Enzymatic Extraction Method Research Articles

Optimised Extraction and Preliminary Characterisation of Mannoproteins from Non-Saccharomyces Wine Yeasts.

The exogenous application of yeast-derived mannoproteins presents many opportunities for the improvement of wine technological and oenological properties. Their isolation from the cell wall of Saccharomyces cerevisiae has been well studied. However, investigations into the efficiency of extraction methods from non-Saccharomyces yeasts are necessary to explore the heterogeneity in structure and composition that varies between yeast species, which may influence wine properties such as clarity and mouthfeel. In this study, nine yeast strains were screened for cell wall mannoprotein content using fluorescence microscopy techniques. Four species were subsequently exposed to a combination of mechanical and enzymatic extraction methods to optimize mannoprotein yield. Yeast cells subjected to 4 min of ultrasound treatment applied at 80% of the maximum possible amplitude with a 50% duty cycle, followed by an enzymatic treatment of 4000 U lyticase per g dry cells weight, showed the highest mannoprotein-rich yield from all species. Furthermore, preliminary evaluation of the obtained extracts revealed differences in carbohydrate/protein ratios between species and with increased enzyme incubation time. The results obtained in this study form an important step towards further characterization of extraction treatment impact and yeast species effect on the isolated mannoproteins, and their subsequent influence on wine properties.

Physicochemical, morphological, functional, and pasting properties of potato starch as a function of extraction methods

The potato starch extracted by physical, chemical, and enzymatic methods were analyzed for physicochemical, morphological, functional, and pasting properties. In the physical extraction methods, T1 (water at 30 °C) was found better in starch yield, water absorption capacity (WAC), and whiteness. In the chemical extraction methods, starch from T5 (NaOH-0.25%) had higher WAC, and lower ash content. The starch from T7 (SDS-ME-2%) had higher amylose, starch purity, and lower amylopectin, moisture, fat content. The starch from T11 (NaOCl-5.25%) had higher starch whiteness value. In the enzymatic extraction methods, starch from T13 (cellulase-0.15%) had higher starch yield. Further, the physical (T1), chemical (T5, T7 and T11) and enzymatic (T13) methods were used in combination and had shown the better physicochemical and morphological characteristics of starch. The starch from combined method (T14) had significantly higher yield (12.4%), WAC (259%), swelling power (40.3%), solubility (23.6%), purity (87.3%), and whiteness (95.2%). The starch from combined method also had significantly lower phosphorus (81.7%), protein (0.30%), fat (0.29%), ash (0.26%), crude fiber (0.15%) content, and syneresis (18.8%). The starch from combined method had resulted in the better starch granule structure, pasting properties than most of the other treatments. So, the physical, chemical, and enzymatic treatments in combination can be used for extraction of high-quality starch from potato.

The Effect of the Aqueous Enzymatic Extraction Method towards Momordica charantia Seed Oil and Its Lignocellulosic Biomass

The development of a sustainable product using natural resources will gain much attention nowadays. This work aimed to study the effect of aqueous enzymatic extraction (AEE) using a different ratio of enzyme cocktails HEL1 and X7 towards the production of Momordica charantia L. (M. charantia) seed oil and to identify the composition of the seeds’ lignocellulosic biomass (extracted-free materials, EFM). The M. charantia seed oil contained omega-3 fatty acids, such as stearidonic acid, that possess hypotriglyceridemic properties. The oil obtained was derivatized to fatty acid methyl esters (FAMEs) before biochemical quantification using gas chromatography (GC). The EFM was subjected to Soxhlet extraction and further analyzed to identify lignin, acid-soluble sugars, and hemicellulose composition. The results showed that the highest seed oil percentage, (6.26±0.53 %) was extracted from M2 sample using a combination of 5 % HEL1 to 1.25 % X7 enzyme cocktails, as compared to the extraction without enzyme (3.32±0.38 %). The percentage of stearidonic acid in the seed oil increased from 13.55 % (without enzyme) to 19.43 % (M2), proving that there were some change in terms of fatty acid composition in the seed oil.

Utilization of Response Surface Methodology in Optimization of Polysaccharides Extraction from Vietnamese Red Ganoderma lucidum by Ultrasound-Assisted Enzymatic Method and Examination of Bioactivities of the Extract.

Red Ganoderma lucidum (G. lucidum) is a popular medicinal herb commonly used in Vietnamese traditional remedies due to its potential value for health. In this study, polysaccharides were extracted from G. lucidum using ultrasound-assisted enzymatic extraction method. The response surface methodology and Box–Behnken design were employed to investigate the effects of pH, extraction temperature, extraction time, and ultrasonic power on the content of polysaccharides. Based on ultraviolet-visible spectroscopy analysis, the highest content of polysaccharides in the extract was 32.08 mg/g under optimum experimental parameters including enzyme concentration of 3%, pH of 5.5, extraction temperature of 45°C, extraction time of 30 min, and ultrasonic power of 480 W. The Fourier-transform infrared spectroscopy was also used to identify the functional groups in the extracts. The molecular weights of polysaccharides were determined by gel permeation chromatography. The obtained extract was then evaluated for anticancer activities by using (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, showing the anticancer activities with the half-maximal inhibitory concentration value of more than 512 μg/mL. This result suggested that UAEE could be considered as an appropriate and effective extraction method for bioactive crude polysaccharides from G. lucidum.

Optimization of Flavonoids Extraction from Vietnamese Male Papaya (Carica papaya, L.) Flowers by Ultrasound‐Asissted Method and Testing Bioactivities of the Extract

AbstractPapaya (Carica papaya Linn.) is popularly cultivated in most tropical countries such as Malaysia, Vietnam, the West Indies, and throughout Africa. In the present work, an ultrasound‐assisted extraction (UAE) method has been developed for flavonoid extraction from Vietnamese male Carica papaya (C. papaya) flowers. Various influential operating parameters such as extraction time, ultrasonic power, solvent–to–material ratio, and extraction temperature on the flavonoid content were examined to optimize the extraction process. The result showed that under optimum extraction conditions including extraction time of 33 min, ultrasonic power of 240 W, solvent–to–material ratio of 27 : 1 mL/g, and extraction temperature of 46 , the flavonoid content obtained was of 10.648±0.228 mg QE/g, which was found to be higher than that obtained by hot water extraction, enzyme‐assisted extraction, and ultrasound‐assisted enzymatic extraction methods. Therefore, UAE could be considered as the most efficient extraction method for flavonoids from Vietnamese male C. papaya flowers. In addition, the extract obtained under optimum conditions was tested bioactivities to prove the medicinal uses of flavonoids extracted from Vietnamese male C. papaya flowers with the half‐maximal inhibitory concentration values for antioxidant effect and anticancer effect against Hep‐G2 cell lines at 179.69 0.03 and 56.63 1.25 μg/mL, respectively.

Study the Effect of Chemical and Enzymatic Extraction Methods on the Kenaf Fibers Properties

ABSTRACT In this paper, two extraction methods for kenaf fiber, The chemical process and the enzymatic one, were performed. These processes were used to extract fibers from the kenaf stem and the chemical, mechanical and morphologic properties of the extracted fibers were investigated. We notice that the fiber diameter decrease when increasing the concentration of the enzyme and the time treatment (101,35 µm for Enz1 and 91,37 µm for Enz4) the same result is obtained for the chemical extraction when increasing the duration, temperature or NaOH concentration. Also the fiber crystallinity index is improved due to the elimination of the noncellulosic materials from the fiber. The results obtained show that we have an increase in this index for all treated fiber however it is more important for the chemical extraction (crude fiber 57.39%, chemical-treated fiber 71.93% and 66.70% for the enzyme-treated fiber). This increase explains also the increase of fiber tenacity and the decrease of strain and we found through the tensile test that kenaf fiber has a pure elastic behavior for all treated fiber. So, the properties of extracted fibers compared to the crude fibers prove that the enzymatic extraction preserves more the fiber structure than the chemical extraction.

Aqueous enzymatic extraction of rice bran oil

An aqueous enzymatic extraction method was developed to extract rice bran oil. Different concentrations of individual enzymes, combinations of enzymes and incubation time were studied. The effect of hexane and aqueous enzymatic extraction on the oil yield was compared. The combination of cellulase (270 U), amylase (216 U) and protease (135 U) for four hours of incubation yielded significantly more oil than the other combinations. Under the optimal reaction conditions, a free oil recovery of 76.48% was obtained. The findings suggest that the critical selection of AEE parameters is key to high oil recovery.

Chemical composition, structural and functional properties of soluble dietary fiber obtained from coffee peel using different extraction methods

This study aimed to evaluate the effects of chemical, enzymatic, chemical–enzymatic, ultrasound-assisted enzymatic, and shear emulsifying-assisted enzymatic extraction methods on the physicochemical, structural, and functional properties of soluble dietary fiber from coffee peel. We found that the highest extraction yield of soluble dietary fiber was obtained using the shear emulsifying-assisted enzymatic method, and that similar protein contents were obtained from coffee peel using the enzymatic and shear emulsifying-assisted enzymatic methods. Compared with the other extraction methods, shear emulsifying-assisted enzymatic processing resulted in a higher water-holding capacity (7.05 g/g) and oil-holding capacity (3.61 g/g), but a lower emulsifying capacity (58.50%) when compared with that of chemical, enzymatic, and chemical-enzymatic processings. The soluble dietary fiber obtained from coffee peel using shear emulsifying-assisted enzymatic extraction was also characterized by the highest glucose absorption activity (228.06 mg/g), and enzymatic processing resulted in the highest nitrite ion absorption capacity (10.09 mg/g, pH = 2). These results indicated that shear emulsifying-assisted enzymatic extraction method was the most appropriate extraction method for coffee peel soluble dietary fiber.

Ultrasound-Assisted Enzymatic Extraction of Adenosine from Vietnamese Cordyceps militaris and Bioactivity Analysis of the Extract

Vietnamese Cordyceps militaris (C. militaris) has long been recognized as one of the most valuable traditional Chinese medicines. In this study, adenosine was extracted from Vietnamese C. militaris by ultrasound-assisted enzymatic extraction method (UAEE) using water as a solvent. Then, the effects of five single factors on adenosine content including pH, enzyme-to-material ratio, ultrasonic power, ultrasonic time, and ultrasonic temperature were determined. After that, three factors consisting of ultrasonic power, ultrasonic time, and ultrasonic temperature were chosen based on their effects on adenosine content. The simultaneous influence of these factors on the adenosine content was investigated by response surface method using central composite design. The adenosine content was evaluated by high-performance liquid chromatography method. Under the optimal conditions, the extract was evaluated for antioxidant and anticancer bioactivities. In addition, different extraction methods including aqueous extraction (AE), ultrasound-assisted extraction (UAE), and enzyme-assisted extraction (EAE) methods were carried out to compare with UAEE. As a result, it can be concluded that UAEE is a promising method for adenosine extraction and further studies regarding isolation and purification need to be conducted.

Ultrasound-Assisted Enzymatic Extraction Method for Multi-element Analysis of Rice

An ultrasound-assisted enzymatic extraction followed by a diluted acid washing was introduced into the multi-element analysis of rice samples. The recoveries of the elements including As, Cd, Mn, and Zn were evaluated by inductively coupled plasma mass spectrometry. The experimental parameters including enzyme type, enzyme dosage, ultrasound power, and duration of ultrasonication were investigated. The optimized condition was pancreatin with the dosage of 6.75 mg enzyme to 30-mg rice and ultrasonication for 5 min with the power of 800 W. The whole extraction procedure can be accomplished in 20 min. Three certified reference rice samples (GBW (E) 100348, GBW10043, and GBW10044) were used for method verification. The recoveries of As, Cd, Mn, and Zn ranged from 82.2 to 97.3%. The relative standard deviations of the four elements were all lower than 10%. Finally, the present method was successfully applied in the multi-element analysis of real samples.

Combination of ultrasound-assisted aqueous enzymatic extraction and cooking pretreatment for date seed oil recovery

Date seed oil is a good source of natural antioxidants due to the presence of phenols, tocopherols, and sterols. The present research is the first study on the extraction of date seed oil by combining the ultrasound-assisted aqueous enzymatic extraction method and the cooking pretreatment. In this study, the effects of two independent variables namely the sample to solvent ratio (at six levels of 1: 2, 1: 3, 1: 4, 1: 6, 1: 8 and 1:10) and the cooking time (at four levels of 0, 20, 40 and 60) were investigated on dependent variables namely the extraction yield, total phenolic compound, peroxide value, and acid value of date seed oil. The results indicated that optimum conditions for extraction of date seed oil were sample to solvent ratio of 1: 2 and the cooking time of 40 min. Furthermore, the Scanning Electron Microscopy images of date seed powder after selective treatments indicated the destruction and fragmentation of residual pulp structures by increasing the cooking time. In general, results of the present study indicated that it was important to select the appropriate sample to solvent ratio and implement the pretreatment of cooking to extract oil with high yield and quality. Combination of the ultrasound-assisted aqueous enzymatic extraction method and the cooking pretreatment can be proposed as a new extraction method in the oil industry. This new extraction method can improve the oil extraction efficiency and the quality of extracted oil.

Standardization and Stabilization of Millet Milk by Enzyme and Its Physicochemical Evaluation

Millets are nutritionally rich and occupy an important place in the diet of people in many regions of the world. Although millets are nutritionally superior to cereals, their utilization as a food is mostly confined to the traditional consumers. So, the present study was undertaken to standardize millet milk from barnyard millet, little millet, kodo millet and finger millet by enzymatic extraction method. Aqueous extract of millet milk was treated with α amylase and pasteurized at 75ºC for 15 minutes. The pasteurized millet milk was evaluated for physical and nutritional parameters. Results showed that the physical properties of developed millet milk have met the requirement of plant-based milk in terms of viscosity (2.32±0.02 to 2.82±0.03). Protein content of millet milk varied from 1.38±.0.03 to 1.12±.0.02 g. Total polyphenols (205.72±0.13 mg/100 ml) and total antioxidant activity (81.64±1.77%) were high for finger millet milk and total flavonoid content was high for barnyard millet milk (96.25±1.88 mg/100 ml). Enzymatic treatment significantly reduced the anti-nutritional factor (phytic acid, tannin and trypsin inhibitor activity) content in millet milk. The enzymatically developed product had high In vitro protein (69.28±0.28 to 85.57±1.39%) and starch digestibility (69.75±0.56 to 63.36±0.12 mg maltose/g). From the results, it was concluded that the current approach provides a convenient way for the production of nutritionally sound millet milk at the household and industrial level.

Simultaneous effect of vacuum and ultrasound assisted enzymatic extraction on the recovery of plant protein and bioactive compounds from sesame bran

The latest studies on the extraction of bio-components from plant materials have elaborated on novel approaches to increase yield. This study evaluated the effect of vacuum operation parameters on the recovery of protein and antioxidant compounds from sesame bran. In this context, vacuum-ultrasound assisted extraction (VU) and vacuum-ultrasound assisted enzymatic extraction (VUE) methods were carried out, and the vacuum parameters were investigated using central composite design by response surface methodology (RSM). The independent parameters were found effective on protein yield, total phenolic content and antioxidant capacity values. Optimum conditions for VU were 21 min vacuum time, 72 min restoration time and 539 mmHg vacuum pressure; and for VUE were 1.82 AU/100 g enzyme concentration, 8 min vacuum time, 68 min restoration time and 238 mmHg vacuum pressure. The protein yields increased by 31.0 and 41.6 % with VU and VUE, respectively compared to standard alkaline extraction. SEM images revealed deteriorative effects of applied extraction methods. SDS-PAGE analysis was conducted to examine detailed protein fractionation and alcalase assisted extraction revealed small protein sub-units (<18 kDa) which implies the presence of potential bioactive peptides. Additionally, LC/Q-TOF/MS analysis was performed and organic compounds in sesame bran extracts were identified.

Profiling Analysis of Fatty Acids and Collagens Obtained from Sea Cucumbers

Investigations of alternative resources for production of functional foods and ingredients containing valuable compounds with biological activities are getting more and more attention. Sea cucumbers are aquatic functional foods with various medical and pharmaceutical effects, such as antioxidant, antibacterial, antifungal, antiviral, anti-inflammatory and neuroprotective activities. This study aimed to conduct profiling analysis of fatty acids and collagens extracted from four different sea cucumbers harvested from Papua New Guinea by using Gas Chromatrography-Mass Spectrometry (GC-MS) and Fourier Transform Infrared Spectrometer (FT-IR). Three different extraction methods in combination with various solvents were used to find the best combination for extracting fatty acids. Enzymatic and chemical extraction methods were applied for collagen extraction. The highest fat recovery in this study was 85.32% of theoretical yield with high proportions of unsaturated fatty acids up to 74.54%, and enriched with omega-3 fatty acid. FT-IR chromatogram of sea cucumber protein extracts showed the characteristic of collagen enriched with glycine and proline. The nutritional analysis of sea cucumber extracts demonstrated the potential use as functional foods and ingredients with high benefits to human health.

Advances in combined enzymatic extraction of ferulic acid from wheat bran

Wheat bran could be utilised as feedstock for innovative and sustainable biorefinery processes. Here, an enzymatic hydrolysis process for ferulic acid (FA) extraction was optimised step by step for total wheat bran (Tritello) and then also applied to the outer bran layer (Bran 1). Proteins, reducing sugars, total phenols and FA were quantified. The highest FA yields (0.82–1.05 g/kg bran) were obtained either by rehydrating the bran by autoclaving (Tritello) or by steam explosion (Bran 1) using a bran/water ratio of 1:20, followed by enzymatic pre-treatment with Alcalase and Termamyl, to remove protein and sugars, and a final enzymatic hydrolysis with Pentopan and feruloyl esterase to solubilise phenol. FA was recovered from the final digestate via solid phase extraction. A 40-fold scale-up was also performed and the release of compounds along all the process steps and at increasing incubation times was monitored. Results showed that FA was initially present at a minimum level while it was specifically released during the enzymatic treatment. In the final optimized process, the FA extraction yield was higher than that obtained with NaOH control hydrolysis while, in comparison with other FA enzymatic extraction methods, fewer process steps were required and no buffers, strong acid/alkali nor toxic compounds were used. Furthermore, the proposed process may be easily scaled-up, confirming the feasibility of wheat bran valorisation by biorefinery processes to obtain valuable compounds having several areas of potential industrial exploitation.

Extraction and characterization of collagens from yak rumen smooth muscle.

This study reports an effective method using enzymatic methods to extract collagen from yak rumen smooth muscle. The enzymatic extraction methods were optimized by response surface methodology. Additionally, the properties of the extracted collagen were analyzed by Fourier transform infrared (FT-IR) spectroscopy and mass spectrometry (MS). The results showed that the optimal conditions were as follows: the pepsin addition was 0.95%, the enzymatic hydrolysis time was 21hr, and the solid-to-solvent ratio was 1:11. Under these conditions, the collagen extraction rate could reach 3.62/100g. The results of FT-IR revealed that the amide A, amide B, amide I, amide II, and amide III bands of the collagen appeared at 3,293.18, 3,068.18, 1654.94, 1,540.58, and 1,236.58cm-1 , respectively. The MS identified seven types of collagen, which were type I, type III, type IV, type V, type VI, type VIII, and type XII. The results demonstrated that the enzymatic method can extract collagen from yak rumen smooth muscle with a considerably high yield and can preserve the intact structure of the collagen.

Physicochemical and Biological Properties of Gelatin Extracted from Marine Snail Rapana venosa.

In this study, we aimed to obtain gelatin from the marine snail Rapana venosa using acidic and enzymatic extraction methods and to characterize these natural products for cosmetic and pharmaceutical applications. Marine gelatins presented protein values and hydroxyproline content similar to those of commercial mammalian gelatin, but with higher melting temperatures. Their electrophoretic profile and Fourier transform infrared (FTIR) spectra revealed protein and absorption bands situated in the amide region, specific for gelatin molecule. Scanning electron microscopy (SEM) analysis showed significant differences in the structure of the lyophilized samples, depending on the type of gelatin. In vitro studies performed on human keratinocytes showed no cytotoxic effect of acid-extracted gelatin at all tested concentrations and moderate cytotoxicity of enzymatic extracted gelatin at concentrations higher than 0.5 mg/mL. Also, both marine gelatins favored keratinocyte cell adhesion. No irritant potential was recorded as the level of IL-1α and IL-6 proinflammatory cytokines released by HaCaT cells cultivated in the presence of marine gelatins was significantly reduced. Together, these data suggest that marine snails are an alternative source of gelatins with potential use in pharmaceutical and skincare products.

Effect of isolation methods on the crystalline, pasting, thermal properties and antioxidant activity of starch from queen sago (Cycas circinalis) seed

Crystalline, pasting and thermal properties and antioxidant activity of starch isolated from queen sago seed by using different isolation methods such as water steeping, alkali steeping and enzymatic extraction methods were evaluated. The properties studied include, chemical composition, morphology, swelling, solubility, color, X-ray diffraction (XRD), texture, pasting (RVA), total phenolic content, antioxidant activity and thermal (DSC) properties. Among the isolation methods, enzymatic extraction method is efficient as compared to alkali and water steeping method. Enzymatically extracted sago starch (ES) showed highest yield, L* value, swelling, solubility and lower non starch components. ES formed harder gel as compared to alkali steeped sago starch (AS) and water steeped sago starch (WS). SEM showed presence of more agglomerated granular structure in AS. ES showed highest peak, trough and break down viscosity. Setback and final viscosity was highest for AS. Isolation methods affected on the total phenolic content and antioxidant activity of queen sago seed starch. Highest gelatinization enthalpy of AS was correlated with higher relative crystallinity and presence of more agglomerated structure. The queen sago seed starch isolated by enzymatic extraction method has better properties and industrial relevance.

Antioxidant properties of a sulfated polysaccharide isolated from an enzymatic digest of Sargassum thunbergii.

The protective effect of sulfated polysaccharides (PS-4) isolated from Sargassum thunbergii against oxidative stress in cells and zebrafish was investigated. To begin, a variety of enzymatic extraction methods were used, including five carbohydrases and four proteases. Among the enzymatic extracts, the S. thunbergii Celluclast-assisted extract (STC) demonstrated the strongest scavenging activities against hydrogen peroxide (H2O2). Five polysaccharide fraction (PS-1 to 5) were isolated from STC and purified using ion exchange column chromatography. The antioxidant activity of these fractions was then assessed. PS-4 demonstrated the most promising free radical scavenging and antioxidant activity. PS-4 had a higher content of sulfated polysaccharide than STC. Thus, increasing the proportion of sulfated polysaccharides may be attributed to protection against H2O2-mediated cell damage and oxidative stress in cell lines and in zebrafish. These results suggest that PS-4 could be a potent antioxidant agent in functional foods and cosmetics.

Characterization of the Chemical Composition of Chinese Moringa oleifera Seed Oil

AbstractThis work focused on physicochemical property assaying, fatty acid composition, triacylglycerol (TAG) profiles, and unsaponifiable matter composition of the Chinese Moringa oleifera seed oil. The results indicated that there was no significant difference in approximate nutritional components between M. oleifera seeds from China and India, while variations in the mineral element contents are significant. Both the Soxhlet extraction method and the aqueous enzymatic extraction method were adopted to extract oil from Chinese M. oleifera seeds. Oil yield obtained using the Soxhlet extraction method was higher than that obtained using the aqueous enzymatic extraction method. While both the iodine value and unsaponifiable matter content of the aqueous enzymatic extracted oil were a little higher than that of the Soxhlet extracted oil. Both oils possess a very low acid value and peroxide value, suggesting their good quality as edible oil. Fatty acid composition results indicated that this oil was especially high in oleic acid. Characterization of the TAG composition was achieved by a two‐dimensional high‐performance liquid chromatography (HPLC) coupling of nonaqueous reverse‐phase and silver ion HPLC with the atmospheric pressure chemical ionization mass spectrometry method. A total of 22 TAG including 16 regioisomers were determined. Composition results of unsaponifiable matters revealed that this oil possesses a number of phytosterols, in which β‐sitosterol and stigmasterol are most predominant.