Search Results (44)

The aqueous enzymatic extraction (AEE) of oil from Thevetia peruviana seeds was studied. The oil extraction yields obtained by treatment with cellulase, Alcalase, and Viscozyme L were compared, finding that the treatment with Alcalase at pH 6 and 50 °C gave the highest yield (63.07%), so it was selected for the optimization of the oil extraction process through a central composite design and a response surface methodology. An oil extraction yield of 78.22% was achieved under the optimum conditions (2% of Alcalase, 4 h of incubation, 250 rpm, and a solid-to-liquid ratio of 1:4.5). The predominant fatty acids in the oil obtained by both AEE and standard solvent extraction (SE) were oleic, linoleic, stearic, and palmitic acids. However, AEE produced an oil presenting lower acidity and peroxide values and greater oxidative stability, indicating that this method produces an oil with better quality. Scanning electron microscopy images of Thevetia peruviana seeds showed a structural alteration of their cell wall caused by AEE, which allowed the release of oil from this seed in the aqueous medium. The results suggest that AEE could be an excellent alternative in terms of efficiency, oil quality, and environmental friendliness for the extraction of this oil. Full article

Walnut (Juglans regia L.) shells are valuable agro-industrial by-products rich in polyphenols. This study investigated traditional (maceration) and advanced extraction techniques—ultrasound-assisted extraction (UAE), enzyme-assisted extraction (EAE), pressurized liquid extraction (PLE), and combined ultrasound–enzyme extraction (US-EAE)—to recover bioactive compounds from walnut shells. Extraction efficiency, total phenolic content (TPC), antioxidant capacity (ABTS^•+, DPPH•), and polyphenol composition were evaluated. UPLC-ESI-MS/MS identified key polyphenols including ellagic acid, 4-hydroxybenzoic acid, vanillin, taxifolin, and quercitrin. The highest TPC (5625 mg GAE/100 g dw) was found in extracts subjected to US-EAE, in which ultrasound pretreatment (200 W, 10 min) was followed by enzymatic extraction using 0.06 mL/g Viscozyme^® L at pH 3.5 and 45 °C. Under the same extraction conditions, UAE alone yielded the second highest TPC (4129 mg GAE/100 g dw). The highest ABTS^•+ scavenging activity (14,478 mg TE/100 g dw) and enhanced DPPH• activity (45.38 mg TE/100 g dw) were also observed in US-EAE extracts. Chemometric techniques (PCA and HCA) revealed meaningful clustering and variation patterns among methods. These findings highlight the potential of walnut shells as a sustainable source of polyphenols and demonstrate the effectiveness of innovative extraction technologies in maximizing bioactive compound recovery for potential functional applications. Full article

Blackcurrant (Ribes nigrum) is valued for its health-promoting compounds, many of which remain in the pomace after juice extraction. Berry pomace can be considered a valuable source of dietary fiber. However, it is typically dominated by insoluble dietary fiber (IDF), and the soluble-to-insoluble fiber ratio is often nutritionally suboptimal. The aim of this study was to evaluate the influence of enzymatic hydrolysis on the composition and technological properties of blackcurrant pomace (BCP). Three commercial enzyme preparations—Viscozyme^® L, Celluclast^® 1.5 L, and Pectinex^® Ultra Tropical (Novozymes A/S, Denmark)—were used for enzymatic hydrolysis, which was conducted at 50 °C for 1 h. The enzymatic treatments altered BCP’s chemical composition and technological properties. Pectinex^® Ultra Tropical and Viscozyme^® L primarily hydrolyzed SDF, while Celluclast^® 1.5 L was more effective on IDF, resulting in increased SDF content and an improved SDF/IDF ratio. Enzymatic hydrolysis reduced the oil retention capacity and impaired stabilizing properties, but it increased both the water retention capacity and the solubility index. It was found that the creaming index of the pomace deteriorated with decreased IDF content. The findings indicate that the effects of enzymatic modification on BCP’s composition and technological properties can vary significantly, supporting its potential application in the development of novel food products. Full article

This research focused on producing water-soluble carbohydrates extracts from the leaves of the wild plant Cacalia firma using commercial enzymatic processes. Different enzymes and conditions were applied to the leaves to determine the optimal method for extracting carbohydrates. Enzymes used were Cellic CTec3 HS, Celluclast 1.5 L, Viscozyme L, Pectinex ultraSP-L, and Amylase AG. Pectinase, cellulase, and other enzymes are isolated from yeast, bacteria, or some higher plants and are commonly used to break down pectin, which is the cell wall or intercellular connective tissue in plant tissues, to soften fruit or vegetable tissues and to make sugars. They are commonly used to soften the tissues of fruits and vegetables, to produce sugars, or to increase the yield of juice in fruit processing. The resulting water-soluble carbohydrates demonstrated significant antioxidant capabilities in vitro, as evidenced by DPPH radical-scavenging and ABTS assays. Furthermore, the carbohydrates exhibited high levels of total polyphenol and flavonoid content. The extraction methodology was fine-tuned using response surface methodology alongside the Box–Behnken design, achieving a maximum carbohydrate yield of 129.7 mg/g, which was very close to the predicted value of 132.4 mg/g. The optimal conditions included an extraction temperature of 47.3 °C, a duration of 63 h, and a pH of 3.7 using Viscozyme L. This study offers a theoretical foundation for the development of natural carbohydrate antioxidants and lays the groundwork for large-scale production and utilization of C. firma leaf carbohydrates. These extracts, showing antioxidant activity, hold potential as functional ingredients in the food industry. Full article

Lingonberry pomace (LP) is a by-product rich in valuable bioactive compounds and can be used in the food industry after various treatments and property characterization. This study aimed to evaluate the impact of commercially available enzymes (Viscozyme^® L, Pectinex^® Ultra Tropical, and Celluclast^® 1.5 L) and supercritical carbon dioxide (SFE-CO₂) extraction technology on the chemical composition and technological properties of treated LP products. The Megazyme kit was used to determine the soluble dietary fiber (SDS) and insoluble dietary fiber (IDF) contents, while the changes in mono-, disaccharide, and oligosaccharides were analyzed by applying high-pressure liquid chromatography with a refractive index detector. The analyzed properties were as follows: the water swelling capacity (WSC), water retention capacity (WRC), water solubility index (WSI), oil retention capacity (ORC), bulk density (BD), and emulsion stability of modified LP. The tested LP contained 8.49 g/100 g of SDF and 65.36 g/100 g of IDF (in dry matter). The partial separation of lipophilic substances during SFE-CO₂ extraction did not significantly affect the enzymatic hydrolysis efficiency. The amount of oligosaccharides in the LP increased using enzymes with pectinolytic activity (Viscozyme^® L and Pectinex^® Ultra Tropical), while cellulolytic enzymes (Celluclast^® 1.5 L) increased the amount of SDF and improved the IDF/SDF ratio. Enzymatic hydrolysis increased the SI, WRC, and ORC of LP powder. Emulsions with LP hydrolyzed with Pectinex^® Ultra Tropical demonstrated the highest stability during storage. This study demonstrates that the modification of LP powders provides diverse technological properties, which could expand the application of such products for further food production. Full article

Enzymes play a vital role in various industrial sectors and are essential components of many products. Hybrid enzyme-polymeric capsules were developed using polysulfone-activated carbon capsules as scaffolds. The polysulfone-activated carbon capsules with an average diameter of 2.55 mm were fabricated by applying a phase inversion precipitation method. An increase in the amount of immobilized enzymes was observed with growth of activated carbon amount in polysulfone matrix. Enzyme immobilization was confirmed by the Bradford method, while Viscozyme^® L activity in carboxymethyl cellulose hydrolysis to glucose was measured by the Reducing Sugar DNS method. The recycling of the hybrid Viscozyme^® L-polysulfone/activated carbon capsules, and their reuse for subsequent cellulose hydrolysis was investigated and demonstrated repeatability of results. Full article

This research presents a novel approach to protein extraction from cauliflower (CL) and broccoli (BL) waste leaves by using enzymatic pretreatment, demonstrating its effectiveness on protein yield. Enzymatic pretreatment (pH 4.5, 10 h, t = 35 °C), was performed using commercial enzyme preparations, Viscozyme^® L and Vinozyme^®, in three different enzyme-to-substrate ratios E/S (0.2%, 2.5%, and 4.8%) of each enzyme. Leaf proteins (LPs) were obtained with alkaline extraction at pH 10–11 and their isoelectric precipitation at pH 4, following the control sample (extraction without enzymes). Protein yield (%), which was used as a parameter to monitor enzymatic efficiency, demonstrated a direct correlation with the enzyme-to-substrate (E/S) ratio. The highest protein yields were obtained at an enzyme concentration of 4.8% for both cellulolytic and pectolytic enzyme preparations, yielding 14.90 ± 0.12% for CL and 29.88 ± 0.86% for BL. The obtained proteins were characterized by FTIR spectroscopy and SDS-PAGE electrophoresis, and these methods confirmed the enzymatic efficiency of protein isolation. Isolated LPs showed high protein content for CLP 4.8% (77.27 ± 0.14%) and BPL 4.8% (84.66 ± 0.51%), and an increase in total amino acids, while the content of essential amino acids was over 40%. Protein solubility was assessed, revealing significant improvements (p < 0.05) in LPs derived from CL and BL at the highest E/S ratio of 4.8%, compared to the control sample C0%. Specifically, the solubility of CLP reached 29.4 mg/mL at pH 11, while BLP achieved 36.4 mg/mL at pH 10. As a result, these leaf proteins not only meet nutritional demands but also open innovative avenues of research in food science and biotechnology. Full article

This study aimed to evaluate the applicational possibilities of enzymatically modified apple pomace (AP) in conjunction with probiotics as value-added ingredients for the production of jelly candies. AP was enzymatically modified with Pectinex^® Ultra Tropical, Viscozyme^® L, and Celluclast^® 1.5 L (Novozyme A/S, Bagsværd, Denmark), and the soluble and insoluble dietary fibre content was determined using the Megazyme kit (Megazyme International Ireland Ltd., Wicklow, Ireland), reducing sugar content using the 3,5-dinitrosalicylic acid assay. The technological properties of the modified AP, such as its swelling capacity, water-retention capacity, oil-retention capacity, bulk density, and static and thermal emulsion stability, were evaluated. Enzymatically modified AP hydrolysed with Celluclast^® 1.5 L was used for the production of jelly candies supplemented with Bifidobacterium animalis DSM 20105. The survival of probiotics in the jelly candies during in vitro digestion, the viability of probiotics during candy storage, and candy quality characteristics were analysed. Enzymatically modified AP had different carbohydrate compositions and technological properties, depending on the enzyme preparation used. Although the viability of probiotics in the jelly candies decreased during storage, a significantly higher viability of B. animalis was determined in jelly candies supplemented with hydrolysed AP compared with control candies made without AP after digestion in the saline, gastric, and intestine phases. This study shows that Celluclast^® 1.5 L can be used for increasing the soluble dietary fibre in AP (18.4%), which can be further applied, in conjunction with B. animalis, for added-value jelly candy production. Full article

Background/Objectives: A variety of phytochemicals from different plants are collected by bees into bee pollen granules. This research focused on evaluating the effects of lactic acid fermentation and enzymatic hydrolysis on the antibacterial activity of bee pollen and its interaction with antibiotics. There is limited knowledge regarding the interactions between treated bee pollen extracts and antibiotics, and this study contributes to the field by providing new insights into the antibacterial activity of pollen subjected to eight distinct treatment methods. Methods: Bee pollen’s bacterial fermentation using a Lacticaseibacillus rhamnosus culture and spontaneous fermentation were performed. Bee pollen hydrolysis was performed using commercial enzymes, including enzyme mixtures as well as pure enzymes. The agar well diffusion assay was employed to assess the antibacterial activity against Staphylococcus aureus, Listeria monocytogenes, and Salmonella enterica serovars Enteritidis and Typhimurium, as well as their interaction with antibiotics (ceftazidime, ciprofloxacin, oxytetracycline dihydrate, and erythromycin). Results: This study showed an enhancement in bee pollen’s antibacterial activity after both fermentation and enzymatic hydrolysis. The increase varied with the pollen’s origin, treatment type, and culture used for antimicrobial tests. More than 77% of bee pollen extracts demonstrated a synergistic effect with antibiotics across all tested bacterial strains, while antagonistic interactions were comparatively rare. Conclusions: The applied treatment methods can improve the antibacterial properties of bee pollen. Bee pollen extracts, in combination with antibiotics, can enhance their effectiveness. These findings provide new insights into the potential use of bee pollen in combating bacterial infections. Full article

Dietary fiber (DF), and especially soluble dietary fiber (SDF), is a nutrient of particular interest today because of its anti-inflammatory role and its ability to reduce cardiovascular risk. Therefore, the enhancement of SDF in foods using different techniques has become a promising field of research. In order to prove the possibility of increasing this SDF content, the effects of different commercial enzymes (Pectinex^® Ultra SP-L, Viscozyme^® L and Celluclast^® 1.5 L) were tested on a variety of carob (CE), artichoke (ARE), apple (APE) and broccoli (BE) by-product extracts. Enzymatic treatment significantly affected SDF content in all by-products, showing the greatest increases for CE, ARE and APE using Celluclast^® 1.5 L, while Viscozyme^® L obtained the best results after application in BE. On the other hand, positive results were reported in the solubility, WHC and FAC of the by-products due to the enzymatic treatment, being increased in all extracts analyzed. Moreover, a general increase in antioxidant capacity (FRAP, ABTS and DPPH) was observed after enzymatic treatment. Finally, high yields were obtained after the application of the enzymatic processes, reaching values of 80–85% for each food by-product. These results evidenced a potential revalorization of carob, artichoke, apple and broccoli by-products after enzymatic treatment, improving its nutritional and physicochemical properties, revealing a possible application as a higher value-added ingredient. Full article

The objectives of the study were to examine the effect of an antibiotic solution applied in the Streptomyces griseus protease method (SGPM) and the effect of carbohydrases in SGPM on the effective crude protein (CP) degradation (ED) with reference to in sacco ED. For this purpose, the ruminal CP degradation of rapeseed meal, dried distillers’ grains with solubles, wheat grain, corn grain, corn silage, grass silage and partial crop field pea silage was determined in sacco using three rumen-fistulated dairy cows and in vitro using SGPM. The impact of the antibiotic solution on CP degradation by S. griseus protease was investigated by supplementing SGPM with Penicillin–Streptomycin solution to reduce microbial mass proliferation during incubation. The carbohydrase α-amylase or Viscozym^® L (cell wall-degrading enzyme mixture) was added to the SGPM at four different doses simultaneously as a co-incubation to improve feed protein accessibility. For most feedstuffs, ED was lower when the antibiotic solution was used in SGPM (p < 0.05). The use of an antibiotic solution in the SGPM is recommended to standardize the SGPM. The in sacco ED values were significantly underestimated by the SGPM and by the SGPM with co-incubated carbohydrase (p < 0.05). Co-incubation of S. griseus protease and carbohydrase was not successful in reducing the differences to the in sacco CP degradation. Full article

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. Full article

Nowadays, the circular economy trend drives researchers in the recovery of various bioactive compounds from agri-food by-products. Enzyme-assisted extraction (EAE) has been shown to be an innovative green technology for the effective extraction of various phytochemicals from agri-food section by-products; therefore, this study aimed to evaluate the application of EAE as green technology to obtain extracts from olive leaves (Olea europaea) for potential industrial production. The used enzymes were Celluclast, Pectinex XXL and Viscozyme L. EAE was conducted under various enzyme dose combinations and an incubation time of 120 min. Obtained extracts were characterized in terms of total polyphenols (TP) and total antioxidant activity (AA). Firstly, the enzyme synergistic effect in the enzymatic extraction of polyphenols was evaluated. TP optimal extraction conditions (468.19 mg GAE (gallic acid equivalent)/L of extract) were achieved after EAE using Pectinex and Viscozyme enzymes (50–50 v/v) and for AA (69.85 AA%). According to the above results, a second experiment investigated the effect of incubation time (min.) and enzyme dose (mL) on the optimal extraction conditions of olive leaves. The final results after optimization were 75% higher than the control sample for the TP content (605.55 mg GAE/L) and 8% higher for the AA (70.14 AA%). These results indicated that EAE is an excellent choice for the green extraction of polyphenols from the olive leaves. Full article

For protein evaluation of feedstuffs for ruminants, the Streptomyces griseus protease test provides a solely enzymatic method for estimating ruminal protein degradation. Since plant proteins are often structured in carbohydrate complexes, the use of carbohydrase during the test might improve its accuracy. It is advisable to co-incubate protease and carbohydrase, risking that the carbohydrase activity is reduced under the influence of the protease. The present study was conducted to investigate this impact by using α-amylase or the multi-enzyme complex Viscozym^® L as carbohydrase. The detection of active protease was determined fluorescence photometrically using internally quenched fluorogenic substrates (IQFS). Cellulose, pectin, and starch degradation were determined spectrophotometrically using 3,5-dinitro salicylic acid as a colorimetric agent. The Streptomyces griseus protease mixture proved to be active for the selected IQFS immediately after the start of measurements (p < 0.05). Starch hydrolysis catalyzed by α-amylase or Viscozym^® L, respectively, was decreased by co-incubation with protease mixture by maximal 3% or 37%, respectively, at 5 h incubation time (p > 0.05). Pectin and cellulose hydrolysis catalyzed by Viscozym^® L, respectively, was not significantly influenced by co-incubation with a protease mixture (p > 0.05). Although a decrease in carbohydrase activity during co-incubation with Streptomyces griseus protease occurred, it was only numerical and might be counteracted by an adapted carbohydrase activity. Full article

Mulberry is renowned for its medicinal properties and bioactive compounds, yet its high moisture content renders it highly perishable and challenging to transport over long distances. This inherent limitation to its shelf life poses sustainability challenges due to potential food waste and the increased carbon footprint associated with transportation. To address this issue sustainably, mulberry vinegar emerges as a biotechnological solution. Utilizing a fermented mixture of crushed mulberries, sugar, and mixed acid, transforms the highly perishable raw material into a more stable product. However, conventional methods of mulberry vinegar production often involve heat-intensive processing, which poses environmental concerns and energy inefficiencies. Recognizing the need for sustainable practices, this review delves into alternative non-thermal technologies (NTTs) that can revolutionize mulberry vinegar production. These technologies, such as ultrasonication, ultra-high-pressure homogenization, pulsed light treatments, enzyme-assisted pretreatment, and membrane filtration, offer eco-friendly alternatives by eliminating the need for excessive heat. NTTs enhance energy efficiency and sustainability in mulberry vinegar production by deactivating the microbes and extending the shelf life, thereby enhancing product stability and quality without using thermal methods. Ultrasonication, for example, plays a pivotal role in improving bioactive compound extraction, contributing to the overall quality enhancement of mulberry juice. Enzyme-assisted pretreatment, specifically with Pectinex Ultra SP-L and Viscozyme L, not only enhances juice quality, but also holds promise for sustainable vinegar production. Furthermore, ultra-high-pressure homogenization and pulsed light treatments positively influence mulberry processing, offering additional sustainable alternatives. Membrane filtration, especially ultrafiltration, not only enhances the phenolic content, but also contributes to stability in mulberry juice, showcasing potential benefits for vinegar production. In conclusion, exploring these NTTs represents a transformative shift from traditional heat treatment methods in mulberry food processing. By providing energy efficient, environmentally friendly, and high-quality alternatives, this review offers valuable insights into sustainable practices, particularly in mulberry vinegar production, thereby contributing to a more sustainable future for the mulberry food industry. Full article