Carbohydrase Enzymes Research Articles

Phenolic Content, Antioxidant and Antimicrobial Properties of Hawthorn (Crataegus orientalis) Fruit Extracts Obtained via Carbohydrase-Assisted Extraction

The enzyme-assisted approaches for plant phenolics extraction are more eco-friendly methods compared to acid or alkaline hydrolysis. Carbohydrase enzymes can release free phenolics from plant materials by cleaving the glycosidic bonds between phenolic compounds and cell wall polymers. In this study, the efficiency of carbohydrase-assisted treatment approaches was evaluated to extract bioactive phenolics from hawthorn (Crataegus orientalis) fruit residues. Enzymatic treatment of the fruits was operated by using a crude cellulolytic enzyme cocktail from Rhizomucor miehei NRRL 5282 and a pectinase preparate from Aspergillus niger. Both cellulase and combined cellulase–pectinase treatments improved the total phenolic content (TPC) and antioxidant activity of extracts. The TPC increased to 1899 ± 27 mg gallic acid equivalents/100 g dry matter during the combined enzyme treatment, showing a strong correlation with the average antioxidant capacity determined by ferric-reducing antioxidant power (1.7-fold increment) and 2,2-diphenyl-1-picrylhydrazyl (1.15-fold increment) reagents. The major phenolics in enzyme-treated extracts were vanillic and ferulic acids, the concentrations of which increased 115.6-fold and 93.9-fold, respectively, during carbohydrase treatment. The planktonic growth of Escherichia coli, Bacillus subtilis, Pseudomonas aeruginosa, and Chromobacterium violaceum was slightly inhibited by the extracts with minimum inhibitory concentration values between 15.0 and 77.9 mg/mL, while the yeasts tested were quite resistant to the samples. B. subtilis and yeast biofilms were sensitive to the enzyme-treated extracts, which also showed quorum-sensing inhibitory effects against C. violaceum. The obtained bioactive hawthorn extracts hold potential as a natural source of antioxidants and antimicrobials.

Dietary manipulation through carbohydrase enzyme supplementation for broiler production in India

Dietary manipulation through carbohydrase enzyme supplementation for broiler production in India

Carbohydrase enzyme and polyethylene glycol altered the chemical composition, growth performance and apparent nutrient digestibility of Prosopis juliflora pods meal diets fed to Boschveld chicken breed

This study was designed to evaluate how treating Prosopis juliflora pods with polyethylene glycol and carbohydrase enzyme will affect its chemical composition and how the inclusion of treated P. juliflora pods in Boschveld chicken's diets will affect growth performance and nutrient digestibility. Before P. juliflora pods were administrated to the diets, they were pre-treated with enzyme, PEG and a combination of both and analysed for their nutrient content. Five diets, namely PJPM1: commercial standard pullet grower diet as a control; PJPM2: commercial standard grower diet with untreated 30 % of P. juliflora; PJPM3: commercial standard grower diet with PEG treated 30 % of P. juliflora; PJPM4: commercial standard grower diet with carbohydrase enzyme treated 30 % of P. juliflora; PJPM5: commercial standard grower diet with PEG and carbohydrase enzyme treated 30 % of P. juliflora. A total of 300 six-week-old Boschveld chickens were used in this trial. After 42 days of feeding trial, three Boschveld chickens per replicate (n = 90) were randomly selected for the determination of nutrient digestibility. Results on the chemical composition of raw P. juliflora pods meal, pre-treated with enzyme, PEG and combination of both show significant difference (P <0.05) in CP, Ash, NDF, CF, EE, Hemicellulose, NFC and NFE. Only overall feed intake shows significant different (P <0.05) in all dietary treatments and all the measured parameters of nutrient digestibility. Enzyme and PEG positively contribute to the chemical composition, growth performance, and nutrient digestibility of diets containing P. juliflora fed to Boschveld chickens.

PSVI-29 Effect of pig weight and exogenous enzyme supplementation on ileal endogenous amino acid losses and standardized ileal amino acid digestibility of soybean meal produced from Kentucky-grown soybeans

Abstract There is increasing interest in using locally sourced soybeans and soybean meal. Ileal amino acid digestibility (IAAD) of soybean meal from Kentucky-grown soybeans was evaluated in 25- and 50-kg cannulated pigs. The experiment used sixteen pigs in a randomized complete block design consisting of four treatments with 4 pigs per treatment. This was repeated in time resulting in eight replicates per treatment for both body weight (BW)groups. After cannulation at about 20 kg BW, all pigs were allowed to recover from the surgery and were fed a corn-SBM-based diet that met all nutrient and energy requirements. The experimental diets consisted of two nitrogen-free diets [NFD] with or without exogenous enzyme supplementation (phytase and a combination of phytase and carbohydrase enzymes) and two semi-purified diets containing soybean meal from Kentucky-grown soybeans as the only source of protein with or without exogenous enzyme supplementation. Each diet was fed for 7 consecutive d with 10-h ileal digesta collection on d 6 and 7. The same diets were fed to the same set of pigs at about 50 kg BW. Data were analyzed as a factorial arrangement of treatments (BW x diet) for ileal endogenous amino acid losses (IEAAL), apparent IAAD and standardized IAAD (SIAAD). Data from the NFD were used to determine the SIAAD from the AIAAD values. Exogenous enzyme supplementation increased (P &amp;lt; 0.05) IEAAL except for Gly and Pro while samples collected during phase 2 had greater (P &amp;lt; 0.05) IEAAL (10 amino acids). Pig BW did not influence IEAAL. Enzyme supplementation did not increase AIAAD except for Arg, Ala, Gly, N, and total AA while apparent ileal N, Met, Thr, and Val digestibility was greater (P &amp;lt; 0.05) in 50 kg pigs. Interactions between BW and diets showed that 50 kg pigs fed the SBM diet containing enzyme supplementation had greater (P &amp;lt; 0.05) N, Arg, and total AA digestibility compared with 50 kg pigs without enzyme supplementation. Digestibility of Val and Asp was greater (P &amp;lt; 0.05) in samples collected during phase 2. After correction for basal IEAAL, 50 kg pigs had greater (P &amp;lt; 0.05) SIAAD values for 12 of the AA analyzed while exogenous enzyme supplementation resulted in greater (P &amp;lt; 0.05) SIAAD for 9 of the AA analyzed. The SIAAD values for four AA (His, Phe, Val, and Asp) were greater (P &amp;lt; 0.05) for samples collected in phase 2. Results from this study showed that exogenous enzyme supplementation increased IEAAL and that IAAD of SBM produced from Kentucky-cultivated soybeans was high and that exogenous enzyme supplementation improved the SIAAD values of several amino acids. The IAAD values of SBM produced from soybeans cultivated in KY was high and are similar to values reported in the literature.

Biochemical characteristics of carbohydrase and serine protease enzymes of beet moth, Scrobipalpa ocellatella (Lepidoptera; Gelechiidae)

Biochemical characteristics of carbohydrase and serine protease enzymes of beet moth, Scrobipalpa ocellatella (Lepidoptera; Gelechiidae)

Evaluation of a carbohydrase (xylanase and glucanase) enzyme complex in diets for Florida pompano Trachinotus carolinus

AbstractFlorida pompano, Trachinotus carolinus, can be fed soy‐based, low‐animal protein diets. However, anti‐nutritional factors and carbohydrates limit digestibility. Dietary carbohydrase enzymes have been used to improve carbohydrate digestibility in several species and may be beneficial for Florida pompano. The present study was designed to evaluate the effects of using a carbohydrase enzyme complex (Natugrain® TS L, BASF Corporation, Ludwigshafen, Germany) composed of endo‐1,4‐ß‐xylanase and endo‐1,4‐ß‐glucanase (Xyl + Glu) on growth performance, feed utilization, and digestibility in diets fed to juvenile Florida pompano. A growth trial was conducted with iso‐nitrogenous (40%) and iso‐lipidic (8%) test diets containing 0, 0.015, 0.030, and 0.045% of Xyl + Glu. The trial was conducted in a recirculating system with 12, 1000‐L tanks. Twenty fish (mean weight: 23.34 g ± 1.49 g) were stocked into each tank in a randomized design. Growth results indicated that there were no significant differences (p &gt; 0.05) in growth and feed conversion; however, thermal growth coefficient displayed significant differences (p = 0.0499) between the 0.015 and 0.030% inclusion. All enzymatic treatments displayed significant differences (p = 0.029) in apparent digestibility of crude protein compared with the basal diet. Results indicate that Xyl + Glu enzyme complex can impact protein digestibility but did not significantly affect growth performance.

51 Application of an energy and nutrient matrix to a mixed cereals-based diet supplemented with a novel phytase and xylanase–β-glucanase achieved a production benefit from wean-to-finish in pigs

Abstract The effect of a novel consensus bacterial 6-phytase variant (PhyG) without or with a xylanase–β-glucanase (XB) when added to an energy and nutrient-reduced diet on growth performance and production benefit was evaluated in pigs from wean-to-finish. A total of 192 weaned pigs [DanBred x Pi; initial body weight (BW) 7.18 ± 0.38 kg] were randomized to 4 treatments with 12 (4 piglets/pen) and 24 pens (2 pigs/pen) per treatment (1:1 male to female) for nursery and grow-finish phases, respectively. Diets were wheat-corn-barley-rye-soybean meal-based and fed in 5 phases (pre-starter, 7–11 kg, starter, 11–25 kg, grower I, 25–55 kg, grower II, 55–85 kg and finisher, 85–110 kg BW). Treatments comprised: 1) a nutritionally adequate positive control (PC); 2) a negative control (NC1) without added inorganic phosphate, reduced (based on specific PhyG dose) in net energy (NE), calcium (Ca), digestible amino acids (AA) and sodium (Na) vs. PC, supplemented with PhyG at 1,000, 1000, 750, 500 and 500 FTU/kg during pre-starter, starter, grower I, II and finisher phase, respectively (NC1+PhyG ‘low’); 3) a negative control (NC2) without added inorganic phosphate, supplemented with PhyG at 2,000, 2,000, 1,000, 750 and 750 FTU/kg in each phase with reduction in NE, Ca, dig AA and Na vs. PC based on the specific PhyG dose (NC2+PhyG ‘high’); and 4) a negative control (NC3) that was similar to 2) but supplemented with XB to provide 2,400 U/kg xylanase and 304 U/kg β-glucanase, with additional reduction in NE and dig AA based on the contribution of XB vs NC1 (NC3+PhyG ‘low’+XB). Data were analyzed by ANOVA, and means separation was by Tukey’s HSD test. Growth performance in all enzyme-supplemented diets was maintained or improved compared with PC. The NC3+PhyG ‘low’+XB treatment significantly improved performance vs. PC; average daily gain (ADG) and BW were greater vs. PC in starter (620 g and 28.2 kg vs. 557 g and 26.4 kg) and grower I (859 g and 65.1 kg vs. 777 g and 60.0 kg) respectively; feed conversion ratio (FCR) was reduced in grower II vs. PC (2.10 vs. 2.25; P &amp;lt; 0.05). The final BW was 114.5, 114.8, 115.6 and 116.7 kg in PC, NC1+PhyG ‘low’, NC2+PhyG ‘high’ and NC3+PhyG ‘low’+XB, respectively (P &amp;gt; 0.05). Carcass parameters were unaffected by treatments. Compared with PC, NC1+PhyG ‘low’, NC2+PhyG ‘high’ and NC3+PhyG ‘low’+XB conferred a 3.8 to 5.8% reduction in total feed cost (ingredients price, October 2023)/kg BWG and 4.4 to 6.8% reduction in the total carbon footprint (FeedPrint, WUR, The Netherland, CO2 eq., g)/kg BWG. The results confirm the appropriacy of the applied matrix values with PhyG supplementation, with or without XB, in mixed cereal diets from wean-to-finish and highlight the potential feed cost and sustainability benefit of using the phytase alone or in combination with the carbohydrase enzymes.

PSV-21 Pediococcus acidilactici survives an in vitro simulated digestion

Abstract In the swine industry, probiotics are recognized for optimizing growth performance and production efficiency by modulating host physiology, and the response across studies is not consistent. One area of focus is probiotic mitigation of post-weaning diarrhea, and further mechanistic understanding of probiotic and host physiology could help with consistent performance outcomes. An essential attribute of a probiotic is the ability to survive the harsh environments of the gastrointestinal tract (GIT). The aim of this study was to determine if Pediococcus Acidilactici PECh3A (PA) that has been previously genome sequenced for determining gut physiology mechanisms can survive an in vitro simulated digestion. In the in vitro digestion PA concentrations that were investigated included 108, 109, and 1010 colony forming units (CFU)/mL. The in vitro digestion (n = 4/concentration) consisted of 5 phases with incubation times simulating the GIT; initial inoculation (0 h), gastric (2 h), upper intestinal (4 h), lower intestinal (9 h) and extended lower intestinal phase (9 h). For the initial inoculation concentrations of PA were added into 25 mL of sterile milk replacer representing dietary requirements of a suckling pig. In the gastric phase pepsin was added and pH was adjusted to 2.5. In the upper intestinal phase bile and pancreatin were added, along with a pH adjustment to 6.8. In the lower intestinal phase, the multi-enzyme complex consisting of carbohydrase enzymes was added and pH adjusted to 4.8. There was no addition during the extended lower intestinal phase. At each sample collection at the end of a phase 1 mL was collected from each flask. For each sample, 100 µL were serially diluted, plated on deMan, Rogosa, and Sharpe (MRS) plates, and incubated at 37°C for 24h. Survivability was determined by counting CFUs and comparing the concentration with the initial CFU/mL at 0 h and reported as percent survival (%S). Data for the in vitro digestion were analyzed according to a randomized complete block design (RCBD) in a split plot using the Repeated Measures Glimmix Procedure of SAS. Additional orthogonal contrasts were analyzed across PA concentrations. Data showed survival for 108,109, and 1010CFU/mL at all sample phases (108 - 44.24, 60.14, 58.36, 55.69%S; 109 - 40.56, 55.60, 49.35, 50.7%S; 1010 - 32.98, 52.55, 46.72, 43.71%S ± 2.98 respectively; phase P &amp;lt; 0.01 and concentration P &amp;lt; 0.01). There is evidence of a phase and concentration effect on survivability, but no interaction. A linear concentration response was observed across concentrations (54.61, 49.05, 43.99%S ± 1.23; P &amp;lt; 0.05) indicating that as concentration increases the survivability decreases. These findings indicate that PA PECh3A can survive a simulated GIT and potentially impact swine intestinal physiology.

Magnetic Multienzyme@Metal-Organic Material for Sustainable Biodegradation of Insoluble Biomass.

Biodegradation of insoluble biomass such as cellulose via carbohydrase enzymes is an effective approach to break down plant cell walls and extract valuable materials therein. Yet, the high cost and poor reusability of enzymes are practical concerns. We recently proved that immobilizing multiple digestive enzymes on metal-organic materials (MOMs) allows enzymes to be reused via gravimetric separation, improving the cost efficiency of cereal biomass degradation [ACS Appl. Mater. Interfaces 2021, 13, 36, 43085-43093]. However, this strategy cannot be adapted for enzymes whose substrates or products are insoluble (e.g., cellulose crystals). Recently, we described an alternative approach based on magnetic metal-organic frameworks (MOFs) using model enzymes/substrates [ACS Appl. Mater. Interfaces 2020, 12, 37, 41794-41801]. Here, we aim to prove the effectiveness of combining these two strategies in cellulose degradation. We immobilized multiple carbohydrase enzymes that cooperate in cellulose degradation via cocrystallization with Ca2+, a carboxylate ligand (BDC) in the absence and presence of magnetic nanoparticles (MNPs). We then compared the separation efficiency and enzyme reusability of the resultant multienzyme@Ca-BDC and multienzyme@MNP-Ca-BDC composites via gravimetric and magnetic separation, respectively, and found that, although both composites were effective in cellulose degradation in the first round, the multienzyme@MNP-Ca-BDC composites displayed significantly enhanced reusability. This work provides the first experimental demonstration of using magnetic solid supports to immobilize multiple carbohydrase enzymes simultaneously and degrade cellulose and promotes green/sustainable chemistry in three ways: (1) reusing the enzymes saves energy/sources to prepare them, (2) the synthetic conditions are "green" without generating unwanted wastes, and (3) using our composites to degrade cellulose is the first step of extracting valuable materials from sustainable biomasses such as plants whose growth does not rely on nonregeneratable resources.

The Comparative Effects of Supplementing Protease Combined with Carbohydrase Enzymes on the Performance and Egg n-3 Deposition of Laying Hens Fed with Corn-Flaxseed or Wheat-Flaxseed Diets.

Flaxseed contains huge quantities of anti-nutritional factors (ANFs), which reduce the performance of livestock. Three different protease and multi-carbohydrase enzymes were included in wheat-flaxseed diets (WFD) and corn-flaxseed diets (CFD) to compare their effects on performance, egg n-3 deposition, and fatty acid transporter genes in laying hens. A total of 540, twenty-week-old, Nongda-3 laying hens (DW brown × Hy-line white) were randomly assigned to six dietary groups, including 10% WFD or 10% CFD plus (i) supplemental enzyme A (alkaline protease 40,000 and neutral protease 10,000 (U/g)), (ii) enzyme B (alkaline protease 40,000, neutral protease 10,000, and cellulase 4000 (U/g)), or iii) enzyme C (neutral protease 10,000, xylanase 35,000, β-mannanase 1500, β-glucanase 2000, cellulose 500, amylase 100, and pectinase 10,000 (U/g)). An interaction (p < 0.05) was found for egg mass, hen day of egg production, and feed conversion ratio on the 9-10th week of the experiment. The WFD with enzyme B was associated with the highest egg weight in the 9-10th week. The deposition of total n-3 was superior with WFD (468.22 mg/egg) compared to CFD (397.90 mg/egg), while addition of enzyme C (464.90 mg/egg) resulted in the deposition of more total n-3 compared to enzymes A and B (411.89 and 422.42 mg/egg). The WFD and enzyme C significantly (p < 0.001) enhanced docosahexaenoic acid (DHA) and reduced the n-6:n-3 ratio in egg yolk compared to the CFD. The hepatic mRNA expression of liver fatty acid binding protein (L-FABP) (p = 0.006), fatty acid desaturase 1 (FADS-1) (p < 0.001), elongase-2 (ELOV-2) (p < 0.001), fatty acid transport protein-1 (FATP1) (p < 0.001), and the intestinal mRNA expression of FATP and FABP genes were increased with WFD compared to CFD. In conclusion, WFD with enzyme C is favorable for optimal performance, results in the deposition of more n-3 and DHA, and increases the expression of fatty acid transporter genes, which helps in n-3 transport.

287 The Effect of Greater Inclusion Levels of Ddgs with or without a Syncra SWI in Swine Grower-Finisher Diets on Performance and Economics

Abstract To evaluate the effect of greater inclusion levels of DDGS with or without a Syncra SWI (SWI; bacillus plus protease; IFF, USA) in swine grower-finisher diets, a trial was conducted consisting of 528 pigs across 6 treatments with 8 replicates consisting of 11 pigs each where weights were balanced at the start of the trial. Inclusion levels of DDGS were 0%, 15%, and 30% all containing phytase and carbohydrase enzymes. Each inclusion level was split to include or not include SWI. At the end of the trial, slaughter weights were measured. While no significant differences were detected, the heaviest slaughter weights were observed in the 0% DDGS diet, 30% DDGS diet with SWI and the 0% DDGS diet with SWI (115.28, 115.16, and 115.15 kg, respectively). For total body weight gain, there were no significant differences between both 0% DDGS diets, the 15% DDGS diet without SWI, and both 30% diets. However, the 15% diet with SWI was significantly lighter than both 0% DDGS diets and the 30% DDGS with SWI diet (P = 0.01, 0.04, 0.008 respectively). Similarly, no significant differences were found between both of the 0% DDGS diets, the 15% DDGS diet without SWI, or both 30% DDGS diets on average daily gain. A significant difference was detected between the 15% DDGS with SWI and both 0% DDGS diets and the 30% DDGS diet with SWI (P = 0.002, 0.0003, and 0.001, respectively) on average daily gain. The lightest cold carcass weight was observed in pigs fed the 15% DDGS diet with SWI, which was significantly (P &amp;lt; 0.05) different from all other diets, while the heaviest cold carcass weight was observed in pigs fed the 0% DDGS diet with SWI (87.22 kg) followed by the 30% DDGS diet with SWI (86.53 kg) but were not significantly different. While there were no significant differences detected in the economics of the trial, the feed with the greatest cost was the 0% DDGS diet with SWI ($299.98) while the least cost was demonstrated in the 30% DDGS with SWI diet ($288.80). The diet that generated the highest revenue was the 0% DDGS diet with SWI ($124.89) followed by the 30% DDGS diet with SWI ($123.94) while the least revenue was seen in the 15% DDGS diet with SWI ($119.65). Margin over feed cost was greatest in the 15% DDGS diet ($66.87) and the 30% DDGS diet with SWI ($66.16) and the least MOFC was seen in the 0% DDGS diet. While no significant differences were detected in performance between the 0% DDGS diets and the 30% DDGS diets, these data shows there is a potential benefit to feeding greater inclusion DDGS diet if an SWI is added.

The effect of enzymatically-modified canola meal on growth performance, nutrient utilization, and gut health and function of broiler chickens

Enzymatically-modified canola meal (ECM) is a product obtained from the incubation of canola meal (CM) with a blend of specific carbohydrase enzymes. Incubation of CM with multi-carbohydrase enzymes resulted in a significant decrease in non-starch polysaccharides (NSP) content, production of potentially bioactive NSP hydrolysis products, and the spontaneous lactic acid bacteria fermentation. The latter resulted in the production of lactic acid, phytate hydrolysis, and the reduction in the product pH from 5.9 to 4.0. The main objective of this study was to evaluate the effects of ECM on growth performance, nutrient utilization, and gut health and function of broiler chickens. Four-day-old as hatched Ross 308 broilers were assigned to 5 dietary treatments, each consisting of 8 cages of 8 birds. Birds were fed the starter (4–14d of age, 100 g/kg CM) and grower (14–21d of age, 200 g/kg of CM) Control diet, and Control diets supplemented with ECM (Low and High) or enzymes (E1 and E2). In the ECM-containing diets, ECM replaced either 50 g/kg (Low ECM diet) or entire CM (High ECM diet) of the Control diets. In the E1 and E2 diets, the enzyme blend was supplemented at 1.25 g/kg and 12.5 g/kg of the starter Control diet and 2.5 g/kg and 25 g/kg of the grower Control diet, respectively. Replacing CM with ECM did not affect growth performance, but significantly increased the apparent total tract NSP digestibility and nitrogen-corrected apparent metabolizable energy content. The ECM-containing diets decreased sialic acids secretion on day 21 and increased the abundance of Lactobacillus (P < 0.01) in ileum and ceca compared to Control, E1, and E2 during the whole trail. The abundance of Escherichia coli was significantly lower in ceca of birds fed the High ECM grower diets compared to the Control. Total short chain fatty acids in cecal digesta of birds fed High ECM starter diet significantly increased compared with the Control. Replacing CM with ECM improved nutrient digestibility and increased the presence of probiotic organisms which was not observed in the enzyme supplemented diets. Therefore, ECM can improve gut health and could be used as a valuable feed ingredient or additive.

In Vitro Digestion of Peanut Skin Releases Bioactive Compounds and Increases Cancer Cell Toxicity.

Peanut skin is a rich source of bioactive compounds which may be able to reduce the risk factors associated with metabolic syndromes. This study aimed to characterize bio-compounds from peanut skin (Arachis hypogaea) and their bioactivity (antioxidant activity, inhibition of lipase, and carbohydrase enzymes) and to evaluate their anti-proliferative properties in colorectal cancer cells (HCT116) upon in vitro digestion. Peanut skin was digested in two sequential phases, and the final content, named phase-1 (P1) and phase-2 (P2) extracts, was evaluated. Several bioactive compounds were positively identified and quantified by liquid chromatography, including quinic acid, released especially after in vitro digestion. The total phenolic content and, regardless of the method, the antioxidant activity of P1 was higher than P2. P1 also showed a lower enzyme inhibitory concentration IC50 than P2, lipase, and α-glucosidase. For cell viability in HCT116 cells, lower concentrations of P1 were found for IC50 compared to P2. In conclusion, bioactive compounds were released mainly during the first phase of the in vitro digestion. The digested samples presented antioxidant activity, enzyme inhibitory activity, and cancer cell cytotoxicity, especially those from the P1 extract. The potential applications of such a by-product in human health are reported.

Effects of carbohydrase and phytase enzymes supplementation within low energy diets on performance and energy utilization of broiler chickens

Two experiments were conducted to evaluate the effects of different enzymes on the performance and gut morphology of broilers fed reduced energy diets, metabolizable energy values of reduced energy level diets. In experiment I - 1,120 male Cobb broilers, aged 1 to 42 days old, were used to evaluate performance parameters and gut morphology. Broilers were distributed into 7 treatments with 8 replicates of 20 broilers each. The treatments consisted of: T1 - positive control (PC) diet with 3050 kcal ME/kg; T2 - negative control (NC) diet with a reduction of 200 kcal ME/kg; T3 - NC + Carbohydrase I (100 g/ton endo-1,4-beta-mannanase); T4 - NC + Carbohydrase II (100 g/ton endo-1,4-beta-xylanase and endo-1,4-beta-glucanase); T5 - NC + phytase (50 g/ton phytase-6); T6 - NC + Carbohydrase II + phytase; T7 - NC + Carbohydrase I + Carbohydrase II + phytase. The evaluated variables were body weight gain (BWG), feed intake (FI), feed conversion ratio (FCR), villus height (VH), crypt depth (CD), and villus height: crypt depth ratio (VH: CD). In experiment II – 360 14-day-old broilers were distributed into six treatments with ten replicates of` six broilers each, to determine the apparent metabolizable energy (AME) and corrected for nitrogen balance (AMEn) of the diets. The same diets were used as treatments, excluding the PC. Comparing the treatments with the NC group, birds fed diets containing enzymes showed better BWG and FCR from 1 to 21 days old (P ≤ 0.05), and better FCR from 1 to 42 days (P ≤ 0.05). Additionally, the addition of Carbohydrase II and phytase promoted an increase in AME and AMEn of feed (P ≤ 0.05). Broilers fed with phytase showed higher VH when compared with the other treatments (P ≤ 0.05), whereas the treatments with phytase and Carbohydrases I, II, and phytase combination showed a better VH:CD (P ≤ 0.05). The inclusion of enzymes (carbohydrases and phytase) in broiler diets makes it possible to reduce feed energy levels without affecting performance parameters, AME corrected for nitrogen, and morphometry characteristics of the jejunum.

Extraction and Purification of Pullulanase from Local Mushrooms

Pullulanase is defined as an extracellular carbohydrase enzyme responsible for the hydrolysis of pullulan into maltotriose. Pullulanase can be extracted from Agaricus bisporus edible mushroom and concentrated by dialysis tube using Tris-HCl buffer. This enzyme was purified by different ratios of ammonium sulphate; the optimal ratio was 30%. It can be purified by ion exchange using a DEAE-cellulose column with a final yield of 5.25 and a purification fold of 4.38. Following Sephadex G-100 gel filtration with a yield of 2.60 and purification fold of 5.0, the total activity of the enzyme reached 160IU and the specific activity was 14.5 U/mg. This study was concerned with estimating the optimum pH and temperature of pullulanase, and it recorded an optimal pH of 7, and the optimal temperature was 60°C. The enzyme was more stable at pH 8 and 70°C; the incubation period was also determined, and it appeared the most appropriate period was 30 minutes. CaCl2 and ZnCl2 were activator metal ions. Mercaptoethanol in different concentrations was the greatest inhibitor of pullulanase. One of the problems arising in the surrounding area is that much of the environmental pollution consists of starchy food waste. Pullulanase can degrade the glycosidic linkage of pullulan or starch present in the starch waste. The objective of this study is to produce enzymes like pullulanases to participate in minimizing environmental pollution through the degradation of starch waste as a biotechnological application. The conclusion of this study is that pullulanase can be produced from readily available, safe, and low-cost sources such as mushrooms.

Effect of temperature and some factors on gut α-amylase of the desert locust; Schistocerca gregaria Forsk. (Orthoptera:Acrididae)

Gut of 4th nymphal instar of the desert locust; Schistocerca gregaria Forsk. was analyzed to study the effect of ambient temperature and some factors controlling α-amylase activity ( in vitro ) as a key carbohydrase enzyme for carbohydrate digestion. The study might provide a basis for locust control in different environments conditions. Crude gut enzymes as α-amylase, cellulase, invertase and trehalase were extracted by gut homogenization and differential centrifugation up to 100,000 Xg. Invertase had the highest activity in the gut followed by amylase and trehalase, while cellulase had the lowest one. Increasing ambient temperature from 10 to 37 °C enhanced, in vivo , α-amylase activity which declined when environmental temperature increased more than 37 °C. In vitro experiments revealed that α-amylase. crude protein extract showed optimal activity at pH 6, 45 °C and 17.9mg/ml soluble starch (substrate) concentration. Amylase was thermostable up to 50 °C, while it did not able to keep its major activity at 70 °C. The enzyme was more stable when exposed to acidic than to alkaline media.V max of α-amylase was 3.73 μmol glucose min −1 mg protein −1 . The results revealed that gut α-amylase had a relatively low affinity to soluble starch (high K m value), and low activation energy (0.451 KJ mol −1 ). Relatively high ambient temperature reduced α-amylase that might affect carbohydrates digestion and reduce locust nutrition.

The effect of multi-carbohydrase preparations on non-starch polysaccharides degradation and growth performance of broiler chickens fed diets containing high inclusion level of canola meal

Although canola meal (CM) contains a significant amount of dietary fiber, including high concentrations of non-starch polysaccharides (NSP), it is still a valuable source of protein for poultry diets. Supplementation of CM-containing poultry diets with multi-carbohydrase enzymes is an effective approach to improve the nutrition value of CM. The enzymatic degradation of NSP will contribute to the production of NSP hydrolysis products, which may improve gut health and growth performance. A series of in vitro incubation studies was carried out to determine if various carbohydrase preparations can target NSP of CM. The most effective enzyme combination was studied further in a growth performance and NSP utilization trial (1–20 d of age) with Ross 308 broiler chickens. Broilers were assigned to 3 dietary treatments each consisting of 8 cages of 5 birds and were fed a control corn/CM-based starter and grower diets (Control) or diets supplemented with an enzyme blend at two levels: 0.3 and 1.5 g/kg. Starter (1–10d) and grower (11–20d) diets contained 270 and 290 g/kg of CM, respectively. In the in vitro studies, the highest NSP depolymerization was observed for a combination of two pectinase-like enzymes fortified with xylanase. No further improvement with other enzyme preparations was observed. In comparison to the Control diets, birds fed the diet supplemented with 1.5 g/kg of multi-carbohydrase cocktail showed higher (P < 0.05) body weight gain during the grower phase and the entire trial. The carbohydrase enzyme combination significantly increased the coefficient of apparent total tract NSP digestibility. Our research suggests that the administration of a carbohydrase cocktail in diets with high inclusion of CM can improve growth performance and NSP utilization in broiler chickens.

Evaluation of the precision-fed rooster assay for detecting effects of supplemental enzymes on metabolizable energy

The precision-fed rooster assay has been used extensively to determine nitrogen-corrected true metabolizable energy (TMEn) of feed ingredients for poultry. However, this assay has not generally been used to evaluate effects of supplemental enzymes for this purpose. Therefore, 2 precision-fed rooster assays were conducted to evaluate several different carbohydrase enzymes on TMEn for a corn/soybean meal diet, a pearled barley diet, and diets containing different inclusion levels of rye/corn. In both rooster assays, Single Comb White Leghorn roosters were fasted for 26 h and then crop intubated with either 25 or 30 g of the test diets, depending on the assay. Excreta were then collected quantitatively for 48 h after feeding. In the first rooster assay with 56 birds, 6 carbohydrase combinations and/or levels (xylanase/alpha-galactosidase were evaluated using a corn/soybean meal control diet. All carbohydrase additions either numerically or significantly (P < 0.05) increased TMEn and the mean increase for the enzyme treatments was 66 kcal/kg DM compared with the corn/soybean meal control diet. The second assay consisted of twenty dietary treatments; 120 roosters were crop-intubated with 25 g of diets that were composed of 100% barley, 100% rye, 50% rye: 50% corn, or 25% rye: 75% corn. The diets were fed with and without inclusion of 2 different levels of either β-glucanase, xylanase, or a multi-carbohydrase combination. Both β-glucanase and the multi-carbohydrase significantly (P < 0.05) increased TMEn of the 100% barley diet, with the multi-carbohydrase increasing it from 3,722 to 4,086 kcal/kg DM at the highest inclusion rate. The xylanase and multi-carbohydrase either numerically or significantly (P < 0.05) increased TMEn of the 100% rye diet, with the multi-carbohydrase increasing it from 3,581 to 3,909 kcal/kg DM at the highest inclusion rate. The magnitude of enzyme response decreased as the level of rye in the diets decreased. Overall, results of this study indicated that the precision-fed rooster assay can detect effects of enzymes, primarily carbohydase, on TMEn of diets containing corn/soybean meal, pearled barley, and/or rye.

Greenhouse gases and performance of growing pigs fed wheat-based diets containing wheat millrun and a multi-carbohydrase enzyme.

The objective of this project was to determine the impact of feeding growing pigs with high wheat millrun diets supplemented with a multi-carbohydrase enzyme (amylase, cellulase, glucanase, xylanase, and invertase activities) on nutrient digestibility, growth performance, and greenhouse gas (GHG) output (carbon dioxide, CO2; nitrous oxide, N2O; methane, CH4). Three experiments were conducted utilizing six treatments arranged as a 3 × 2 factorial (0%, 15%, or 30% wheat millrun; with or without enzyme) for the digestibility experiment or as a 2 × 2 factorial (0% or 30% wheat millrun; with or without enzyme) for the performance and GHG experiments. The digestibility, performance, and GHG experiments utilized 48 individually housed pigs, 180 pigs housed 5 per pen, or 96 pigs housed 6 per chamber, respectively. Increasing wheat millrun up to 30% in the diet of growing pigs resulted in decreased energy, nitrogen (N) and phosphorus (P) apparent total tract digestibility and net energy content (P < 0.01). Overall, average daily gain (ADG) and gain to feed ratio were reduced in pigs fed wheat millrun (P < 0.05). Enzyme supplementation had minimal effects on the digestibility or performance parameters measured. Feeding diets with 30% millrun did not affect GHG output (CH4: 4.7 and 4.9; N2O: 0.45 and 0.42; CO2: 1,610 and 1,711 mg/s without or with millrun inclusion, respectively; P > 0.78). Enzyme supplementation had no effect on GHG emissions (CH4: 4.5 and 5.1; N2O: 0.46 and 0.42; CO2: 1,808 and 1,513 mg/s without or with enzymes, respectively; P > 0.51). Overall, the carbohydrase enzyme had minimal effects on parameters measured, regardless of wheat millrun inclusion (P > 0.10). Although energy, N and P digestibility, and ADG were reduced, the inclusion of up to 30% wheat millrun in the diet has no effect on GHG emissions from growing pigs (P > 0.10).

Comparative study on yields and quality parameters of argan oils extracted by conventional and green extraction techniques

Argan oil is most frequently sold as pure oil, which can be directly applied topically due to its cosmetological proprieties or ingested in order to provide several health benefits. It's also commonly mixed into a number of cosmetic products like shampoos, soaps and conditioners. In this study we aimed to improve the argan oil extraction yield and quality parameters by comparing the effects of different extraction technologies. Argan kernel oils were extracted using four methods: mechanical cold press, Soxhlet extraction with n -hexane, supercritical fluid extraction (SFE), and enzyme assisted extraction with three different enzyme solutions cellulase (cellulast), pectinase (Pectinex) and a mixture of carbohydrase enzymes (Viscozyme®). The quality parameters was evaluated by determining the acid, peroxide and iodine values as well as the extinction coefficients K 232 and K 270 as measures of conjugated dienes and trienes, respectively. The results showed that the highest yield (66.37% ± 3.3%) was obtained by enzyme assisted extraction using the carbohydrases enzymes mixture (Viscozyme®), followed by Soxhlet extraction (59.5% ± 3.1%) and pectinase extraction (52.03% ± 3.55%). All argan oils samples obtained by the different methods showed a good oxidation stability, with acid, peroxide and iodine values lower than 0.8 mg/g, 15 meq/kg and 110 g/100 g according to the official argan oil norm, respectively. The results of argan oils quality parameters demonstrated that the enzyme extracted argan oils showed low oxidation, with the following quality parameters: acid values (0.4–0.6 mg/g), iodine values (95–100 g/100 g), dienes ( K 232 < 2), trienes ( K 270 < 0.35), and peroxide values (<1.5 meq/kg). The results proved that the enzyme assisted extraction method can be applied to improve the argan oil yield without affecting the oil quality. The enzyme extraction method may be a great alternative to solvent and cold press extractions for this eco-friendly processing approach.