with the highest total phenolic content. In a second step, the optimised extracts were characterised. In total, twenty-four phenolic compounds were identified through LC-ESI-MS/MS. The EAE in optimised conditions showed a higher amount of total phenolic compounds and antioxidant potential. It was possible to note an analogous potential of antibacterial activity of the extracts, which showed action mainly against Gram-positive bacteria. These findings suggest that the aqueous extracts of uvaia leaves are feasible, economic, and sustainable alternatives for adding value to uvaia leaves, which are an agricultural residue that is generally underutilised.

Optimized Single-Step Recovery of Lipophilic and Hydrophilic Compounds from Raspberry, Strawberry and Blackberry Pomaces Using a Simultaneous Ultrasound-Enzyme-Assisted Extraction (UEAE).

An ultrasound-enzyme-assisted extraction (UEAE) was optimized to extract, simultaneously, the hydrophilic and lipophilic compounds from three berry pomaces (raspberry, strawberry and blackberry). First, an enzyme screening designated a thermostable alkaline protease as the most suitable enzyme to recover, in an aqueous medium, the highest yields of polyphenols and oil in the most efficient way. Secondly, the selected enzyme was coupled to ultrasounds (US) in sequential and simultaneous combinations. The simultaneous US-alkaline enzyme combination was selected as a one-single-step process and was then optimized by definitive screening design (DSD). The optimized parameters were: US amplitude, 20% (raspberry pomace) or 70% (strawberry and blackberry pomaces); pH, 8; E/S ratio, 1% (w/w); S/L ratio, 6% (w/v); extraction time, 30 min; temperature, 60 °C. Compared to conventional extractions using organic solvents, the UEAE extracted all the polyphenols, with around 75% of the active polyphenols (measured by the DPPH● method) and up to 75% of the initial oil from the berry pomaces. Characterized lipophilic compounds were rich in polyunsaturated fatty acids (PUFAs), tocols and phytosterols. The polyphenolics were analyzed by UPLC-MS/MS; characteristic ellagitannins of the Rosaceae family (sanguiin H-6 or agrimoniin, sanguiin H-10, …) and ellagic acid conjugates were found as the major components.

Optimization of Enzyme-Assisted Extraction of Bioactive Compounds from Sea Buckthorn (Hippophae rhamnoides L.) Leaves: Evaluation of Mixed-Culture Fermentation.

Hippophae rhamnoides L. leaves possess a remarkable amount of polyphenols that could serve as a natural remedy in various applications. In comparison, numerous techniques, such as conventional and high-pressure techniques, are available for extracting the bioactive fractions from sea buckthorn leaves (SBL). However, enzyme-assisted extraction (EAE) of SBL has not been comprehensively studied. The aim of this study was to optimize critical EAE parameters of SBL using the cellulolytic enzyme complex, Viscozyme L, to obtain a high-yield extract with a high concentration of bioactive compounds. In order to determine the optimal conditions for EAE, the study employed a central composite design and response surface methodology to analyze the effects of four independent factors (pH, temperature, extraction time, and enzyme concentration) on two different responses. Our findings indicated that under optimal conditions (3:15 h extraction, temperature 45 °C, pH 4.9, and 1% Viscozyme L v/w of leaves DW), EAE yielded 28.90 g/100 g DW of the water-soluble fraction. Furthermore, the EAE-optimized liquid extract was continuously fermented using an ancient fermentation starter, Tibetan kefir grains, which possess lactic acid bacteria (LAB) and have significant potential for use in biopreservation. Interestingly, the results indicated various potential prebiotic characteristics of LAB. Additionally, alterations in the cell wall morphology of the SBL residue after EAE were examined using scanning electron microscopy (SEM). This study significantly optimized EAE parameters for sea buckthorn leaves, providing a promising natural source of bioactive compounds for various applications, such as nutraceuticals, functional foods, and high-value products.

Applications of Enzyme Technology to Enhance Transition to Plant Proteins: A Review.

As the plant-based food market grows, demand for plant protein is also increasing. Proteins are a major component in foods and are key to developing desired structures and textures. Seed storage proteins are the main plant proteins in the human diet. They are abundant in, for example, legumes or defatted oilseeds, which makes them an excellent candidate to use in the development of novel plant-based foods. However, they often have low and inflexible functionalities, as in nature they are designed to remain densely packed and inert within cell walls until they are needed during germination. Enzymes are often used by the food industry, for example, in the production of cheese or beer, to modify ingredient properties. Although they currently have limited applications in plant proteins, interest in the area is exponentially increasing. The present review first considers the current state and potential of enzyme utilization related to plant proteins, including uses in protein extraction and post-extraction modifications. Then, relevant opportunities and challenges are critically discussed. The main challenges relate to the knowledge gap, the high cost of enzymes, and the complexity of plant proteins as substrates. The overall aim of this review is to increase awareness, highlight challenges, and explore ways to address them.

Green extraction and biological activity of phenolic extracts from cashew nut testa using a combination of enzyme and ultrasound-assisted treatments.

BACKGROUND: A combination of enzymes and ultrasound treatment was employed to extract bioactive compounds from cashew nut testa, a by-product of the food industry. The total catechin, flavonoid, and phenolic content of extracts was investigated together with their biological activity. RESULTS: Enzyme and ultrasound-assisted extraction (E-UAE) was performed by incubation with Viscozyme L (20 mL kg-1 of testa powder, v/w) for 60 min before sonication for 40 min. Ultrasound and enzyme-assisted extraction (U-EAE) was carried out using sonication for 40 min before incubation with Viscozyme L (20 mL kg-1 of testa powder) for 60 min. Under appropriate conditions, the total phenolic, flavonoid, catechin, and epigallocatechin gallate content of the extracts from cashew nut testa obtained from a combination method (U-EAE or E-UAE) was significantly higher than that obtained using a single method (EAE or UAE). Extracts of cashew nut testa obtained from E-UAE displayed significantly higher antioxidant and α-amylase inhibitory activity than those from the U-EAE. The E-UAE extract at a concentration of 100 µg mL-1 had a greater impact on the cell viability of MCF-7 after treatment (22% cell viability) than did the doxorubicin (DOX) at 4 µg mL-1 (39% cell viability), and the E-UAE extract at 100 µg mL-1 was considered to be safe for healthy cells because the viability of the bovine aerotic endothelial cells treated with this extract was 91%, which was similar to the DOX treatment. CONCLUSION: The extract of cashew nut testa obtained from E-UAE is valuable and promising for the development of anti-inflammatory therapeutic drugs. © 2023 Society of Chemical Industry.

High-Purity Bioactive Ingredient-3S,3'S-Astaxanthin: A New Preparation from Genetically Modified Kluyveromyces marxianus without Column Chromatography and Gel Filtration.

A highly efficient methodology for bioactive ingredient 3S,3'S-astaxanthin (3S,3'S-AST) preparation from genetically modified yeast (Kluyveromyces marxianus) with a combination of enzyme-assisted extraction and salt-assisted liquid-liquid extraction (SALLE) was achieved. The highest yield of 3S,3'S-AST indicated that FoodPro® CBL for yeast cell walls hydrolysis could significantly enhance extraction and obtain, with the help of SALLE procedure, quantified 3S,3'S-AST over 99% in purity through cation chelation. In the oxygen radical antioxidant capacity (ORAC) assay, the antioxidant capacity of high-purity 3S,3'S-AST products were 18.3 times higher than that of the original raw material extract. This new combination preparation may replace previous methods and has the potential to be scaled up in the manufacture of high-purity 3S,3'S-AST from low-value bioresources of raw materials to high-value products in the food and/or drug industries with lower cost and simple equipment.

Insights into the Oxidative Stress Alleviation Potential of Enzymatically Prepared Dendrobium officinale Polysaccharides.

(1) Background: The extraction parameters can dramatically alter the extraction rate and biological activity of polysaccharides. (2) Methods: Here, an enzyme-assisted extraction (EAE) was employed to extract D. officinale polysaccharides (DOPs), and its optimal extraction conditions were established by single-factor and Box-Behnken design (BBD) experiments. Further, on the basis of in vitro antioxidant capacity, the paraquat (PQ)-induced oxidative stress of Caenorhabditis elegans (C. elegans) was chosen as a research model to explore the antioxidant activity of DOPs. (3) Results: The results showed that the extraction yield of DOPs reached 48.66% ± 1.04% under the optimal condition. In vitro experiments had shown that DOPs have considerable ABTS+ radical scavenging capacity (EC50 = 7.27 mg/mL), hydroxyl radical scavenging capacity (EC50 = 1.61 mg/mL), and metal chelating power (EC50 = 8.31 mg/mL). Furthermore, in vivo experiments indicated that DOPs (0.25 mg/mL) significantly prolonged the lifespan, increased antioxidant enzyme activity, and upregulated the expression of daf-16 (>5.6-fold), skn-1 (>5.2-fold), and sir-2.1 (>2.3-fold) of C. elegans. (4) Conclusions: DOPs can be efficiently extracted by EAE and are effective in the reduction of oxidative stress levels in C. elegans.

Effect of hot water, ultrasound, microwave, and pectinase-assisted extraction of anthocyanins from black goji berry for food application.

Lycium ruthenicum, commonly known as black goji berry, is a rich anthocyanin source containing a high amount of monoacylated anthocyanins. This study investigates the effect of different extraction methods to extract anthocyanins from black goji berry for food application. Different hot water extraction conditions were applied to investigate the effect of specific substrate: solvent ratio (1:15 and 1:20 (w/v)), extraction time (30 and 60 min) and extraction temperature (40, 50 and 60 °C) on the extraction yield, total anthocyanin content (TAC) and the total phenolic content (TPC) of the anthocyanin extracts. Best hot water extraction conditions for obtaining an anthocyanin extract with high TAC (13.8 ± 1.14 mg CGE/g), TPC (69.7 ± 2.50 mg of GAE/g), and extraction yield (48.3 ± 3.25%) consuming less solvent, time and heat were substrate: solvent ratio of 1: 15 (w/v), extraction temperature of 50 °C, and extraction time of 30 min. The effect of pectinase, ultrasound, and microwave on hot water extraction of anthocyanins from black goji berry was investigated using the best conditions for hot water extraction. Pectinase-assisted extraction [1.5% (w/v) pectinase, substrate: solvent ratio of 1:15 (w/v) at 50 °C for 30 min] was the best extraction method to extract black goji berry anthocyanins demonstrating higher extraction yield, TAC, TPC, and the highest percentage of petunidin-3-O-(trans-p-coumaroyl)-rutinoside-5-O-glucoside.

Optimization of enzyme-assisted extraction of anthocyanins from eggplant (Solanum melongena L.) peel.

Enzyme-assisted extraction (EAE) of total yield (TY), total anthocyanin content (TA), antioxidant activity (AOA) and total phenolic content (TPC) from eggplant peel was optimized using response surface methodology. Enzyme concentration (5-15 %), extraction temperature (35-60 °C) and maceration time (1-4.5 hrs) were used to optimize the process of extraction using a Central-Composite design. Optimum values of TY (71.45 %), TAC (578.66 mg C3G L-1), TPC (2040.87 mg GAE L-1), DPPH (79.92 %) and FRAP (29.90 mmol AAE/100 g) were obtained for the optimized extraction parameters viz., temperature (37.32 °C), enzyme concentration (5%) and extraction time (1 h). Further, a comparative study was also done between conventional extraction and enzyme-assisted extraction of eggplant peel. It was observed that the responses of the extract obtained by conventional method showed significant variation from that obtained by EAE indicating the superiority of latter.

Novel strategies for extraction, purification, processing, and stability improvement of bioactive molecules.

Bioactive molecules gain significance in pharmaceutical and nutraceutical industries for showcasing various beneficial biological properties including but not limited to anticancer, antimicrobial, antioxidant, antifungal, anti-inflammatory, cardioprotective, neuroprotective, and antidiabetic. However, the practice of using traditional approaches to produce bioactive molecules is gradually declining due to various limitations such as low product quality, high toxicity, low product yield, low efficiency, and product degradation. Thus, with the escalating demand for these bioactive molecules and active agents in food and other food-related industries, it has become a dire need for the scientific world to come up with novel approaches and strategies that cannot just improve the quality of these bioactives but also prepare them in a comparatively shorter time span. This review includes the latest approaches and techniques used either independently or in combinations for the extraction, purification, processing, and stability improvement of general bioactive molecules. Different parameters of these versatile techniques have been discussed with their effectiveness and work principles.

A Case Study for the Extraction, Purification, and Co-Pigmentation of Anthocyanins from Aronia melanocarpa Juice Pomace.

Chokeberry (Aronia melanocarpa) pomace is a by-product from the juice industry very rich in anthocyanins and other bioactive components. Recovery and purification of anthocyanins from the pomace is a viable valorization strategy that can be implemented to produce high-value natural food colorants with antioxidant properties. In this study, chokeberry pomace was subjected to enzyme-assisted extraction using commercial pectinases. The extracts were further purified by adsorption-desorption using an acrylic resin and stabilized by co-pigmentation with ferulic acid. The anthocyanin concentration and antioxidant activity of the extracts were unaffected by the enzymatic treatment at the conditions tested. The total phenolic content of the extracts suffered minor variations depending on the enzyme formulation used, whereas the dissolved solid content increased in all cases. The adsorption-desorption strategy allowed a 96% recovery of the anthocyanins initially present in the extract, whereas the co-pigmentation treatment magnified the intensity of the color in terms of absorbance, and improved the stability during storage up to one month.