Enzymatic Extraction Technique Research Articles

Structural characteristics of neutral polysaccharides purified from coix seed and its anti‐insulin resistance effects on HepG2 cells

AbstractCoix seed is recognized as a functional medicinal food due to its valuable biological activities, with polysaccharides being the primary active compounds. In this study, an ultrasonic‐assisted enzymatic extraction technique was employed, and response surface methodology was used to optimize the yield of polysaccharides to 9.55 ± 0.26%. A novel neutral polysaccharide, CSPsN‐1, was purified with a molecular weight of 7.75 kDa. CSPsN‐1 was composed of arabinose, galactose, glucose, xylose, and mannose in molar ratios of 0.48: 7.92: 86.39: 2.42: 2.79. Its backbone composed of →4)‐α‐D‐Glcp‐(1→ and →3,4)‐α‐D‐Glcp‐(1→ units, with terminal residues of α‐D‐Glcp. In vitro experiments, CSPsN‐1 enhanced glucose consumption in insulin‐resistant HepG2 cells and upregulated GLUT4 expression by activating the PI3K/AKT signaling pathway. These findings suggest that CSPsN‐1 holds significant promise as a functional ingredient for treating insulin resistance and related metabolic disorders.

Eumelanin: A natural antioxidant isolated from squid ink by new enzymatic technique and prediction of its structural tetramer

The investigation into natural eumelanin found in squid ink is an emerging research topic due to its antioxidative properties. However, the methodologies for acquiring pure eumelanin and establishing the intricate relationship between its structure and antioxidative efficacy remain limited. Novel enzyme extraction technique, termed response surface optimization, was employed to synthesize high-purity eumelanin, and its antioxidant activity was evaluated at free radical scavenging and cellular levels and correlated with detailed tetrameric structure prediction for the first time. Results demonstrated that enzymatic extraction technique yielded eumelanin (ANF) nanostructure with stability, boasting particle size of 112.35 nm, a maximum absorption wavelength of 224 nm, a ΔHpp value of 0.4802 mT, and a purity of 92%. Compared with melanin standard (MelS), ANF had superior free radical scavenging activity, eliminating 89.62% of DPPH radicals, 96.79% of superoxide ion radicals, and 93.20% of hydroxyl radicals, and better repair of cellular oxidative damage. Based on molecular energy, molecular electrostatic potential (MEP), and molecular orbitals by Gaussian calculations, the tetrameric structure (DHICA3-DHI1‴ and “DHICA3-DHI1”) with significant antioxidant activity contained inside eumelanin was anticipated. Our findings reveal its potential as a natural antioxidant, offering broad prospects for antioxidant food additives or therapeutic applications in nutritional and pharmaceutical ingredients.

Effect of combined ultrasonic and enzymatic extraction technique on the quality of noni (Morinda citrifolia L.) juice.

In order to obtain noni juice with high yield and good quality, the effect of combined extraction technique of enzymatic treatment (EZ) and ultrasonication (US) on the overall quality of noni juice was investigated. Moreover, the extraction performance of the EZ-US combined extraction technique was compared with that of EZ-based extraction and the US-based extraction. Response surface methodology (RSM) was designed to optimize the parameters of ultrasonic treatment, by taking consideration of the extraction efficiency, quality parameters and bioactive ingredients of noni juice. The results indicated that combined ultrasonic and enzymatic treatment achieved a synergistic effect on promoting the quality of noni juice. The maximum juice yield of 67.95% was obtained under ultrasonication for 10min at 600W after enzymatic treatment (EZU). In addition, EZU-treated juice exhibited the highest contents of total phenolic and flavonoid, which were 148.19±2.53mg gallic acid/100mL and 47.19±1.22mg rutin/100mL, respectively, thus contributing to better antioxidant activity. Moreover, the EZU treatment significantly reduced the particle size of noni juice, and improved its suspension stability and rheological properties. FTIR results indicated that the treatments did not bring major changes in the chemical structure and the functional groups of compounds in noni juice. Therefore, EZU treatment can be successfully applied to the extraction of noni juice with better nutritional properties and overall quality.

Extraction of high purity fucoidans from brown seaweeds using cellulases and alginate lyases

Fucoidans are fucose rich sulfated polysaccharides that are found in the cell wall of brown seaweeds and have been shown to have several beneficial bioactivities. In the present study, we report a new enzymatic extraction technique for the production of pure and intact fucoidans from the two brown seaweeds Saccharina latissima and Alaria esculenta. This new extraction protocol uses the commercial cellulase blend Cellic® CTec2 in combination with endo- and exo-acting thermophilic alginate lyases. The fucoidans obtained by this extraction technique are compared to traditionally extracted fucoidans in terms of chemical compositions and molecular weights and are shown to contain significantly higher amounts of fucose and sulfate, the main components of fucoidans, while cellulose, laminarin, and alginate contamination is low. Thus, by using this combination of enzymes, the extracted fucoidans do not undergo depolymerization during extraction and additional purification steps are not needed. The high purity fucoidans isolated by this new enzymatic extraction technique can be used to provide insight into the different fucoidan structures and biological activities.

Microwave‐assisted enzymatic extraction of plant protein with antioxidant compounds from the food waste sesame bran: Comparative optimization study and identification of metabolomics using LC/Q‐TOF/MS

This study aimed to investigate the effects of microwave-assisted and combined microwave-assisted enzymatic extraction techniques for the recovery of plant-based protein and antioxidant compounds from the food waste sesame bran. The effects of independent variables, that is, process temperature (25–55 °C), time (10–120 min), and enzyme concentrations (0.12–2.40 AU/100 g) were examined using a central composite design by the response surface methodology and were all found to have significant effects on protein yield, total phenolic content, and antioxidant capacity values. Optimal process conditions for microwave-assisted extraction were 51 °C process temperature and 29 min time; and for combined microwave-assisted enzymatic extraction were 1.94 AU/100 g enzyme concentration, 49 °C, 98 min. Protein fractions of extracts by SDS-PAGE, cellular structures of sesame brans by the scanning electron microscope, and identification of protein subunits and phenolic compounds by LC/Q-TOF/MS analyses were also inspected and compared with a standard alkaline method. Practical applications Cereal and oilseed by-products like brans have gained great importance in food technology due to their valuable protein contents. Recently, several attempts were performed to recover plant-based protein from various food wastes. Sesame bran is an unevaluated agro-industrial by-product and is a potential source of plant protein with phenolic compounds possessing functional properties. Latest studies showed that sesame processing plants reveal a huge amount of bran which have valuable compounds such as sulphur-containing amino acids. This study shows that highest recovery rates of plant-based protein with bioactive compounds from sesame bran could be achieved by the microwave-assisted enzymatic extraction technique. Obtained protein isolates could be evaluated in functional food developments. Results of this study could be an insight into researchers and food technologists who like to carry out studies on similar subjects.

Improvements in the extraction of bioactive compounds by enzymes

Bioactives from plants are always in high demand in nutraceutical, pharmaceutical, and functional food sectors due to their health benefits. This intensifies the need of extraction of bioactives by different methods that can improve the yield and purity of the compound. Enzymes help in the release of bioactives from the plant material under optimized conditions so as to make the extraction process efficient. Though enzymatic extraction of bioactives have been used since long, it needs improvement to further enhance the yield, reduce the process time, and make the process cost competitive. The combination of enzymatic extraction with other green techniques such as ultrasound extraction, supercritical fluid extraction, three-phase partitioning, ionic liquid extraction, and microwave extraction can boost the advantages of enzymatic extraction. This review focuses on the improvement in the enzymatic extraction techniques for the bioactives in detail.

Extraction and Characterization of Omega-3 Fatty Acid from Catfish Using Enzymatic Hydrolysis Technique

Extraction of Omega-3 fatty acid from Catfish using enzymatic hydrolysis techniques are expected to be more economically possible techniques due to the uses of enzyme with the characteristic of reusable, environmental friendly and less energy required for large-scale production. Malaysian like to consume Catfish in their daily diet. Thus, it is very popular among farmers in Malaysia. Omega-3 polyunsaturated fatty acid have many benefit to human health. Based on research, EPA and DHA can reduce the risk of heart disease by regulate the blood coagulation. The main purpose of this research is to extract the Omega-3 polyunsaturated fatty acid by using enzymatic hydrolysis method. Enzymatic extraction technique by using Alcalase was used to extract the Catfish oil. Then, lipase was used to concentrate the omega-3 polyunsaturated fatty acid. Peroxide value and acid value were used to determine the Catfish oil quality. Optimization using DOE verify that the highest percentage of oil yields at 51°C, 0.91% enzyme concentration and 155 minute incubation time. Finally, there are two types of omega-3 polyunsaturated fatty acid can be found in Catfish oil which is EPA and ALA.

Improved antioxidant activities of brown seaweed Ecklonia radiata extracts prepared by microwave-assisted enzymatic extraction

Seaweeds are important sources of bioactive compounds with potential use in functional foods and nutraceutical products. This study aims to investigate the extraction efficiency of phlorotannins and antioxidant compounds of a South Australian brown seaweed Ecklonia radiata by enzymatic and microwave-assisted enzymatic extraction in order to evaluate their uses as potential functional food ingredients. A selected group of carbohydrases (Viscozyme, Celluclast, and Ultraflo) and proteases (Alcalase, Neutrase, and Flavourzyme) has been applied to improve the extraction efficiency, alone and intensified with microwave heating, using conventional acid-base and water extractions as controls. The antioxidant activities of the extracts were evaluated using both ferric reducing ability of plasma (FRAP) and oxygen radical absorbance capacity (ORAC) assays. Significantly higher yields in total phlorotannin content (TPC) and antioxidant activities of the extracts were achieved by enzymatic and microwave-assisted enzymatic extraction. Microwave-assisted Viscozyme extraction for 5 to 30 min was the most effective process with an extraction yield achieved of 52 %. The extract had a TPC of 4.4 g phloroglucinol equivalents (PGE).100 g−1 dry weight (DW) and antioxidant activities of 29.7 mmol FeSO4 equivalents.100 g−1 DW and 740.1 μmol Trolox equivalents (TE).g−1 DW. In contrast, the conventional acidic extraction for 24 hours resulted in a TPC of 3.4 g PGE.100 g−1 DW and antioxidant activities of 21.1 mmol FeSO4 equivalents.100 g−1 DW and 512.4 μmol TE.g−1 DW. Extracts of brown seaweed E. radiata have potential for use in value-added products for nutritional purposes, using the microwave-assisted enzymatic extraction techniques.

Ultrasonic-assisted enzymatic extraction of phenolics from broccoli (Brassica oleracea L. var. italica) inflorescences and evaluation of antioxidant activity invitro.

An efficient ultrasonic-assisted enzymatic extraction technique was applied to extracting phenolics from broccoli inflorescences without organic solvents. The synergistic model of enzymolysis and ultrasonication simultaneously was selected, and the enzyme combination was optimized by orthogonal test: cellulase 7.5 mg/g FW (fresh weight), pectinase 10 mg/g FW, and papain 1.0 mg/g FW. The operating parameters in ultrasonic-assisted enzymatic extraction were optimized with response surface methodology using Box-Behnken design. The optimal extraction conditions were as follows: ultrasonic power, 440 W; liquid to material ratio, 7.0:1 mL/g; pH value of 6.0 at 54.5 ℃ for 10 min. Under these conditions, the extraction yield of phenolics achieved 1.816 ± 0.0187 mg gallic acid equivalents/gram FW. The free radical scavenging activity of ultrasonic-assisted enzymatic extraction extracts was determined by 1,1-diphenyl-2-picrylhydrazyl·assay with EC50 values of 0.25, and total antioxidant activity was determined by ferric reducing antioxidant power assay with ferric reducing antioxidant power value of 0.998 mmol FeSO4/g compared with the referential ascorbic acid of 1.184 mmol FeSO4/g.

Molecular characteristics and anti-inflammatory activity of the fucoidan extracted from Ecklonia cava

Enzymatic extraction has been successfully used for extracting numerous biologically active compounds from a wide variety of seaweeds. In this study, we found that enzymatic extraction of the fucoidan from Ecklonia cava may be more advantageous than water extraction. Therefore, we studied the E. cava fucoidans extracted by the enzymatic extraction technique and used ion-exchange chromatography to determine their molecular characteristics and anti-inflammatory activities. The crude and fractionated fucoidans (F1, F2, and F3) consisted mostly of carbohydrates (47.1–57.1%), uronic acids (9.0–15.8%), and sulfates (16.5–39.1%), as well as varying levels of proteins (1.3–8.7%). The monosaccharide levels significantly differed, and the composition included fucose (53.1–77.9%) and galactose (10.1–32.8%), with a small amount of rhamnose (2.3–4.5%), xylose (4.0–8.2%), and glucose (0.8–2.2%). These fucoidans contained one or two subfractions with an average molecular weight (Mw) ranging from 18 to 359×103g/mol. These fucoidans significantly inhibited NO production in lipopolysaccharide (LPS)-induced Raw 264.7 macrophage cells by down-regulating the expression of iNOS, COX-2, and pro-inflammatory cytokines such as TNF-α, IL-6, and IL-1β. Thus, the present results suggest that E. cava fucoidan may be a potentially useful therapeutic approach for various inflammatory diseases.

Selenium Species in Selenium-Enriched and Drought-Exposed Potatoes

The aim of this work was to study selenium (Se) speciation in the potato ( Solanum tuberosum L.) cultivar Desiree, enriched in Se by foliar spraying with a water solution containing 10 mg of Se/L in the form of sodium selenate. Four combinations of treatments were used: well-watered plants with and without Se foliar spraying and drought-exposed plants with and without Se foliar spraying. Water-soluble Se compounds were extracted from potato tubers by water or enzymatic hydrolysis with the enzyme protease XIV, amylase, or a combination of protease XIV and amylase. Extraction was performed using incubation at a constant temperature and stirring (37 degrees C at 200 rpm) or by ultrasound-assisted extraction (300 W), using different extraction times. Separation of soluble Se species (SeCys2, SeMet, SeMeSeCys, selenite, and selenate) was achieved by ion-exchange chromatography, and detection was performed by inductively coupled plasma-mass spectrometry (ICP-MS). Results showed that the concentration of selenate extracted was independent of the enzymatic extraction technique (approximately 98 ng/g for drought-exposed and 308 ng/g for well-watered potato tubers), whereas the extraction yield of SeMet changed with the protocol used (10-36%). Selenate and SeMet were the main soluble Se species (representing 51-68% of total Se) in potato tubers, regardless of the growth conditions.

Radical scavenging capacity and cytoprotective effect of enzymatic digests of Ishige okamurae

In order to obtain natural antioxidants of good quality from a marine bioresource, we selected the brown marine alga, Ishige okamurae, which is an abundant and unutilized source of biomass and grows near Jeju Island, South Korea. An enzymatic extraction technique was used in order to determine the antioxidant effects of I. okamurae using commercially available food-grade digestive enzymes. In the evaluation of the radical scavenging capacity of the enzymatic extracts from I. okamurae using an electron spin resonance (ESR) spectrophotometer, all of the enzymatic extracts showed profound dose-dependent radical scavenging abilities. The majority of enzymatic extracts generated by proteases (except for Neutrase) had stronger hydrogen peroxide scavenging effects than the carbohydrase extract. In particular, the cytoprotective effects of the Kojizyme extract against H2O2 - induced DNA damage increased significantly with increasing extract concentrations in our comet assay tests. These results showed that I. okamurae may prove to be a valuable natural source of antioxidants.

Collagen polymorphism in the lung: An immunochemical study of pulmonary fibrosis

The localization of type I, III, IV, and AB 2 (V) collagens in normal and fibrotic human lungs was studied by indirect immunofluorescence techniques with affinity purified antibodies specific for these types of collagen. In the normal lung type I collagen appears to be localized to the interstitium of alveolar septa in irregular patterns. Type III appears to have a more prominent, irregular localization in septa and a perivascular localization. Types IV and V codistribute in linear patterns on alveolar and capillary basement membranes. Type V is also present in the interstitium. Dramatic changes in relative amounts and localization were noted in fibrotic lung samples by fluorescence microscopy. In fibrotic lung samples there is a marked increase in type I in thickened septae. Type III is markedly reduced and seen only in a perivascular localization. Type V is markedly increased in the interstitium and located in areas of smooth muscle cell proliferation. No apparent change is noted in amount or localization of type IV collagen. These findings correlate well with biochemical data relating to the amounts of various collagens isolated by enzymatic extraction techniques. These results are discussed with respect to staging and evolution of the fibrotic process.