Search Results (54)

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

Efficient hydrolysis of cellulose in agricultural waste (e.g., coconut fiber) is critical for biorefining processes such as second-generation bioethanol (2G ethanol) production. However, free cellulases suffer from low thermal stability and challenges in recovery. To address this, we developed cross-linked enzyme aggregates (CLEAs) combined with magnetic nanoparticles (magnetic CLEAs, m-CLEAs) to enhance enzyme stability and reusability. In this context, solutions of ethanol, acetone, and ammonium sulfate were used to prepare enzymatic aggregates, with subsequent use of glutaraldehyde and magnetic nanoparticles to obtain the biocatalysts. The addition of bovine serum albumin (BSA) protein was also tested to improve immobilization. Biocatalysts with ethanol and acetone performed better. Acetone (AC) and BSA yielded the highest enzymatic activities (287.27 ± 42.59 U/g for carboxymethyl cellulase (CMCase) with Celluclast; 425.37 ± 48.11 U/g for CMCase with Cellic CTec2). Magnetic nanoparticles were incorporated to expand the industrial applicability, producing m-CLEAs with excellent thermal stability and high catalytic activities. The m-CLEA–Celluclast–AC–BSA–GA 5% maintained 58% of its activity after 72 h at 70 °C. The m-CLEA–Celluclast-AC–BSA–GA 2.5% proved effective in hydrolyzing coconut fiber and isolated cellulose, producing up to 0.91 ± 0.01 g/L of glucose and 2.7 ± 0.15 g/L of glucose, respectively, after 72 h. Therefore, this approach supports sustainability by using coconut fiber, which is often discarded into the environment. Full article

Water-soluble cellulosic oligomers (WCOs) are increasingly recognized for their prebiotic benefits, but their efficient enzymatic production is hindered by the high crystallinity of cellulose, which limits enzyme accessibility. This study introduced an efficient and scalable strategy combining phosphoric acid pretreatment with enzymatic hydrolysis to produce high-purity WCOs. Microcrystalline cellulose treated with 85 wt% phosphoric acid at 10 °C exhibited significantly reduced crystallinity and crystallite size, improving its susceptibility to enzymatic degradation. Subsequent hydrolysis of the hydrated regenerated cellulose (HRC85-10) using Celluclast^® at pH 7.0 for 1 h resulted in a WCO selectivity of 93.5%, with cellobiose and cellotriose identified as major oligomeric products via electrospray ionization mass spectrometry. Maintaining cellulose in a hydrated form significantly improved both the yield and selectivity of WCOs. In vivo studies further confirmed the prebiotic potential, with a significant increase in fecal Lactobacillus spp. and Bifidobacterium spp. (p < 0.05) following WCO supplementation. These findings demonstrated a practical and effective approach for producing functional WCOs for use in dietary and gut health applications. 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

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

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

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

Mandarin peel, a main by-product from the processing of citrus juice, has been highlighted for its various bioactivities and functional ingredients. Our previous study proved the inhibitory effects of Celluclast extract from mandarin peel (MPCE) on lipid accumulation and differentiation in 3T3-L1 adipocytes. Therefore, the current study aimed to evaluate the anti-obesity effect of MPCE in high-fat diet (HFD)-induced obese mice. The high-performance liquid chromatography (HPLC) analysis exhibited that narirutin and hesperidin are the main active components of MPCE. Our current results showed that MPCE supplementation decreased adiposity by reducing body and organ weights in HFD-induced obese mice. MPCE also reduced triglyceride (TG), alanine transaminase (ALT), aspartate transaminase (AST), and leptin contents in the serum of HFD-fed mice. Moreover, MPCE significantly inhibited hepatic lipid accumulation by regulating the expression levels of proteins associated with lipid metabolism, including sterol regulatory element-binding protein (SREBP1c), fatty acid synthase (FAS), and acetyl-CoA carboxylase (ACC). Furthermore, MPCE administration significantly inhibited both adipogenesis and lipogenesis, with modulation of energy metabolism by activating 5′ adenosine monophosphate-activated protein kinase (AMPK) and lipolytic enzymes such as hormone-sensitive lipase (HSL) in the white adipose tissue (WAT). Altogether, our findings indicate that MPCE improves HFD-induced obesity and can be used as a curative agent in pharmaceuticals and nutraceuticals to alleviate obesity and related disorders. Full article

A biocompatible, heterogeneous, fucose-rich, sulfated polysaccharide (fucoidan) is biosynthesized in brown seaweed. In this study, fucoidan was isolated from Padina arborescens (PAC) using celluclast-assisted extraction, purified, and evaluated for its anti-inflammatory potential in lipopolysaccharide (LPS)-induced RAW 264.7 cells. Structural analyses were performed using Fourier transform infrared (FTIR) and scanning electron microscopy. Among the purified fucoidans, fucoidan fraction 5 (F5) exhibited strong inhibitory activity against LPS-induced nitric oxide (NO) production and pro-inflammatory cytokine generation through the regulation of iNOS/COX-2, MAPK, and NF-κB signaling in LPS-induced RAW 264.7 cells. Determination of the structural characteristics indicated that purified F5 exhibited characteristics similar to those of commercial fucoidan. In addition, further analyses suggested that F5 inhibits LPS-induced toxicity, cell death, and NO generation in zebrafish models. Taken together, these findings imply that P. arborescens fucoidans have exceptional anti-inflammatory action, both in vitro and in vivo, and that they may have prospective uses in the functional food sector. Full article

Ecklonia cava, a brown seaweed native to the East Asian coast, is known for its unique composition, including polysaccharides, polyphenols, and phlorotannins. Fucoidan is a sulfated polysaccharide widely used as a functional ingredient in foods. This study obtained crude polysaccharides (ECC_CPS) from E. cava celluclast enzymatic hydrolysate using ethanol precipitation. ECC_CPS increased cell viability during the proliferation of Hanwoo muscle satellite cells (HMSCs). The effect of ECC_CPS on the expression of proliferation-related markers was confirmed as MYF5 and MYOD expression significantly increased, whereas PAX7 expression was maintained. The evaluation of cell migration activity has a major impact on cell proliferation and differentiation, and the cell migration index significantly increased with ECC_CPS treatment (p < 0.01). This was related to the HGF/MET pathway and FAK pathway. Treatment with ECC_CPS promoted differentiation at the cell differentiation stage, thereby increasing the expression of differentiation markers, such as MYH2, MYH7, and MYOG (p < 0.001 or p < 0.01). Therefore, our findings imply that crude polysaccharide obtained from E. cava can be an additive ingredient that enhances the proliferation and differentiation of muscle satellite cells used in the manufacture of cultured meat products. Full article

The current study investigated the feasibility of developing and adopting a few state-of-the-art fermentation techniques to maximize the efficiency of the lignocellulosic waste bioconversion. There have been various efforts towards utilizing the fermentable sugars released from the specific parts of lignocellulose, i.e., cellulose and hemicellulose. However, complete utilization of carbon sources derived from lignocellulosic biomass remains challenging owing to the generated glucose in the presence of β-glucosidase, which is known as glucose-induced carbon catabolite repression (CCR). To overcome this obstacle, a novel simultaneous saccharification and fermentation (SSF) of lactic acid was designed by using Celluclast 1.5L as a hydrolytic enzyme to optimize the generation and utilization of pentose and hexose. Under the optimal enzyme loading and pH condition, 53.1 g/L optically pure L-lactic acid with a maximum volumetric productivity of 3.65 g/L/h was achieved during the SSF from the brewer’s spent grain without any nutrient supplementation. This study demonstrated the potential of lactic acid production from the designed lignocellulosic substrate. Full article

Cellulases are a class of enzymes of great industrial interest that present several strategic applications. However, the high cost of enzyme production, coupled with the instabilities and complexities of proteins required for hydrolytic processes, still limits their use in several protocols. Therefore, enzyme immobilization may be an essential tool to overcome these issues. The present work aimed to evaluate the immobilization of cellulolytic enzymes of the commercial enzyme cocktail Celluclast^® 1.5 L in comparison to the cellulolytic enzyme cocktail produced from the wild strain Trichoderma harzianum I14-12 in Accurel^® MP1000. Among the variables studied were temperature at 40 °C, ionic strength of 50 mM, and 72 h of immobilization, with 15 m·L ⁻¹ of proteins generated biocatalysts with high immobilization efficiencies (87% for ACC-Celluclast biocatalyst and 95% for ACC-ThI1412 biocatalyst), high retention of activity, and specific activities in the support for CMCase (DNS method), FPase (filter paper method) and β-glucosidase (p-nitrophenyl-β-D-glucopyranoside method). Presenting a lower protein concentration (0.32 m·L⁻¹) than the commercial Celluclast^® 1.5 L preparation (45 m·L⁻¹), the ACC-ThI1412-derived immobilized biocatalyst showed thermal stability at temperatures higher than 60 °C, maintaining more than 90% of the residual activities of FPase, CMCase, and β-glucosidase. In contrast, the commercial-free enzyme presented a maximum catalytic activity at only 40 °C. Moreover, the difference in molecular weight between the component enzymes of the extract was responsible for different hydrophobic and lodging interactions of proteins on the support, generating a robust and competitive biocatalyst. Full article

Chitosan is a non-cytotoxic polysaccharide that, upon hydrolysis, releases oligomers of different sizes that may have antioxidant, antimicrobial activity and the inhibition of cancer cell growth, among other applications. It is, therefore, a hydrolysis process with great biotechnological relevance. Thus, this study aims to use a crude enzyme concentrate (C_EC) produced by a filamentous fungus to obtain oligomers with different molecular weights. The microorganism was cultivated in a liquid medium (modified Czapeck—with carboxymethylcellulose as enzyme inducer). The enzymes present in the C_EC were identified by LC-MS/MS, with an emphasis on cellobiohydrolase (E.C 3.2.1.91). The fungus of the Aspergillus genus was identified by amplifying the ITS1-5.8S-ITS2 rDNA region and metaproteomic analysis, where the excreted enzymes were identified with sequence coverage greater than 84% to A. nidulans. Chitosan hydrolysis assays compared the C_EC with the commercial enzyme (Celluclast 1.5 L^®). The ability to reduce the initial molecular mass of chitosan by 47.80, 75.24, and 93.26% after 2.0, 5.0, and 24 h of reaction, respectively, was observed. FTIR analyses revealed lower absorbance of chitosan oligomers’ spectral signals, and their crystallinity was reduced after 3 h of hydrolysis. Based on these results, we can conclude that the crude enzyme concentrate showed a significant technological potential for obtaining chitosan oligomers of different sizes. Full article

The food industry’s increasing demand for new functional ingredients that meet both organoleptic and healthy requirements has driven the exploration of new sources of functional ingredients in agro-industrial by-products. The aim of this work was to valorize grape pomace (Vitis vinifera L. garnacha) as a source of pectins using food-grade extracting agents. Obtained pectins were evaluated for monomeric composition, methyl esterification, molecular weight, water retention, oil-holding capacity, and antioxidant properties. The relatively soft extraction conditions used permitted obtaining low methoxyl pectin (10–42%) enriched in homogalacturonan (38–45%) or rhamnogalacturonan (33–41%) with different branching degrees, molecular weight, and fewer impurities than those found in the scarce previous literature. The relationship between structure and functionality was studied. Among the different pectins obtained, the sample derived from the extraction with sodium citrate could resume the best characteristics, such as pectin purity and higher water retention and oil holding capacity. These results underscore the relevance of grape pomace as a viable alternative source of pectin. Full article