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In the context of biorefinery, researchers have been looking for lignocellulosic biomasses and ideal treatments to produce economically viable biofuels. In this scenario, the bamboo culm appears as a plant matrix of great potential, given the high cellulose content of low crystallinity. Thus, the objective and differential of this work was to determine the best conditions for enzymatic hydrolysis of cellulose extracted from bamboo culm and to evaluate its potential application in the production of bioethanol through Separate Hydrolysis and Fermentation (SHF) and Saccharification and Simultaneous Fermentation (SSF) by Saccharomyces cerevisiae modified via CRISPR/Cas9. The average cellulose extraction yield was 41.87 % with an extraction efficiency of 86.76 %. In general, as the hydrolysis time increased, an increase in glucose production was observed in almost all assays, with higher hydrolysis efficiency values at 72 h. The results ranged from 2.09 to 19.8 g/L of glucose obtained with efficiency values of 10.47 to 99 %. The best conditions were found in test 5 (temperature of 36 °C and pH 5.0, with only 10 FPU/g of substrate Cellic Ctec2 Novozymes ® cocktail). It is observed that for all hydrolysis times the independent variables pH and temperature were significant under the hydrolysis efficiency, showing a negative effect, indicating that higher values of the same promote lower values of the response variable. For bioethanol production, a maximum concentration of 7.84 g/L was observed for the SSH process after 4 h of fermentation, while for the SSF process it was 12.6 g/L after 24 h of fermentation, indicating the large potential of the simultaneous process together with the application of bamboo culm biomass for high production of biofuel.

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BACKGROUND: Protein-derived peptide fractions can play a key role in the physiological and metabolic regulation and modulation of the body, which suggests that they could be used as functional ingredients to improve health and to reduce the risk of disease. This work aimed to evaluate the in vitro antithrombotic and anticariogenic bioactivity of hydrolysates and protein fractions obtained from cowpea (Vigna unguiculata) by biocatalysis. RESULTS: Cowpea protein concentrate was hydrolyzed by sequential action with two enzyme systems, Pepsin-Pancreatin or Alcalase-Flavourzyme. There was extensive enzymatic hydrolysis, with degrees of hydrolysis of 34.94% and 81.43% for Pepsin-Pancreatin and Alcalase-Flavourzyme, respectively. The degree of hydrolysis for the control treatments, without the addition of the enzymes Pepsin-Pancreatin and Alcalase-Flavourzyme was 1.1% and 1.2%, respectively. The hydrolysates were subjected to fractionation by ultrafiltration, with five cut-off points according to molecular weight (<1, 1-3, 3-5, 5-10 and >10 kDa). The Alcalase-Flavourzyme hydrolysate led to 100% inhibition of platelet aggregation, while the Pepsin-Pancreatin hydrolysate showed 77.41% inhibition, but this was approximately 100% in the ultrafiltered fractions. The highest anticariogenic activity was obtained with the Pepsin-Pancreatin system, with 61.55% and 56.07% for calcium and phosphorus demineralization, respectively. CONCLUSION: Hydrolysates and their peptide fractions from Vigna unguiculata exhibited inhibition of platelet aggregation and protection of tooth enamel and have the potential for use in the development of functional products with beneficial health effects. © 2024 Society of Chemical Industry.

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The hydrothermal pretreatment process stands out as a pivotal step in breaking down the hemicellulosic fraction of lignocellulosic biomasses, such as sugarcane bagasse and eucalyptus sawdust. This pretreatment step is crucial for preparing these materials for subsequent processes, particularly in food applications. This technique aims to disintegrate plant wall components like cellulose, hemicellulose, and lignin, and facilitating access in later phases such as enzymatic hydrolysis, and ultimately making fermentable sugars available. In this study, sugarcane bagasse and eucalyptus sawdust biomass underwent hydrothermal pretreatment at specific conditions, yielding two key components: dry biomass and hemicellulose liquor. The primary focus was to assess the impact of hydrothermal pretreatment followed by enzymatic hydrolysis, using the Celic Ctec III enzyme cocktail, to obtain fermentable sugars. These sugars were then transformed into membranes via strain Gluconacetobacter xylinus bacterial biosynthesis. Notably, the addition of a nitrogen source significantly boosted production to 14.76 g/ in hydrolyzed sugarcane bagasse, underscoring its vital role in bacterial metabolism. Conversely, in hydrolyzed eucalyptus, nitrogen source inclusion unexpectedly decreased yield, highlighting the intricate interactions in fermentation media and the pivotal influence of nitrogen supplementation. Characterization of membranes obtained in synthetic and hydrolyzed media through techniques such as FEG-SEM, FTIR, and TGA, followed by mass balance assessment, gauged their viability on an industrial scale. This comprehensive study aimed not only to understand the effects of pretreatment and enzymatic hydrolysis but to also evaluate the applicability and sustainability of the process on a large scale, providing crucial insights into its feasibility and efficiency in practical food-related scenarios, utilizing nanocellulose bacterial (BNC) as a key component.

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The ß-glucosidase gene from Aspergillus nidulans FGSC A4 was cloned and overexpressed in the A. nidulans A773. The resulting purified ß-glucosidase, named AnGH3, is a monomeric enzyme with a molecular weight of approximately 80 kDa, as confirmed by SDS-PAGE. Circular dichroism further validated its unique canonical barrel fold (ß/α), a feature also observed in the 3D homology model of AnGH3. The most striking aspect of this recombinant enzyme is its robustness, as it retained 100% activity after 24 h of incubation at 45 and 50 ºC and pH 6.0. Even at 55 °C, it maintained 72% of its enzymatic activity after 6 h of incubation at the same pH. The kinetic parameters Vmax, KM, and Kcat/KM for ρ-nitrophenyl-ß-D-glucopyranoside (ρNPG) and cellobiose were also determined. Using ρNPG, the enzyme demonstrated a Vmax of 212 U mg - 1, KM of 0.0607 mmol L - 1, and Kcat/KM of 4521 mmol L - 1 s - 1 when incubated at pH 6.0 and 65 °C. The KM, Vmax, and Kcat/KM using cellobiose were 2.7 mmol L - 1, 57 U mg - 1, and 27 mmol -1 s - 1, respectively. AnGH3 activity was significantly enhanced by xylose and ethanol at concentrations up to 1.5 mol L - 1 and 25%, respectively. Even in challenging conditions, at 65 °C and pH 6.0, the enzyme maintained its activity, retaining 100% and 70% of its initial activity in the presence of 200 mmol L - 1 furfural and 5-hydroxymethylfurfural (HMF), respectively. The potential of this enzyme was further demonstrated by its application in the saccharification of the forage grass Panicum maximum, where it led to a 48% increase in glucose release after 24 h. These unique characteristics, including high catalytic performance, good thermal stability in hydrolysis temperature, and tolerance to elevated concentrations of ethanol, D-xylose, furfural, and HMF, position this recombinant enzyme as a promising tool in the hydrolysis of lignocellulosic biomass as part of an efficient multi-enzyme cocktail, thereby opening new avenues in the field of biotechnology and enzymology.

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Essential oils show several biological properties, such as antimicrobial activity, but have limitations regarding their availability and stability. To maximize their antimicrobial effect and protection against environmental conditions, Pickering-type emulsions were used to vehiculate oregano essential oil (OEO) using cellulose nanofibers (CNF) as emulsion stabilizer. Enzymatic hydrolysis was used to produce CNF from a food industry waste (cassava peel), obtaining an environmentally sustainable emulsion stabilizer. It was evaluated how the different properties of the nanofibers affected the stability of the emulsions. Furthermore, the composition of the dispersed phase was varied (different ratios of OEO and sunflower oil-SO) in view of the target application in biodegradable active coatings. Even at very low concentration (0.01 % w/w), CNF was able to form kinetically stable emulsions with small droplet sizes using oil mixtures (OEO + SO). The stabilization mechanism was not purely Pickering, as there was a reduction in interfacial tension. Excellent antimicrobial activity was observed against bacteria and the fungus Alternaria alternata, demonstrating the ability to apply these emulsions in active systems such as coatings and films. An improvement in the stability of emulsions was observed when using a mixture of oils, which is extremely advantageous considering costs and stability to heat treatments, since the desired antimicrobial activity is maintained for the final application.

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Cellulose nanocrystals (CNCs) are crystalline domains isolated from cellulosic fibers. They have been utilized in a wide range of applications, such as reinforcing fillers, antibacterial agents and manufacturing of biosensors. Whitin this context, the aim of this work was to obtain and analyze CNCs extracted from bacterial nanocellulose (BNC) using two distinct methods combined with milling pre-treatment: an acidic hydrolysis using 64 % sulfuric acid and an enzymatic hydrolysis using a commercial cellulase enzyme mixture. The CNCs obtained from the enzymatic route (e-CNCs) were observed to be spherical nanoparticles with diameter of 56 ± 11 nm. In contrast, the CNCs from the acid hydrolysis (a-CNCs) appeared as needle-shaped nanoparticles with a high aspect ratio with lengths/widths of 158 ± 64 nm/11 ± 2 nm. The surface zeta potential (ZP) of the a-CNCs was -30,8 mV, whereas the e-CNCs has a potential of +2.70 ± 3.32 mV, indicating that a-CNCs consisted of negatively charged particles with higher stability in solution. Although the acidic route resulted in nanocrystals with a slightly higher crystallinity index compared to the enzymatic route, e-CNCs was found to be more thermally stable than BNC and a-CNCs. Here, we also confirmed the safety of a-CNCs and e-CNCs using L929 cell line. Lastly, this article describes two different CNCs synthesis approaches that leads to the formation of nanoparticles with different dimensions, morphology and unique physicochemical properties. To the best of our knowledge, this is the first study to yield spherical nanoparticles as a result of BNC enzymatic treatment.

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Cellulose nanofibers (CNF) have been widely studied for their biodegradability and for their unique advantages as a stabilizer in Pickering-type emulsions. However, it is challenging to produce cellulose nanofibers from agroindustry waste with good techno-functional properties, without the use of harsh process conditions. Green alternatives (eco-friendly) have been studied to obtain nanofibers, such as enzymatic hydrolysis and/or application of mechanical processes. In this work, we used acid hydrolysis (as a control and example of an efficient method), enzymatic hydrolysis and a mechanical process (ultrasound) to obtain cellulose nanofibers. We also evaluated the effect of the presence of ethyl groups in the cellulosic matrix (ethylcellulose) on the stabilizing mechanism of emulsions. All cellulose nanofibers were able to produce Pickering emulsions at concentrations of 0.01-0.05% (w/w), although showing differences in emulsion stability and digestibility. Morphology of the different cellulose nanofibers affected the viscosity of the aqueous suspensions used as continuous phase. Emulsions with nanofibers obtained from cassava peel (without the presence of ethyl groups) were stabilized only by the Pickering-type mechanism, while ethylcellulose nanofibers also showed surface activity that contributed to the stability of the emulsion. Furthermore, these latter emulsions showed greater release of free fatty acids in in vitro digestion compared to emulsions stabilized by cellulose nanofibers. Despite these differences, in vitro digestion showed the potential of applying cellulose-stabilized emulsions to control the rate of lipid digestion, due to the low amount of free fatty acids released (<20%).

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In light of the growing demand for alternative protein sources, laboratory-grown meat has been proposed as a potential solution to the challenges posed by conventional meat production. Cultured meat does not require animal slaughter and uses sustainable production methods, contributing to animal welfare, human health, and environmental sustainability. However, some challenges still need to be addressed in cultured meat production, such as the use of fetal bovine serum for medium supplementation. This ingredient has limited availability, increases production costs, and raises ethical concerns. This review explores the potential of non-animal protein hydrolysates derived from agro-industrial wastes as substitutes for critical components of fetal bovine serum in cultured meat production. Despite the lack of standardization of hydrolysate composition, the potential benefits of this alternative protein source may outweigh its disadvantages. Future research holds promise for increasing the accessibility of cultured meat.

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Inulin is a fructose-based polysaccharide that can be found in several plant species, from grass and onions to chicory roots; thus, it has the potential to be an excellent renewable source of fructose for several industrial applications. Among them, inulin hydrolysis can be coupled to a fermentation operation to produce polyhydroxybutyrate (PHB) using Cupriavidus necator H16. This work reports the PHB production process involving chicory root inulin hydrolysis using inulinase Novozym 960 followed by a C. necator fermentation. It was found that the maximum saccharification (95% wt.) was reached at 269 U/ginulin after 90 min. The hydrolysates obtained were then inoculated with C. necator, leading to a biomass concentration of 4 g/L with 30% (w/w) polymer accumulation. Although PHB production was low, during the first hours, the cell growth and polymer accumulation detected did not coincide with a fructose concentration decrease, suggesting a simultaneous saccharification and fermentation process, potentially alleviating the product inhibition inherent to the inulinase-fructose system. The characterization of the obtained PHB showed a polymer with more homogeneous values of Mw, and better thermal stability than PHB produced using pure fructose as a fermentation substrate. The results obtained demonstrate a viable alternative carbon substrate for PHB production, opening the possibility for inulin-rich renewable feedstock valorization.

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Β-Carotene is a red-orange pigment that serves as a precursor to important pharmaceutical molecules like vitamin A and retinol, making it highly significant in the industrial sector. Consequently, there is an ongoing quest for more sustainable production methods. In this study, glucose and xylose, two primary sugars derived from sugarcane bagasse (SCB), were utilized as substrates for ß-carotene production by Rhodotorula glutinis CCT-2186. To achieve this, SCB underwent pretreatment using NaOH, involved different concentrations of total solids (TS) (10%, 15%, and 20%) to remove lignin. Each sample was enzymatically hydrolyzed using two substrate loadings (5% and 10%). The pretreated SCB with 10%, 15%, and 20% TS exhibited glucose hydrolysis yields (%wt) of 93.10%, 91.88%, and 90.77%, respectively. The resulting hydrolysate was employed for ß-carotene production under batch fermentation. After 72 h of fermentation, the SCB hydrolysate yielded a ß-carotene concentration of 118.56 ± 3.01 mg/L. These findings showcase the robustness of R. glutinis as a biocatalyst for converting SCB into ß-carotene.

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Whey is a by-product that represents a cheap source of protein with a high nutritional value, often used to improve food quality. When used as a raw material to produce hypoallergenic infant formulas (HIF), a processing step able to decrease the allergenic potential is required to guarantee their safe use for this purpose. In the present paper, thermal treatments, high hydrostatic pressure (HHP), and enzymatic hydrolysis (EH) were assessed to decrease the antigenicity of whey protein solutions (WPC). For monitoring purposes, a competitive ELISA method, able to detect the major and most allergenic whey protein ß-lactoglobulin (BLG), was developed as a first step to evaluate the efficiency of the processes. Results showed that EH together with HHP was the most effective combination to reduce WPC antigenicity. The evaluation method proved useful to monitor the processes and to be employed in the quality control of the final product, to guarantee the efficiency, and in protein antigenicity reduction.

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The objective of this study was to investigate the dual modification of red rice starch using pulsed electric field (PEF) and α-amylase, focusing on morpho-structural, thermal, and viscoamylographic properties. Native starch (Control) underwent various treatments: PEF at 30 kV cm-1 (PEF30), α-amylase at 9.0 U mg-1 (AA0), and a combination of both (PEF30 + α and α + PEF30). The PEF30 + α treatment exhibited the highest degree of digestion (10.66 %) and resulted in morphological changes in the starch granules, which became elongated and curved, with an increased average diameter of 50.49 µm compared to the control. The starch was classified as type A, with a maximum reduction in crystallinity of up to 21.17 % for PEF30. The deconvolution of FT-IR bands indicated an increase in the double helix degree (DDH) for PEF30 and AA0, while the degree of order (DO) was reduced for PEF30, AA0, and PEF30 + α. DSC analysis revealed significant modifications in gelatinization temperatures, particularly for PEF30, and these changes were supported by a reduction in gelatinization enthalpy (ΔH) of up to 28.05 % for AA0. These findings indicate that both individual and combined treatments promote a decrease in starch gelatinization and facilitate the process, requiring less energy. Differences were observed between the formulations subjected to single and alternating dual treatments, highlighting the influence of the order of PEF application on the structural characteristics of starch, especially when applied before the enzymatic treatment (PEF + α). Regarding the viscoamylographic parameters, it was observed that AA0 presented higher values than the control, indicating that α-amylase enhances the firmness of the paste. The double modification with PEF + α was more effective in reducing syneresis and starch retrogradation, leading to improvements in paste properties. This study provided significant insights into the modification of red rice starch using an efficient and environmentally friendly approach.

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The amylolytic action of α-amylase and amyloglucosidase has been directly implemented in native cassava starches for the formation of cassava microporous granules with unsatisfactory results, however, its incidence in hydrothermally treated granules has never been evaluated. The effect of hydrothermal processes and simultaneous enzymatic hydrolysis on the physicochemical, morphological and structural properties of native cassava starch was evaluated. Native cassava starch presented a rigid, smooth surface, and was exempt from porosities, whereas hydrothermal processes altered the semicrystalline order and increasing the size and number of pores and increasing the size (4.11 ± 0.09 nm) and volume of pores (0.82 ± 0.00 cm3/g × 10-3). The hydrothermal action followed by enzymatic processes with α-amylase and amyloglucosidase, augmented the processes of internal degradation (endo-erosion) and pore widening (exo-erosion), improving the hydrophilic properties compared to the hydrothermal treatment. Likewise, the hydrothermally process followed by enzymatic hydrolysis for 24 h (HPS + EMS-24) increased the degradation of the amorphous lamellae, consistent with a significant decrease in amylose content. This same dual treatment increased the pore size at 17.68 ± 0.13 nm relative to the native counterpart; therefore, they are considered an effective method in the development of modified cassava starches with porous surfaces.

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A medida que como sociedad vamos dando más importancia a lograr una economía circular, se hace importante encontrar fuentes renovables aptas para la producción de biocombustibles y bioquímicos. En los últimos años, diversas fuentes de biomasa lignocelulósica han sido estudiadas para estos propósitos. Dentro de estas fuentes de biomasa se encuentra el cáñamo (Cannabis sativa L.), siendo parte de una industria que ha crecido a pasos agigantados en las últimas décadas, en Colombia, desde su legalización. Específicamente, la industria del cannabis medicinal es responsable de generar una enorme cantidad de residuos en forma de los tallos de la planta, considerados un subproducto de bajo valor. En esta revisión se compila la información de diferentes estudios sobre el aprovechamiento de la fracción de polisacáridos de biomasa cáñamo, mediante transformaciones químicas y bioquímicas, para la obtención de productos de valor agregado. Se encontró que la mayoría de estudios están enfocados en la obtención de bioetanol o biogás; se encontraron también reportes de otras moléculas como ácido succínico, ácido láctico, furfural, polihidroxialcanoatos y bisaboleno. La viabilidad a nivel industrial de todos estos procesos permanece siendo una incógnita, pues los pasos de pretratamiento, hidrólisis y de conversión final utilizados suelen ser costosos. Es necesario que los estudios que realicen en el futuro se enfoquen en optimizar las condiciones de estos procesos y hacerlos verdes y así asegurar que puedan ser escalados.

As we as a society, give more importance to achieving a circular economy, it becomes important to find renewable sources suitable for the production of biofuels and biochemicals. In the last years, several different sources of lignocellulosic biomass have been studied for these purposes. One of these biomass sources is hemp (Cannabis sativa L), being part of an industry that has grown through giant steps in the last decades, in Colombia, since its legalization. Specifically, the industry of medicinal hemp is responsible for the generation of huge amounts of residues in the form of the plant stalks, considered a low value subproduct. This review compiles the information of several studies about the exploitation of the polysaccharide portion of hemp biomass through chemical and biochemical transformations, obtaining value-added products. It was found that most of these studies focus on the production of bioetanol or biogas; reports of other molecules such as succinic acid, furfural, polyhydroxyalkanoates and bisabolene were also found. Industrial viability of these processes remains a question, since pretreatment, hydrolysis and final conversion steps are usually expensive. It necessary that future studies focus on optimizing conditions of these processes as well as making them green, ensuring that they can be scaled.

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Concern over environmental impacts has spurred many efforts to replace fossil fuels with biofuels such as ethanol. However, for this to be possible, it is necessary to invest in other production technologies, such as second generation (2G) ethanol, in order to raise the levels of this product and meet the growing demand. Currently, this type of production is not yet economically feasible, due to the high costs of the enzyme cocktails used in saccharification stage of lignocellulosic biomass. In order to optimize these cocktails, the search for enzymes with superior activities has been the goal of several research groups. For this end, we have characterized the new ß-glycosidase AfBgl1.3 from A. fumigatus after expression and purification in Pichia pastoris X-33. Structural analysis by circular dichroism revealed that increasing temperature destructured the enzyme; the apparent Tm value was 48.5 °C. The percentages of α-helix (36.3%) and ß-sheet (12.4%) secondary structures at 25 °C were predicted. Biochemical characterization suggested that the optimal conditions for AfBgl1.3 were pH 6.0 and temperature of 40 °C. At 30 and 40 °C, the enzyme was stable and retained about 90% and 50% of its activity, respectively, after pre-incubation for 24 h. In addition, the enzyme was highly stable at pH between 5 and 8, retaining over 65% of its activity after pre-incubation for 48 h. AfBgl1.3 co-stimulation with 50-250 mM glucose enhanced its specific activity by 1.4-fold and revealed its high tolerance to glucose (IC50 = 2042 mM). The enzyme was active toward the substrates salicin (495.0 ± 49.0 U mg-1), pNPG (340.5 ± 18.6 U mg-1), cellobiose (89.3 ± 5.1 U mg-1), and lactose (45.1 ± 0.5 U mg-1), so it had broad specificity. The Vmax values were 656.0 ± 17.5, 706.5 ± 23.8, and 132.6 ± 7.1 U mg-1 toward p-nitrophenyl-ß-D-glucopyranoside (pNPG), D-(-)-salicin, and cellobiose, respectively. AfBgl1.3 displayed transglycosylation activity, forming cellotriose from cellobiose. The addition of AfBgl1.3 as a supplement at 0.9 FPU/g of cocktail Celluclast® 1.5L increased carboxymethyl cellulose (CMC) conversion to reducing sugars (g L-1) by about 26% after 12 h. Moreover, AfBgl1.3 acted synergistically with other Aspergillus fumigatus cellulases already characterized by our research group-CMC and sugarcane delignified bagasse were degraded, releasing more reducing sugars compared to the control. These results are important in the search for new cellulases and in the optimization of enzyme cocktails for saccharification.

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The high hydrostatic pressure (HHP) process has been studied for several applications in food technology and has been commercially implemented in several countries, mainly for non-thermal pasteurization and shelf-life extension of food products. HHP processing has been demonstrated to accelerate proteolytic hydrolysis at a specific combination of pressure and pressure-holding time for a given protein source and enzyme. The enzymatic hydrolysis of proteins is a well-known alternative to producing biologically active peptides, with antioxidant and antihypertensive capacity, from different food protein sources. However, some of these protein sources contain allergenic epitopes which are often not degraded by traditional hydrolysis. Moreover, the peptide profile and related biological activity of a hydrolysate depend on the protein source, the enzymes used, the parameters of the proteolysis process (pH, temperature, time of hydrolysis), and the use of other technologies such as HHP. The present review aims to provide an update on the use of HHP for improving enzymatic hydrolysis, with a particular focus on studies which evaluated hydrolysate antihypertensive and antioxidant capacity, as well as residual allergenicity. Overall, HHP has been shown to improve the biological properties of hydrolysates. While protein allergenicity can be reduced with traditional hydrolysis, HHP can further reduce the allergenicity. Compared with traditional hydrolysis methods, HHP-assisted protein hydrolysis offers a greater opportunity to add value to protein-rich products through conversion into high-end hydrolysate products with enhanced nutritional and functional properties.

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In the context of a biorefinery, lignocellulosic materials represent an important source of raw material for the bioconversion of cellulose, hemicellulose, and lignin into value-added products, such as xylose for fermentation, oligosaccharides, and bioplastics for packaging. Among the most abundant lignocellulosic materials in Brazil, sugarcane bagasse biomass stands out, as it is rich in cellulose and hemicellulose. In this context, through an experimental design, this study developed a robust enzyme cocktail containing xylanases and accessory enzymes to complete the hydrolysis of xylan from sugarcane bagasse, obtaining a low xylose yield and concentration (9% and 1.8 g/L, respectively, observed in experiment number 16 from the complete hydrolysis of a xylan assay), a fermentable sugar that is important in the production of second-generation ethanol, and a high xylooligosaccharides (XOS) yield and concentration (93.1% and 19.6 g/L, respectively, obtained from a xylooligosaccharides production assay); in general, xylan has prebiotic activities that favor an improvement in intestinal functions, with immunological and antimicrobial actions and other benefits to human health. In addition to completely hydrolyzing the sugarcane bagasse xylan, this enzymatic cocktail has great potential to be applied in other sources of lignocellulosic biomass for the conversion of xylan into xylose and XOS due to its enzymes content, involving both main chain and pendant groups hydrolysis of hemicelluloses.

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The INFOGEST protocol creation was a watershed for phenolic bioaccessibility studies. Because of this important initiative to standardize bioaccessibility studies, data comparisons between different laboratories are now expedited. It has been eight years since the INFOGEST protocol creation, and three from the latest update. However, the current status in terms of phenolic bioaccessibility and how far different laboratories are from reaching a consensus are still unrevealed. In this sense, this narrative review considered an evaluation of different studies that applied the INFOGEST protocol to investigate the bioaccessibility of phenolic compounds. The central objective was to compile the main findings and consensus and to identify possible gaps and future opportunities. This approach intends to further facilitate the use of this protocol by professionals in the field of food science and technology and related areas, generating a reflection on the actual level of standardization of the method. Despite the differences in phenolic compounds from diverse food matrices, and their peculiar behavior, some trends could be elucidated, in terms of phenolic release, stability, and/or transformation upon in vivo digestion. In contrast, there was no general consensus regarding sample preparation, how to report results and the form to calculate bioaccessibility, making it difficult to compare different studies. There is still a long road to effectively standardize the results obtained for phenolic bioaccessibility using the INFOGEST protocol, which is also an opportunity in terms of food analysis that can impact the food industry, especially for the development of nutraceuticals and functional foods.

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The high costs of bioplastics' production may hinder their commercialization. Development of new processes with high yields and in biorefineries can enhance diffusion of these materials. This work evaluated the production of polyhydroxybutyrate (PHB) from the combination of milled corn starchy fraction hydrolysate and crude glycerol as substrates by the strain Cupriavidus necator LPB 1421. After optimization steps, maximum accumulation of 62 % of PHB was obtained, which represents 11.64 g.L-1 and productivity of 0.162 g.Lh-1. In a stirred tank bioreactor system with 8 L of operational volume, 70 % of PHB accumulation was reported, representing 14.17 g.L-1 of the biopolymer with 0.197 g.Lh-1 productivity. PHB recovery was conducted using a chemical digestion method, reaching >99 % purity. Therefore, the potential application of milled corn as substrate for PHB production was confirmed. The developed bioplastic process could be coupled to a bioethanol producing unit creating the opportunity of a sustainable and economic biorefinery.

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Specific kinds of enzymes have been used as an eco-friendly pre-treatment for mechanical extraction of cellulose nanofibrils (CNFs) from vegetal pulps. Another well-established pre-treatment is the 2,2,6,6-tetramethylpiperidine 1-oxyl (TEMPO)-mediated oxidation, which has gained considerable attention. Pre-treatments assist in fiber swelling, facilitating mechanical fibrillation, and reducing energy consumption; however, some of these methods are extremely expensive. This work aimed to evaluate the influence of enzymatic pre-treatment with endoglucanase on the energy consumption during mechanical fibrillation of cellulose pulps. Bleached pulps from Eucalyptus sp. and Pinus sp. were pre-treated with endoglucanase enzyme compared to TEMPO-meditated oxidation. Average diameters of CNFs pre-treated with enzymes were close to that found for TEMPO-oxidized nanofibrils (TOCNFs). Results showed that enzymatic pre-treatment did not significantly modify the pulp chemical and morphological characteristics with efficient stabilization of the CNFs suspension at higher supernatant turbidity. Energy consumption of pulps treated with endoglucanase enzymes was lower than that shown by pulps treated with TEMPO, reaching up to 58% of energy savings. The enzyme studied in the pulp treatment showed high efficiency in reducing energy consumption during mechanical fibrillation and production of films with high mechanical quality, being an eco-friendly option for pulp treatment.

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