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Simple Summary: Ruminal protein degradation of seven feedstuffs was estimated in sacco and in vitro by using a protease assay. In sacco protein degradation data were used as reference. The accuracy of the protease assay is reduced by two methodological aspects. During in vitro incubation, microbial activity induced by microbial colonization of the feedstuff may increase, requiring antibiotic solutions in protease assays. The protease alone cannot sufficiently hydrolyze protein–carbohydrate complexes. Therefore, a carbohydrase of fiber- or starch-hydrolyzing activity was added to the protease assay as a simultaneous incubation. The antibiotic solution reduced protein degradation by protease. The antibiotic solution is recommended to prevent microbial activity and improve standardization of the protease assay. Differences between in sacco and in vitro degradation data were not essentially reduced by additional carbohydrases. Unfavorable incubation conditions and the inhibitory effect of protease on the carbohydrase activity during simultaneous incubation may be responsible for the insufficient hydrolysis of protein–carbohydrate complexes by the carbohydrases. It does not seem promising to incubate protease and carbohydrase simultaneously. 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. [ABSTRACT FROM AUTHOR]

Agroindustrial processes have increasingly focused on sustainability and pressing coffee beans generates a waste known as the residual cake. This study evaluated the effects of aqueous enzymatic extraction with carbohydrases applied in coffee bean and its residual cake. Optimized conditions for enzymatic extraction were 75 min, pH 4.5 and 49°C (CB); 51 min, pH 5.0 and 46 °C(RC). The extracts obtained from aqueous enzymatic extraction, when compared to their respective control groups, showed statistically significant increases in the extraction of caffeine (0.32 ± 0.02 and 0.73 ± 0.10 g/100 g for CB RC, respectively) phenolic compounds (3.06 ± 0.30 and 3.85 ± 0.51 g/100 g, for CB and RC, respectively), oil content and improved antioxidant activity by the DPPH and ABTS methods (184.1 ± 2.5 for CB and 153.6 ± 8.4 µmolTrolox/g for RC). These results also showed the potential of the residual cake as a raw material for the production of extracts rich in bioactive compounds and the use of clean technology. [ABSTRACT FROM AUTHOR]

The present study investigated the effect of supplementing a multi-carbohydrase enzyme complex (MCE) in corn–soy-based diets of Pekin ducks. The treatments were as follows: positive control (PC, 2980 and 3120 kcal/kg for starter and grower, respectively); negative control (NC, −132 kcal/kg energy reduction to PC achieved by reduction of fat and wheat middlings as filler); NC + MCE at 75 ppm (E75); 100 ppm (E100); and 125 ppm (E125) randomly distributed in 10 replicate pens with 25 birds each. Performance was evaluated after 14 and 35 d. On day 36 of the trial, five ducks/pen were processed to evaluate carcass traits. During days 1–14, the PC had a lower (p < 0.01) feed intake compared to all other treatments. At 14 and 35 d of age the NC decreased (p < 0.001) the body weight (BW) of the ducks compared to the PC by −8.3% and −5.3%, respectively. The NC BW was lower (p < 0.001) compared to all MCE-supplemented treatments at 14 and 35 d. The BW of E75, E100, and E125 treatments was not different (p > 0.05) from the PC at both evaluation periods. Cumulatively (1–35 d), the NC resulted in a weight-adjusted FCR increase (p = 0.001) of 9.9% compared to the PC, and the FCR of E75, E100, and E125 were able to recover 72.3%, 66.4%, and 63.5%, respectively, compared to the PC. The carcass and breast weights were lower (p < 0.001) in the NC compared to all other treatments, and no differences (p > 0.05) were observed between the MCE-supplemented treatments and the PC. In conclusion, these results suggest that the MCE supplementation can maintain duck growth performance with no negative effects on carcass traits in energy-reduced corn–soybean meal-based diets. From the results of the trial, 75 ppm MCE delivered the best performance recovery and 125 ppm MCE supplementation had the best % breast yield. [ABSTRACT FROM AUTHOR]

The actuality of processing of substandard by external signs fruit and vegetables is caused by their higher volumes in the processing industry and retail chains, and additionally by their high biopotentials. The aim of research is to ground complex processing of such raw materials to obtain food supplements containing natural biologically active components, and assessment of their nutritional functionality. The experiments used apples, bananas, oranges, potatoes, carrots, tomatoes and cucumbers that did not meet the freshness criteria of GOST 57976-2017, but without any signs of microbial damage. Three types of experimental mixtures were made from this raw material: fruit, vegetable and fruit-vegetable. The key processing operation was the enzymatic hydrolysis of the mixtures with various carbohydrases (cellulase, ?-glucanase, amylase and glucoamylase). The hydrolyzed mixtures were fractionated, followed by dehydration of the fractions to obtain water-soluble and water-insoluble powder supplements. During the fermentation process, the accumulation of simple sugars, dry substances in water-soluble fractions, and the yield of finished products were studied. The highest concentrations of sugars are found in water-soluble fractions from fruit mixtures. In the resulting supplements from three types of mixtures, organoleptic characteristics, chemical composition, and the content of functional food ingredients – vitamin C, substances with P-vitamin activity, and dietary fiber have been studied. The qualitative composition of dietary fiber and other polysaccharides in a water-insoluble supplement has been established using IR-spectra. Taking into account the obtained data, calculations have been made to satisfy the daily physiological needs for established biologically active substances when consuming 100 g of supplements for ordinary people and athletes. It is recommended to use supplements as enriching agents for functional purposes in specialized antioxidant, immunomodulatory and prebiotic nutrition, including as part of dietary supplements for athletes.

Cocoa bean shells (CBSs) are a byproduct of the chocolate production process, representing the external layer of the cocoa bean. CBSs exhibit many interesting chemical and nutritional characteristics resulting in a very rich content of dietary fiber (DF) and antioxidant compounds such as phenolic acids and flavan-3-ols. The DF fraction of CBSs is notably rich in soluble dietary fibers (SDFs), which may be associated with fermentability and prebiotic properties. The objective of this study was the valorization of CBSs through enzymatic treatments, thereby increasing the solubility of DF and potentially augmenting fermentability. CBSs were treated both raw and defatted. Three sets of carbohydrases were used in order to impact the dietary fiber profile. Cellulase, xylanase, pectinase and their combinations were used to perform enzymatic treatments. The application of cellulase, xylanase and a combination of both enzymes proved effective in achieving a high SDF destructuring of the insoluble dietary fiber (IDF) fraction in both defatted and raw CBSs. Notably, the SDF/IDF ratio was significantly elevated in the enzymatically hydrolyzed samples (1.13–1.33) compared to the untreated CBSs (0.33). Furthermore, the various treatments did not affect the antioxidant activity or the content of the main bioactive compounds. These results provide a foundation for new opportunities in the biovalorization of CBSs through green techniques for a range of potential industrial applications in the food and nutraceutical sectors. [ABSTRACT FROM AUTHOR]

Red seaweed Gelidium is used for agar extraction, generating large quantities of by-products (red seaweed by-product - GBP). Its carbohydrate and protein-rich composition makes it a suitable substrate to produce value-added compounds through solid-state fermentation (SSF). For the first time, GBP was used in SSF with Aspergillus ibericus and Aspergillus niger for carbohydrases production and protein enrichment. SSF was performed with unsupplemented GBP, supplemented with Mandel salt solution, and mixed at equal dry mass proportion with plant feedstuffs and with the seaweed Ulva rigida. A. niger CECT 2915 was the best strain under SSF obtaining the maximum xylanase activity (498 ± 49Ug-1), in GBP mixed with sunflower cake and the maximum of cellulase activity (382 ± 37Ug-1) in GBP mixed with rapeseed cake. In these mixtures, protein content increased 1.2-fold by SSF, as well as in GBP mixed with rice bran (1.3-fold). Scale-up of SSF using GBP mixed with sunflower cake from flasks to tray-type bioreactors demonstrated SSF reproducibility, achieving similar xylanase and cellulase activities. In the stirred-drum bioreactor, aeration and low agitation favored both enzymes production. Bioprocessing of GBP with plant feedstuffs using SSF was a cost-effective strategy to produce high-value enzymes, valorizing this seaweed by-product. [ABSTRACT FROM AUTHOR]

Extraction strategies impact the efficiency and nature of extracted compounds. This work assessed the chemical composition and antioxidant capacity of ethanolic, hydroethanolic, and aqueous versus enzyme-assisted extracts (isolated or with the sequential use of alcalase®, cellulase®, and viscozyme®) of the macroalgae Fucus vesiculosus (brown, Phaeophyceae) and Porphyra dioica (red, Rhodophyta. For both macroalgae, enzyme-assisted extraction (EAE) was the most efficient process compared to solvent-assisted extraction (SAE), independent of solvent. Fucus vesiculosus extraction yields were higher for EAE than for SAE (27.4% to 32.2% and 8.2% to 30.0%, respectively). Total phenolics content (TPC) was at least 10-fold higher in EAE extracts (229.2 to 311.3 GAE/gextract) than in SAE (4.34 to 19.6 GAE/gextract) counterparts and correlated well with antioxidant capacity (ABTS and ORAC methods), with EAE achieving values up to 8- and 2.6-fold higher than those achieved by SAE, respectively. Porphyra dioica followed F. vesiculosus's trend for extraction yields (37.5% to 51.6% for EAE and 5.7% to 35.1% for SAE), TPC, although of a lower magnitude, (0.77 to 8.95 GAE/gextract for SE and 9.37 to 14.73 GAE/gextract for EAE), and antioxidant capacity. Aqueous extracts registered the highest DPPH values for both macroalgae, with 2.3 µmol TE/gextract and 13.3 µmol TE/gextract for F. vesiculosus and P. dioica, respectively. EAE was a more efficient process in the extraction of soluble protein and reducing sugars in comparison to SAE. Furthermore, an improved effect of enzyme-assisted combinations was observed for almost all analyzed parameters. This study shows the promising application of enzyme-assisted extraction for the extraction of valuable compounds from F. vesiculosus and P.dioica, making them excellent functional ingredients for a wide range of health and food industrial applications. [ABSTRACT FROM AUTHOR]

Enzymatic solid-state fermentation can improve the nutritional quality of feed materials. The current study aimed to determine the effect of the solid-state fermentation of rapeseed meal (RSM) with carbohydrase/s and phytases in various combinations. RSM was fermented for 24 h at 25 °C with eight commercial preparations and mixtures thereof to prepare eleven products (PHYL—liquid-6-phytase; RON NP—6-phytase; RON HI—6-phytase; RON R—α-amylase; RON WX—β-xylanase; RON VP—β-glucanase; RON A—α-amylase, β-glucanase; RON M—xylanase, β-glucanase; RON NP+M; RON NP+A; RON NP+M+R). Afterward, the enzymes were deactivated at 70 °C within 15 min, and the biomass was dried for 24 h at 55 °C. Carbohydrase and/or phytase additives did not positively affect crude or true protein content or reduce crude fiber (p > 0.05). Among the products after fermentation, a significant reduction in the content of the raffinose family oligosaccharides, glucosinolates, and phytate was found. In the presence of phytase, the phytate reduction was more significant (p < 0.01) than that in the presence of carbohydrases only. The addition of carbohydrases together with phytases did not improve the results in comparison with phytases alone (p > 0.05). The most valuable effect was found for liquid-6-phytase (PHYL). [ABSTRACT FROM AUTHOR]

Simple Summary: Cereal grains and their by-products incorporated into non-ruminant animal feeds contain non-starch polysaccharides and phytate, which are anti-nutritional factors. The animals' endogenous digestive enzymes do not break down non-starch polysaccharides and phytate. Thus, exogenous enzymes such as carbohydrases and phytase, are supplemented to increase dietary energy and nutrient availability, reducing dietary cost and improving growth performance. Nevertheless, recent studies reveal that the improvement in growth performance goes beyond the release of indigestible nutrients. The positive impacts extend to the intestinal microbiota, immune system, and antioxidant status. However, it is unclear whether the benefits of enzyme action can be translated into better animal growth on commercial farms that face environmental, immunological, and management challenges. In this review, we observed that the supplementation of carbohydrases and phytase under poor sanitary conditions aligns with the energy and nutritional valorization matrices. This suggests that enzymes have properties that promote overall intestinal health, because they expedite post-sanitary challenge recovery and, at the same time, maintain improved utilization of the nutritional matrix. Additionally, studies under commercial conditions demonstrate that matrices containing carbohydrases and phytase sustain the growth and general health of broiler chickens and pigs. However, future research is needed to determine the extent of energy and nutrient savings at the commercial farm level. This review aimed to clarify the mechanisms through which exogenous enzymes (carbohydrases and phytase) influence intestinal health, as well as their effects on the nutrients and energy matrix in diets fed to poultry and pigs reared under sanitary challenging conditions. Enzyme supplementation can positively affect intestinal microbiota, immune system, and enhance antioxidant status. Although enzymes have been shown to save energy and nutrients, their responses under sanitary challenging conditions are poorly documented. Immune system activation alters nutrient partitioning, which can affect the matrix values for exogenous enzymes on commercial farms. Notably, the carbohydrases and phytase supplementation under sanitary challenging conditions align with energy and nutritional valorization matrices. Studies conducted under commercial conditions have shown that matrices containing carbohydrases and phytase can maintain growth performance and health in poultry and pigs. However, these studies have predominantly focused on assessing a single level of reduction in energy and/or available phosphorus and total calcium, limiting our ability to quantify potential energy and nutrient savings in the diet. Future research should delve deeper into determining the extent of energy and nutrient savings and understanding the effects of alone or blended enzymes supplementation to achieve more specific insights. [ABSTRACT FROM AUTHOR]

Various studies have shown that the microbial proteins are often more stable than belongs to other sources like plant and animal origin. Hence, the interest in microbial enzymes has gained much attention due to many potential applications like bioenergy, biofuel production, biobleaching, bioconversion and so on. Additionally, recent trends revealed that the interest in isolating novel microbes from harsh environments have been the main focus of many scientists for various applications. Basically, industrially important enzymes can be categorized into mainly three groups: carbohydrases, proteases, and lipases. Among those, the enzymes especially carbohydrases involved in production of sugars. Carbohydrases include amylases, xylanases, pectinases, cellulases, chitinases, mannases, laccases, ligninases, lactase, glucanase, and glucose oxidase. Thus, here, an approach has been made to highlight five enzymes namely amylase, cellulase, laccase, pectinase, and xylanase from different sources with special emphasis on their properties, mechanism, applications, production optimization, purification, molecular approaches for its enhanced and stable production, and also biotechnological perspectives of its future development. Also, green and sustainable catalytic conversion strategies using nanoparticles of these enzymes have also been discussed. This review will provide insight into the carbohydrases importance and their usefulness that will help to the researchers working in this field. [ABSTRACT FROM AUTHOR]

Current understanding of site-specific digestion of carbohydrates in mammals is that whereas starch can be degraded by mammalian enzymes, other complex carbohydrates (e.g. pectin, inulin, xylan or cellulose) are typically digested by the enzymes of symbiotic microbes. To test whether the previously reported presence of complex carbohydrate-digesting enzymes in the small intestine of sheep represents actual small intestinal enzyme activity or just outflux of microbial enzymes from the forestomach, we applied the same methodology of isolating enzymes from gastrointestinal contents and applying them in vitro to test substrates in the nutria (Myocastor coypus), a hindgutfermenting rodent without a forestomach. No enzymatic activity against carbohydrates was detected in the stomach, excluding an effect of coprophagy on the presence of the investigated enzymes. While - as expected - starch digestion was highest in the small intestine, and that of the other carbohydrates was highest in the caecum, there was nevertheless detectable enzymatic activity against pectin, inulin, xylan and cellulose in the small intestine. While further research is warranted, we suggest that these results indicate a certain degree of unspecific carbohydrase activity by small intestinal enzymes that plays no relevant role in vivo due to the short residence time of digesta in the small intestine. [ABSTRACT FROM AUTHOR]

Extraction strategies impact the efficiency and nature of extracted compounds. This work assessed the chemical composition and antioxidant capacity of ethanolic, hydroethanolic, and aqueous versus enzyme-assisted extracts (isolated or with the sequential use of alcalase®, cellulase®, and viscozyme®) of the macroalgae Fucus vesiculosus (brown, Phaeophyceae) and Porphyra dioica (red, Rhodophyta. For both macroalgae, enzyme-assisted extraction (EAE) was the most efficient process compared to solvent-assisted extraction (SAE), independent of solvent. Fucus vesiculosus extraction yields were higher for EAE than for SAE (27.4% to 32.2% and 8.2% to 30.0%, respectively). Total phenolics content (TPC) was at least 10-fold higher in EAE extracts (229.2 to 311.3 GAE/gextract) than in SAE (4.34 to 19.6 GAE/gextract) counterparts and correlated well with antioxidant capacity (ABTS and ORAC methods), with EAE achieving values up to 8- and 2.6-fold higher than those achieved by SAE, respectively. Porphyra dioica followed F. vesiculosus’s trend for extraction yields (37.5% to 51.6% for EAE and 5.7% to 35.1% for SAE), TPC, although of a lower magnitude, (0.77 to 8.95 GAE/gextract for SE and 9.37 to 14.73 GAE/gextract for EAE), and antioxidant capacity. Aqueous extracts registered the highest DPPH values for both macroalgae, with 2.3 µmol TE/gextract and 13.3 µmol TE/gextract for F. vesiculosus and P. dioica, respectively. EAE was a more efficient process in the extraction of soluble protein and reducing sugars in comparison to SAE. Furthermore, an improved effect of enzyme-assisted combinations was observed for almost all analyzed parameters. This study shows the promising application of enzyme-assisted extraction for the extraction of valuable compounds from F. vesiculosus and P.dioica, making them excellent functional ingredients for a wide range of health and food industrial applications.

Simple Summary: Sustainable animal nutrition and feeding are critical to meeting the growing population's food needs, requiring more resource-efficient feed production with less waste. This work aims at the nutritional valorization of plant feedstuffs using one of the most ecological, inexpensive, and convenient large-scale industrial and technological treatments, as well as solid-state fermentation. Fermentation proved useful in adding value-added compounds to the plant feedstuffs as proteins and enzymes, increasing protein digestibility, and reducing fiber, creating a novel feedstuff that could help reduce waste and the use of unsustainable ingredients when applied to animal nutrition. Plant feedstuffs are the main ingredients of animal feed. Owing to food–feed competition, increasing the utilization efficiency of these feedstuffs is important for animal nutrition. This can be achieved via solid-state fermentation (SSF). SSF of a plant feedstuff mixture (PFM) (25% rapeseed meal, soybean meal, rice bran, and sunflower meal) by three fungi (Aspergillus ibericus MUM 03.29, Aspergillus niger CECT 2088, and Aspergillus niger CECT 2915) resulted in an increase in protein content by 5%, irrespective of fungi, a reduction in cellulose content by 9 to 11%, and of hemicellulose content by 21 to 34%, relative to unfermented PFM. Enzyme production was measured: the highest cellulase (123.7 U/g), xylanase (431.8 U/g), and beta-glucosidase (117.9 U/g) activity were achieved with A. niger CECT 2088. Principal component analysis showed a positive correlation between all fermented PFMs and enzyme production, protein content, digestibility, and fiber reduction. Bioprocessing of the PFM by SSF increased its nutritional value and digestibility, making it more appealing for animal feeds. [ABSTRACT FROM AUTHOR]

One of the important problems of current biotechnology is the usage of enzymes of microbial origin for destruction of poorly soluble compounds and synthesis of new drugs. In recent years a great deal of researchers’ attention has been given to such technologically promising carbohydrases as O-glycosylhydrolases catalyzing the hydrolysis of O-glycoside links in glycosides, oligo- and polysaccharides, glycolipids, and other glycoconjugates. Aim. The review provides data on the position of α-L-rhamnosidases in the modern hierarchical classification of glycosidases and presents data available in the literature on the features of the enzyme structure in various microorganisms. Methods. The publications from the following databases were analyzed: PubMed (https://pubmed. nsbi.nlm.nih.gov/), the Carbohydrate-Active enZYmes (http://www.cazy.org/), the BRENDA Enzyme Database (https://www.brenda-enzymes.org/). Results. Data on the physicochemical, catalytic, and kinetic properties of α-L-rhamnosidases in microorganisms of different taxonomic groups have been systematized. The peculiarities of the substrate specificity of the enzyme depending on the nature of the protein and the growing conditions of the producer are characterized. Conclusions. Functional properties and specificity action of microbial α-L-rhamnosidases suggest their broad-range applicability for food and animal feed processing, as well as obtaining biologically active compounds for the pharmaceutical industry and medicine. [ABSTRACT FROM AUTHOR]

This study aimed at investigating the effects of genotype and temperatures on the proteolytic activity in green malt of 48 barley genotypes, including 19 mutants, 15 hulled, 4 hulless, and 10 wild using enzyme assays based on casein, as substrate. During malting, insoluble barley protein must be hydrolyzed into soluble peptides and free amino acids to supply the brewing yeast with sufficient nutrients to grow rapidly and metabolize glucose and other sugars into alcohol through fermentation. However, the relatively hot temperatures employed during kilning usually denature the proteolytic enzymes due to their thermolabile nature. Even though the hydrolytic activity of most of the proteases is destroyed during the kilning process, the malt includes a small fraction of thermostable proteases that can further degrade protein in the subsequent mashing process. Considering the higher temperature range employed in industrial kilning and mashing, three temperatures (37, 50, and 70°C) were selected to identify the genotypes possessing high activity at the higher range of temperatures as well as thermostable variant of the enzyme. The proteolytic activity in all the genotypes declined after 50°C depicting its optimum temperature. Overall proteolytic activity was observed to be positively correlated with the amino acids and negatively correlated with protein content. Three mutant (BL2086, BL2091, and BL2079) and one wild (WS 237) genotypes possessing proteolytic activity in a higher range at all the studied temperatures have the potential to be exploited in the breeding programs for incorporating trait of thermostable proteolytic activity into low malting efficiency cultivars. Practical Application: The optimal hydrolytic activities of carbohydrases and proteases during mashing are essential for producing high‐quality wort from malted barley to ensure that hydrolyzed molecules are available to brewers' yeast to support fermentative metabolism. In this study, several barley cultivars were grown under identical environmental conditions but assayed at different temperatures. As result, four genotypes had been obtained that possessed optimal proteolytic activities at a higher temperature range and can be of great interest to breeders and maltsters for altering wort amino acid profiles and better exposure of starch to mashing enzymes, thereby increasing the fermentable sugar yield from the malt. [ABSTRACT FROM AUTHOR]

Although several enzymatic approaches have been applied with this intent, the sequential use of enzymes covering both cellulolytic and proteolytic activity has never been performed in seaweeds. Sequential use of these enzymes improved the overall extraction yield by up to 160%, 30% and 80% in the different seaweeds when compared to the control condition (water extraction), use of a carbohydrases' cocktail alone and use of proteases alone, respectively. Regarding the proximate composition of extracts, it proved to be an efficient approach for the solubilization of carbohydrates (up to 28% in G. vermiculophylla, 66% in P. dioica, 77% in U. rigida and 35% in F. vesiculosus) and protein (up to 55% in G. vermiculophylla, 47% in P. dioica, 52% in U. rigida and 42% in F. uesiculosus). For all biomasses, the combination of enzymes induced a significant increase in antioxidant activity, not only by the increase of phenolic compounds but also by the hydrolysis of protein to peptides. Moreover, extracts from red seaweeds displayed prebiotic activity which can be ascribed to their increased content in oligosaccharides and protein/peptides. Overall, the sequential use of enzymes with different activities demonstrated to be an efficient approach for extracting functional fractions to be used as functional ingredients to improve the nutritional value (e.g. in proteins) and/or to include antioxidant and prebiotic features in the food product. [ABSTRACT FROM AUTHOR]

This review aimed to clarify the mechanisms through which exogenous enzymes (carbohydrases and phytase) influence intestinal health, as well as their effects on the nutrients and energy matrix in diets fed to poultry and pigs reared under sanitary challenging conditions. Enzyme supplementation can positively affect intestinal microbiota, immune system, and enhance antioxidant status. Although enzymes have been shown to save energy and nutrients, their responses under sanitary challenging conditions are poorly documented. Immune system activation alters nutrient partitioning, which can affect the matrix values for exogenous enzymes on commercial farms. Notably, the carbohydrases and phytase supplementation under sanitary challenging conditions align with energy and nutritional valorization matrices. Studies conducted under commercial conditions have shown that matrices containing carbohydrases and phytase can maintain growth performance and health in poultry and pigs. However, these studies have predominantly focused on assessing a single level of reduction in energy and/or available phosphorus and total calcium, limiting our ability to quantify potential energy and nutrient savings in the diet. Future research should delve deeper into determining the extent of energy and nutrient savings and understanding the effects of alone or blended enzymes supplementation to achieve more specific insights.

Simple Summary: There are numerous feed additives that can be used to enhance grow–finish pig growth performance and carcass characteristics, which can potentially lead to a higher economic return. Therefore, we collected scientific data to summarize the results of these additives' effects on grow–finish pigs. The feed additive categories chosen were acidifiers, betaine, chromium, conjugated linoleic acid, copper, direct-fed microbials, carbohydrases, proteases, phytases, multi-enzymes, essential oils, L-carnitine, yeasts, and zinc. We found that most of the results for each feed additive showed positive outcomes. For daily gain, direct-fed microbials, copper, L-carnitine, and multi-enzymes showed relatively large (>2.1% improvement) and positive effects. For feed efficiency, acidifiers, betaine, conjugated linoleic acid, multi-enzymes, direct-fed microbials, L-carnitine, and yeasts showed relatively large (>2.5% improvement) and positive effects. For backfat thickness, except for betaine, chromium, conjugated linoleic acid, and L-carnitine, most feed additives showed little effect (<1.7% improvement). This review provides information on the feed additives that have the most evidence in improving the performance of grow–finish pigs for pork producers around the world. Feed additives have shown benefits throughout the literature in improving grow–finish pigs' growth performance and carcass characteristics. However, the results have not been well summarized. Therefore, this review summarizes the available research (402 articles) on 14 feed additive categories fed to grow–finish pigs. The categories were acidifiers, betaine, Cr, conjugated linoleic acids, Cu, direct-fed microbials, carbohydrases, proteases, phytases, multi-enzymes, essential oils, L-carnitine, yeasts, and Zn. Qualified articles were collected and selected based on inclusion and exclusion criteria from online databases. The percentage difference for each response variable between the treatment and control group was calculated and summarized. Most results were positive for each feed additive; however, the magnitude of improvement varied, and most were not statistically significant. For ADG, DFM, Cu, L-carnitine, and multi-enzymes showed relatively large positive effects (>2.1% improvement) across a reasonable number of articles. Acidifiers, betaine, CLA, multi-enzymes, DFM, L-carnitine, and yeasts showed relatively large positive effects (>2.5% improvement) on improving G:F. Moreover, except for betaine, Cr, CLA, and L-carnitine, most feed additives showed little and non-significant effects on BF thickness (<1.7% improvement). This review provides a descriptive analysis for commonly used feed additives in the hope of better understanding feed additives' effects on grow–finish pigs. [ABSTRACT FROM AUTHOR]

Two novel Bacillus glycoside hydrolase family 53 (GH53) endo-1,4-β-galactanases (Bs936 and Bs4828) were identified. The recombinant Bs936 and Bs4828 displayed maximal activities at pH 5.5/55 °C and pH 6.5/65 °C, respectively. Stability analyses revealed that the enzymes were stable between pH 4.5–10, retaining over 80% activities after 12 h incubation at 25 °C. Moreover, Bs936 maintained about 75% activity after being treated at 45 °C for 2 h, while Bs4828 kept full activity after 4 h of incubation at 50 °C. Importantly, Bs936 and Bs4828 exhibited good activity towards β-1,4-galactan, showing specific activities of 1859.46- and 3110.79 U/mg towards potato galactan, respectively. The corresponding K m and k cat /K m were 4.93 mg/mL and 296.35 mL/mg/s for Bs936, and 6.60 mg/mL and 707.12 mL/mg/s for Bs4828. Extensive synergy assays revealed that each endo-1,4-β-galactanase acted synergistically with the other β-galactosidase (Bs937 or Bs4829), endo-1,5-α-L-arabinosidase (BsAbn2) and an endoxylanase/endoglucanase+cellulase mixture (BaXynA + LsCel) towards the hydrolysis of galactan, debranched arabinan/arabinan, and pretreated corn stover, respectively. The corresponding degrees of synergy were 1.28, 1.17/1.09, and 1.15 for Bs486 and 1.55, 1.04/1.08, and 1.11 for Bs4828, respectively. This study expands the repertoire of GH53 enzymes, and demonstrates the role of two highly active endo-1,4-β-galactanases in polysaccharides hydrolysis. [ABSTRACT FROM AUTHOR]

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Seaweeds are valuable feedstocks with the potential to be used as ingredients in aquafeeds. However, their use are still limited, given their recalcitrant polysaccharide structure. To break this structure, a biotechnological approach such as solid-state fermentation (SSF) by filamentous fungi can be used, which simultaneously increases the nutritional value of the biomass. However, SSF has hardly been studied in seaweeds; thus, in this study, five different seaweeds (Gracilaria sp., Porphyra dioica, Codium tomentosum, Ulva rigida, and Alaria esculenta) were used as substrates in SSF with Aspergillus ibericus MUM 03.49 and A. niger CECT 2915. Firstly, the seaweeds were fully characterized, and, then, changes in the crude protein and carbohydrate contents were assessed in the fermented biomass, as well as any carbohydrases production. The SSF of U. rigida with both fungi resulted in the maximum xylanase and β-glucosidase activities. The maximum cellulase activity was achieved using Gracilaria sp. and U. rigida in the SSF with A. niger. The protein content increased in C. tomentosum after SSF with A. ibericus and in U. rigida after SSF with both fungi. Moreover, U. rigida's carbohydrate content decreased by 54% and 62% after SSF with A. ibericus and A. niger, respectively. Seaweed bioprocessing using SSF is a sustainable and cost-effective strategy that simultaneously produces high-value enzymes and nutritionally enhanced seaweeds to be included in aquafeeds. [ABSTRACT FROM AUTHOR]

Rice bran (RB) is a by-product with limited application due to technological constraints. Enhancing its technological functionality as potential food ingredient will improve the sustainability of rice production. The aim was to study the impact of enzymatic and thermal treatments on defatted rice bran using six distinct commercial enzymes (carbohydrases and proteases) and dry heating by evaluating its technological, nutritional and functional properties. Enzymatic treatment increased up to 208% the soluble dietary fiber content (8.19 g/100 g) of defatted RB. Moreover, the solvent retention capacity, including water, oil, sodium carbonate, and sucrose, exhibited a noteworthy increase across all treatments (p < 0.05). Bran color changed after treatments, increasing its luminosity (L*) and decreasing the value of a* in all cases, but b* decreased when treated with protein-acting enzymes while increased with carbohydrate-acting enzymes. Proteases played a pivotal role in reducing particle size and forming gels requiring minimal force for application. Microscopic analysis revealed that carbohydrases-treated samples exhibited prominent cell wall breakage, while protease-treated ones showed a gel-like surface with less distinct protein bodies and layered walls. These comprehensive study sheds new transformations brought about by these enzymatic interventions, offering valuable insights into the optimization of rice bran functionality. [Display omitted] • Carbohydrases, proteases and thermal treatments modify defatted rice bran. • Enzymatic treatment shifts hemicellulose from an insoluble to a soluble fraction. • Alcalase significantly reduces the particle size distribution of defatted rice bran. • Enzymatic treatment increases water and oil binding capacity of defatted rice bran. [ABSTRACT FROM AUTHOR]

Plant feedstuffs are the main ingredients of animal feed. Owing to food–feed competition, increasing the utilization efficiency of these feedstuffs is important for animal nutrition. This can be achieved via solid-state fermentation (SSF). SSF of a plant feedstuff mixture (PFM) (25% rapeseed meal, soybean meal, rice bran, and sunflower meal) by three fungi (Aspergillus ibericus MUM 03.29, Aspergillus niger CECT 2088, and Aspergillus niger CECT 2915) resulted in an increase in protein content by 5%, irrespective of fungi, a reduction in cellulose content by 9 to 11%, and of hemicellulose content by 21 to 34%, relative to unfermented PFM. Enzyme production was measured: the highest cellulase (123.7 U/g), xylanase (431.8 U/g), and beta-glucosidase (117.9 U/g) activity were achieved with A. niger CECT 2088. Principal component analysis showed a positive correlation between all fermented PFMs and enzyme production, protein content, digestibility, and fiber reduction. Bioprocessing of the PFM by SSF increased its nutritional value and digestibility, making it more appealing for animal feeds.

ABSTRACT: The aim of this study was to determine the effect of emulsifier and multicarbohydrase enzyme supplementation on performance, nutrient utilization, and apparent metabolizable energy—nitrogen (AMEN) value of broiler diets containing rapeseed meal (RSM) as well as their influence on the gut morphological structures, excretion of total and free sialic acid, and cecum concentration of short-chain fatty acids (SCFAs) in broiler chickens. A total of 384 male broiler chicks were assigned to four dietary treatments. The diet of the control treatment (CON) consisted of soybean, maize, and RSM (5% in starter, 7% in grower, 15% in finisher) with soybean and palm oils. The diets used for the experimental treatments were the control diet supplemented with an emulsifier (EMU), enzyme (ENZ), or both (EMU + ENZ). The duodenum (n = 10/treatment) and ileum (n = 10/treatment) digesta samples were assessed to determine nutrient digestibility: crude protein (CP), ether extract (EE), starch, Ca. Throughout the experimental period, EMU + ENZ treatment indicated the lowest total average feed intake and feed conversion ratio, with the highest average weight gain among the studied treatments (P < 0.05). The EMU + ENZ treatment also resulted in higher (P < 0.05): apparent prececal digestibility (APD) of CP, total tract neutral detergent fibre (NDF) degradation, apparent total tract digestibility (ATTD) of EE, villus height to crypt depth ratio (P < 0.1). The highest APD of EE was noted in the EMU treatment (P < 0.05). No significant differences were found in the AMEN values of the diets. A greater jejunum villi surface area was found in groups supplemented by enzyme compared to CON (P < 0.05). The EMU + ENZ treatment presented lower sialic acid excretion in the ileum and concentration of cecum SCFAs compared to the CON treatment (P < 0.05). The obtained results indicate that simultaneous usage of additives had beneficial effect on production parameters, nutrient digestibility, NDF degradation, as well as gut mucosa morphology. Based on the SCFAs concentration results, separate or simultaneous addition of emulsifier or/and enzyme did not provoke excessive fermentation activity of cecal bacteria.

Wheat is rich in non-starch polysaccharides (NSP) and their degradation in poultry diets is promoted by exogenous carbohydrases. The objective here was to evaluate the effect of adding an intrinsically thermostable xylanase on wheat-based diets for laying hens in yolk color, carotenoid and fatty acid profiles of eggs. A total of 128 laying hens were used for 12 weeks. They were randomly allocated to four dietary treatments with different levels of xylanase: T1: control (no xylanase), T2: 30,000 U/g, T3: 45,000 U/g and T4: 90,000 U/g, with 32 birds, 16 replicates per treatment (2 birds/replicate). At the end of the experimental period, egg yolk color index, redness (a*) and yellowness (b*) of egg yolks were found significantly higher in all the enzyme supplemented diet groups (T2, T3, T4) compared with the control (T1). Canthaxanthin levels were significantly higher in T3 than T1 (p < 0.05). Total n-3, n-6 and total polyunsaturated fatty acids (FAs) were significantly higher in T4 compared with the control (p < 0.01), while the reverse trend was evidenced for monounsaturated FAs. Additionally, total n-3 FAs were higher in the T2 than T1 (p < 0.005). Overall, the results showed that exogenous xylanase enzyme supplementation in wheat-based diets for laying hens contribute to maintaining egg yolk color. Overall, exogenous xylanase enzyme supplemented at all levels in wheat-based laying hens' diets improved egg yolk color compared to the control diet. The enzyme supplemented at the higher level (90,000 U/g) improved polyunsaturated and reduced monounsaturated egg yolk fatty acid content. [ABSTRACT FROM AUTHOR]

Simple Summary: Macroalgae have been increasingly exploited worldwide for feed, food and biofuel applications, due to their nutritive and bioactive compounds. Green seaweeds belonging to the genus Ulva have high growth rates, which makes them suitable for being cultured in sustainable algae production, such as an integrated multi-trophic aquaculture system. This is expected to increase the use of Ulva sp. as an alternative source to conventional feedstuffs (e.g., cereals and soybean meal) for poultry. The objective of the current study was to assess if the incorporation of 15% Ulva lactuca in broiler chickens' diet, combined or not with carbohydrate-active enzymes, would enhance meat nutritional quality without compromising animal growth performance. Overall, U. lactuca led to an accumulation of antioxidant carotenoids, n-3 PUFA and macrominerals, including magnesium, potassium and phosphorus, in the breast muscle, with likely health benefits, without significantly impairing growth performance. The supplementation of macroalgae with a recombinant ulvan lyase reduced ileal viscosity with possible beneficial effects on broiler digestibility. Although dietary U. lactuca showed potential to increase meat quality, it reduced meat overall acceptability, which suggests the use of a lower algae inclusion level to prevent a negative meat sensory perception for consumers. The aim of the study was to test if feeding 15% U. lactuca to broilers, alone or combined with carbohydrases, enhanced meat nutritional quality, without compromising growth performance. One hundred and twenty 22-day-old broilers were allocated to the following diets and replicated 10 times for 14 days: (1) maize and soy-based diet (control); (2) control with 15% U. lactuca (UL); (3) UL diet with 0.005% commercial carbohydrase mixture (ULC); and (4) UL diet with 0.01% ulvan lyase (ULE). Final body weight and average daily gain decreased (p < 0.050) with the ULE diet compared with the control, but no significant differences were found for the other diets. The intestinal viscosity increased (p < 0.001) with all alga diets but was lowered (p < 0.050) in the ileum with the ULE diet, relative to UL and ULC diets. Meat lightness and redness values, off-flavours, and total carotenoids increased (p < 0.001), while yellow values, tenderness, juiciness, overall acceptability, α- and γ-tocopherol, and total lipids decreased (p < 0.001) with alga diets. The n-3 polyunsaturated fatty acids (PUFA) increased (p < 0.050), and the n-6/n-3 PUFA ratio decreased (p < 0.001) with the ULE diet. Total minerals in meat increased (p < 0.001) with alga diets, conversely to sodium and zinc (p < 0.001). Feeding 15% of U. lactuca to broilers did not impair growth but increased meat nutritional value through the accumulation of health-promoting antioxidant carotenoids, n-3 PUFA and total minerals, although reducing overall meat acceptability. [ABSTRACT FROM AUTHOR]

Pink bollworm (PBW), Pectinophora gossypiella (Saunders) is a notorious monophagous pest of cotton. Any changes in the nutritional quality of cotton have direct influences on the life cycle of the pink bollworm. In this regard, the host-mediated effect of elevated carbon dioxide (CO2) and temperature on the life cycle of pink bollworm, P. gossypiella on Bt cotton was studied in the open top chambers at the Center for Agro-climatic Studies, UAS, Raichur. The results revealed that variations in phyto-chemistry of Bt cotton particularly the Crytoxin expression and nitrogen have reduced in elevated CO2 and temperature conditions and also reduced the larval, pupal period, adult longevity, fecundity and incubation period. However, there was no significant difference in the biology of PBW among the climate change treatments. Similar results were noticed when PBW was reared on Bt cotton collected from different open top chambers and reared separately in growth chambers and the same larvae were analyzed for their enzyme activity. The results showed that there was significant increase in the activity of midgut proteases (trypsin and chymotrypsin) and carbohydrases (a-amylase) and decrease in the activity of lipase enzyme was noticed. [ABSTRACT FROM AUTHOR]

Herbivorous surgeonfishes are an ecologically successful group of reef fish that rely on marine algae as their principal food source. Here, we elucidated the significance of giant enteric symbionts colonizing these fishes regarding their roles in the digestive processes of hosts feeding predominantly on polysiphonous red algae and brown Turbinaria algae, which contain different polysaccharide constituents. Using metagenomics, single-cell genomics, and metatranscriptomic analyses, we provide evidence of metabolic diversification of enteric microbiota involved in the degradation of algal biomass in these fishes. The enteric microbiota is also phylogenetically and functionally simple relative to the complex lignocellulose-degrading microbiota of terrestrial herbivores. Over 90% of the enzymes for deconstructing algal polysaccharides emanate from members of a single bacterial lineage, "Candidatus Epulopiscium" and related giant bacteria. These symbionts lack cellulases but encode a distinctive and lineage-specific array of mostly intracellular carbohydrases concurrent with the unique and tractable dietary resources of their hosts. Importantly, enzymes initiating the breakdown of the abundant and complex algal polysaccharides also originate from these symbionts. These are also highly transcribed and peak according to the diel lifestyle of their host, further supporting their importance and host-symbiont cospeciation. Because of their distinctive genomic blueprint, we propose the classification of these giant bacteria into three candidate genera. Collectively, our findings show that the acquisition of metabolically distinct "Epulopiscium" symbionts in hosts feeding on compositionally varied algal diets is a key niche-partitioning driver in the nutritional ecology of herbivorous surgeonfishes.

This study provides a comprehensive chemical characterization of blueberry (Vaccinium corymbosum L.) pomace subjected to hydrolysis using carbohydrases (xylanase, cellulase and Extralyve®). Blueberry pomace obtained from the juice industry represents a rich source of insoluble dietary fiber, which holds potential for the liberation of oligosaccharides and phenolic compounds. Oligosaccharides were determined using HPLC-RID and change in phenolics were monitored by HPLC-DAD and HPLC-ESI-MS. Out of three enzymatic preparations, Extralyve®, which represents a commercial blend of xylanase, cellulase and polygalacturonase, demonstrated high hydrolytic activity towards the insoluble fiber present in blueberry pomace. This reduction led to an increased concentration of oligosaccharides, peaking after 2 h of hydrolysis (59.8 mg/g). Additionally, the levels of phenolic acids and non-anthocyanin flavonoids reached their maximum after 4 h of hydrolysis (1.22 mg/g), enhancing the antioxidant capacity of the hydrolyzed product (FRAP and ABTS assays). Finally, phenolic extracts from treated and untreated blueberry pomace were tested to evaluate cytotoxic potential in cell viability assays (MTT) using gastric carcinoma (NCI–N87) and two colorectal adenocarcinoma (Caco-2 and HT29-MTX) cells. Extracts obtained from Extralyve® treatment, rich in isorhamnetin, quercetin and kaempferol, resulted in interesting antiproliferative actions compared with the other treated and untreated samples. • Carbohydrases broke down the insoluble dietary fiber (IDF) present in blueberry pomace (BP). • The combination of enzymes facilitated the release of phenolic acids and non-anthocyanin flavonoids. • The release of phenolics enhances the antioxidant and antiproliferative activities of blueberry pomace (BP) extracts. • The breakdown of insoluble dietary fiber (IDF) led to an increased generation of oligosaccharides. [ABSTRACT FROM AUTHOR]

In this study, proteins were extracted from cress seeds using enzymatic pretreatment and ultrasound. Optimization of enzymatic pretreatment, employing a multi-enzyme complex rich in carbohydrases, aimed to maximize extraction yield and reduce sugar content. Ultrasound replaced classical extraction under optimal conditions by comparing functional properties between enzyme-treated and untreated samples. Combined enzymatic pretreatment and ultrasound significantly increased extraction yield from 39.56% to 94.03%. Optimal enzymatic pretreatment conditions included pH 3.84, 45.16 °C, enzyme concentration 2.32 FBGU/g seed, and 3.71 h of treatment time. Ultrasound-treated isolates exhibited superior functionality and minimal structural changes, with molecular weights ranging from ∼22 kDa to ∼59 kDa. These findings underscore the potential of cress seed protein isolate for diverse applications, enhanced through enzymatic pretreatment and ultrasound, owing to its high nutritional quality. [ABSTRACT FROM AUTHOR]

Enzymes, also known as biocatalysts, display vital properties like high substrate specificity, an eco-friendly nature, low energy inputs, and cost-effectiveness. Among their numerous known applications, enzymes that can target biofilms or their components are increasingly being investigated for their anti-biofouling action, particularly in healthcare, food manufacturing units and environmental applications. Enzymes can target biofilms at different levels like during the attachment of microorganisms, formation of exopolymeric substances (EPS), and their disruption thereafter. In this regard, a consortium of carbohydrases that can target heterogeneous polysaccharides present in the EPS matrix may provide an effective alternative to conventional chemical anti-biofouling methods. Further, for complete annihilation of biofilms, enzymes can be used alone or in conjunction with other antimicrobial agents. Enzymes hold the promise to replace the conventional methods with greener, more economical, and more efficient alternatives. The present article explores the potential and future perspectives of using carbohydrases as effective anti-biofilm agents. [ABSTRACT FROM AUTHOR]

The purpose of this study was to evaluate the impact of Chlorella vulgaris (5% in the diet), supplemented or not with two exogenous carbohydrase mixtures on piglets' performance, nutrient digestibility and gut morphology, fermentation products and microbiota. Forty-four male piglets weaned at 28 days of age, with 11.2 ± 0.46 kg of live weight, were used and assigned to 1 of 4 dietary treatments: cereal and soybean meal based-diet (control, n = 11), control diet with 5% of C. vulgaris (CH, n = 10), CH diet supplemented with 0.005% of Rovabio® Excel AP (CH + R, n = 10) and CH diet supplemented with 0.01% of a recombinant 4-carbohydrase mixture (CH + M, n = 11). Growth performance was not changed by the of C. vulgaris inclusion during 21 days of trial. However, total tract apparent digestibility of nutritional fractions was negatively impacted by the inclusion. In addition, the viscosity of duodenum plus jejunum contents slightly increased in all groups fed with the microalga. In contrast, dietary microalga increased duodenum villus height and promoted a healthier gut microbiota, with higher abundance of some specific bacterial taxa (Colidextribacter, Oscillospira and Lactobacillus). This study indicates that the dietary inclusion of 5% C. vulgaris improves piglets' gut health without impairing performance. Data also indicate that C. vulgaris reduces nutrient digestibility but promotes compensatory developments of gut mucosa and prebiotic effects. Dietary supplementation with exogenous carbohydrases does not seem to be necessary for this inclusion level. Therefore, the incorporation of CH as a sustainable feed ingredient in piglets' nutrition is a viable alternative approach. [ABSTRACT FROM AUTHOR]

This study was designed to examine the growth performance, nutrient digestibility, blood components and carcass cuts of broiler chickens fed cassava distillers' waste meal (CDWM) as a replacement for maize in a 6-week feeding trial. A total of 300 one-day-old Abor-Acre chicks were assigned to five groups using a completely randomized design. Each group contained 60 chicks with 6 replicates. The CDWM was incorporated in the diets at 25%, 50%, 75%, and 100% as a replacement for the maize used in the control diet. CDWM had higher protein and fibre content than maize. Significant differences (p < 0,05) were observed in growth parameters, nutrient digestibility, packed cell volume and breast weight. Final weight and daily weight gain decreased as CDWM increased in the diet, while average daily feed consumed and the conversion ratio increased linearly; birds fed the control diet had the lowest and best feed conversion ratio, while those fed 100% CDWM in place of maize had the highest feed conversion ratio. Thus, the optimal inclusion level of CDWM in broiler diets as a replacement for maize should not exceed 25%. Further studies are needed to determine whether the addition of a multi-carbohydrase enzyme will allow for a higher percentage of CDWM in place of maize with in broiler diets. [ABSTRACT FROM AUTHOR]

Usefulness of red grape pomace (GP) by-product in ruminant rations is restricted by its high fibre content. Pre-treatment with exogenous carbohydrases may partly resolve this problem. Therefore, the effects of various levels of an exogenous fibrolytic enzyme mixture (Viscozyme® L) on chemical nutrient content and in vitro ruminal fermentation of GP were studied. This product, with an enzyme activity of =100 fungal betaglucanase per gram (FBG/g) and a density of 1.2 g/mL, was applied to GP at 0 (ENZ0), 20 (ENZ20), 30 (ENZ30), 35 (ENZ35), and 45 g/kg DM (ENZ45). Neutral detergent fibre (NDF), acid detergent fibre (ADF) and insoluble condensed tannins (ICT) declined linearly as enzyme levels increased. Significant negative linear trends were observed for the rate of gas production at 36 and 48 hours, whereas quadratic trends were observed at 12- and 24-hours incubation in response to enzyme levels. Cumulative gas production at 12 hours and 24 hours and in vitro organic matter degradability at 96 hours post incubation increased linearly as treatment levels increased. Effective gas production declined linearly (P =0.041; R2 =0.085), whereas the slowly degradable fraction (P =0.042, R2 =0.084), the fractional degradation rate (P =0.003, R2 =0.175) and potential gas production (P =0.043, R2 =0.083) showed quadratic trends as enzyme levels increased. Pretreatment with Viscozyme reduced the fibre levels of GP and increased in vitro ruminal organic matter degradability. The optimum level for inclusion of Viscozyme was 5 g/kg. [ABSTRACT FROM AUTHOR]

Purpose: Okara is a highly perishable food processing by-product from soymilk and tofu manufacture. The present study aimed to optimize the pre-treatment of okara on the extraction of reducing sugars by applying a multi-enzyme complex, Viscozyme, containing a variety of carbohydrases to hydrolyze the okara cell walls. The chemical composition and the morphological structure of enzymatic-treated okara (ETO) were scrutinized. Methods: Enzymatic hydrolysis pre-treatment was carried out under different enzyme concentrations (5–8%), temperatures (50–60 °C) and pH values (3–4) according to a central composite rotatable design, in which the ranges were chosen based on preliminary study. Results: The optimal conditions for maximum reducing sugars extraction (4.78 mg/mL) were 8.0% (w/w) of enzyme, 50 °C and pH 3.0 for 4 h hydrolysis time, representing a 158.33 times increment, compared to the sample without enzymatic pre-treatment. The chromatogram from high performance liquid chromatography (HPLC) revealed that the carbohydrates such as glucose, fructose, raffinose and stachyose increased after hydrolysis. Surprisingly, arabinose which was not detected in raw okara (RO) was present in ETO. Scanning electron microscopy (SEM) confirmed the highly structured matrix of okara was unfolded and broken by the action of enzyme pre-treatment while energy-dispersive X-ray spectroscopy (EDS) imparted the changes of elemental components in ETO. The ETO showed higher antioxidant capacity than the control by DPPH, ABTS and FRAP tests. Conclusion: Enzymatic hydrolysis proved to be an efficient way to enhance the physicochemical properties of okara for possible uses, making this residue into a newly added value product. [ABSTRACT FROM AUTHOR]

Abstract Biofilm formation on both biotic and abiotic surfaces accounts for a major factor in spread of antimicrobial resistance. Due to their ubiquitous nature, biofilms are of great concern for environment as well as human health. In the present study, an integrated process for the co-production of a cocktail of carbohydrases from a natural variant of Aspergillus niger was designed. The enzyme cocktail was found to have a noteworthy potential to eradicate/disperse the biofilms of selected pathogens. For application of enzymes as an antibiofilm agent, the enzyme productivities were enhanced by statistical modelling using response surface methodology (RSM). The antibiofilm potential of the enzyme cocktail was studied in terms of (i) in vitro cell dispersal assay (ii) release of reducing sugars from the biofilm polysaccharides (iii) the effect of enzyme treatment on biofilm cells and architecture by confocal laser scanning microscopy (CLSM). Potential of the enzyme cocktail to disrupt/disperse the biofilm of selected pathogens from biopolymer surfaces was also assessed by field emission scanning electron microscopy (FESEM) analysis. Further, their usage in conjunction with antibiotics was assessed and it was inferred from the results that the use of enzyme cocktail augmented the efficacy of the antibiotics. The study thus provides promising insights into the prospect of using multiple carbohydrases for management of heterogeneous biofilms formed in natural and clinical settings.

The Moroccan locust, Dociostaurus maroccanus (Thunberg), is an agricultural pest which has caused considerable agricultural damage and economic importance. α-/β-Glucosidases and α-/β-Galactosidases are essential enzymes in the carbohydrate digestion of insect pest. Characterization of digestive enzymes is essential for gathering knowledge to production of resistant plants, in this research α-/β-Glucosidases and α-/β-Galactosidase from D. maroccanus. Results showed that the activities of α-/β-Glucosidases were higher than those of α-/β-Galactosidase in digestive system Moroccan locust. Maximum activity for α-Glucosidase was at pH 4. Optimum activity for β-Glucosidases and α-/β-Galactosidase were found at pH 5. The D. maroccanus α-/β-Glucosidases and α-/β-Galactosidases have an optimum temperature activity at 45, 55, 45, and 55 oC, respectively. As calculated using Lineweaver-Burk plots, the Km were about 3.96 ± 0.005, 1.927 ± 0.04, 1.051 ± 0.07, 0.558 ± 0.02 mM for α-/β-Glucosidases and α-/β-Galactosidase, respectively. Fe2+, Hg+, Zn2+, and EDTA inhibited carbohydrases activity, whereas the addition of Mg2+ and K+ increased enzyme activity. The zymogram pattern in the native gel revealed that α-/β-Glucosidases and α-/β-Galactosidases in the digestive system showed 3, 2, 1 and 1 major bands, respectively. Biochemical characterization of digestive carbohydrases is essential for achieving new and safe methods for pest control.

Brewer’s spent grain (BSG) is the main brewery industry by-product, with potential applications in the feed and food industries due to its carbohydrate composition. In addition, the lignocellulosic nature of BSG makes it an adequate substrate for carbohydrases production. In this work, solid-state fermentation (SSF) of BSG was performed with Aspergillus ibericus, a non-mycotoxin producer fungus with a high capacity to hydrolyze the lignocellulosic matrix of the agro-industrial by-products. SSF was performed at different scales to produce a crude extract rich in cellulase and xylanase. The potential of the crude extract was tested in two different applications: -(1) - the enzymatic hydrolysis of the fermented BSG and (2) - as a supplement in aquafeeds. SSF of BSG increased the protein content from 25% to 29% (w/w), while the fiber content was reduced to 43%, and cellulose and hemicellulose contents were markedly reduced to around 15%. The scale-up of SSF from 10 g of dry BSG in flasks to 50 g or 400 g in tray-type bioreactors increased 55% and 25% production of cellulase and xylanase, up to 323 and 1073 U g−1 BSG, respectively. The optimum temperature and pH of maximal activities were found to be 55°C and pH 4.4 for xylanase and 50°C and pH 3.9 for cellulase, cellulase being more thermostable than xylanase when exposed at temperatures from 45°C to 60°C. A Box–Behnken factorial design was applied to optimize the hydrolysis of the fermented BSG by crude extract. The crude extract load was a significant factor in sugars release, highlighting the role of hydrolytic enzymes, while the load of fermented BSG, and addition of a commercial β-glucosidase were responsible for the highest phenolic compounds and antioxidant activity release. The lyophilized crude extract (12,400 and 1050 U g−1 lyophilized extract of xylanase and cellulase, respectively) was also tested as an enzyme supplement in aquafeed for European seabass (Dicentrarchus labrax) juveniles. The dietary supplementation with the crude extract significantly improved feed and protein utilization. The processing of BSG using biological treatments, such as SSF with A. ibericus, led to the production of a nutritionally enriched BSG and a crude extract with highly efficient carbohydrases capable of hydrolyzing lignocellulosic substrates, such as BSG, and with the potential to be used as feed enzymes with remarkable results in improving feed utilization of an important aquaculture fish species.

Copyright of Revista Brasileira de Saúde e Produção Animal (RBSPA) is the property of Revista Brasileira de Saude e Producao Animal (RBSPA) and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Evidence shows that polyphenols can attenuate postprandial blood glucose responses to meals containing digestible carbohydrate. Polyphenol-rich plant extracts are emerging as potential ingredients in functional foods and/or beverages despite limited understanding of their physiological effects. Many studies have investigated the mechanisms of polyphenol-rich fruit extracts on inhibition of digestive enzymes. However, the evidence available has yet to be critically evaluated systematically. This report reviews the in vitro literature to quantify the effect of fruit polyphenol extracts on the activities of digestive carbohydrases. A systematic literature search was conducted using six science databases. Included studies, totaling 34 in number, were in vitro digestion models which quantified gut digestive enzyme(s) activity on starch digestion in the presence of fruit polyphenol extracts. Most studies assessed the effects of fruit extracts on either α-amylase (n = 30) or α-glucosidase (n = 30) activity. Studies were consistent overall in showing stronger inhibition of α-amylase compared to α-glucosidase by proanthocyanidin- and/or ellagitannin-rich fruit extracts. Recommendations are proposed for future reporting of this type of research to enable meaningful synthesis of the literature as a whole. Such knowledge could allow effective choices to be made for development of novel functional foods and beverages. [ABSTRACT FROM AUTHOR]

The objective of this study was to evaluate the effects of exogenous enzymes on nutrient intake and digestibility, rumen fermentation, and productivity of mid-lactating cows. Experiment 1 was designed to test increasing doses [0, 0.5, 1.0, or 1.5 g/kg of dry matter (DM)] of a combination of 2 enzyme products with xylanase and β-glucanase activities (Ronozyme Wx and Ronozyme VP, respectively; DSM Nutritional Products) on rumen fermentation and total apparent digestibility. Enzyme combinations had a ratio of endo-1,3(4)-β-glucanase to endo-1,4-β-xylanase of 8:2 (wt/wt). For experiment 1, 8 rumen cannulated lactating cows were used into a double 4 × 4 Latin square design experiment with 14 d of diet adaptation and 7 d of sampling. Despite no differences in feed intake, carbohydrases linearly increased neutral detergent fiber digestibility. Treatments marginally affected rumen fermentation, where a linear trend for lower rumen pH and a linear trend for greater isobutyrate concentration were observed with increasing enzyme dose. A trend for lower rumen NH 3 -N concentration was observed for cows receiving carbohydrases in comparison with control group. When comparing all enzyme treatments against control group, cows fed enzymes tended to produce more 3.5% fat-corrected milk (FCM), produced more milk fat, and had greater blood glucose concentration. Experiment 2 evaluated 3 doses (0, 0.5, or 0.75 g/kg of DM) of the same combination of enzyme products on performance of cows (n = 36) in a complete randomized block (n = 12) design. Cows received treatments for 9 wk. No interaction effects between treatments and time were observed for all variables assessed in this study. In agreement with experiment 1, no differences were detected for feed intake, but cows fed the enzyme products tended to produce more 3.5% FCM and milk fat compared with control. In addition, cows fed enzymes exhibited greater efficiency of FCM production (FCM ÷ DM intake) compared with control. No differences were detected for intake and productivity when comparing the 2 doses of carbohydrases. In summary, the enzyme products tested in this study may improve feed efficiency due to greater milk fat concentration. [ABSTRACT FROM AUTHOR]

Abstract Extracellular polymeric substance (EPS) produced by the microorganisms provides protection and stability to them when they are encased within biofilms. Heterogeneous polysaccharides form a major constituent of the EPS and are crucial for the formation and integrity of the biofilms/slime. Thus, breakdown of polysaccharides might help in dispersion of biofilms from abiotic surfaces. In the present study we isolated a fungus, Aspergillus niger APS, capable of concurrently producing a cocktail of carbohydrases and optimized the conditions for higher yields of all the enzymes by one variable at a time (OVAT) approach. The optimization studies resulted in 1.5 to 12 fold augmentation in the enzyme yields using biodegradable waste. Further, keeping in view the heterogeneous nature of polysaccharides in biofilm matrix, the in-house produced enzyme cocktail was used for the dispersal of biofilms formed by Salmonella enterica serovar Typhi, Escherichia coli and Staphylococcus aureus. Treatment with enzyme preparation caused 90.23 ± 4.0, 82.64 ± 5.0 and 76.32 ± 5.0% reduction of the biofilms formed by these organisms respectively which was also evidenced by Field emission scanning electron microscopy (FESEM) revealing the loss of biofilm architecture. Interestingly, the enzyme cocktail could also remove viscous slime formed under natural conditions in the kitchen drainage pipe (KDP). To the best of our knowledge, this is the first report on biotreatment of abiotic surfaces for removal of biofilms/slime formed under natural conditions. The study thus indicates the prospects of using multiple carbohydrases as an anti-biofouling agent on abiotic surfaces like equipments as well as implants/prostheses and pipelines.

Brewer's spent grain (BSG) is the largest by-product originated from the brewery industry with a high potential for producing carbohydrases by solid-state fermentation. This work aimed to test the efficacy of a carbohydrases-rich extract produced from solid-state fermentation of BSG, to enhance the digestibility of a plant-based diet for European seabass (Dicentrarchus labrax). First, BSG was fermented with A. ibericus to obtain an aqueous lyophilized extract (SSF-BSG extract) and incorporated in a plant-based diet at increasing levels (0—control; 0.1%, 0.2%, and 0.4%). Another diet incorporating a commercial carbohydrases-complex (0.04%; Natugrain; BASF) was formulated. Then, all diets were tested in in vitro and in vivo digestibility assays. In vitro assays, simulating stomach and intestine digestion in European seabass, assessed dietary phosphorus, phytate phosphorus, carbohydrates, and protein hydrolysis, as well as interactive effects between fish enzymes and dietary SSF-BSG extract. After, an in vivo assay was carried out with European seabass juveniles fed selected diets (0—control; 0.1%, and 0.4%). In vitro digestibility assays showed that pentoses release increased 45% with 0.4% SSF-BSG extract and 25% with Natugrain supplemented diets, while amino acids release was not affected. A negative interaction between endogenous fish enzymes and SSF-BSG extract was observed in both diets. The in vivo digestibility assay corroborated in vitro data. Accordingly, the dietary supplementation with 0.4% SSF-BSG increased the digestibility of dry matter, starch, cellulose, glucans, and energy and did not affect protein digestibility. The present work showed the high potential of BSG to produce an added-value functional supplement with high carbohydrases activity and its potential contribution to the circular economy by improving the nutritional value of low-cost and sustainable ingredients that can be included in aquafeeds. [ABSTRACT FROM AUTHOR]

The present study aimed to explore the optimal zymogram of combination of 6 carbohydrases (glucoamylase, pullulanase, maltase, thermostable α-amylase, medium temperature α-amylase, and cold-active α-amylase) supplemented in corn-soybean based diet of broilers aged 1 to 3 wk for the maximum starch digestibility, by using in vitro simulated gastrointestinal digestion and response surface method. The third generation of simulated monogastric animal digestion system was used for in vitro digestion experiment. By using single factor completely random design, the optimal supplement levels of single carbohydras were determined by the reducing sugar release amount and improved dry matter digestibility, which were the parameters representing the starch digestibility of the diet. Additionally, Box-Behnken response surface method was used to predict the optimal combination of 6 carbohydrases. The results showed that the optimistic zymogram of 6 carbohydrases in corn-soybean based diet for broilers aged 1 to 3 wk were 297.39 U/g glucoamylase, 549.72 U/g pullulanase, 3.01 U/g maltase, 1,455.73 U/g thermostable α-amylase, 278.64 U/g medium temperature α-amylase, and 1,985.97 U/g cold-active α-amylase, and the associated reduced sugar release amount and improved dry matter digestibility were 215.98 mg/g, and 6.23%, respectively. Furthermore, we conducted in vitro digestion experiments with diets supplemented with the predicted optimistic zymogram and found that the experimental reduced sugar release amount and improved dry matter digestibility were 219.26 mg/g and 6.31% respectively, whose errors to the predicted optimistic reducing sugar release amount and the improved dry matter digestibility were 1.05% and 1.02%. To sum up, the predicted optimal zymogram of 6 carbohydrases in the present study were capable to improve the starch digestibility in diet for broilers aged 1 to 3 wk, which were represented by increased reduced sugar release amount and improved dry matter digestibility. [ABSTRACT FROM AUTHOR]

The efficacy of exogenous carbohydrases in pig diets has been suggested to depend on enzyme activity and dietary fiber composition, but recent evidence suggests other factors such as ambient temperature might be important as well. Therefore, we investigated the effect of heat stress (HS) on the efficacy of a multienzyme carbohydrase blend in growing pigs. Ninety-six (barrows: gilts; 1:1) growing pigs with initial body weight (BW) of 20.15 ± 0.18 kg were randomly assigned to six treatments, with eight replicates of two pigs per pen in a 3 × 2 factorial arrangement: three levels of carbohydrase (0, 1X, or 2X) at two environmental temperatures (20 °C or cyclical 28 °C nighttime and 35 °C day time). The 1X dose (50 g/tonne) provided 1,250 viscosimetry unit (visco-units) endo-β-1,4-xylanase, 4,600 units α-l-arabinofuranosidase and 860 visco-units endo-1,3(4)-β-glucanase per kilogram of feed. Pigs were fed ad libitum for 28 d and 1 pig per pen was sacrificed on day 28. There was no enzyme × temperature interaction on any response criteria; thus, only main effects are reported. Enzyme treatment quadratically increased (P < 0.05) BW on day 28, average daily gain (ADG) (P < 0.05), and average daily feed intake (ADFI) (P < 0.05) with the 1X level being highest. HS reduced the BW at day 14 (P < 0.01) and day 28 (P < 0.01), ADG (P < 0.01), and ADFI (P<0.001). There was a trend of increased feed efficiency (G:F) (P < 0.1) in the HS pigs. HS increased apparent jejunal digestibility of energy (P < 0.05) and apparent ileal digestibility of calcium (P < 0.01). At day 1, HS reduced serum glucose (P < 0.001) but increased nonesterified fatty acid (P < 0.01). In the jejunum, there was a trend of increased villi height by carbohydrases (P < 0.1), whereas HS reduced villi height (P < 0.05). HS increased the jejunal mRNA abundance of IL1β in the jejunum (P < 0.001). There was a trend for a reduction in ileal MUC2 (P < 0.1) and occludin (P < 0.1) by HS, and a trend for increased PEPT1 (P < 0.1). There was no effect of HS on alpha diversity and beta diversity of the fecal microbiome, but there was an increase in the abundance of pathogenic bacteria in the HS group. In conclusion, HS did not alter the efficacy of carbohydrases. This suggests that carbohydrases and HS modulate pig performance independently. [ABSTRACT FROM AUTHOR]

The modern context in which broiler nutrition is inserted makes it possible to formulate diets containing commercial enzymes in order to improve the metabolizability of nutrients and the utilization of food of plant origin when acting on the breakdown of phytates and non-starch polysaccharides (NSP’s) soluble and insoluble. Enzymes acting on these substrates, phytase and xylanase, respectively, are not produced or have insignificant endogenous production by birds. The effects of phytase and carbohydrases on antinutritional factors can be considered additive since the phytase efficiency added to the diet can be reduced by the restricted access to the substrate in the presence of NSP’s. Similarly, insufficiency of phytase could reduce the action of carbohydrases on nutrients that may be complexed to the phytate molecule. Thus, the association between these enzymes in broiler feed can contribute to increasing yields and favoring the reduction of costs by the producer by providing the same performance for birds fed a traditional diet, as well as minimizing the environmental impacts due to the better metabolizability of the nutrients of the diets. Thus, this review was developed with the objective of approaching the use of phytase and xylanase in the diet of broilers [ABSTRACT FROM AUTHOR]