Enzyme-assisted Extraction Research Articles

Correction to: Enzyme-assisted extraction of apricot polysaccharides: process optimization, structural characterization, rheological properties and hypolipidemic activity

Correction to: Enzyme-assisted extraction of apricot polysaccharides: process optimization, structural characterization, rheological properties and hypolipidemic activity

Enzyme-assisted extraction of anthocyanins and other phenolic compounds from blackcurrant (Ribes nigrum L.) press cake: From processing to bioactivities

The effects of commercial enzymes (pectinases, cellulases, beta-1-3-glucanases, and pectin lyases) on the recovery of anthocyanins and polyphenols from blackcurrant press cake were studied considering two solid:solvent ratios (1:10 and 1:4 w/v). β-glucanase enabled the recovery of the highest total phenolic content – 1142 mg/100 g, and the extraction of anthocyanins was similar using all enzymes (∼400 mg/100 g). The use of cellulases and pectinases enhanced the extraction of antioxidants (DPPH − 1080 mg/100 g; CUPRAC – 3697 mg/100 g). The freeze-dried extracts presented antioxidant potential (CUPRAC, DPPH), which was associated with their biological effects in different systems: antiviral activity against both non-enveloped viruses (enterovirus coxsackievirus A-9) and enveloped coronaviruses (HCoV-OC43), and cytotoxicity towards cancer cells (A549 and HCT8). No cytotoxic effects on normal human lung fibroblast (IMR90) were observed, and no anti-inflammatory activity was detected in lipopolysaccharides-treated murine immortalised microglial cells.

The compound-independent calibration of five selenium species in rice using ion-pairing reversed phase chromatography coupled to inductively coupled plasma tandem mass spectrometry

An enzyme-assisted extraction and an ion pairing reversed phase chromatography (IP-RPC) coupled to inductively coupled plasma tandem mass spectrometry (ICP-MS/MS) method were established for the simultaneous analysis of five selenium species in rice: selenious acid (SeIV), selenic acid (SeVI), selenocystine (SeCys2), methylselenocysteine (SeMeCys) and selenomethionine (SeMet). Optimal extraction efficiencies were obtained by using 15 mg protease XIV, water bath temperature of 45°C, pH of 6.5 and incubation of six hours. The total extracted Se species account for 92.5-109.3 % of the total Se in rice. The instrumental limits of detection for five selenium species ranged from 0.04 to 0.12 ng Se g−1. Spike recovery experiments performed on rice samples were in the range of 96.1-102.9 % for all analytes except for SeCys2 (66.1-77.1 %). A consistency of Se elemental response was observed among Se species analyzed in this study as the ratio of linear equation slope ranged from 1.020 to 1.036 (SeIV as reference) in rice matrix. The developed compound-independent calibration method was applied to detect Se species in eleven rice samples from China. Both selenium-enriched rice and regular rice were predominated with SeMet, accounting for ∼89.4 % of total selenium.

The influence of enzymatic pretreatment of chickpea on properties of protein nanoparticles prepared by heat treatment

Protein nanoparticles from chickpea protein isolates were prepared by heat treatment and their properties were evaluated. Protein was extracted from defatted chickpea seed under alkaline conditions after pretreatment with single arabinofuranosidase or combination of cellulase and xylanase, or without it. Both enzymatic pretreatments delivered protein isolates with enhanced ratio between α-helices and β-sheets/β-strands in their secondary structure comparing to alkaline isolate. Applied heat treatment was performed for 10 min or 20 min at 90 °C, and at pH 7 or pH 9.3. Particle size of prepared nanoparticles varied from 28 to 290 nm, with the smallest particles fabricated from isolate from (cellulase + xylanase)-assisted alkaline extraction at pH 9.3. Protein isolate extracted with the assistance of arabinofuranosidase enabled preparation of nanoparticles with the highest linoleic acid binding capacity - 58% at pH 7, and 69% at pH 9.3. Generally, nanoparticles with smaller particle sizes, and higher linoleic acid binding capacity and ABTS scavenging activity were fabricated from protein extracted by enzyme-assisted alkali protocols and at higher investigated pH. Chickpea protein isolates from enzyme-assisted extractions can be great source for preparation of nanoparticles with advanced properties for the application in food sector.

Enhanced Growth Inhibition and Apoptosis Induction in Human Colon Carcinoma HT-29 Cells of Soluble Longan Polysaccharides with a Covalent Chemical Selenylation.

The selenylated polysaccharides chemically belong to the organic Se-conjugated macromolecules and have recently been attracting more and more attention due to their potential to promote body health or prevent cancers. Longan (Dimocarpus longan L.), as a subtropical fruit, contains soluble and non-digestible polysaccharides that are regarded with health care functions in the body. In this study, the longan polysaccharides (LP) were obtained via enzyme-assisted water extraction, and then chemically selenylated using a reaction system composed of HNO3–Na2SeO3 to yield two selenylated products, namely, SeLP1 and SeLP2, with Se contents of 1.46 and 4.79 g/kg, respectively. The anti-cancer effects of the three polysaccharide samples (LP, SeLP1, and SeLP2) were thus investigated using the human colon cancer HT-29 cells as the cell model. The results showed that SeLP1 and SeLP2 were more able than LP to inhibit cell growth, alter cell morphology, cause mitochondrial membrane potential loss, increase intracellular reactive oxygen and [Ca2+]i levels, and induce apoptosis via regulating the eight apoptosis-related genes and proteins including Bax, caspases-3/-8/-9, CHOP, cytochrome c, DR5, and Bcl-2. It was thereby proven that the selenylated polysaccharides could induce cell apoptosis via activating the death receptor, mitochondrial-dependent, and ER stress pathways. Collectively, both SeLP1 and SeLP2 showed higher activities than LP in HT-29 cells, while SeLP2 was consistently more active than SeLP1 in exerting these assessed anti-cancer effects on the cells. In conclusion, this chemical selenylation covalently introduced Se into the polysaccharide molecules and caused an enhancement in their anti-cancer functions in the cells, while higher selenylation extent was beneficial to the activity enhancement of the selenylated products.

Bioactive compounds and its optimization from food waste: review on novel extraction techniques

Purpose The amount of food wasted every year is 1.3 billion metric tonne (MT), out of which 0.5 billion MT is contributed by the fruits processing industries. The waste includes by-products such as peels, pomace and seeds and is a good source of bioactive compounds like phenolic compounds, flavonoids, pectin lipids and dietary fibres. Hence, the purpose of the present study is to review the novel extraction techniques used for the extraction of the bio active compounds from food waste for the selection of suitable extraction method. Design/methodology/approach Novel extraction techniques such as ultrasound-assisted extraction, microwave-assisted extraction, enzyme-assisted extraction, supercritical fluid extraction, pulsed electric field extraction and pressurized liquid extraction have emerged to overcome the drawbacks and constraints of conventional extraction techniques. Hence, this study is focussed on novel extraction techniques, their limitations and optimization for the extraction of bioactive compounds from fruit and vegetable waste. Findings This study presents a comprehensive review on the novel extraction processes that have been adopted for the extraction of bioactive compounds from food waste. This paper also summarizes bioactive compounds' optimum extraction condition from various food waste using novel extraction techniques. Research limitations/implications Food waste is rich in bioactive compounds, and its efficient extraction may add value to the food processing industries. Hence, compressive analysis is needed to overcome the problem associated with the extraction and selection of suitable extraction techniques. Social implications Selection of a suitable extraction method will not only add value to food waste but also reduce waste dumping and the cost of bioactive compounds. Originality/value This paper presents the research progress on the extraction of bioactive active compounds from food waste using novel extraction techniques.

Natural colorants from vegetable food waste: Recovery, regulatory aspects, and stability-A review.

Worldwide, approaches inspired by the Circular Economy model have been increasing steadily, generating new business opportunities such as the recovery of high-added value molecules (e.g., pigments) from vegetable food waste that may be applied as food additives (e.g., colorants). Indeed, food waste is a global problem that does not seem to be decreasing, leading to economic, environmental, and social issues. Moreover, synthetic dyes have been associated with adverse effects on human health, encouraging research to explore much safer, natural, and eco-friendly pigments. This state-of-the-art review gives a brief overview of the regulatory aspects concerning food waste, Circular Economy, and natural versus synthetic colorants. We have critically reviewed the recent advances in pigment recovery from vegetable food waste bringing back the green/unconventional extraction methods. Among them, enzyme-assisted extraction as a depth feature technique is highlighted, given that it allows the recovery of pigments in a mild, selective, efficient, and sustainable way. Furthermore, the stability issue of the different natural colorants has been critically discussed in relation to the extraction and application conditions. Several and tailored stabilization methods have been described and reported for each pigment although additional research is necessary on their long-lasting stabilization and utilization in food matrices. PRACTICAL APPLICATION: This review focuses on the main types of natural pigments in vegetable food waste, their legislative framework, extraction technologies and strategies to improve the stability , as well as their possible applications.

Enzyme-assisted extraction of apricot polysaccharides: process optimization, structural characterization, rheological properties and hypolipidemic activity

Enzyme-assisted extraction of apricot polysaccharides: process optimization, structural characterization, rheological properties and hypolipidemic activity

PH-controlled reversible deep-eutectic solvent based enzyme system for simultaneous extraction and in-situ separation of isoflavones from Pueraria lobata

In this study, we developed an efficient and green pH-controlled reversible deep-eutectic solvents (DESs) based enzyme system for efficient extraction, transformation and in-situ separation of natural products from herbs. As a case study, seven isoflavones in the root of Pueraria lobata were selected as target compounds. A series of DESs were firstly prepared and the extraction efficiency for target compounds with or without cellulase catalysis was investigated. We found that [N4444][Cl]:ethylene glycol (1:2) with cellulase addition show the highest extraction efficiency for isoflavones. More importantly, the phase behavior of [N4444][Cl]:ethylene glycol (1:2) based cellulase system was found to be pH-controlled. When pH is 5.0, the system is a homogeneous single-phase system, which is conducive to the extraction of natural products. When pH is 6.4, the system changes from the single-phase to the heterogeneous two-phase, to realize the in-situ separation of the compounds. Therefore, by adjusting the pH of the system, the in-situ separation of isoflavones was successfully realized with high recovery yields. The pH-controlled DESs based enzyme-assisted extraction system overcomes the incompatibility between enzyme catalysis and conventional organic solvent in extraction, which could efficiently extract seven isoflavones from Pueraria lobata root.

Application of Enzyme-Assisted Extraction for the Recovery of Natural Bioactive Compounds for Nutraceutical and Pharmaceutical Applications

Enzyme-assisted extraction (EAE) involves the use of hydrolytic enzymes for the degradation of the cell wall or other cell components. This supports the diffusion of the solvent into the plant or fungal material, leading to easier elution of its metabolites. This technique has been gaining increasing attention, as it is considered an eco-friendly and cost-effective improvement on classical or modern extraction methods. Its promising application in improving the recovery of different classes of bioactive metabolites (e.g., polyphenols, carotenoids, polysaccharides, proteins, components of essential oil, and terpenes) has been reported by many scientific papers. This review summarises information on the theoretical aspects of EAE (e.g., the components of the cell walls and the types of enzymes used) and the most recent discoveries in the effective involvement of enzyme-assisted extraction of natural products (plants, mushrooms, and animals) for nutraceutical and pharmaceutical applications.

Effects of five extraction methods on total content, composition, and stability of flavonoids in jujube

The present study investigated the effects of different extraction methods including water-water bath (W-WB), ethanol-water bath (E-WB), deep eutectic solvent (DES) combined with ultrasound-assisted extraction (DES-UAE), microwave-assisted extraction (DES-MAE), and enzyme-assisted extraction (DES-EAE) on flavonoids (total flavonoid content, flavonoid composition, and stability) in jujube. The highest total flavonoid content of 8.03mg/g was obtained by the DES-MAE extraction. Fifteen types of flavonoids were identified from jujube. The amount of rutin produced by the E-WB and DES-UAE methods was 66.88±1.58μg/g and 45.23±3.22μg/g, respectively. The retention of flavonoids in DES-UAE extracts were 98.15±0.51%, 64.25±2.21% after 2h of high temperature treatment at 90°C and 21days of dark storage, respectively. The flavonoids extracted by different methods were suitable for dark storage under different light contrasts, where the retention of flavonoids extracted by DES-UAE method was 86.44±2.45%. In conclusion, DES-UAE would be an efficient method for flavonoid extraction from jujube.

A Concise Review on Taro Mucilage: Extraction Techniques, Chemical Composition, Characterization, Applications, and Health Attributes.

Taro (Colocasia esculenta) is an important source of carbohydrates as an energy source and is used as a staple food throughout the world. It is rich in mucilage and starch granules, making it a highly digestible ingredient. Mucilage can act as a matrix and a thickening, binding, emulsifying, or foaming agent in food, pharmaceutical, and several other fields of research. Moreover, mucilage can be extracted from several living organisms and has excellent functional properties, such as water-holding, oil-holding, and swelling capacities. Therefore, these remarkable functional properties make mucilage a promising ingredient with possible industrial applications. Furthermore, several extraction techniques, including enzyme-assisted, ultrasonication, microwave-assisted, aquatic, and solvent extraction methods, are used to obtain quantitative amounts of taro mucilage. Coldwater extraction with ethanol precipitation can be considered an effective and cost-effective technique to obtain high-quality mucilage with suitable industrial applications, whereas the ultrasonication method is more expensive but results in a higher amount of mucilage than other emerging techniques. Mucilage can also be used as a fat replacer or reducer, dye remover, coating agent, and antioxidating agent. Therefore, in this review, we detail the key properties related to the extraction techniques, chemical composition, and characterization of taro mucilage, along with its suitable applications and health benefits.

Enhanced Liberation of Soluble Sugar, Protein, and R-Phycoerythrin Under Enzyme-Assisted Extraction on Dried and Fresh Gracilaria gracilis Biomass

This study aims to investigate the bio-refinery process through an enzyme-assisted extraction (EAE) on freeze-dried and fresh macroalgae Gracilaria gracilis for the release of water-soluble components (R-phycoerythrin, proteins, and sugar). Three enzymes, cellulase, protease, and enzyme cocktail (mixture of cellulase and protease), were applied in the study. Results showed that freeze-dried biomass yielded the highest target components in the presence of enzyme cocktail while a single enzyme was better with fresh biomass, either protease for the release of R-PE and protein or cellulase for sugar. The extraction of protein and sugar was improved by 43% and 57%, respectively, from fresh biomass compared to dried biomass. The difference of biomass status was shown to affect the required enzyme and recovery yield during the extraction process. Employing an enzyme cocktail on freeze-dried biomass boosted the extraction yield, which was probably due to the complementary effect between enzymes. On the other hand, single enzyme worked better on fresh biomass, giving economic benefits (enzyme limitation and drying stage) for further implementation of the bio-refinery process. Thus, biomass treatment (fresh or freeze-dried) and enzyme-type determined the efficiency of enzyme-assisted extraction according to the target components.

A green ultrasound-assisted enzymatic extraction method for efficient extraction of total polyphenols from Empetrum nigrum and determination of its bioactivities

In this study, a green and efficient ultrasound-assisted enzymatic extraction (UAEE) method was established for total polyphenols (TPP) extraction from E. nigrum aerial parts. Fractional Factorial design (FFD) was initially implemented for the selection of significant parameters affecting TPP yield and then Box-Behnken design-response surface methodology (BBD-RSM) was employed to optimize the parameters. The optimum extraction conditions for UAEE were as follows: incubation temperature of 38.39 ℃, incubation time of 3.39 h, tannase of 386.53 U/g of sample (U/g), and cellulase of 224.42 U/g. Under the optimum conditions, the TPP yield was 52.17 ± 0.39 mg/g, which was 1.62 and 1.73 times those of the methods of ultrasound-assisted extraction (UAE) and enzyme-assisted extraction (EAE), respectively. The crude extract was purified by HPD-600 microporous resin, and the purity was improved from 8.11% to 22.56%. In addition, the results of antioxidant and antibacterial activity assays showed that the purified extract has stronger bioactivities in contrast with the crude extract. This study indicated that UAEE was an effective technique for the extraction of TPP from the aerial parts of E. nigrum, and TPP extracted by this extraction method could be applied in pharmaceuticals, cosmetics, and food industries.

Depolymerization of fucoidan with endo-fucoidanase changes bioactivity in processes relevant for bone regeneration

Fucoidans are polysaccharides from brown macroalgae, showing multiple bioactivities important for bone regeneration and bone health. However, the use of fucoidans in medical applications remains sparse due to the heterogeneity in their chemical properties and unclear structure-function relationships. Innovations in extraction techniques and post processing steps are needed to produce homogeneous fucoidan molecules with tailorable bioactivities. Here, we applied enzyme-assisted extraction coupled with enzymatic hydrolysis by Fhf1 fucoidanase to generate low (LMW) and medium molecular weight (MMW) fucoidans from Fucus evanescens. In contrast to the anti-angiogenic properties of the high molecular weight fucoidan, LMW and MMW no longer suppressed the production of pro-angiogenic molecules by bone stem cells, nor impaired the formation of prevascular structures in vitro. In contrast to LMW, a pro-inflammatory response of OEC was observed after treatment with high concentrations of MMW. Thus, fucoidanase hydrolysis could be a useful tool to tailor the bioactivity of fucoidans.

Improving Enzyme-Assisted Extraction of Brazilin from Sappanwood (Caesalpinia Sappan L.) Extract by Fungal Cellulase

Brazilin was one of phytoconstituent from sappanwood that widely used as textile-colouring agent and found in traditional concoction for therapeutic purposes. Due to obtain its maximum level in sappanwood (Caesalpinia sappan L.) extract, the extraction method of brazilin has been developed. Enzyme-assisted extraction (EAE) is one of green chemistry methods to achieve that outcome. This study aims to enhance brazilin level by optimize the EAE condition with fungi cellulase. The cellulase are produced by monoculture Aspergillus niger UICC371 in carboxymethyl cellulose submerged fermentation’s media. Sappanwood extracted with fungi cellulase through variation conditions: enzyme concentrations (2.0; 4,0; 6.0%); temperature (45, 50, 55℃); and time (1, 2, 3 hrs). The optimization are provided by response surface method-BoxBehnken design and brazilin level was carried out through High Performance Liquid Chromatography (HPLC) with asetonitril : 0,3% acetic acid in water (14.5 : 85.5) as eluents. The study showed that cellulase from monoculture of Aspergillus niger UICC371 showed optimum condition of cellulase-EAE method at 6.0% concentration enzyme at 50℃ for 3 hours extraction time which provide an increase in brazilin level to 5.014% compare to reflux method. Key Words: Caesalpinia sappan L., Cellulase, Enzyme assisted extraction, Fungi, Response surface method.

Enzyme-assisted extraction of red seaweed Solieria chordalis (C.Agardh) J. Agardh 1842—the starting point for the production of biostimulants of plant growth and biosorbents of metal ions

The underexploited biomass of red seaweed Solieria chordalis (C.Agardh) J.Agardh 1842 represents a potential source for biotechnological development. Extracts obtained from S. chordalis by enzyme-assisted extraction using Protamex® and Neutrase® were evaluated as potential biostimulants of plant growth. Additionally, this alga, as well as post-extraction residues, were examined as biosorbents of metal ions. The biostimulant properties of different concentrations of algal extracts (20, 40, 60, 80, 100%) were tested in vitro on radish seeds. Chlorophyll content, plant weight, and height were measured for each experimental group. For all tested concentrations, the plants demonstrated higher chlorophyll content and were higher and heavier than the control group, showing that enzyme extracts could be used as efficient biostimulants. The biosorption properties of seaweed and post-extraction residues were tested on Cr(III) ions for different experimental conditions—pH, initial metal ion concentration, and biosorbent dosage. The maximum biosorption capacity of S. chordalis was 48.1 mg/g, for the post-extraction residue obtained with Protamex®, 47.6 mg/g, and Neutrase®, 50.5 mg/g. The binding of Cr(III) ions to the surface of biosorbents was confirmed by FT-IR analysis. Good biosorption properties of tested materials can be used in the production of novel components of fertilizers or biosorbents for wastewater treatment. In this paper, it was shown that waste biomass of S. chordalis can be turned into valuable bio-products using environmental-friendly technologies.

Effects of Novel Extraction Strategies on the Recovery of Phenolic Compounds and Associated Antioxidant Properties from Buckwheat Hull (Fagopyrum esculentum)

This study investigated the effects of novel extraction technologies, including ultrasound-assisted extraction (UAE), microwave-assisted extraction (MAE), pulsed electric field (PEF), high-pressure processing (HPP), enzyme-assisted extraction (EAE), and conventional extraction, on the recovery of phenolic compounds and associated antioxidant properties from buckwheat hull (Fagopyrum esculentum). Initially, twenty-four extraction strategies were investigated. Based on the results of the total phenolic content and antioxidant properties (DPPH and FRAP), twelve strategies (i.e., US (n = 2), PEF (n = 1), MW (n = 4), HPP (n = 4), and a control method) were selected for phenolic profiling carried out using liquid chromatography-mass spectrometry (LC-MS). Forty-one phenolic compounds were identified in the extracts, and a scanning electron microscope (SEM) analysis was also carried out on the treated residues to analyze the surface damage post-treatments. The results showed that samples treated with US (16.14 ± 0.06), PEF (9.94 ± 0.02), MW (12.63 ± 0.13), and HPP (21.76 ± 0.78) contained the highest total phenolic content (mg GAE/100 mg of DW). In the case of the antioxidant activities, the highest DPPH activities were obtained using HPP, MAE, and UAE, while no clear pattern was recorded in the case of FRAP activities. The highest DPPH and FRAP activities observed were 80.91 ± 0.22% and 23.98 ± 0.2 mg Trolox equivalents/100 mg, respectively. Additionally, the LC-MS results identified eleven different groups of phenolic compounds in buckwheat hull extracts, including anthocyanin, flavanol, flavanones, flavones, flavonol, phenolic acids, isoflavones, lignans, and quinones.

Effects of High-Pressure, Hydrothermal, and Enzyme-Assisted Treatment on the Taste and Flavor Profile of Water-Soluble Ginger (Zingiber officinale) Extract.

Ginger, a plant widely consumed worldwide, is used as a spice or to enhance the flavor of foods. In this study, the taste characteristics (gingerol, shogaol, and amino acid) of extracts treated with various solubilizing methods were objectively compared. In addition, an E-nose confirmed the flavor pattern combined with principal component analysis (PCA) between each extract gas chromatogram-tandem mass spectrometry was performed to compare and analyze volatile compounds between extraction methods. As a result, high-pressure enzyme-assisted extraction (HPE) and hydrothermal enzyme-assisted extraction (HWE) treatment effectively improved the extraction yield of ginger and the contents of gingerol and shogaol and removed the bitter taste. In addition, radar charts of both E-nose and PCA provided the distribution of flavor substances in HPE and HWE products of ginger. After enzyme-assisted treatment, a strong fruity and piquant flavor was noted. In conclusion, it is suggested that ginger extract of enzyme-assisted treatment has increased flavor compounds and can be an excellent food material.

A STUDY ON CHARACTERIZATION OF PECTINASE ASSISTED AQUEOUS EXTRACTION OF RICE BRAN OIL

The Rice bran oil is known as wonder oil because of its numerous health benefits. It has a number of advantages over other edible oils because of the presence of antioxidant known as oryzanol. The rice bran contains 10-26% oil depending on the factors like variety, milling process, and other conditions. Various solvents are currently used for rice bran oil extraction process at industrial sectors. These solvents are carcinogenic and also cause adverse effects to the environment. The enzyme-assisted aqueous extraction process has been regarded as an eco-friendly process. Pectinase enzyme is used to break down complex carbohydrates like pectin present in rice bran and allows easy oil extraction in aqueous solution. The present study aims to perform the aqueous extraction of rice bran oil using crude pectinase enzyme extracted from the pectinase producing bacteria Bacillus paramycoides and to study the oil characteristics. Out of the various solvents used in the study, hexane yielded highest percentage of oil. The oil yield was found to be 49.8% in 6 h. The total activity of enzyme was found to be 11 U/mL/min. 500 Units of enzyme was taken for rice bran oil extraction. The Average yield of oil was found to be 80.5% in 3 h and 76.9% in 6 h. The acid value, saponification value, iodine value and oryzanol content of the oil was found to be11.22, 84.15, 5.0 and 0.05 percentage respectively. The percentage of radical scavenging activity of the extracted rice bran oil was found to be 49.51%. The fatty acids present in the oil sample were analyzed by GC MS analysis.