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1.
Appl Biochem Biotechnol ; 190(1): 270-292, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31342308

RESUMO

The enzyme phytase has important applications in animal feed, because it favors the bioavailability of phosphorus present in phytate, an antinutritional compound widely found associated with plant proteins. However, for feed applications, the phytase must withstand high temperatures during the feed pelleting process, as well as the gastrointestinal conditions of the animal. This work evaluates the feasibility of immobilizing phytase on hydroxyapatite (HA) nanoparticles, in order to improve its properties. HA is a material with excellent physicochemical characteristics for enzyme immobilization, and it can also act as an inorganic source of phosphorus and calcium in animal feed. The strong affinity of the phytase for the support resulted in rapid adsorption, with total immobilization yield and recovered activity greater than 100%. After immobilization, the phytase showed a broader activity profile in terms of pH and temperature, together with considerably higher thermoresistance at 80 and 90 °C. As a proof of concept, it was shown that the phytase immobilized on HA presented good resistance to acidic conditions and resistance to proteolysis when passing through simulated gastrointestinal conditions of fish. The findings showed that phytase immobilized onto HA presents suitable properties and has great potential for use in animal feed.


Assuntos
6-Fitase/metabolismo , Ração Animal , Durapatita/química , Enzimas Imobilizadas/metabolismo , Nanopartículas/química , Animais , Peixes , Temperatura Alta , Concentração de Íons de Hidrogênio , Concentração Osmolar , Proteólise
2.
Food Chem ; 306: 125620, 2020 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-31606627

RESUMO

The lactic acid bacteria (LAB) and yeast strains with phytate degrading ability were isolated from Iranian traditional sourdough, and based on the acid and bile tolerance, three LAB and three yeast strains were selected and molecularly identified. In this study, baker's yeast (Saccharomyces cerevisiae) was considered as a positive control strain to investigate the nutritional and technological properties of the isolated strains. All of the identified microorganisms were characterized based on additional probiotic properties and were evaluated for nutritional and technological characteristics. The functional features are associated with degradation of phytate, antioxidant capacity, exopolysaccharides, phenolic compound content and in vitro starch digestion. Among all the tested strains the highest amount of phytase production capacity (1.64 Unit/ml) and lowest phytate content (17.49 mg/5 g) belonged to Kluyveromyces marxianus. According to the results, the bread prepared by using Kluyveromyces aestuarii possessed the highest porosity percentage (70.43%), and the lowest hardness (508.71 g).


Assuntos
Pão/análise , Lactobacillales/metabolismo , Ácido Fítico/metabolismo , Saccharomyces cerevisiae/metabolismo , Triticum/química , 6-Fitase/metabolismo , Pão/microbiologia , Irã (Geográfico) , Lactobacillales/isolamento & purificação , Probióticos , Saccharomyces cerevisiae/isolamento & purificação , Triticum/metabolismo
3.
J Agric Food Chem ; 67(41): 11396-11402, 2019 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-31537068

RESUMO

Phytase is commonly used as a feed enzyme in monogastric animals to increase the bioavailability of phytate phosphorus and other nutrients. The accumulation of myo-inositol phosphate intermediates during phytate degradation in various segments of the gastrointestinal tract (GIT) is poorly understood. The aim of this study was to determine the efficacy of Buttiauxella spp. phytase in degrading the phytate in corn, soybean meal, and complete corn-soybean meal diet to myo-inositol phosphate esters (IP1-IP5) and completely dephosphorylated myo-inositol rings using an in vitro model of the poultry upper GIT. Our results show that the phytase hydrolyzes phytate efficiently to small IP esters, whereas the myo-inositol level remains constant between control and phytase treatments. Although the in vitro digestion model does not incorporate all factors that govern phytate hydrolysis, it is a valuable tool for evaluating phytase efficacy at various enzyme doses and with different feed ingredients.


Assuntos
6-Fitase/química , Ração Animal/análise , Enterobacteriaceae/enzimologia , Ésteres/química , Fosfatos de Inositol/química , Ácido Fítico/química , 6-Fitase/metabolismo , Animais , Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo , Galinhas , Digestão , Ésteres/metabolismo , Aditivos Alimentares/química , Aditivos Alimentares/metabolismo , Trato Gastrointestinal/metabolismo , Hidrólise , Fosfatos de Inositol/metabolismo , Modelos Biológicos , Ácido Fítico/metabolismo , Soja/química , Soja/metabolismo , Zea mays/química , Zea mays/metabolismo
4.
Poult Sci ; 98(11): 5789-5800, 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31265114

RESUMO

The anti-nutritional effects of dietary inositol phosphates (IP6 through IP3) have been recognized in broiler chickens; however, inositol hexaphosphate (IP6) is more potent than the lower IP esters. The efficacies of 2 commercial phytases, a Buttiauxella sp. phytase (BSP) and a Citrobacter braakii phytase (CBP) at 500 and 1,000 FTU/kg, were studied on IP6-3 concentrations in the crop, proventriculus + gizzard, and distal ileum digesta, and ileal IP6 disappearance in broilers at day 22. Apparent ileal P and Ca digestibility, and bone quality at days 22 and 33 were also measured. Female Ross 308 broilers (n = 1,890; 30 birds × 7 diets × 9 replicates) were fed corn-soy-based crumbled diets. The 7 diets included a primary breeder recommendation-based positive control diet (PC); the PC marginally reduced in available P by 0.146% and Ca by 0.134% of the diet, (NC1) or moderately reduced by 0.174 and 0.159% of the diet, respectively (NC2). Other diets were the NC1 + BSP or CBP at 500 FTU/kg (NC1+500BSP and NC1+500CBP) and the NC2 + BSP or CBP at 1,000 FTU/kg (NC2+1,000BSP and NC2+1,000CBP). Each of the NC1 and NC2 had distal ileum IP6 disappearance similar to that of PC, but each had lower P digestibility and the majority of measured bone quality parameters than the PC. The ileal IP6 levels were decreased by 52.0 and 32.7% for NC1+500BSP and NC1+500CBP, respectively, relative to NC1 and by 73.6 and 50.9% for NC2+1,000BSP and NC2+1,000CBP, respectively, relative to NC2 (P < 0.001), with a similar effect for distal ileum IP6 disappearance. Overall, phytase in the NC diets increased P digestibility, and femur breaking strength and cortical bone mineral density at days 22 and 33. Overall, each of the phytases at each dose degraded IP6-3 across the gastrointestinal tract segments to increase P digestibility and the P and Ca utilization in bone. However, dietary BSP at 1,000 FTU/kg was most effective. Supplemental phytase degrades phytate to decrease the anti-nutritional effects in a dose- and phytase-dependent manner.


Assuntos
6-Fitase/metabolismo , Densidade Óssea/efeitos dos fármacos , Galinhas/fisiologia , Digestão/efeitos dos fármacos , Enterobacteriaceae/química , Fosfatos de Inositol/metabolismo , 6-Fitase/administração & dosagem , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal/efeitos dos fármacos , Animais , Proteínas de Bactérias/administração & dosagem , Proteínas de Bactérias/metabolismo , Cálcio na Dieta/metabolismo , Citrobacter/química , Dieta/veterinária , Suplementos Nutricionais/análise , Trato Gastrointestinal/metabolismo , Fósforo na Dieta/metabolismo , Especificidade da Espécie
5.
Poult Sci ; 98(10): 4729-4744, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31329980

RESUMO

In a 2 × 2 × 2 factorial design, the effects of dietary non-phytate phosphorus (NPP) levels, 0.17% (low) and 0.33% (moderate), diet moisture (dry and wet), and diet particle size (coarse and fine), were studied on egg production, characteristics of gastrointestinal tract (GIT) and tibia, digesta pH, and phytase activity in layer pullets (16 to 28 wk of age). The low NPP diet increased average daily water intake (ADWI) to ADFI ratio (4.2%) from 16 to 17 wk, but decreased this ratio (2.8%) from 23 to 27 wk. It decreased ADFI (1.5%) and egg mass production (3.8%) from 19 to 22 wk. It decreased egg weight (0.29 g) and ADWI (2.1%) from 23 to 27 wk. At 22 wk, the GIT relative empty organ weights were (g/kg BW) higher for proventriculus + gizzard (0.96), duodeneum (0.94), and jejunum + ileum (1.95) with the low vs. moderate NPP diet. The low NPP diet decreased digesta phytase activity in crop and proventriculus+gizzard at 28 wk. The wet diet increased ADFI, ADWI, and ADWI/ADFI ratio from 16 to 27 wk, egg mass production (3.0%) from 19 to 22 wk, and egg weight (0.45 g) from 23 to 27 wk. The wet diet also increased digesta phytase activity in proventriculus+gizzard. The coarse diet decreased ADFI from 19 to 22 wk (1.7%) and 23 to 27 wk (1.2%). The coarse diet caused reduced egg mass production (2.6%) from 23 to 27 wk. Egg shell breaking strength was increased on the coarse diet (0.9 Newton). The coarse diet increased ADWI/ADFI ratio from 16 to 27 wk, and increased relative gizzard weight by 1.95 and 0.81 g/kg BW at 22 and 28 wk, respectively. The coarse diet increased jejunal/ileal pH with 0.16 units at 28 wk. None of the tested parameters affected tibia characteristics. It was concluded that a low NPP diet did not clearly affect the studied parameters. The wet diet increased ADFI, ADWI, and egg production. The coarse diet increased ADWI, egg shell breaking strength, relative gizzard weight, and reduced ADFI and egg production.


Assuntos
6-Fitase/metabolismo , Galinhas/fisiologia , Trato Gastrointestinal/efeitos dos fármacos , Fósforo na Dieta/metabolismo , Fósforo/metabolismo , Tíbia/efeitos dos fármacos , Ração Animal/análise , Animais , Dieta/veterinária , Fezes/química , Feminino , Trato Gastrointestinal/fisiologia , Fósforo na Dieta/administração & dosagem , Tíbia/fisiologia
6.
Appl Microbiol Biotechnol ; 103(16): 6435-6448, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31254000

RESUMO

Phytases are important industrial enzymes able to catalyze the release of up to six phosphates from phytate in a stepwise hydrolysis reaction. Phytases are almost exclusively used as a feed supplement. However, phytases are also used in human nutrition, food processing, non-food industrial products, and emerging applications like enzymatic phosphate recovery from renewable resources. Phytate, the main phosphorus storage form in seeds, and its hydrolysis products act as a chelator and reduce protein and mineral bioavailability in intestinal absorption. Full phosphate hydrolysis from the common storage compound phytate remains a challenge. Phytate hydrolysis patterns of tailored phytases and their protein engineering campaigns are discussed. The aim of our review is to give an overview on developed and emerging application areas (animal nutrition, food processing, and environmental resource management) and thereby generate an awareness for the importance of phosphorus stewardship in a circular bioeconomy. Emphasis will be given to processes using organic-bound phosphorus and related recycling strategy of this valuable resource. In detail, the main challenge in designing phytases to completely hydrolyze phosphate from phytate to inositol and the need for engineering campaigns to broaden their industrial use are described.


Assuntos
6-Fitase/genética , 6-Fitase/metabolismo , Biotecnologia/métodos , Fosfatos/metabolismo , Ácido Fítico/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Humanos , Hidrólise , Engenharia de Proteínas/métodos
7.
Poult Sci ; 98(11): 5700-5713, 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31250002

RESUMO

The purpose of this study was to investigate the effects of phytase and protease supplementation on prececal (pc) amino acid (AA) digestibility, phytate (InsP6) degradation, and MEn concentration in diets using 3 oilseed meals as main protein sources in broiler chicken feed. The broiler chicken diets, which lacked mineral phosphorus, contained either soybean meal (SBM), SBM and rapeseed meal (SBM/RSM), or SBM and sunflower meal (SBM/SFM) as main protein sources. Diets were not supplemented with enzymes or supplemented with 1,500 or 3,000 FTU phytase/kg, or with 1,600 mg protease/kg. For diets containing SBM as the main protein source, the effects of phytase supplementation with and without monocalcium phosphate were also investigated. Data were obtained during 2 subsequent runs from days 14 to 22 and from days 23 to 31. Each diet was tested using 8 replicates with 4 replicates per run. For pc AA digestibility, no significant interactions were observed between main protein sources, enzyme supplementation, or addition of monocalcium phosphate except for Cys. Supplementation of 1,500 FTU phytase/kg increased pc digestibility of all AA. No differences in pc AA digestibility were observed between 1,500 and 3,000 FTU phytase/kg supplementation treatments. Prececal disappearance of InsP6 and pc P digestibility were greater in the high phytase supplementation treatment. Protease supplementation increased pc digestibility of all AA except for Cys when SBM/RSM was the main protein source. Supplementation of protease and 3,000 FTU phytase/kg increased MEn concentrations. The effect of phytase on pc AA digestibility was fully expressed at a lower supplementation level than needed for a maximized pc InsP6 disappearance and MEn concentration.


Assuntos
6-Fitase/metabolismo , Aminoácidos/fisiologia , Galinhas/fisiologia , Digestão/fisiologia , Peptídeo Hidrolases/metabolismo , Ácido Fítico/metabolismo , 6-Fitase/administração & dosagem , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Animais , Brassica rapa/química , Dieta/veterinária , Suplementos Nutricionais/análise , Intestinos/fisiologia , Peptídeo Hidrolases/administração & dosagem , Soja/química
8.
Poult Sci ; 98(11): 5525-5532, 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31180123

RESUMO

Two experiments were conducted to evaluate 3 novel proteases in broilers. In experiment 1, 600 male, Cobb 500 broilers were allocated to 1 of 12 experimental diets (5 birds/pen and 10 replicates/diet). A control (C) diet was formulated to be adequate in all nutrients. Proteases 1, 2, or 3 were added to this diet at 3 doses (1x, 3x, or 9x) in a 3 × 3 factorial arrangement of treatments. The factorial was augmented with 2 treatments of phytase at 500 or 1,500 FTU/kg added to the C diet. In experiment 2, 2,050 male Ross 308 broilers were allocated to 1 of 10 experimental diets (25 birds/pen and 9 replicates/diet). A C diet was formulated to be adequate in all nutrients. Protease 1, 2, or 3 was then added to the C diet at 3 doses (1x, 2x, or 4x) in a 3 × 3 factorial arrangement of treatments plus the C. In experiment 1, birds fed phytase gained more (P < 0.05) than birds fed protease, but neither were different than birds fed the C. Supplementation of 9x dose of any protease resulted in a reduction (P < 0.05) in BWG when compared with birds fed 1x dose of protease or phytase at 500 or 1,500 FTU/kg. Feed conversion ratio was improved (P < 0.05) in birds fed phytase compared with birds fed the C diet. Nitrogen digestibility was greater (P < 0.05) in birds fed protease 1 when compared with birds fed protease 2. Birds fed the 1x dose of protease or 500 FTU/kg of phytase had a greater (P < 0.05) N digestibility than birds fed 3x dose of protease. In experiment 2, protease supplementation significantly reduced (P < 0.05) BWG when compared with birds fed the C from hatch to 35 D post-hatch. Protease supplementation did not improve broiler growth performance or N digestibility above that of a nutrient adequate control diet or a diet supplemented with 500 FTU/kg of phytase.


Assuntos
6-Fitase/metabolismo , Galinhas/fisiologia , Proteínas na Dieta/metabolismo , Digestão/efeitos dos fármacos , Peptídeo Hidrolases/metabolismo , Fatores Etários , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal/efeitos dos fármacos , Animais , Galinhas/crescimento & desenvolvimento , Dieta/veterinária , Proteínas na Dieta/administração & dosagem , Suplementos Nutricionais/análise , Relação Dose-Resposta a Droga , Masculino , Nutrientes/fisiologia , Peptídeo Hidrolases/administração & dosagem , Distribuição Aleatória
9.
Poult Sci ; 98(11): 5562-5570, 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31189179

RESUMO

Gnotobiotic broiler chickens were used to study interactive effects of supplemented phosphorus, calcium (PCa), and phytase (Phy) on myo-inositol 1,2,3,4,5,6-hexakis (dihydrogen phosphate) (InsP6) degradation and release of myo-inositol in the digestive tract. In 2 subsequent runs, the chickens were subjected to 1 of 4 dietary treatments with and without PCa and Phy supplementation. Sanitized eggs were hatched in 8 germfree isolators, and a minimum of 9 male Ross 308 chickens were placed in each pen (total 16 pens). Treatments implemented on day 10 included gamma-irradiated diets without (PCa-; 4.1 g P and 6.2 g Ca/kg DM) or with (PCa+; 6.9 g P and 10.4 g Ca/kg DM) monosodium phosphate and limestone supplementation and without (Phy-) or with (Phy+) 1,500 FTU Phy/kg feed in a factorial arrangement. On day 15, digesta was collected from different sections of the intestinal tract and analyzed for InsP isomers and myo-inositol. The isolators did not remain germfree, but analysis of contaminants and results of InsP degradation indicated no or minor effects of the identified contaminants. Prececal InsP6 disappearance was 42% with the PCa-Phy- treatment and 17% with PCa+Phy-. No InsP3-4 isomers were found in the digesta of the terminal ileum in PCa-Phy-. The concentration of myo-inositol in the ileal digesta from PCa-Phy- (6.1 µmol/g DM) was significantly higher than that from PCa+Phy- (1.7 µmol/g DM), suggesting rapid degradation of the lower InsP isomers by mucosal phosphatases and their inhibition by PCa. Phytase supplementation increased InsP6 disappearance and prevented inhibitory effects of PCa supplements (72% in PCa-Phy+ and 67% in PCa+Phy+). However, PCa supplementation reduced the degradation of lower InsP isomers mainly in the posterior intestinal sections in the presence of Phy, resulting in significantly lower myo-inositol concentrations. It is concluded that mucosa-derived phosphatases might significantly contribute to InsP6 degradation in broiler chickens. The potential of mucosa-derived phosphatases to degrade InsP6 and lower InsP is markedly reduced by dietary PCa supplementation.


Assuntos
6-Fitase/metabolismo , Cálcio na Dieta/metabolismo , Galinhas/metabolismo , Vida Livre de Germes , Fósforo na Dieta/metabolismo , Ácido Fítico/metabolismo , Ração Animal/análise , Animais , Dieta/veterinária , Suplementos Nutricionais/análise , Masculino
10.
J Anim Sci ; 97(8): 3451-3459, 2019 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-31190064

RESUMO

Phytase is added to swine diets to improve the utilization of phytate-bound P in swine diets. This provides financial and environmental benefits to the pig industry. However, it is unclear if phytase works equally well in all dietary circumstances. The objective of this experiment was to determine if insoluble fiber affects the efficacy of the phytase enzyme in nursery pigs when fed diets limiting in P content. A total of 480 pigs (initial BW 5.48 ± 0.14 kg) were blocked by BW and randomly assigned (10 pigs per pen) to treatment within the block. A common nutrient-adequate diet was fed from days -14 to -5, and two basal P deficient diets (either a corn-soy diet containing 0.16% standardized total tract digestible [STTD] P [low insoluble fiber [LF]], or a corn-soybean meal plus 20% corn bran containing 0.14% STTD P [high insoluble fiber [HF]]) were fed from days -5 to 0 to acclimate pigs to a P deficient diet. From days 0 to 21, pigs received eight dietary treatments (six pens per treatment: n = 6). Experimental diets consisted of LF supplemented with one of four levels of added phytase (0, 109, 218, and 327 phytase units [FTU]/kg; Quantum Blue 5 G, AB Vista, Wiltshire, United Kingdom) expected to provide 0.16, 0.21, 0.26, and 0.31% STTD P, respectively, or HF supplemented with one of the same four levels of added phytase expected to provide 0.14, 0.19, 0.24, and 0.29% STTD P. Titanium dioxide was added to the diet at 0.4% as an indigestible marker. On day 21, one pig representing the average BW for each pen was euthanized, and fibulae were collected and analyzed for bone ash. Fecal samples were collected from each pen on days 19-20. Data were analyzed using PROC MIXED of SAS. There were no interactions between insoluble fiber and phytase for any of the variables evaluated. For days 0-21, adding phytase increased ADG (P < 0.001) with the response being linear (P < 0.001), whereas insoluble fiber decreased ADG (P = 0.033). There were no effects of phytase or insoluble fiber on ADFI (P = 0.381 and P = 0.632, respectively). Phytase improved G:F ratio (P < 0.001) with the response being linear (P < 0.001). Insoluble fiber tended to decrease G:F ratio (P = 0.097). Phytase increased bone ash (P = 0.005) with the response being linear (P = 0.001), but there was no effect of insoluble fiber (P = 0.949). Phytase did not affect the apparent total tract digestibility of DM, NDF, or ADF (P > 0.050), whereas insoluble fiber decreased the ATTD of DM (P < 0.001), NDF (P < 0.001), and ADF (P < 0.001). In conclusion, the addition of insoluble fiber did not affect the ability of phytase to improve growth performance and bone mineralization in nursery pigs fed a P deficient diet.


Assuntos
6-Fitase/metabolismo , Ração Animal/análise , Fibras na Dieta/farmacologia , Suplementos Nutricionais , Fósforo/metabolismo , Animais , Dieta/veterinária , Digestão , Fezes/química , Feminino , Trato Gastrointestinal/metabolismo , Masculino , Minerais , Fósforo/deficiência , Ácido Fítico/metabolismo , Distribuição Aleatória , Soja , Suínos , Zea mays
11.
Int J Mol Sci ; 20(10)2019 May 18.
Artigo em Inglês | MEDLINE | ID: mdl-31109025

RESUMO

Phytases are pro-nutritional enzymes that hydrolyze phytate and make associated nutrients, such as phosphorous, iron, and zinc, bioavailable. Single-stomached animals and humans depend on phytase supplied through the diet or the action of phytase on the food before ingestion. As a result, phytases-or lack thereof-have a profound impact on agricultural ecosystems, resource management, animal health, and public health. Wheat, barley and their Triticeae relatives make exceptionally good natural sources of phytase. This review highlights advances in the understanding of the molecular basis of the phytase activity in wheat and barley, which has taken place over the past decade. It is shown how the phytase activity in the mature grains of wheat and barley can be ascribed to the PAPhy_a gene, which exists as a single gene in barley and in two or three homeologous copies in tetra- and hexaploid wheat, respectively. It is discussed how understanding the function and regulation of PAPhy_a may support the development of improved wheat and barley with even higher phytase activity.


Assuntos
6-Fitase/genética , 6-Fitase/metabolismo , Hordeum/enzimologia , Hordeum/genética , Triticum/enzimologia , Triticum/genética , 6-Fitase/química , 6-Fitase/classificação , Fenômenos Bioquímicos , Ativação Enzimática , Regulação da Expressão Gênica de Plantas , Desenvolvimento Vegetal
12.
PLoS One ; 14(5): e0217490, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31125379

RESUMO

The objective of this study was to evaluate the effect of increasing doses of bacterial phytase (RONOZYME HiPhos) on performance and carcass characteristics of growing and finishing pigs. The study included 120 castrated males with initial weight of 23.21 ± 1.91 kg and 68 days of age, distributed in a randomized block design with five treatments and eight replicates with three animals each. The pigs were fed five corn-soybean meal-based diets: positive control (PC), supplemented with inorganic phosphorus and calcium; negative control (NC), with 0.13% reduction in available phosphorus and 0.11% in calcium; and three NC diets supplemented with 1,000, 2,000, and 3,000 phytase units (FYT)/kg in the feed. Compared with the NC diets without phytase, diets with 1,000, 2,000, and 3,000 FYT/kg inclusion increased the daily weight gain by +12% (quadratic, p<0.05) during the growing I period; +2.9, +2.9, and +10.5% (linear, p<0.01), respectively, during the growing II period; and +4.1, +5.1, and +8.2% (linear, p<0.001), respectively, over the entire experimental period. The daily feed intake increased by 0, +2.8, and +4.3% (linear, p<0.05), respectively, considering the entire experimental period; and the final live weight increased by +3.2, +4.2, and +6.1% (linear, p<0.001), respectively. The phytase treatments did not influence feed conversion ratio, carcass weight and yield, backfat thickness, loin depth and carcass lean meat. According to the European Carcass Classification (SEUROP), however, the animals fed the PC diet and the three phytase levels had more carcasses classified as E (between 55-60% lean meat) when compared to carcasses of pigs fed the NC. Supplementing increasing levels of phytase to a corn- and soybean meal-based diet with inorganic P and Ca reduction improved daily weight gain and feed intake of growing pigs, and such effects were maintained until slaughter age.


Assuntos
6-Fitase/metabolismo , Ração Animal , Cálcio na Dieta/metabolismo , Citrobacter/enzimologia , Fósforo na Dieta/metabolismo , Suínos/crescimento & desenvolvimento , 6-Fitase/administração & dosagem , Ração Animal/análise , Criação de Animais Domésticos , Fenômenos Fisiológicos da Nutrição Animal , Animais , Cálcio na Dieta/análise , Dieta , Masculino , Carne/análise , Fósforo na Dieta/análise , Suínos/fisiologia
13.
Poult Sci ; 98(10): 4848-4859, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31032860

RESUMO

Adequate dietary Ca and available phosphorus (avP) are essential to long-term egg production and bone health in laying hens. The effects of dietary Ca and avP levels and Buttiauxella sp. phytase (BSP) were studied in Lohmann LSL Lite hens from 30 to 70 wk of age (woa). Hens (n = 456; 4 per cage) were fed either a primary breeder recommendation-based diet (positive control; PC); the PC with avP and Ca levels reduced by 0.146 and 0.134% of the diet, respectively, without (NC) or with 300 FTU/kg BSP (NC+BSP). Egg production, BW, feed intake, FCR, and eggshell quality from 30 to 70 woa, and apparent ileal digestibility of P (AIDP) and Ca (AIDCa), and bone quality at 32, 48, and 70 woa were measured. The avP and Ca levels in the NC diet were not clinically deficient, as most parameters were unaffected by diet. Hen BW from 34 to 70 woa tended to be 2.9% greater (P = 0.076) for PC and NC+BSP compared to NC. Mid-diaphysis cortical bone mineral content (CBMC) tended to be 10% and 9% higher (P = 0.065) in the NC+BSP hens than in NC hens at 48 and 70 woa, respectively. AIDP of NC+BSP was 24% greater (P = 0.034) than of NC at 32 woa and tended to be 18% greater (P = 0.082) than AIDP of PC at 48 woa, and 25% lower than of NC and PC at 70 woa (P = 0.028). AIDCa was 25% lower for NC+BSP than PC at 48 woa only (P = 0.037). The avP and Ca sufficiency in the NC diet limited the opportunity to determine a phytase effect. Although the supplemental BSP tended to increase BW and 48 and 70 woa CBMC, and increased 32 woa AIDP, the efficacy of BSP could not be determined due to the lack of an NC effect on most parameters. Commercial laying hens can maintain health and productivity at lower than recommended levels of dietary Ca and avP; phytase supplementation may allow for even further reductions.


Assuntos
6-Fitase/metabolismo , Cálcio/metabolismo , Galinhas/fisiologia , Digestão/efeitos dos fármacos , Casca de Ovo/efeitos dos fármacos , Enterobacteriaceae/química , Fósforo/metabolismo , 6-Fitase/administração & dosagem , Ração Animal/análise , Animais , Osso e Ossos/química , Osso e Ossos/efeitos dos fármacos , Dieta/veterinária , Suplementos Nutricionais/análise , Casca de Ovo/fisiologia , Feminino , Íleo/efeitos dos fármacos , Íleo/fisiologia
14.
Poult Sci ; 98(9): 3926-3936, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-30938806

RESUMO

This study was conducted to determine the effects of residual superdoses of phytase on growth performance, tibia mineralization, and relative organ weight in ducks fed phosphorus-deficient diets. In Exp. 1, 4 kinds of commercial phytase were used to determine retention rate of phyatse with the phytase C being the highest via both high water-bath temperature (90%) and pelleting (50%), followed by phytase A, B, and D. In Exp. 2, a total of 560 male ducks were blocked based on body weight, and then allocated randomly to 7 treatments (5 replicates with 16 birds per replicate). Treatments included a maize-soybean meal-based diet with recommended calcium and 4.0 g non-phytate phosphorus (nPP)/kg starter diet or 3.8 g nPP/kg grower diet (positive control; PC), an nPP-deficient diet with 1.3 g nPP/kg starter diet or 1.1 g nPP/kg grower diet (negative control; NC), NC diets with increasing levels of residual phytase C (500, 1,000, 2,000, 3,000, and 4,000 units/kg feed) after pelleting. Birds fed NC diets had lower (P < 0.05) average daily gain (ADG) and average daily feed intake (ADFI) throughout the experiment compared with those fed PC diet. Supplementing NC diet with increasing residual superdoses of phytase improved (P < 0.05) ADG and ADFI quadratically in the entire experiment, while reduced feed-to-gain ratio (P < 0.05) quadratically during day 0 to 14. On day 14 and 35, birds fed NC diet had lower (P < 0.05) tibia length, weight, ash, calcium, phosphorus, and manganese contents than those fed PC diet. Increasing residual superdoses of phytase in NC diet increased (P < 0.05) tibia weight and ash, calcium, phosphorus contents quadratically on day 14 and 35. NC treatment increased (P < 0.05) the duodenum, jejunum, ileum, and cecum index compared with other treatments on day 14 and 35. Taken together, feeding increasing residual superdoses of phytase could counteract or exceed the negative effects of NC diet on growth performance, tibia mineralization, and relative organ weight in ducks.


Assuntos
6-Fitase/metabolismo , Calcificação Fisiológica/efeitos dos fármacos , Patos/fisiologia , Fósforo na Dieta/análise , Tíbia/efeitos dos fármacos , 6-Fitase/administração & dosagem , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal/efeitos dos fármacos , Animais , Dieta/veterinária , Suplementos Nutricionais/análise , Patos/crescimento & desenvolvimento , Masculino , Tamanho do Órgão/efeitos dos fármacos , Distribuição Aleatória , Tíbia/fisiologia
15.
Enzyme Microb Technol ; 126: 69-76, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31000166

RESUMO

The beta-propeller phytase (BPP) is an enzyme that hydrolyzes phyate to release inorganic phosphorus. The BPP produced by Pseudomonas sp. FB15 (PSphy) possesses an additional N-terminal domain that is not present in BPP produced by other Bacillus species. In this study, BPP produced by Bacillus sp. SJ-10 (SJ-10phy) was fused with the N-terminal domain of PSphy and the enzymatic properties of the resulting fusion protein (FUSJ-10phy) were investigated. FUSJ-10phy exhibited an optimal temperature that was 10 °C lower than that of the wild-type enzyme. A comparison of kinetic parameters showed that the turnover rate of FUSJ-10phy was 2.39-fold higher than that of SJ-10phy, representing a 1.79-fold increase in catalytic efficiency. In addition, BPP produced by Bacillus sp. SJ-48 has relatively low sequence similarity with SJ-10phy, was fused with N-terminal domain (FUSJ-48phy). FUSJ-48phy also increased catalytic efficiency and changed the optimal temperature. These results indicate that, when fused to other BPPs, the N-terminal domain of PSphy increases catalytic efficiency and enzyme activity at lower temperatures.


Assuntos
6-Fitase/metabolismo , Bacillus/enzimologia , Proteínas de Bactérias/metabolismo , Pseudomonas/enzimologia , Proteínas Recombinantes de Fusão/metabolismo , Temperatura Ambiente , 6-Fitase/química , 6-Fitase/genética , Sequência de Aminoácidos , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Catálise , Estabilidade Enzimática , Concentração de Íons de Hidrogênio , Hidrólise , Cinética , Conformação Proteica , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Homologia de Sequência
16.
Br Poult Sci ; 60(4): 439-448, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30966791

RESUMO

1.The objective of the present study was to rank the importance of the following dietary factors; canola meal, wheat, whole barley, digestible lysine, phytate-P, calcium, available P, sodium and three NSP-degrading feed enzymes. Their influence on growth performance, gastro-intestinal tract parameters, energy utilisation, ileal N digestibility and disappearance rates were determined via the Plackett-Burman design in broiler chickens offered phytase-supplemented diets. 2. The eleven dietary factors were assigned two levels in the Plackett-Burman design matrix. The resulting twelve dietary treatments were offered to six replicates per treatment (six birds per cage) with a total of 468 male Ross 308 broiler chicks from 7 to 28 d post-hatch. 3. Increasing digestible lysine levels improved weight gain by 15.6% (P < 0.001) and gain:feed by 9.36% (P < 0.001). Increasing calcium levels reduced weight gain by 6.36% (P < 0.001) and gain:feed by 2.60% (P < 0.001). The high calcium level increased gizzard pH from 2.78 to 3.01 (P < 0.005). Whole barley significantly increased relative gizzard weights and contents, pancreas weights and both ileal N digestibility coefficients (0.774 versus 0.803; P < 0.001) and ileal N disappearance rates (23. 3 versus 24.5 g/bird/day; P < 0.001). 4. Overall, digestible lysine level and calcium level were identified as the most influential dietary factors to influence growth performance of broilers offered phytase-supplemented diets, which hold implications for practical diet formulations.


Assuntos
6-Fitase/metabolismo , Galinhas/fisiologia , Metabolismo Energético , Nitrogênio/metabolismo , 6-Fitase/administração & dosagem , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal/efeitos dos fármacos , Animais , Galinhas/crescimento & desenvolvimento , Dieta/veterinária , Suplementos Nutricionais/análise , Metabolismo Energético/efeitos dos fármacos , Íleo/efeitos dos fármacos , Íleo/metabolismo , Masculino
17.
FEMS Microbiol Lett ; 366(5)2019 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-30869784

RESUMO

In this study, production of fungal phytase in thermotolerant methylotrophic yeast Ogataea thermomethanolica TBRC656 employing methanol-inducible OtAOX promoter and sucrose-inducible OtMal promoter was investigated in a high cell density fed-batch fermentation. Although a similar maximum cell concentration was obtained in both expression systems, the OtMal system gave ~2-fold higher phytase activity, specific yield, production yield, volumetric productivity and specific productivity rate compared with the OtAOX system. In addition to being more efficient, the OtMal system is more flexible because sucrose or sugarcane molasses can be utilized as less expensive carbon sources instead of glycerol in batch and fed-batch stages. Phytase yields from the OtMal system produced using sucrose or sugarcane molasses are comparable with those obtained with glycerol. We estimate the cost of phytase production by the OtMal system using sucrose or sugarcane molasses to be ~85% lower than the OtAOX system.


Assuntos
6-Fitase/genética , Proteínas Fúngicas/genética , Regulação Fúngica da Expressão Gênica , Microbiologia Industrial/métodos , Proteínas Recombinantes/genética , Saccharomycetales/genética , Sacarose/farmacologia , 6-Fitase/metabolismo , Aspergillus niger/enzimologia , Aspergillus niger/genética , Contagem de Células , Fermentação , Proteínas Fúngicas/metabolismo , Regulação Fúngica da Expressão Gênica/efeitos dos fármacos , Metanol/metabolismo , Melaço , Regiões Promotoras Genéticas , Proteínas Recombinantes/metabolismo , Saccharomycetales/efeitos dos fármacos , Saccharomycetales/crescimento & desenvolvimento , Saccharomycetales/metabolismo , Sacarose/metabolismo , Termotolerância
18.
Poult Sci ; 98(9): 3884-3893, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-30877747

RESUMO

An experiment was conducted to evaluate graded doses of phytase in high-phytate diets. Ross 308, male broilers (n = 600) were assigned to one of 4 diets, with 10 replicate pens/diet and 15 birds/replicate pen. Diets were a nutrient adequate positive control (PC), a negative control (NC) diet with a reduction of Ca by 0.22%, available P by 0.20%, energy by 120 kcal/kg, and amino acids by 1 to 5% compared with the PC. The NC diet was supplemented with 0, 2,000, or 4,000 phytase units (FTU)/kg. Phytase increased (linear, P < 0.05) weight gain from hatch to day 18. Birds fed the NC + 4000 FTU/kg ate and gained more (P < 0.05) than birds fed the PC. The apparent ileal digestibility (AID) of all nutrients and amino acids were reduced (P < 0.05) in birds fed the NC compared with birds fed the PC. Phytase increased (linear, P < 0.10) AID of most nutrients. Digestibility was lower (P < 0.10) in birds fed the NC + 4000 FTU/kg compared with birds fed the PC. Using daily intake and AID to determine digestible nutrient intake resulted in no differences between birds fed the PC or NC + 4000 FTU/kg diets. Digestible intake of methionine or glutamate was better correlated with BW gain (P < 0.0001) than AID (P > 0.10). Phytase reduced (linear, P < 0.01) phytate concentration and increased inositol (linear, P < 0.01), phytate hydrolysis (linear, P < 0.05), and jejunal expression (linear, P < 0.05) of SNAT-1 and LAT-4 transporters. Supplementation of increasing doses of phytase in high-phytate, low-nutrient dense diets improved gain and digestibility through nearly complete phytate destruction. Digestible nutrient intake may be a better indication of broiler gain than AID alone.


Assuntos
6-Fitase/metabolismo , Aminoácidos/metabolismo , Galinhas/fisiologia , Digestão/efeitos dos fármacos , Ácido Fítico/metabolismo , 6-Fitase/administração & dosagem , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal/efeitos dos fármacos , Animais , Galinhas/crescimento & desenvolvimento , Dieta/veterinária , Suplementos Nutricionais/análise , Relação Dose-Resposta a Droga , Íleo/efeitos dos fármacos , Íleo/fisiologia , Masculino
19.
Poult Sci ; 98(9): 3870-3883, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-30877748

RESUMO

This study was conducted to evaluate the effect of microbial phytase and myo-inositol supplementation in low non-phytate phosphorus (nPP) diets on pH and the solubility of minerals in an in vitro digestion procedure (IVDP) and to compare this with digesta from birds fed different diets (grower diets) compared to the in vitro test (starter diets). A total of 660 1-day-old broilers were randomly allotted into 11 dietary treatments and fed a corn-soybean-meal-based diet with recommended nPP (positive control; PC), an nPP-deficient diet (negative control; NC), NC diets supplemented with phytase (500; 1,000; 2,000; 3,000; 4,000; 5,000; and 6,000 FTU/kg), an NC diet plus 0.15% myo-inositol, and an NC diet with reduced Ca level (Ca: nPP ratio same as PC) from 1 to 23 D of age. The pH and Ca solubility of the NC diet was increased compared with the PC when subjected to IVDP (P < 0.05). P solubility in the gizzard and jejunal digesta was reduced in the NC compared with the PC diet and this was also reflected in the IVDP. Phytase addition to the NC diets linearly increased (P < 0.05) the pH value and Ca and P solubilities in both digesta and diets subjected to IVDP. Higher doses of microbial phytase increased (P < 0.05) Zn and Fe solubilities in both digesta and IVDP. Myo-inositol supplementation of the NC diet had no effect on mineral solubility, but decreased (P < 0.05) the pH of the IVDP. Lowering the Ca content of the NC diet decreased (P < 0.05) the pH of the in vitro digested diets and Ca solubility in both broiler digesta and IVDP and also increased (P < 0.05) P solubility in both the jejunal digesta and IVDP. Correlations were noted between the solubility of P in the in vitro assay and that in the gizzard and jejunal digesta, and also with bird performance, confirms the usefulness of in vitro assay.


Assuntos
6-Fitase/metabolismo , Galinhas/fisiologia , Digestão/efeitos dos fármacos , Inositol/metabolismo , Minerais/metabolismo , Fósforo na Dieta/metabolismo , 6-Fitase/administração & dosagem , Ração Animal/análise , Animais , Galinhas/crescimento & desenvolvimento , Dieta/veterinária , Suplementos Nutricionais/análise , Relação Dose-Resposta a Droga , Concentração de Íons de Hidrogênio , Inositol/administração & dosagem , Distribuição Aleatória , Solubilidade
20.
Int J Biol Macromol ; 131: 1117-1124, 2019 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-30910675

RESUMO

A preferable phytase for use in animal feeds for industrial applications should have high, optimal activity at low pH in the monogastric gut environment and high thermostability. To obtain enzymes with enhanced catalytic efficiency (pH 5.5) and excellent activity in acidic pH range, we performed structure-based rational design of a thermostable phytase (PhyAn). For this, six mutants based on different rational design strategies were constructed and heterologously expressed in Pichia pastoris. Particularly, the extracellular enzymatic activity was assessed to ensure that the produced enzymes met requirements of further analyses. Several positive mutants with enhanced catalytic efficiency or pH-profile shifts were carefully examined. Biochemical and kinetic investigations of purified mutants revealed that E79K, E80K, E79K + E80K and D68K had higher catalytic efficiency than the parent enzyme by approximately 49%, 67%, 86% and 15%, respectively. Moreover, the optimum pH of mutant Y65H was shifted from 5.0 to 3.0, and the peak of D68K shifted to pH 5.5. Analysis of the structural-functional relationships revealed that changes in amino acid charges, structural flexibility and space hindrance could significantly influence certain enzyme characteristics. Our results illustrate the feasibility and present a structural foundation for enhancing the phytase-catalytic efficiency and acid resistance by assembling mutations derived using rational design.


Assuntos
6-Fitase/química , 6-Fitase/metabolismo , Aspergillus niger/enzimologia , 6-Fitase/genética , 6-Fitase/isolamento & purificação , Aspergillus niger/genética , Catálise , Estabilidade Enzimática , Proteínas Fúngicas/química , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Concentração de Íons de Hidrogênio , Cinética , Modelos Moleculares , Conformação Molecular , Mutagênese Sítio-Dirigida , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , Solventes
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