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1
Chuiko, N. V., A. Yu Chobotarov, and I. K. Kurdish. "Growth and Phytase Activities of Bacillus subtilis IMV B-7023 During Cultivation with Sodium Phytate." Mikrobiolohichnyi Zhurnal 83, no. 6 (December 17, 2021): 13–19. http://dx.doi.org/10.15407/microbiolj83.06.013.
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Bacteria of the genus Bacillus are known for their ability to mineralize organic phosphorus compounds. Phytates constitute up to 60–80% of the total plant phosphorus and almost 50% of soil organic phosphorus. Phytates phosphorus is unavailable for plants. Bacillus can synthesize phosphatases both wide spectrum of action, and highly specific phytases that catalyze the hydrolysis of phytates. Therefore, the aim of this work was to study the growth and phytase activity of Bacillus subtilis IMV B-7023, which is the component of the ”Azogran” complex bacterial preparation for plant growing. Methods. The growth activity of bacteria was studied by cultivation methods, the phytase activity – by measuring the amount of phosphate released from sodium phytate during the enzymatic reaction. Results. It was shown that B. subtilis IMV B-7023 assimilated phytate as the source of phosphorus nutrition during cultivation in media with 0.5, 1.0 and 2.0 g/L of sodium phytate. The highest growth activity of these bacteria was observed after two days of cultivation in medium with 1.0 g/L of phytate. The number of bacteria was (3.91±0.32)×109 CFU/mL under these conditions. At the same time, B. subtilis IMV B-7023 demonstrated a low level of phytate assimilation as a source of carbon nutrition. Thus, after two days of cultivation the number of bacteria increased from (4.12±0.09)×106 CFU/mL to (1.07±0.07–3.11±0.51)×107 CFU/mL in the presence of 0.5–2.0 g/L phytate in the medium and the absence of another carbon source. It was determined that strain B. subtilis IMV B-7023 had phytase activity, the highest activity (221.85±0.12 U/g) was on the first day of their cultivation in medium with inorganic phosphates. It should be noted that B. subtilis IMV B-7023 phytase activity was lower during cultivating in medium with sodium phytate as a source of phosphorus nutrition, than in medium with inorganic phosphates. The obtained fact may be due to phytate hydrolysis by widespecific phosphatases. Higher rates of phytase activity obtained on the first and third days compared to the second and fourth days of bacterial cultivation may indicate the expression of phosphatases genes only in the period required for maximum bacterial development, in the absence of these proteins in the media. At the same time, the phytase activity of B. subtilis IMV B-7023 after 2 days cultivation in a media with 0.5 and 1.0 g/L of sodium phytate (194.80±0.15 U/g and 160.90±0.13 U/g, respectively) as the source of carbon and phosphorus was higher compared to the activity of bacteria on medium with inorganic phosphates (137.79±0.10 U/g). This may be caused by the synthesis of a larger number of highly specific phosphatases (phytases) in bacterial cells at the presence of only phytate in the medium as a substrate. Conclusions. B. subtilis IMV B-7023 strain is characterized by growth on nutrient medium with sodium phytate and phytase activity. Because they are soil microorganisms used as the component of the ”Azogran” complex bacterial preparation for plant growing, the ability to hydrolyze and assimilate phytate is important for functioning of this strain in the rhizosphere. The obtained results extend the understanding of B. subtilis IMV B-7023 influence on phosphorus nutrition and development of plants.
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Williams, P. J., and T. G. Taylor. "A comparative study of phytate hydrolysis in the gastrointestinal tract of the golden hamster (Mesocricetus auratus) and the laboratory rat." British Journal of Nutrition 54, no. 2 (September 1985): 429–35. http://dx.doi.org/10.1079/bjn19850128.
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1. The role of bacterial, dietary and intestinal phytases (EC 3. 1. 3. 8) in the hydrolysis of phytate was investigated in the golden hamster and rat by assaying phytase in the small intestine and by measuring the disappearance of phytate from the stomach and large intestine, using chromium oxide as an insoluble solid-phase marker.2. It was confirmed that an active phytase was present in the proximal third of the small intestine of the rat but the enzyme was undetectable in the hamster.3. Extensive bacterial breakdown of phytate occurred in the pregastric pouch and true stomach of the hamster with both phytase-containing and phytase-free diets, with phytate digestibilities in the true stomach ranging from 0.69–0.90, confirming that the hamster can be regarded as a pseudo-ruminant.4. With a phytase-free diet, the digestibility of phytate in the stomach of the rat was very low (0.05) but with a wheat-based diet substantial breakdown of phytate occurred (digestibility up to 0.49), presumably under the influence of the cereal phytase.5. Intestinal phytase did not appear to be of great significance in the rat but some further hydrolysis of the residual phytate probably occurred in the large intestine of both species by bacterial phytase.
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Moss, Amy F., Sonia Yun Liu, and Peter H. Selle. "Progress in comprehending the phytate–phytase axis in chicken-meat production." Animal Production Science 58, no. 10 (2018): 1767. http://dx.doi.org/10.1071/an17594.
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After an extended delay, the level of acceptance of exogenous phytases by the global chicken-meat industry is now almost complete. Contemporary bacterial phytases degrade phytate primarily in the gizzard. The extent of phytate degradation determines the extent to which phytate-bound phosphorus (P) is liberated; however, studies designed to investigate phytate degradation along the digestive tract have generated some confusing outcomes. This may be related to the reactivity of the phytate moiety, coupled with problems with inert dietary markers and perhaps a lack of complete and uniform extractions of phytate from digesta due to variations in digesta pH and phytate solubility. Quite recently, phytase was shown to have profound impacts on sodium (Na) digestibility coefficients in four segments of the small intestine. This has obvious implications for intestinal uptakes of glucose and amino acids via their respective Na+-dependent transport systems and it is possible that phytate and phytase have reciprocal impacts on ‘sodium pump’ (Na+, K+-ATPase) activity. It has been recently demonstrated unequivocally that phytase has the capacity to increase amino acid digestibility coefficients to the extent that phytase may generate a ‘proximal shift’ in the sites of amino acid absorption. The impact of phytase on starch digestibility is more equivocal and phytase responses may stem more from enhanced glucose absorption rather than starch digestion. The acceptance of phytase is hardly surprising, given its capacity to increase P utilisation coupled with numerous other positive influences that are still being properly realised.
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Chuiko, N. V., A. Yu Chobotarov, and I. K. Kurdish. "Abiotic Factors Influence on Bacillus subtilis IMV B-7023 Phytase Activity." Mikrobiolohichnyi Zhurnal 84, no. 6 (February 28, 2023): 3–9. http://dx.doi.org/10.15407/microbiolj84.06.003.
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Bacteria of the Bacillus genus can synthesize specific phytase enzymes. This property is especially important for soil bacteria. It helps to mineralize phytin and phytates and to provide these bacteria and plants (in the root zone of which they live) with the available phosphorus. Our previous studies have demonstrated that the Bacillus subtilis IMV B-7023 strain exhibits a phytase activity and can use phytate as a nutrition source. It is a component of the «Azogran» complex bacterial preparation for crop production. As known, abiotic environmental factors can influence the phytase activity of bacteria. In particular, the phytase activity changes significantly under different pH and temperatures. Solid soil particles, including nanosized minerals, can also influence bacteria’s enzymatic activity. The influence of abiotic factors on Bacillus subtilis IMV B-7023 phytase activity has not previously been studied, so this was the aim of our research. Methods. The phytase activity of bacteria was studied by measuring the amount of phosphate released from sodium phytate during the enzymatic reaction, and the nanomaterials’ influence on growth — by cultivation methods. Results. The highest B. subtilis IMV B-7023 phytase activity was observed at 28°C. Also, there was no B. subtilis IMV B-7023 phytase activity at pH 4—6. However, this activity increased at pH 7 and did not change significantly with increasing the buffer system pH to 12. Silicon dioxide influence on the B. subtilis IMV B-7023 growth activity during cultivation in a media with phytate as a phosphorus source depended on the nanomaterial concentration. Thus, at 0.05 and 0.5 g/L of silicon dioxide in the medium, this strain growth activity increased by 8—18%, and at 5.0 g/L of these nanoparticles, bacteria growth inhibition by 19% was observed. At the same time, clay mineral bentonite did not affect the B. subtilis IMV B-7023 growth under the studied cultivation conditions. In addition, silicon dioxide and bentonite stimulated B. subtilis IMV B-7023 phytase activity at all studied concentrations. So, phytase activity increased by 1.82—3.34 times upon adding silicon dioxide and by 2.54—5.83 times upon adding bentonite into the medium. Since the optimal values for phytase activity of most genus Bacillus bacteria are within neutral pH values and temperatures within 50—55°C, a property of B. subtilis IMV B-7023 to show maximum phytase activity at alkaline pH and lower temperatures (28°C) and also stimulation of this activity by soil minerals increases competitiveness of this strain as a component of a bacterial preparation for crop production. Conclusions. Abiotic environmental factors influence the B. subtilis IMV B-7023 supernagrowth and phytase activity. Optimal physicochemical factors for the phytase activity of these bacteria are temperature 28°C and pH 7—12. The concentrations 0.05, 0.5, and 5.0 g/L of silicon dioxide and bentonite increase B. subtilis IMV B-7023 phytase activity. The effect of these nanoscale minerals on the B. subtilis IMV B-7023 growth depends on their type and concentration during cultivation in the medium with phytate as a phosphorus source. The obtained results indicate the potential ability of the B. subtilis IMV B-7023 strain to effectively assimilate phytates in neutral and alkaline soils, especially due to the interaction of these bacteria with bentonite and silicon dioxide nanoparticles. This expands the possibility of using B. subtilis IMV B-7023 in agricultural technologies.
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Menezes-Blackburn, Daniel, Nahad Al-Mahrouqi, Buthaina Al-Siyabi, Adhari Al-Kalbani, Ralf Greiner, and Sergey Dobretsov. "Bacterial Communities Associated with the Cycling of Non-Starch Polysaccharides and Phytate in Aquaponics Systems." Diversity 13, no. 12 (November 30, 2021): 631. http://dx.doi.org/10.3390/d13120631.
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Aquaponics are efficient systems that associate aquatic organisms’ production and plants by recirculating water and nutrients between aquaculture and hydroponic tanks. In this study, we characterised the bacterial communities in the freshwater aquaponics system that can mineralise polysaccharides and phytate by producing carbohydrate-degrading enzymes and phytases, by 16S rRNA gene sequencing and in vitro culture techniques. Around 20% of the operational taxonomic units (zOTUs) identified were previously reported to carry fibre-degrading enzyme putative genes, namely β-glucanase (1%), xylanase (5%), or cellulases (17%). Ten % of the zOTUs were previously reported to carry putative genes of phytases with different catalytic mechanisms, namely β-propeller (6%), histidine acid phytases (3%), and protein tyrosine phytase (<1%). Thirty-eight morphologically different bacteria were isolated from biofilms accumulated in fish and plant compartments, and identified to belong to the Bacilli class. Among these, 7 could produce xylanase, 8 produced β-glucanase, 14 produced cellulase, and 11 isolates could secrete amylases. In addition, Staphylococcus sp. and Rossellomorea sp. could produce consistent extracellular phytate-degrading activity. The PCR amplification of β-propeller genes both in environmental samples and in the isolates obtained showed that this is the most ecologically relevant phytase type in the aquaponics systems used. In summary, the aquaponics system is abundant with bacteria carrying enzymes responsible for plant-nutrient mineralisation.
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Suleimanova, Aliya, Daria Bulmakova, and Margarita Sharipova. "Heterologous Expression of Histidine Acid Phytase From Pantoea sp. 3.5.1 in Methylotrophic Yeast Pichia Pastoris." Open Microbiology Journal 14, no. 1 (July 30, 2020): 179–89. http://dx.doi.org/10.2174/1874285802014010179.
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Background and Objective: The major storage form of phosphorus in plant-derived feed is presented by phytates and not digested by animals. Phytases are able to hydrolyze phytates and successfully used as feed additives. Nevertheless, nowadays, there is a constant search of new phytases and expression systems for better production of these enzymes. In this study, we describe cloning and expression of gene encoding histidine acid phytase from Pantoea sp. 3.5.1 using methylotrophic yeast Pichia pastoris as the host. Methods: The phytase gene was placed under the control of the methanol-inducible AOX1 promoter and expressed in P. pastoris. Experiments of small-scale phytase expression and activity assays were used to test recombinant colonies. Four different signal peptides were screened for better secretion of phytase by P. pastoris. After 36 h of methanol induction in shake flasks, the maximum extracellular phytase activity (3.2 U/ml) was observed in P. pastoris strain with integrated construct based on pPINK-HC vector and Kluyveromyces maxianus inulinase gene signal sequence. This phytase was isolated and purified using affinity chromatography. Results: Recombinant phytase was a glycosylated protein, had a molecular weight of around 90 kDa and showed maximum activity at pH 4.0 and at 50°C. Recombinant phytase had excellent thermal stability – it retained high residual activity (100% ± 2%) after 1 hour of heat treatment at 70°C. Conclusion: The enhanced thermostability of the recombinant phytase, its expression provided by strong inducible promotor and the effectively designed expression cassette, the simple purification procedure of the secreted enzyme, and the possibility of large-scale expression make the foundation for further production of this bacterial phytase in P. pastoris at an industrial scale.
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Gauchan, Dhurva Prasad, Shishir Pandey, Bikash Pokhrel, Nabin Bogati, Puja Thapa, Ashesh Acharya, Bishnu Maya KC, and Janardan Lamichhane. "Growth Promoting Role of Phytase Producing Bacteria Isolated from Bambusa tulda Roxb. Rhizosphere in Maize Seedlings Under Pot Conditions." Journal of Nepal Biotechnology Association 4, no. 1 (March 22, 2023): 17–26. http://dx.doi.org/10.3126/jnba.v4i1.53442.
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Phytase enzymes have focused on their role in agriculture to generate bioavailable phosphorus (P) requires for plant nutrition. Herein, the feasibility of phytase-producing microbes as biofertilizers was studied. Bacteria with greater potential for hydrolysing calcium phytate based on the halo-to-colony ratio from Bambusa tulda Roxb. rhizosphere was isolated. Phytase activity by incorporating wheat bran, phytase screening and Luria Bertani (LB) medium after acetone precipitation was measured. Bacterial genomes were screened for the presence of β-propeller phytase gene corresponding to the Bacillus spp. using polymerase chain reaction (PCR). Finally, the effect of the isolates on the growth of maize seedlings under pot conditions in P-deficient loamy soil was evaluated. Ten distinct bacterial isolates collected from B. tulda rhizosphere were capable of mineralizing phytate and the maximum effect was observed for designated SRBR-04. Most isolates solubilized Ca3(PO4)2 as a sole P source in Pikovskaya’s agar. Five isolates selected for the study synthesized auxin in the LB broth supplemented with 1 mg mL-1 L-Tryptophan (1.63 to 4.5 μg mL-1). Phytase production was highest in wheat bran with isolate SRBR-04 producing a maximum of 0.34 U mg-1. Two isolates (SRBR-01, SRBR-04) screened positive for the presence of Bacillus phytase gene. Pot assay in P-deficient soil showed significant (p < 0.05) biomass promotion for the isolate SRBR-07 in shoot height (57%), dry shoot weight (178%), dry root weight (104%) and leaf area (113%) over the untreated control. Amendment of P-deficient agricultural soils with phytase-producing bacteria would provide a sustainable approach for P nutrition management in Zea mays.
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Christensen, Trine, Yueming Dersjant-Li, Vincent Sewalt, Rie Mejldal, Svend Haaning, Sina Pricelius, Igor Nikolaev, Robin A. Sorg, and Arno de Kreij. "In Vitro Characterization of a Novel Consensus Bacterial 6-Phytase and One of its Variants." Current Biochemical Engineering 6, no. 3 (December 28, 2020): 156–71. http://dx.doi.org/10.2174/2212711906999201020201710.
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Background:: Microbial phytases are added to animal feed to hydrolyze phytic acid (myoinositol hexakisphosphate, IP6) and phytate (salt of phytic acid) increasing phosphorus bioavailability. Novel phytases with enhanced bio-efficacy are being developed. Objective:: To characterize the biochemical and enzymatic properties of a novel consensus bacterial 6- phytase and its variant (PhyG), produced in Trichoderma reesei. Methods:: The in vitro specific activity, kinetic parameters, pH-activity profiles (relative to pH5.5), IP6 degradation, hydrolysis products and phosphate release of the phytases were determined using sodium phytate substrate. Melting point (Tm) was determined by differential scanning calorimetry and thermostability assessed by measuring residual activity at different temperatures. In vivo effects of PhyG supplementation at 0 to 1,000 FTU/kg on ileal IP6 digestibility and IP ester concentrations were determined in piglets. Results:: Both phytases exhibited pH optima of 3.5-4.5, high relative activity over a wide pH range (pH2.0-5.0), and substantial relative activity at pH1.5. At pH3.0, the specific activity of the PhyG variant was 1487 U/mg protein and at pH3.5 the kinetic constants were 240 μM (Km) and 1873 s-1 (Kcat). The hydrolysis of IP6 by both phytases was rapid. The major initial hydrolysis product was DLI( 1,2,3,4,5)P5, designating the phytases as bacterial 6-phytases (EC 3.1.3.26). Hydrolysis occurred at the D-3 (L-1) position in ~30% of instances, indicating a dual-specificity. Conclusion: Both phytases showed high thermostability compared to wild type and existing commercial bacterial 6-phytases; PhyG exhibited 95% residual activity after 20 min incubation at 85.4ºC (pH5.5), Tm50 of ~93.2ºC and Tm of 98.8ºC. In vivo, PhyG at 1,000 FTU/kg achieved an ileal digestibility of IP6 of 89.3%.
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Espinosa, Charmaine D., Deepak E. Velayudhan, Yueming Dersjant-Li, and Hans H. Stein. "60 Effect of a Novel Consensus Bacterial 6-phytase Variant on Mineral Digestibility and Bone Ash in Young Growing Pigs Fed Diets with Different Concentrations of Phytate." Journal of Animal Science 99, Supplement_1 (May 1, 2021): 45–46. http://dx.doi.org/10.1093/jas/skab054.079.
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Abstract An experiment was conducted to test the hypothesis that increasing levels of phytase increases mineral digestibility and bone ash by pigs fed diets containing 0.23%, 0.29%, or 0.35% phytate-P. Within each level of phytate, 5 diets were formulated based on corn, soybean meal, and canola meal to contain 0, 500, 1,000, 2,000, or 4,000 phytase units (FTU)/kg of a novel consensus bacterial 6-phytase variant (PhyG). In addition, 3 reference diets were formulated by adding a commercial Buttiauxella phytase (PhyB) at 1,000 FTU/kg to the 3 diets containing no PhyG. A randomized complete block design with 144 pigs (12.70 ± 4.01 kg), 18 diets, and 8 replicate pigs per diet was used. Pigs adapted to diets for 15 d followed by 4 d of fecal collection. Femurs were collected on the last day. Data were analyzed as a 3 × 5 factorial with 3 levels of phytate-P and 5 levels of phytase, and contrast statements were used to compare 1,000 FTU of PhyG with PhyB. Pig was the experimental unit. Diets containing 0.35% phytate-P had reduced (P < 0.01) apparent total tract digestibility (ATTD) of Ca, P, Mg, and K compared with diets containing 0.23% or 0.29% phytate-P, but inclusion of phytase increased (P < 0.01) ATTD of Ca, Na, and K (Table 1). Phytase increased ATTD of P and Mg, but to a greater extent in diets with 0.23 or 0.29% phytate-P than in diets with 0.35% phytate-P (interaction, P < 0.05). Phytase increased bone ash, but to a greater extent if there was 0.35 rather than 0.23 or 0.29% phytate-P in the diets (interaction, P < 0.05). PhyG increased ATTD of P more (P < 0.05) than PhyB. In conclusion, the novel consensus phytase is effective in increasing bone ash and ATTD of Ca, P, Na, Mg and K.
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Suleimanova, Aliya D., Astrid Beinhauer, Liia R. Valeeva, Inna B. Chastukhina, Nelly P. Balaban, Eugene V. Shakirov, Ralf Greiner, and Margarita R. Sharipova. "Novel Glucose-1-Phosphatase with High Phytase Activity and Unusual Metal Ion Activation from Soil Bacterium Pantoea sp. Strain 3.5.1." Applied and Environmental Microbiology 81, no. 19 (July 24, 2015): 6790–99. http://dx.doi.org/10.1128/aem.01384-15.
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ABSTRACTPhosphorus is an important macronutrient, but its availability in soil is limited. Many soil microorganisms improve the bioavailability of phosphate by releasing it from various organic compounds, including phytate. To investigate the diversity of phytate-hydrolyzing bacteria in soil, we sampled soils of various ecological habitats, including forest, private homesteads, large agricultural complexes, and urban landscapes. Bacterial isolatePantoeasp. strain 3.5.1 with the highest level of phytase activity was isolated from forest soil and investigated further. ThePantoeasp. 3.5.1agpPgene encoding a novel glucose-1-phosphatase with high phytase activity was identified, and the corresponding protein was purified to apparent homogeneity, sequenced by mass spectroscopy, and biochemically characterized. The AgpP enzyme exhibits maximum activity and stability at pH 4.5 and at 37°C. The enzyme belongs to a group of histidine acid phosphatases and has the lowestKmvalues toward phytate, glucose-6-phosphate, and glucose-1-phosphate. Unexpectedly, stimulation of enzymatic activity by several divalent metal ions was observed for the AgpP enzyme. High-performance liquid chromatography (HPLC) and high-performance ion chromatography (HPIC) analyses of phytate hydrolysis products identifydl-myo-inositol 1,2,4,5,6-pentakisphosphate as the final product of the reaction, indicating that thePantoeasp. AgpP glucose-1-phosphatase can be classified as a 3-phytase. The identification of thePantoeasp. AgpP phytase and its unusual regulation by metal ions highlight the remarkable diversity of phosphorus metabolism regulation in soil bacteria. Furthermore, our data indicate that natural forest soils harbor rich reservoirs of novel phytate-hydrolyzing enzymes with unique biochemical features.
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Hardy, Rachael, Yueming Dersjant-Li, Trine Christensen, Deepak Velayudhan, and Janet C. Remus. "115 Functionality of a novel consensus bacterial 6-phytase variant on ileal phytate degradation in weaned piglets fed diets without inorganic phosphate." Journal of Animal Science 98, Supplement_4 (November 3, 2020): 104–5. http://dx.doi.org/10.1093/jas/skaa278.191.
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Abstract Microbial phytases are used in animal feed to hydrolyze phytic acid (myo-inositol hexakisphosphate, IP6) and phytate (salt of phytic acid), to improve phosphorus (P) bioavailability. This study evaluated the efficacy of a novel consensus bacterial 6-phytase variant (PhyG) on ileal IP6 degradation in piglets. A positive control diet was formulated based on corn and soybean meal with adequate digestible P (2.9g/kg) and calcium (Ca, 7.0g/kg). A negative control (NC) diet was formulated without inorganic P (1.1 g digestible P/kg), reduced in Ca (5.0 g/kg) with analyzed phytate P of 2.6g/kg. This diet was supplemented with PhyG at 250, 500 and 1,000 FTU/kg. A total of 90 crossed Pietrain x (large White x Landrace) 21-d-old piglets were fed adaptation diets until 42 d old and then assigned to pens (2 pigs/pen and 9 pens/treatment) in a completely randomized block design. Piglets were fed mash diets ad libitum for 28 d, at the end of the study 1 piglet/pen was euthanized for collecting ileal digesta samples for determination of IP6 digestibility, using TiO2 as a marker. Data was analyzed by one-way ANOVA using JMP 14.0 with treatment comparison by Tukey test and dose response by curve fitting. The ileal IP6 content was reduced with increasing phytase dose in a dose dependent manner (Plinear < 0.0001; Pexponential = 0.08). Similarly, the ileal IP6 digestibility increased exponentially (P < 0.05) with increasing phytase dose (49.1% for NC and 89.3% for PhyG at 1,000 FTU/kg). Improved ileal IP6 digestibility is closely related to improved bone ash, body weight gain and feed conversion ratio (R2 > 0.91). The in vitro IP6 degradation profile supports these in vivo observations. The results demonstrated that the novel consensus phytase variant is highly efficient in hydrolyzing phytate and increasing the bio-availability of phytate bound P in piglets fed diets without inorganic phosphate.
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Velayudhan, Deepak, Rachael Hardy, Leon Marchal, and Yueming Dersjant-Li. "116 Meta-analysis of 3 trials: the effect a novel consensus bacterial 6-phytase variant on phosphorus digestibility and retention in piglets fed inorganic phosphate free diets." Journal of Animal Science 98, Supplement_4 (November 3, 2020): 104. http://dx.doi.org/10.1093/jas/skaa278.190.
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Abstract A meta-analysis was conducted to investigate the effect of a novel consensus bacterial 6-phytase variant on apparent total tract digestibility (ATTD) of phosphorus (P) and P retention in piglets (initial body weight of 9.6 ± 1.8 kg). Database was constructed using 3 studies (unpublished), contributing 189 data points. All studies had a positive control (PC) diet with adequate levels of calcium (Ca; 0.57–0.85%) and digestible P (0.25–0.29%), a negative control (NC) diet with low Ca and digestible P levels (total removal of inorganic P). Piglets were fed a corn or wheat soybean meal-based diet with rice bran or sunflower meal for 7–10 d, followed by 4 d fecal and urine collection. Total analyzed Ca, total P, calculated digestible P and phytate P levels in NC varied from 0.48 to 0.79%, 0.37 to 0.40%, 0.16 to 0.18%, and 0.24 to 0.28%, respectively, among studies. The NC diet was supplemented with a novel consensus bacterial 6-phytase variant (PhyG) or a commercial Buttiauxella sp. phytase (PhyB), at 500 and 1,000 FTU/kg. Data were analyzed using Mixed Model by JMP 14.0. Addition of both phytases improved (P < 0.05) ATTD P compared to NC diets. The NC reduced digestible P (g/kg) vs PC, both phytases showed greater (P < 0.05) digestible P (g/kg) when compared to NC and recovered to the level of PC. Digestible P improvement (g/kg) over NC showed a significant phytase effect, where PhyG supplementation had 17% greater (P < 0.05) digestible P improvement when compared to PhyB. Phosphorus retention improved (P < 0.05) with phytase supplementation, with 1,000 FTU/kg showing greater (P < 0.05) P retention when compared to 500 FTU/kg. It was concluded that both PhyG and PhyB are efficient in improving ATTD P and P retention, PhyG showed greater digestible P improvement when compared to PhyB, and 1,000 FTU/kg showed greater effect than 500 FTU/kg.
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Castillo Villamizar, Genis Andrés, Heiko Nacke, Marc Boehning, Kristin Herz, and Rolf Daniel. "Functional Metagenomics Reveals an Overlooked Diversity and Novel Features of Soil-Derived Bacterial Phosphatases and Phytases." mBio 10, no. 1 (January 29, 2019): e01966-18. http://dx.doi.org/10.1128/mbio.01966-18.
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ABSTRACTPhosphatases, including phytases, play a major role in cell metabolism, phosphorus cycle, biotechnology, and pathogenic processes. Nevertheless, their discovery by functional metagenomics is challenging. Here, soil metagenomic libraries were successfully screened for genes encoding phosphatase activity. In this context, we report the largest number and diversity of phosphatase genes derived from functional metagenome analysis. Two of the detected gene products carry domains which have never been associated with phosphatase activity before. One of these domains, the SNARE-associated domain DedA, harbors a so-far-overlooked motif present in numerous bacterial SNARE-associated proteins. Our analysis revealed a previously unreported phytase activity of the alkaline phosphatase and sulfatase superfamily (cl23718) and of purple acid phosphatases from nonvegetal origin. This suggests that the classical concept comprising four classes of phytases should be modified and indicates high performance of our screening method for retrieving novel types of phosphatases/phytases hidden in metagenomes of complex environments.IMPORTANCEPhosphorus (P) is a key element involved in numerous cellular processes and essential to meet global food demand. Phosphatases play a major role in cell metabolism and contribute to control the release of P from phosphorylated organic compounds, including phytate. Apart from the relationship with pathogenesis and the enormous economic relevance, phosphatases/phytases are also important for reduction of phosphorus pollution. Almost all known functional phosphatases/phytases are derived from cultured individual microorganisms. We demonstrate here for the first time the potential of functional metagenomics to exploit the phosphatase/phytase pools hidden in environmental soil samples. The recovered diversity of phosphatases/phytases comprises new types and proteins exhibiting largely unknown characteristics, demonstrating the potential of the screening method for retrieving novel target enzymes. The insights gained into the unknown diversity of genes involved in the P cycle highlight the power of function-based metagenomic screening strategies to study Earth’s phosphatase pools.
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Jlali, Maamer, Sarper Ozbek, and Estelle Devillard. "54 Effects of a new biosynthetic 6-phytase supplementation on nutrient digestibility, phytate disappearance and growth efficiency in weaning piglets fed low in calcium and digestible phosphorus diets." Journal of Animal Science 102, Supplement_2 (May 1, 2024): 85–86. http://dx.doi.org/10.1093/jas/skae102.097.
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Abstract The objective of this study was to investigate the influence of adding graded levels of a new biosynthetic bacterial 6-phytase on nutrient digestibility, phytate disappearance and growth performance in weaned piglets. A total of 192 male weaned piglets with initial body weight of 8.70 ± kg, distributed in a randomized block design with four treatments and 24 replicate pens with 2 animals each for 42 d. The experimental diets included a phosphorus and Ca deficient diet (STTD of P/Ca = 0.21%/0.58% and 0.19%/0.54%, respectively, in post-weaning 1 and 2 diets) containing 0 (a control diet), 500, 1,000 or 1,500 FTU/kg diet. The diets were fed into 2 phases: post-weaning 1 and 2 from 1 to 14 and 15 to 42 d, respectively. Body weight and feed intake were measured at the end of each phase, and average daily gain (ADG), average daily feed intake (ADFI) and feed conversion ratio (FCR) were calculated for the overall experimental period. On d 42, ileal digesta samples were collected from 1 piglet/pen (n = 12 samples/treatment) for apparent ileal digestibility (AID) of Ca, P, and amino acids and phytate disappearance determination. Supplementation of diet with phytase from 0 to 1500 FTU/kg diet linearly improved (P < 0.01) AID of Ca. Both linear and quadratic effects (P < 0.001) was observed for AID of P and phytate-P. Increasing level of phytase supplementation resulted in linear and quadratic increases of Arg, His, Ile, Leu, Lys, Phe, Thr, Trp, Val, Ala, Asp, Cys, Glu, Ser, Tyr and total amino acids. Phytase supplementation increased (P < 0.0001) linearly Met and Pro, with an AID of Met showing a tendency (P = 0.09) for a quadratic increase. In addition, increasing phytase dose linearly increased (P = 0.006) the overall ADFI and ADG and decreased (P < 0.001) F:G ratio. Also, a tendency (P = 0.07) for quadratic effect of phytase was also observed on F:G ratio. These findings highlight that the supplementation of diets with the new bacterial 6-phytase significantly enhanced the utilization of nutrients, disappearance of phytate and performances of weaned piglets in a dose dependent manner.
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Vijayaraghavan, Ponnuswamy, R. Raja Primiya, and Samuel Gnana Prakash Vincent. "Thermostable Alkaline Phytase from Alcaligenes sp. in Improving Bioavailability of Phosphorus in Animal Feed: In Vitro Analysis." ISRN Biotechnology 2013 (February 13, 2013): 1–6. http://dx.doi.org/10.5402/2013/394305.
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A bacterial isolate, Alcaligenes sp. secreting phytase (EC 3.1.3.8), was isolated and characterized. The optimum conditions for the production of phytase included a fermentation period of 96 h, pH 8.0, and the addition of 1% (w/v) maltose and 1% (w/v) beef extract to the culture medium. This enzyme was purified to homogeneity and had an apparent molecular mass of 41 kDa. The optimum pH range and temperature for the activity of phytase were found to be 7.0-8.0 and 60°C, respectively. This enzyme was strongly inhibited by 0.005 M of Mn2+, Mg2+, and Zn2+. In vitro studies revealed that the phytase from Alcaligenes sp. released inorganic phosphate from plant phytates. Phytase released 1930 ± 28, 1740 ± 13, 1050 ± 31, 845 ± 7, 1935 ± 32, and 1655 ± 21 mg inorganic phosphate/kg plant phytates, namely, chick pea, corn, green pea, groundnut, pearl pea, and chick feed, respectively.
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Pirgozliev, V., and M. R. Bedford. "Energy utilisation and growth performance of chicken fed diets containing graded levels of supplementary bacterial phytase." British Journal of Nutrition 109, no. 2 (April 13, 2012): 248–53. http://dx.doi.org/10.1017/s0007114512000943.
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A total of 364 female Ross 308 chicks (1 d old) were used in the present study conducted in floor pens to investigate the effects of graded levels of supplementary bacterial phytase on dietary energy utilisation and growth performance. For this purpose, four maize–soyabean-based diets were offered to the birds from 0 to 21 d of age. These included a suboptimal P negative control (NC, 3·0 g/kg non-phytate P), NC+250 phytase units (FTU)/kg feed, NC+500 FTU and NC+2500 FTU. The effect of phytase activity on bird growth performance was best described as a linear relationship between increasing dose and increased feed intake (P< 0·001), but was quadratic for body-weight gain (P= 0·002) and feed efficiency (P= 0·023). There was no significant response (P>0·05) of dietary apparent metabolisable energy (AME) to supplementary phytase. The birds fed phytase increased their retention of total carcass energy in a linear fashion (P= 0·009) with increased phytase dose. The efficiency of dietary AME used for overall carcass energy retention also improved (P= 0·007) in a linear manner with increased dietary phytase activity. Dietary net energy for production (NEp) increased (P= 0·047) with an increase in phytase dose following a linear pattern, as an increase of 100 FTU increased dietary net energy by 15·4 J (estimated within the range of doses used in the present experiment). Dietary NEp was more highly correlated with performance criteria than dietary AME, and it seems to be a more sensitive way to evaluate broiler response to phytase supplementation.
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Jorquera, Milko, Oscar Martínez, Fumito Maruyama, Petra Marschner, and Maria de la Luz Mora. "Current and Future Biotechnological Applications of Bacterial Phytases and Phytase-Producing Bacteria." Microbes and Environments 23, no. 3 (2008): 182–91. http://dx.doi.org/10.1264/jsme2.23.182.
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Zhilochkina, Tatiana I., Yelena N. Andrianova, Georgy Yu Laptev, and Larisa A. Ilyina. "L-asparaguinates effect on broiler intestine microbiome at bacterial families level." Poultry and Chicken Products 25, no. 1 (2023): 45–48. http://dx.doi.org/10.30975/2073-4999-2023-25-1-45-48.
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At the results of experiment at four groups of “Smena 8” cross broilers it has been established that organic form of mineral additive usage in combination with phytase promotes changes of bacterial community in poultry intestine. For example in groups 1 and 3 without phytase adding more high content has been detected of bacteria that produce car- bohydrate-active enzymes: cellulose and hemicellulase. In 2 and 4 groups poultry that receive phytase and mineral addi- tive as L-asparaginates (4 group) in their diet there were content growth of Clostridia UCG-014 and bacterial families Veil- lonellaceae, Lactobacillaceae, Pseudomonadaceae, Enterobacteriaceae and Ruminococcaceae. The data received have confirmed some assumptions on phytase introduction in diet promotes intestine microbiota increasing and phytase ad- dition together with feed additive being researched effects on poultry feed digestion and increase poultry production and activates osteohistogenesis processes.
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Nurul Izyan Che Mohamood, Nadiawati Alias, and Nurul Asma Hasliza Zulkifly. "Effect of Carbon Sources on Different Strains of Phytase-Producing Bacteria Isolated from Malaysia’s Hot Spring." Journal Of Agrobiotechnology 12, no. 2 (August 17, 2021): 29–38. http://dx.doi.org/10.37231/jab.2021.12.2.255.
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Animal feed from cereal grains and oilseed meals mainly containing phytic acid which has adverse effects on animal nutrition and its environment. Ruminants can easily digest the phytic acid as they have fungi and bacteria in their guts which can produce phytase to degrade the phytic acid. Meanwhile, phytic acid in non-ruminant animals is poorly digested due to the lack of sufficient phytase in their guts. Thus, the feed must be supplemented with inorganic phosphate to ensure it can absorb adequate nutrients. This study aimed to determine the effects of using different carbon sources to the growth of different strains of phytase producing bacteria based on optical density (OD), colony forming unit (CFU), and their phytase production. All four strains of potentially producing-phytase bacteria have been isolated from several hot springs in Malaysia. The bacteria were grown in modified Phytase Screening Medium (PSM) with glucose and lactose as a carbon source and under optimum culture conditions (pH 5.5, 37˚C, 200 rpm) for 72 hours. For quantitative screening of phytase production, the bacterial cultures were harvested to obtain the supernatants that were used to measure the amount of inorganic phosphorus released by the bacterial strains. Among these carbon sources, glucose has shown consistency between their CFU counts and the observed ODs whereas lactose shown inconsistency. Meanwhile, the maximum phytase activity was recorded for all strains in the presence of glucose in which bacteria strain L3 (0.0404 U/mL), RT (0.0359 U/mL), B9 (0.0262 U/mL), and A (0.0263 U/mL). As for the overall, strain L3 (Labis, Johor) gave a promising rate of inorganic phosphate released with optimum phytase activity value of 0.0404 U/mL in presence of glucose and lactose. The optimisation of the fermentation medium can contribute to more economical production of industrial enzyme as phytase has the potential to produce feed additives for poultry feeding.
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Moita, Vitor Hugo C., and Sung Woo Kim. "317 Awardee Talk: Functional Roles of Phytase and Xylanase for Nursery Pigs and Broiler Chickens." Journal of Animal Science 101, Supplement_2 (October 28, 2023): 119–20. http://dx.doi.org/10.1093/jas/skad341.133.
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Abstract The objective of this research was to investigate the functional roles of phytase and xylanase in nursery pigs and broiler chickens. These enzymes have been widely studied and have shown consistent results improving nutrient digestibility and growth performance. However, recent studies have raised the hypothesis that phytase and xylanase could also have potential effects on intestinal and bone health, and intestinal microbiota. The first study studied at the effects of a bacterial 6-phytase on nutrient digestibility and retention of nursery and growing pigs. The results showed that supplementing a bacterial 6-phytase in pig diets can be a potential alternative for reducing high levels of inorganic phosphorus sources while still increasing the digestibility and utilization of phosphorus by the animals. The second study investigated the efficacy and optimal level of a bacterial 6-phytase supplemented beyond traditional dose levels on bone and intestinal health, nutrient digestibility, and growth performance of nursery pigs. The results revealed that the supplementation of the bacterial 6-phytase enhanced bone health, nutrient digestibility, and growth performance in nursery pigs. The third study aimed to determine the effects of phytase supplementation on the intestinal microbiota and morphology, bone health, nutrient digestibility, and growth performance of broiler chickens. The results showed that phytase supplementation had potential benefits on the microbiota by reducing potential harmful bacteria and increasing beneficial bacteria. Additionally, phytase had positive effects on bone health, intestinal morphology, and nutrient digestibility of broiler chickens. The fourth study examined the functional roles of xylanase on the intestinal health and growth performance of nursery pigs. The results showed that xylanase supplementation had beneficial effects on intestinal health by reducing the digesta viscosity, the relative abundance of potentially harmful bacteria, and the oxidative stress in the jejunal mucosa. All of these factors collectively reflected in improvements on the intestinal morphology, nutrient digestibility, and growth performance of nursery pigs. In conclusion, the results of these studies reaffirmed the benefits of phytase and xylanase related to nutrient digestibility and growth performance in nursery pigs and broiler chickens, as well as showed potential benefits on intestinal health and microbiota of the animals. Supplementing these enzymes could also contribute to a reduction in feed costs and environmental concerns associated with a poor hydrolysis and breakdown of antinutritional factors in animal diets.
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Sardar, Rozina, Muhammad Javaid Asad, Muhammad Sheeraz Ahmad, Tanveer Ahmad, and Syed Muhammad Kamal Nasir. "Evaluation of different levels of Bacillus sp. (HCYL03) phytase in broiler chickens fed maize-soyabean meal based diets with a low non-phytate phosphorus content." Veterinarski arhiv 92, no. 4 (September 19, 2022): 483–95. http://dx.doi.org/10.24099/vet.arhiv.1662.
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The objective of the present study was to evaluate the impact of various levels of phytase derived from Bacillus sp. (HCYL03) in corn-soy diets fed to broilers. Experimental treatments included a positive control (PC) with a calculated non-phytate phosphorus (nPP) level of 4.0g/kg for the 35 days of trial. The negative control (NC) diet included a reduction in nPP to 3.0g/kg during the experiment, and commercially available phytase (@500FTU/kg), as well as new bacterial phytase added to the NC diet in increasing amounts of 500, 800, and 1100FTU/kg. Treatment effects on growth performance, the apparent digestibility of P, tibia mineralization, and Ca and P status in blood plasma were evaluated on day 35. The NC diet decreased feed intake (P
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Itkina, Daria L., Aliya D. Suleymanova, and Margarita R. Sharipova. "Expression of Bacillus ginsengihumi M2.11 bacterial phytase by recombinant Pichia pastoris strains." BIO Web of Conferences 36 (2021): 07013. http://dx.doi.org/10.1051/bioconf/20213607013.
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Phytic acid is the main storage form of organic phosphorus. Due to its structural features, phosphorus in phytate is inaccessible for assimilation by animals. Moreover, remaining inaccessible reservoir of phosphorus for animal nutrition, phytic acid is capable of forming insoluble complex salts, which lead to soil and water pollution. Мicrobial enzymes - phytases, capable of decomposing phytic acid to organic phosphorus are being used as feed additives in animal nutrition to solve this problem. Thus, search and development of technologies for the production of enzymes on an industrial scale are the most urgent. Methylotrophic yeast P. pastoris are widely used in biotechnology, as an efficient system for the recombinant proteins expression. They have many advantages, including rapid growth on inexpensive media, a wide range of molecular tools for genetic manipulation in optimizing production processes, they are safe for humans and animals, carry-out many post-translational modifications and produce recombinant proteins intracellularly or extracellularly within a short period of time. It was found that the recombinant P. pastoris strains pPINK-LC-α-MF -phyC, pPINK-HC-α-amyl -phyC, pPINK-LC-α-amyl -phyC, pPINK-HC-α-MF -phyC are able to produce and to secrete B. ginsengihumi bacterial phytase M 2.11 phyC. The maximum activity was observed in the pPINK-LC-α-MF strain – 2.6 (U / mg). Recombinant B. ginsengihumi M 2.11 phytases exhibited high activity in a wide pH range from 2.5 to 9.0. The MF-phyC-HC construction is pH stable. The temperature optimum of all recombinant phytases corresponds to 37 ° C; recombinant phytases retain their activity in the range from -80 to 90C.
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Tang, Hooi Chia, Chin Chin Sieo, Norhani Abdullah, Chun Wie Chong, and Yin Wan Ho. "Preservation of phytase enzyme produced by anaerobic rumen bacteria, Mitsuokella jalaludinii." Journal of Biochemistry, Microbiology and Biotechnology 5, no. 1 (July 31, 2017): 13–17. http://dx.doi.org/10.54987/jobimb.v5i1.334.
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Poultry feed consists of feed ingredients like soybean meal and corn, which contain high levels of phytate that is poorly utilised especially by the monogastric animals that lack of phytase. Hence, phytase has been extensively applied as a feed supplement in poultry production due to the efficiency of this enzyme in improving phosphorous (P) availability, thus reducing P excretion to the environment as well as reducing the feed cost by reducing inorganic P supplementation. Mitsuokella jalaludinii, an obligate anaerobe, Gram-negative rumen bacterium, produces high phytase activity. Birds supplemented with bacterial preparation of M. jalaludinii showed comparable performance to that of commercial phytase. However, the anaerobic nature of this bacterium renders difficulty in the use of live cells as feed supplement in commercial poultry production. Therefore, this study was conducted to determine a suitable method to preserve phytase activity of M. jalaludinii regardless of cells viability. Mitsuokella jalaludinii was grown in MF medium under anaerobic condition and the cells were subjected to various treatments to preserve the enzyme, including bead beating, compressed air, moist heat, dry heat and freeze-drying under aerobic condition. The results showed that the total number of viable cells were significantly (p<0.05) reduced when the cells were subjected to bead beating, whereas no viable cells were detected for compressed air, moist heat, dry heat and freeze-drying. Bead beating, compressed air, moist heat and dry heat treatments resulted in the reduction of phytase activity. However, only freeze-drying method was able to preserve high level of phytase activity significantly (p<0.05).
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Hu, Jun, Chao Ran, Suxu He, Yanan Cao, Bin Yao, Yuantu Ye, Xuezhen Zhang, and Zhigang Zhou. "Dietary microbial phytase exerts mixed effects on the gut health of tilapia: a possible reason for the null effect on growth promotion." British Journal of Nutrition 115, no. 11 (April 15, 2016): 1958–66. http://dx.doi.org/10.1017/s0007114516001240.
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AbstractThe present study evaluated the effects of dietary microbial phytase on the growth and gut health of hybrid tilapia (Oreochromis niloticus♀×Oreochromis aureus♂), focusing on the effect on intestinal histology, adhesive microbiota and expression of immune-related cytokine genes. Tilapia were fed either control diet or diet supplemented with microbial phytase (1000 U/kg). Each diet was randomly assigned to four groups of fish reared in cages (3×3×2 m). After 12 weeks of feeding, weight gain and feed conversion ratio of tilapia were not significantly improved by dietary microbial phytase supplementation. However, significantly higher level of P content in the scales, tighter and more regular intestinal mucosa folds were observed in the microbial phytase group and the microvilli density was significantly increased. The adhesive gut bacterial communities were strikingly altered by microbial phytase supplementation (0·41<similarity coefficient<0·54). Stimulated intestinal inflammation and stress status were observed in the fish fed diet supplemented with microbial phytase, as indicated by the up-regulated intestinal expressions of the cytokine genes (tnf-αandtgf-β) andhsp70. In addition, the gut microvilli height was significantly decreased in the phytase group. These results indicate that dietary microbial phytase may exert mixed effects on hybrid tilapia, and can guide our future selection of phytases as aquafeed additives – that is, eliminating those that can stimulate intestinal inflammation.
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Saleh, Ahmed A., Mohammed Elsawee, Mohamed M. Soliman, Reyad Y. N. Elkon, Mohammed H. Alzawqari, Mustafa Shukry, Abdel-Moneim Eid Abdel-Moneim, and Hatem Eltahan. "Effect of Bacterial or Fungal Phytase Supplementation on the Performance, Egg Quality, Plasma Biochemical Parameters, and Reproductive Morphology of Laying Hens." Animals 11, no. 2 (February 19, 2021): 540. http://dx.doi.org/10.3390/ani11020540.
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Catalytic and physicochemical properties of microbial phytase sources may differ, affecting phosphorus (P) release and subsequently the productive and reproductive performance of layers. The current study aimed to evaluate the impact of bacterial and fungal phytase sources on layer productivity, egg production, biochemical blood indices, and reproductive morphology. For this purpose, 360 Bovans brown hens at 42 weeks of age were randomly allocated into 4 experimental groups, each with 15 replicates of 6 hens. The first group (control) was fed a basal diet with 4.6 g/kg available P. In contrast, the second, third, and fourth groups were fed diets treated with 3.2 g/kg available P, supplemented with either 5000 FTU/kg of bacterial E. coli (QuantumTM Blue 5G), fungal Aspergillus niger (VemoZyme® F 5000 Naturally Thermostable Phytase (NTP)), or fungal Trichodermareesei (Yemzim® FZ100). Dietary supplementation of bacterial and fungal phytases did not affect the productive performance or egg quality criteria, except for increased shell weight and thickness (p < 0.05). Serum hepatic function biomarkers and lipid profiles were not altered in treated hens, while calcium and P levels were increased (p < 0.05) related to the controls. Ovary index and length, and relative weight of oviduct and its segments were not influenced. The contents of cholesterol and malondialdehyde in the yolks from treated birds were lower compared to control hens, while calcium and P content increased (p < 0.05). Conclusively, bacterial and fungal phytase sources can compensate for the reduction of available P in layers’ diets and enhance shell and yolk quality without affecting productive performance, and no differences among them were noticed.
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Choi, Hyunjun, and Sung Woo Kim. "272 Effects of Increasing Dose of a Novel Hybrid Bacterial 6-Phytase on Apparent Total Tract Nutrient Digestibility, Release of Free Myoinositol, and Retention of Calcium and Phosphorus in Pigs." Journal of Animal Science 101, Supplement_3 (November 6, 2023): 190–91. http://dx.doi.org/10.1093/jas/skad281.231.
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Abstract The objective of this study was to investigate the effects of increasing dose of a novel hybrid bacterial 6-phytase on apparent total tract digestibility (ATTD) of energy and nutrients, plasma concentrations of free myoinositol, and retention of calcium (Ca) and phosphorus (P). A total of 15 barrows (21.5 ± 1.4 kg BW) were allotted to a triplicate 5 × 5 Latin square design with 5 dietary treatments and 5 periods per each square. Pigs were individually housed in a metabolism crate. Five dietary treatments consisted of 5 levels of phytase-supplemented diets with increasing phytase units (0, 500, 1,000, 1,500, and 2,000 FTU/kg). Diets contained nutrients meeting the requirement except for Ca and P. Degree of deficient Ca and P content in the NC diet were 0.22 and 0.17%, respectively. Each period lasted 10 days consisting of a 5-d adaptation period and 5-d collection period for total collection. Data were analyzed using orthogonal polynomial contrasts using the Proc MIXED and optimal phytase levels were determined using the NLMIXED of SAS. The increasing phytase supplementation linearly increased (P < 0.05) average daily gain (651 to 695 g/d) and tended to linearly increase (P = 0.072) gain to feed ratio (0.53 to 0.56). Increasing phytase supplementation linearly increased (P < 0.01) ATTD of Ca and P and retention of Ca and P and also showed quadratic effects (P < 0.01). Increasing phytase supplementation linearly increased (P < 0.01) myoinositol content in plasma and also showed quadratic effects (P < 0.05). Broken line analyses showed that ATTD of Ca (P < 0.01) and retention of Ca (P < 0.01) were increased until phytase supplementation were increased (79.4 to 92.1% and 77.4 to 91.4%) from 0 to 671 FTU/kg (SE = 67; P < 0.01) and 0 to 668 FTU/kg (SE = 72; P < 0.01), respectively. Broken line analyses showed that ATTD of P (P < 0.01) and retention of P (P < 0.01) were increased (59.0 to 80.1% and 55.7 to 78.2%) until phytase contents were increased from 0 to 783 FTU/kg (SE = 91; P < 0.01) and 0 to 776 FTU/kg (SE = 76; P < 0.01), respectively. The broken line analyses showed that myoinositol contents in plasma (P < 0.01) were increased (13.2 to 23.2 µmol/L) until phytase supplementation were increased from 0 to 624 FTU/kg (SE = 174; P < 0.01). In conclusion, increasing phytase supplementation up to 2,000 FTU/kg feed improved ATTD of Ca and P, retention of Ca and P, complete hydrolysis of phytate, and growth performance of pigs compared with a P-deficient diet. Phytase supplementation at a range of 600 to 800 FTU/kg feeds provided the optimal improvement of Ca, P, and phytate utilization.
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Leeson, S., H. Namkung, M. Cottrill, and C. W. Forsberg. "Efficacy of new bacterial phytase in poultry diets." Canadian Journal of Animal Science 80, no. 3 (September 1, 2000): 527–28. http://dx.doi.org/10.4141/a99-123.
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Phytase produced by Escherichia coli was fed to broiler chicks fed graded levels of available phosphorus (0.25–45%). A positive control involved Natuphos® a fungal phytase commonly used in poultry nutrition. Feeding diets with only 0.25% available phosphorus from 1 to 13 d of age resulted in reduced feed intake and weight gain, with associated loss in tibia weight, tibia ash and tibia phosphorus content (P < 0.01). Adding 600 IU phytase kg−1 diet corrected these problems, and both sources were equally efficacious for most parameters studied. Key words: Phosphorus, phytase, chicken
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Muñoz-Muñoz, Patricia L. A., Celina Terán-Ramírez, Rosa E. Mares-Alejandre, Ariana B. Márquez-González, Pablo A. Madero-Ayala, Samuel G. Meléndez-López, and Marco A. Ramos-Ibarra. "Surface Engineering of Escherichia coli to Display Its Phytase (AppA) and Functional Analysis of Enzyme Activities." Current Issues in Molecular Biology 46, no. 4 (April 17, 2024): 3424–37. http://dx.doi.org/10.3390/cimb46040215.
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Escherichia coli phytase (AppA) is widely used as an exogenous enzyme in monogastric animal feed mainly because of its ability to degrade phytic acid or its salt (phytate), a natural source of phosphorus. Currently, successful recombinant production of soluble AppA has been achieved by gene overexpression using both bacterial and yeast systems. However, some methods for the biomembrane immobilization of phytases (including AppA), such as surface display on yeast cells and bacterial spores, have been investigated to avoid expensive enzyme purification processes. This study explored a homologous protein production approach for displaying AppA on the cell surface of E. coli by engineering its outer membrane (OM) for extracellular expression. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of total bacterial lysates and immunofluorescence microscopy of non-permeabilized cells revealed protein expression, whereas activity assays using whole cells or OM fractions indicated functional enzyme display, as evidenced by consistent hydrolytic rates on typical substrates (i.e., p-nitrophenyl phosphate and phytic acid). Furthermore, the in vitro results obtained using a simple method to simulate the gastrointestinal tract of poultry suggest that the whole-cell biocatalyst has potential as a feed additive. Overall, our findings support the notion that biomembrane-immobilized enzymes are reliable for the hydrolysis of poorly digestible substrates relevant to animal nutrition.
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Dhiman, Sandhya, Ramesh Chand Dubey, Dinesh Kumar Maheshwari, and Sandeep Kumar. "Sulfur-oxidizing buffalo dung bacteria enhance growth and yield of Foeniculum vulgare Mill." Canadian Journal of Microbiology 65, no. 5 (May 2019): 377–86. http://dx.doi.org/10.1139/cjm-2018-0476.
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This study aimed to harness the benefits of sulfur-oxidizing beneficial bacteria from buffalo dung to improve crop yields of Foeniculum vulgare. A total of 61 bacterial isolates were screened from buffalo dung, of which 40 isolates exhibited plant-growth-promoting attributes, such as phosphate solubilization, indole-3-acetic acid production, and hydrogen cyanide production. Of these 40, four bacterial isolates, viz., BUFF12, BUFF14, BUFF23, and BUFF38, were the most potent, having plant-growth-promoting and sulfur-oxidizing properties. These four isolates produced phytase by solubilizing calcium phytate and sodium phytate. They solubilized potassium besides oxidizing the sulfur, causing an increase in soil fertility and crop production. All four isolates were nonpathogenic in nature, as demonstrated by a negative haemolysis test. According to the 16S rRNA gene sequence, the isolate BUFF14 was identified as Proteus mirabilis. Proteus mirabilis BUFF14 maximized seed germination with enhanced vegetative and reproductive parameters during pot and field trial studies, compared with the other isolates.
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Velayudhan, Deepak, Xandra Benthem de Grave, Katie Waller, Leon Marchal, and Yueming Dersjant-Li. "117 Functionality of a novel consensus bacterial 6-phytase variant in enhancing phosphorus digestibility in gestating and lactating sows." Journal of Animal Science 98, Supplement_4 (November 3, 2020): 109. http://dx.doi.org/10.1093/jas/skaa278.200.
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Abstract Two experiments were conducted to determine the efficacy of phytase on apparent total tract digestibility (ATTD) of phosphorus (P) in gestating and lactating sows. In Exp. 1, a total of 40 sows (parity 2–6) at d 78 of gestation were randomly assigned, based on parity, over 4 experimental diets. In Exp. 2, 40 sows (parity 2.0–6.0) at d 4 of lactation were randomly assigned based on parity, over 4 experimental diets. Both studies had a positive control (PC) diet with adequate levels of calcium (Ca) and digestible P (6.5 and 2.5 g/kg and 7.0 and 3.2 g/kg for gestation and lactation diets, respectively), a negative control (NC) diet with low Ca and digestible P levels (5.0 and 1.0 g/kg and 5.0 and 1.5 g/kg for gestation and lactation diets, respectively), and NC supplemented with a novel consensus bacterial 6-phytase variant (PhyG) or a commercial Buttiauxella sp. phytase (PhyB), both at 500 FTU/kg. Diets were corn, soybean meal and sunflower meal based (phytate P of 3.0 g/kg) with restricted feeding during gestation and ad libitum during lactation. In both studies, fecal samples were collected after an adaptation of 14 d on 4 successive days to determine ATTD of P. Data were analyzed using ANOVA, treatment mean comparison using Tukey test using JMP 14. In addition, data from the Exp. 1 and 2 were pooled to further investigate their combined effect, using trial as random effect. In both studies, supplementation of both phytases improved (P < 0.05) ATTD of P vs NC. With data combined from two trials, sows fed PhyG showed greater (P < 0.05) ATTD of P when compared those fed PhyB. In conclusion, PhyG at 500 FTU/kg showed a greater ATTD of P in sows when compared to the PhyB at 500 FTU/kg, when data combined from two trials.
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Nelson, Megan E., Su A. Lee, Yueming Dersjant-Li, Deepak Velayudhan, Janet C. Remus, and Hans H. Stein. "PSVI-8 Effects of Phosphorus Level and Increasing Phytase Dose on Basal Endogenous Loss of Calcium and Balance of Phosphorus in Pigs Fed Diets Containing Phytate P at Commercial Levels." Journal of Animal Science 100, Supplement_2 (April 12, 2022): 165–66. http://dx.doi.org/10.1093/jas/skac064.282.
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Abstract The objective was to test the hypothesis that dietary P concentration and level of phytase influences basal endogenous loss of Ca in pigs. Seventy barrows (body weight: 17.66 kg) were housed in metabolism crates and allotted to 7 Ca-free diets containing corn, potato protein concentrate, and full-fat rice bran (0.27% phytate P). A positive control (PC) diet contained P at the requirement for digestible P for 11 to 25 kg pigs. Negative control (NC) diets were formulated by reducing concentration of digestible P by 0.15% and adding a novel consensus bacterial 6-phytase variant at 0, 250, 500, 1,000, 2,000, or 4,000 phytase units/kg diet. Feces and urine samples were collected separately for 4 d after 5 d of adaptation. Dried and ground fecal samples were analyzed for dry matter, Ca, and P and urine was analyzed for P. Data were analyzed using a model that included diet as fixed effect. Positive control and NC were compared using a contrast statement and linear and exponential effects of dietary phytase were tested. Results indicated that apparent total tract digestibility of dry matter was not affected by dietary P or phytase (Table 1). The basal endogenous loss of Ca was not affected by dietary P, but exponentially (P = 0.030) decreased as dietary phytase increased. Phosphorus retention (g/d) was greater (P < 0.001) in pigs fed PC compared with NC without phytase. The standardized total tract digestibility (STTD) of P exponentially (P < 0.001) increased as phytase increased; however, retention of P as % of absorbed P decreased (linear, P = 0.006) as phytase increased because the extra P absorbed as a result of phytase could not be retained due to the lack of Ca. In conclusion, increasing the phytase dose decreased basal endogenous loss of Ca and increased STTD of P.
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Zailan, Nur Dinie, Nurul Asma Hasliza Zulkifly, Afnani Alwi, Siti Noor Syuhada Muhammad Amin, and Nadiawati Alias. "Effects of Nitrogen Sources in Phytase Production on Bacterial Strains Isolated from Malaysia’s Hot Spring." Journal Of Agrobiotechnology 12, no. 1S (September 29, 2021): 31–39. http://dx.doi.org/10.37231/jab.2021.12.1s.268.
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Efficient strategies for phytase production gained increasing importance as more applications require high amounts of phytase for the market. Four phytase-producing bacterial strains isolated from Malaysia’s hot springs were used in this study to determine the effect of nitrogen sources on phytase production. All of the strains were screened out by applying halozone method which shows all of the strains were definitely positive phytase producer. Phytase Screening Medium (PSM) with soybean extract as substrate was used as a cultivation medium. Optimised condition with 1.0 % (w/v) of glucose (as carbon source), pH 5.5 and 37°C temperature was applied. Yeast extract and peptone were used to identify optimum nitrogen source in maximum phytase production. Quantitative analysis observed were optical density, colony forming unit, pH values and phytase activity to identify the effect of nitrogen source in phytase production. The finding was bacterial strain L3 as the best producer in producing maximum phytase (0.2162 U/mL) with optimised condition using yeast extract as nitrogen source. Findings in this study proved that yeast extract act as the optimum nitrogen source which contribute to maximum phytase production as supported by previous studies. This study can provide an efficient strategy to produce maximum phytase as few studies stated that phytase is an application tool in functional food production that consists of myo-inositol phosphates that is believed to have important pharmacological effects.
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Garrett, James B., Keith A. Kretz, Eileen O'Donoghue, Janne Kerovuo, William Kim, Nelson R. Barton, Geoffrey P. Hazlewood, Jay M. Short, Dan E. Robertson, and Kevin A. Gray. "Enhancing the Thermal Tolerance and Gastric Performance of a Microbial Phytase for Use as a Phosphate-Mobilizing Monogastric-Feed Supplement." Applied and Environmental Microbiology 70, no. 5 (May 2004): 3041–46. http://dx.doi.org/10.1128/aem.70.5.3041-3046.2004.
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ABSTRACT The inclusion of phytase in monogastric animal feed has the benefit of hydrolyzing indigestible plant phytate (myo-inositol 1,2,3,4,5,6-hexakis dihydrogen phosphate) to provide poultry and swine with dietary phosphorus. An ideal phytase supplement should have a high temperature tolerance, allowing it to survive the feed pelleting process, a high specific activity at low pHs, and adequate gastric performance. For this study, the performance of a bacterial phytase was optimized by the use of gene site saturation mutagenesis technology. Beginning with the appA gene from Escherichia coli, a library of clones incorporating all 19 possible amino acid changes and 32 possible codon variations in 431 residues of the sequence was generated and screened for mutants exhibiting improved thermal tolerance. Fourteen single site variants were discovered that retained as much as 10 times the residual activity of the wild-type enzyme after a heated incubation regimen. The addition of eight individual mutations into a single construct (Phy9X) resulted in a protein of maximal fitness, i.e., a highly active phytase with no loss of activity after heating at 62°C for 1 h and 27% of its initial activity after 10 min at 85°C, which was a significant improvement over the appA parental phytase. Phy9X also showed a 3.5-fold enhancement in gastric stability.
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Velayudhan, Deepak, Rosil Lizardo, Boris Villca, Janet C. Remus, Leon Marchal, and Yueming Dersjant-Li. "90 Efficacy of Different Phytases on Growth Performance, Energy and Nutrient Digestibility and Bone Quality of Weaned Piglets Fed a Wheat-Corn Soybean Meal Based Complex Diet." Journal of Animal Science 100, Supplement_2 (April 12, 2022): 38–39. http://dx.doi.org/10.1093/jas/skac064.061.
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Abstract The study determined efficacy of different phytases in diets without inorganic phosphorus (P) in piglets. Piglets (initial body weight 9.7 ± 1.2 kg) were randomly assigned (2 piglets/pen), based on sex and body weight over 6 diets. Positive control (PC) corn-wheat based diet was formulated to meet NRC specification. Negative control (NC) diet was formulated without inorganic P (-0.16% STTD P), with reduced calcium (Ca; -0.15%), net energy (-33 kcal/kg), and standardized ileal digestible amino acids (average -0.02% unit). The NC diets were supplemented with 4 different phytases at 1,500 FTU/kg feed including a novel consensus bacterial 6-phytase variant expressed in Trichoderma reesei (PhyG), a Citrobacter braakii phytase expressed in Aspergillus oryzae (PhyC), a hybrid phytase (Hafnia sp., Yersinia sp. and Buttiauxella sp.) expressed in Aspergillus niger (PhyH) and an Escherichia coli phytase expressed in Pichia pastoris (PhyE). Piglets were fed ad libitum for 42 d. Fecal samples were collected during last 3 d. On d 42, 1 piglet per pen was euthanized and metacarpi collected for bone strength and ash analysis. Data were analysed by one-way ANOVA and treatment mean comparison by Tukey test using JMP 14. Pigs fed NC diets showed reduced (P < 0.05) growth, P digestibility, bone ash and bone strength vs PC and all phytases treatments. Overall feed conversion ratio (FCR) was less (P < 0.05) with PhyG (-22 points) and PhyE (-8 points) vs NC, while PhyC and PhyH showed intermediate FCR between PhyG and PhyE. PhyG improved (P < 0.05) bone strength and P digestibility vs PhyC, PhyE and PC. Based on FCR and bone strength, ranking of the phytases are: PhyG >PhyC >PhyH >PhyE. In conclusion, all phytases improved growth, nutrient digestibility and bone strength vs NC; however, only PhyG fully compensated nutrients and energy reduction, fully recovered performance to the level of PC and resulted in improved bone strength compared with PC.
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Riaño-Castillo, Edna Rocio, Juan Carlos Rodríguez-Ortiz, Hye-Ji Kim, María de la Luz Guerrero González, María Fernanda Quintero-Castellanos, and Pablo Delgado-Sánchez. "Isolation and Identification of Lysinibacillus sp. and Its Effects on Solid Waste as a Phytate-Mineralizing Bacterium in an Aquaponics System." Horticulturae 10, no. 5 (May 11, 2024): 497. http://dx.doi.org/10.3390/horticulturae10050497.
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Sedimentable solids generated in aquaponic systems are mainly composed of organic waste, presenting molecules such as phytate, which can be a potential source of inorganic nutrients through mineralization. This work aimed to isolate and identify phytase-producing bacteria and evaluate the inoculation effects of pure strains on mineralization and nutrient release from solid waste generated in aquaponic systems at different oxygen and temperature conditions. The bacteria were isolated from the settleable solids of a commercial aquaponic system and molecularly identified by amplifying the 16S rRNA gene. Subsequently, two tests were carried out: 1. Test for the biochemical identification of phytase-producing bacteria; 2. In vitro mineralization test, where the ability to mineralize phytate and release nutrients under different oxygen conditions [0 rpm (2.1 mg L−1) and 200 rpm (7.8 mg L−1)] and temperatures (24 and 37 °C) were evaluated. Our findings show that two pure strains of Lysinibacillus mangiferihumi can mineralize phytate under conditions of 200 rpm and 24 °C, mainly increasing the mineralization of PO4- and Ca, a property that has not yet been reported for this species. On the other hand, at 0 rpm and 24 °C, an increase in K was observed (control conditions), while the conditions of 200 rpm and 24 °C, regardless of bacterial inoculation, favored a rise in S, Mg, and Fe. The Lysinibacillus strains obtained in this investigation are of great importance due to their application in agriculture and the optimization of mineralization in aquaponic systems. A proper combination of oxygen and temperature will lead to a greater availability of nutrients for the growth and development of vegetables.
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Osman, AA, and BM El-Boussairi. "Isolation of phytate-degrading microorganisms with inorganic phosphate solubilizing activity (biofertilizers) for plant growth enhancement." IOP Conference Series: Earth and Environmental Science 1055, no. 1 (July 1, 2022): 012017. http://dx.doi.org/10.1088/1755-1315/1055/1/012017.
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Abstract Phosphorus (P) shortage in the soil is a key problem in agricultural output. Phytate (Myo-inositol phosphate), which is stable and unavailable to plants, is abundant in soils, accounting for up to 50% of all organic P. The conversion of phytate to Myo-inositol and phosphate is catalyzed by phytases. Microorganisms that solubilize P, biofertilizers, are common in soils and may play a major role in providing P to plants. Secreted microbial phytases enhancing plant development in terms of phosphorus availability. This study aimed to identify possible phytate degrading microorganisms from the rhizosphere, animal dung, soil, and livestock farms samples collected from farms in Khartoum, Sudan. Modified Luria Bertani and phytase screening media supplemented with wheat bran as a source of phytate were used as selection media. Each sample was serially diluted 10 times, and 100 µl from dilution 103 was distributed onto each of two screening medium plates, cultured aerobically at 37°C for 1 to 4 days, A putative phytate degrading bacterium was detected in isolates with a clear zone around their growing colonies. A total of 17 isolates were obtained, with 15/17 (88%) being bacterial isolates and 2/17 being fungal isolates. Phosphate solubilizing activity was performed on Pikovskaya’s agar plate, and plates were incubated at 37°C for 7 days. The plates were examined for the presence of a halo zone around each isolated culture. out of the 17 isolates, only one Gram-positive Bacilli (isolate AB13) was found to have phosphate solubilizing activity and the ability to liberate P from phytate, suggesting that it could be used as a biofertilizer.
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Suldina, E. V., I. I. Bogdanov, N. A. Feoktistova, and N. G. Bart. "PROTEIN PROFILING OF CANDIDATE STRAINS OF BACTERIAL COMPOSITION." Vestnik of Ulyanovsk state agricultural academy 167, no. 4 (60) (December 23, 2022): 102–10. http://dx.doi.org/10.18286/1816-4501-2022-4-102-110.
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Organic farming and the application of organic and biological fertilizers and protective equipment is not only a promising, but also a safe way to obtain food products. Various bacterial compositions are widely used to increase the microbial activity of the soil, which increases the availability of nutrients that plants can easily absorb. They increase soil fertility by fixing nitrogen and dissolve insoluble phosphates in the soil, resulting in the formation of chemicals that promote plant growth. The aim of this work was to determine the presence of phytase, nitrogenase and alkaline phosphatase enzymes in strains of candidate bacteria of bacterial composition by proteomic profiling in polyacrylamide gel. The molecular weights of phytase, nitrogenase and phosphotase of candidate strains were determined in the system http://molbiol.ru/ based on amino acid sequences. After that, proteins with the corresponding molecular weights were detected on the forogram. According to the results of proteomic analysis of Bacillus subtilis, proteins with molecular weights of 50.5, 16.5 and 74.9 kDa corresponding to the enzymes phytase, nitrogenase and phosphatase were identified in 10 strains. Proteins with molecular weights of 41.8, 16.7 and 24.8 kDa corresponding to the enzymes phytase, nitrogenase and phosphatase were detected in 6 strains of Bacillus megaterium. 4 strains of Pseudomonas stutzeri contained proteins with molecular weights of 69, 31.8 and 73.5 kDa corresponding to the required enzymes. As a result of the carried out experiments, candidates bacterial strains were identified for the development of a biocomposition to increase the coefficient of assimilation of mineral components of fertilizers.
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Ahmed, Bulbul, Jean-Baptiste Floc’h, Zakaria Lahrach, and Mohamed Hijri. "Phytate and Microbial Suspension Amendments Increased Soybean Growth and Shifted Microbial Community Structure." Microorganisms 9, no. 9 (August 25, 2021): 1803. http://dx.doi.org/10.3390/microorganisms9091803.
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Phytate represents an organic pool of phosphorus in soil that requires hydrolysis by phytase enzymes produced by microorganisms prior to its bioavailability by plants. We tested the ability of a microbial suspension made from an old growth maple forest’s undisturbed soil to mineralize phytate in a greenhouse trial on soybean plants inoculated or non-inoculated with the suspension. MiSeq Amplicon sequencing targeting bacterial 16S rRNA gene and fungal ITS was performed to assess microbial community changes following treatments. Our results showed that soybean nodulation and shoot dry weight biomass increased when phytate was applied to the nutrient-poor substrate mixture. Bacterial and fungal diversities of the root and rhizosphere biotopes were relatively resilient following inoculation by microbial suspension; however, bacterial community structure was significantly influenced. Interestingly, four arbuscular mycorrhizal fungi (AMF) were identified as indicator species, including Glomus sp., Claroideoglomus etunicatum, Funneliformis mosseae and an unidentified AMF taxon. We also observed that an ericoid mycorrhizal taxon Sebacina sp. and three Trichoderma spp. were among indicator species. Non-pathogenic Planctobacteria members highly dominated the bacterial community as core and hub taxa for over 80% of all bacterial datasets in root and rhizosphere biotopes. Overall, our study documented that inoculation with a microbial suspension and phytate amendment improved soybean plant growth.
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Suliasih and S. Widawati. "Inorganic and organic phosphate solubilization potential of Stenotrophomonas maltophilia." IOP Conference Series: Earth and Environmental Science 948, no. 1 (December 1, 2021): 012054. http://dx.doi.org/10.1088/1755-1315/948/1/012054.
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Abstract Most soils contain Phosphorus (P) in insoluble compounds as organic and inorganic forms which is unavailable to plants. Furthermore, phosphate solubilizing bacteria (PSB) play an important role in converting insoluble P to a bioavailable form through solubilizing and mineralizing inorganic or organic P. Therefore, this study aims to determine the ability of isolate a phosphate solubilizing bacterium (PSB), Stenotrophomonas maltophilia to solubilize inorganic and organic P. The bacterium was isolated from peat soil of West Kalimantan using Pikovskaya medium added with Ca3 (PO4)2 as the P source. The activity of organic P (acid phosphatase and phytase) was measured using p-nitrophenyl phosphate disodium (pNPP 0.115 M) and sodium phytate as the substrate of bacterial broth culture. Also, the optimization for inorganic and organic P solubilization by the bacterium was conducted. The maximum values of inorganic and organic phosphate solubilization was recorded to be 52.26 μg/mL and 44.51 U/mL (acid phosphatase), 0.13 U/mL (phytase) respectively. Optimum conditions found were temperature at 30 °C, pH 6.0 and in the presence of sucrose and beef extract, which serve as carbon and nitrogen sources, respectively. Therefore, it can probably be used as a biological fertilizer for plants.
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Munmun, S., MA Rahman, KMS Islam, and R. Chowdhury. "Efficacy of bacterial phytase, citric acid and their combination in broiler fed inorganic phosphorus free diet." Bangladesh Journal of Animal Science 51, no. 3 (September 28, 2022): 107–14. http://dx.doi.org/10.3329/bjas.v51i3.61786.
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Present study was conducted to assess the efficacy of Escherichia Coli derived phytase alone or with citric acid (CA) given inorganic phosphorus (P) free broiler diet. Ninety six one-day old COBB-500 male broilers were divided into four groups and fed one of the following diets for a period of 35 days: positive control (PC) diet formulated based on the NRC (1994) recommendations, negative control (NC) diet (inorganic P free diet; containing 0.20% lower P than that in the PC diet), and two other diets were formulated by adding only phytase (500 FTU/kg of feed) or phytase with CA (500 FTU/kg of feed with 2 % CA) in the NC diet. Growth performance, serum minerals concentration, tibia and shank characteristics, and nutrient retention were measured. Impaired growth performance (final BW, BW gain, and FCR) in broilers (NC group) fed inorganic P free diet was restored with the addition of phytase in NC diet. Best FCR (1.83) was recorded in phytase added group and worst in NC group (2.03), however, further addition of CA with phytase did not show any significant variation. Lowest concentration of serum P (mg/dl), tibia P (%) and shank P (%) were ameliorated (P<0.05) by the addition of phytase, and the restoration magnitude was non-significantly greater in phytase with CA group. Retention of total P (%) in broilers given phytase added diet was comparable with broilers given PC diet, although the former diet contained lower level of available P in diet than the later one. Addition of CA further increased this retention (%) numerically. Broilers fed inorganic P free diet deficient in available P, with Escherichia Coli derived phytase showed the growth performance (P<0.05) and relative retention of P comparable with broilers fed NRC (1994) recommended one. Therefore, relying on Escherichia Coli derived phytase alone in available P deficient diet may be viable to ensure sufficient supply of P in broilers diets.
Bangladesh Journal of Animal Science 51 (3): 107-114, 2022
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Farias, Nathálya, Isabela Almeida, and Carlos Meneses. "New Bacterial Phytase through Metagenomic Prospection." Molecules 23, no. 2 (February 17, 2018): 448. http://dx.doi.org/10.3390/molecules23020448.
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Guerrero-Olazarán, Martha, Lilí Rodríguez-Blanco, Jesús G. Carreon-Treviño, Juan A. Gallegos-López, Miguel Castillo-Galván, and José M. Viader-Salvadó. "Bacterial phytase produced in Pichia pastoris." Journal of Biotechnology 131, no. 2 (September 2007): S233—S234. http://dx.doi.org/10.1016/j.jbiotec.2007.07.425.
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Castillo Villamizar, Genis Andrés, Heiko Nacke, Laura Griese, Lydia Tabernero, Katrina Funkner, and Rolf Daniel. "Characteristics of the First Protein Tyrosine Phosphatase with Phytase Activity from a Soil Metagenome." Genes 10, no. 2 (January 29, 2019): 101. http://dx.doi.org/10.3390/genes10020101.
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Protein tyrosine phosphatases (PTPs) fulfil multiple key regulatory functions. Within the group of PTPs, the atypical lipid phosphatases (ALPs) are known for their role as virulence factors associated with human pathogens. Another group of PTPs, which is capable of using inositol-hexakisphosphate (InsP6) as substrate, are known as phytases. Phytases play major roles in the environmental phosphorus cycle, biotechnology, and pathogenesis. So far, all functionally characterized PTPs, including ALPs and PTP-phytases, have been derived exclusively from isolated microorganisms. In this study, screening of a soil-derived metagenomic library resulted in identification of a gene (pho16B), encoding a PTP, which shares structural characteristics with the ALPs. In addition, the characterization of the gene product (Pho16B) revealed the capability of the protein to use InsP6 as substrate, and the potential of soil as a source of phytases with so far unknown characteristics. Thus, Pho16B represents the first functional environmentally derived PTP-phytase. The enzyme has a molecular mass of 38 kDa. The enzyme is promiscuous, showing highest activity and affinity toward naphthyl phosphate (Km 0.966 mM). Pho16B contains the HCXXGKDR[TA]G submotif of PTP-ALPs, and it is structurally related to PtpB of Mycobacterium tuberculosis. This study demonstrates the presence and functionality of an environmental gene codifying a PTP-phytase homologous to enzymes closely associated to bacterial pathogenicity.
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Metzler-Zebeli, Barbara U., Jutamat Klinsoda, Julia C. Vötterl, and Doris Verhovsek. "Maturational Changes Alter Effects of Dietary Phytase Supplementation on the Fecal Microbiome in Fattening Pigs." Microorganisms 8, no. 7 (July 18, 2020): 1073. http://dx.doi.org/10.3390/microorganisms8071073.
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Age-related successions in the porcine gut microbiome may modify the microbial response to dietary changes. This may especially affect the bacterial response to essential nutrients for bacterial metabolism, such as phosphorus (P). Against this background, we used phytase supplementation (0 or 650 phytase units/kg complete feed) to alter the P availability in the hindgut and studied the dietary response of the fecal bacterial microbiome from the early to late fattening period. Fecal DNA were isolated after 0, 3, 5 and 10 weeks and the V3-V4 region of the 16S rRNA gene was sequenced. Permutational analysis of variance showed distinct bacterial communities for diet and week. Alpha-diversity and taxonomy indicated progressing maturation of the bacterial community with age. Prevotellaceae declined, whereas Clostridiaceae and Ruminococcaceae increased from weeks 0 to 3, 5, and 10, indicating changes in fiber-digesting capacities with age. Phytase affected all major bacterial taxa but reduced species richness (Chao1) and diversity (Shannon and Simpson). To conclude, present results greatly support the importance of available P for bacterial proliferation, including fibrolytic, lactic acid- and butyrate-producing genera, in pigs. Results also emphasize the necessity to assess bacterial responses to dietary manipulation at several time points throughout the fattening period.
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Velayudhan, Deepak E., Ester Vinyeta-Punti, Rachael Hardy, Leon Marchal, Charles Martin Nyachoti, Hans H. Stein, and Yueming Dersjant-Li. "154 The Effect of a Novel Consensus Bacterial 6-Phytase Variant on Ileal Digestibility of Energy in Growing Pigs: Meta-Analysis of 4 Trials." Journal of Animal Science 101, Supplement_2 (October 28, 2023): 117–18. http://dx.doi.org/10.1093/jas/skad341.131.
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Abstract This meta-analysis evaluated the effect of a novel consensus bacterial 6-phytase variant (PhyG) on the apparent ileal digestibility (AID) of energy in growing pigs. It was hypothesized that increasing phytase doses improves energy digestibility. The database was constructed using 4 studies (PIC, L 800 × Camborough sows or TN Tempo × TN70 breeds; average body weight of 26.5 kg at sampling), contributing 220 data points. Pigs were fed corn or corn-wheat based diets with soybean meal, canola meal, rice bran, wheat middlings or DDGS. All studies had a negative control (NC) diet deficient in total Ca, dig P, standardized ileal digestible amino acids and without feed phosphates. The analyzed total Ca, total P, phytate P and calculated net energy ranged from 0.46 to 0.53%, 0.37 to 0.43%, 0.24 to 0.30%, and 2,441 to 2,472 kcal/kg, respectively. The NC was supplemented with PhyG at 250, 500, 1,000 or 2,000 FTU/kg. Digesta were collected using ileal T-cannulation (9-10 replicates/treatment, Latin square design). Data were analyzed using the Fit Model platform of JMP 14.0 with trial code as a random effect. Phytase dose response (using analyzed phytase activity) was analyzed with exponential curve fitting. The AID of gross energy and the ileal digestible energy (DE, kcal/kg) improved with 1,000 or 2,000 FTU/kg PhyG treatments vs. NC (P < 0.05, Table 1). Increasing the PhyG dose exponentially increased (P < 0.1) the AID of gross energy (%) and the ileal DE. Ileal DE improvement above NC increased exponentially (P < 0.06) up to 120.8 and 132.6 kcal/kg at 1,000 and 2,000 FTU/kg, respectively. In conclusion, PhyG supplementation improved ileal DE in growing pigs in a dose-dependent manner.
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Suldina, E. V., A. V. Masilenko, N. A. Feoktistova, and I. I. Bogdanov. "INDICATION OF ENZYME GENE FRAGMENTS OF BACILLUS MEGATERIUM BACTERIA." Vestnik of Ulyanovsk state agricultural academy 230 (September 25, 2021): 74–78. http://dx.doi.org/10.18286/1816-4501-2021-3-74-78.
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The soil microbial component plays a critical role in maintaining the soil functionality, performing several vital functions such as soil formation, decomposition of dead and decayed organic matter, circulation of macro and micro elements and transformation of toxic chemicals into non-toxic forms. Various Bacillus species, including B. megaterium, are known to fix atmospheric nitrogen. To ensure nitrogen fixation and diversity analysis of diazotrophs, the most widely used gene marker is nifH, which is responsible for synthesis of proteins of the nitrogenase complex. Various soil microorganisms, including B. megaterium, are endowed with the ability to solubilize phosphorus inaccessible form to accessible one and / or mineralize organic phosphorus to accessible phosphorus form. Phosphatases and phytases are two groups of enzymes that catalyze transformation of organic phosphates to inorganic ones. It was found that Bacillus megaterium exhibits acidic and alkaline phosphatase activity and produces phytases. The aim of this work was to determine by RT-PCR method the presence of genes responsible for enzyme synthesis of phytase, nitrogenase, and alkaline phosphatase in strains of previously isolated bacteria of Bacillus megaterium species. To specify the presence of genes encoding synthesis of the desired enzymes in Bacillus megaterium, an in-silico analysis of the annotated genomes of this bacterial species, presented in the NCBI information database, was carried out. Further on, the selection of primers for screening the target parts was made. According to the results of the study, 7 out of 16 isolated B. megaterium strains contained all three required DNA parts responsible for synthesis of phytase, nitrogenase, and alkaline phosphatase enzymes.
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Dessimoni, Gabriel Villela, Nilva Kazue Sakomura, Daniella Carolina Zanardo Donato, Fábio Goldflus, Nayara Tavares Ferreira, and Felipe Santos Dalólio. "Effect of supplementation with Escherichia coli phytase for broilers on performance, nutrient digestibility, minerals in the tibia and diet cost." Semina: Ciências Agrárias 40, no. 2 (April 15, 2019): 767. http://dx.doi.org/10.5433/1679-0359.2019v40n2p767.
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Although phytase has been researched, new enzymes have been produced, leading to different animal responses. In this scenario, the present study proposes to evaluate the inclusion of a bacterial phytase produced by Escherichia coli in broiler diets based on corn and soybean meal, with or without nutrient reductions, on the performance, nutrient digestibility, phosphorus bioavailability, and bone minerals of those animals and on the economic viability of this practice. A total of 896 male broiler chickens were distributed into 32 experimental units, each housing 28 broilers. The experiment was set up as a completely randomized design with four treatments (Positive Control (PC)- diet meeting the nutritional requirements of the broiler chickens; Negative Control (NC) with reductions of 100kcal/kg of ME, 0.14% avP and 0.11% tCa; NC + phytase (500 FTU/kg); PC + phytase (500FTU/kg)) and 8 replicates. Phytase increased (p < 0.05) feed intake and body weight gain and improved feed conversion ratio in starter (1 to 21 days) and total (1 to 42 days) phases, respectively, compared with Negative Control diet without supplementation. The Negative Control + phytase diet also led to a feed intake similar to Positive Control in the starter and total phases. The inclusion of phytase without nutrient reductions improved (p < 0.05) feed conversion ratio in the starter phase, compared to Positive Control diet. There was an increase (p < 0.05) in the digestibility of dry matter, crude protein and phosphorus and in apparent digestible energy in the Negative Control + phytase diet compared to the Negative Control diet. Phytase supplementation increased (p < 0.05) the digestibility of crude protein, calcium and phosphorus, and apparent digestible energy compared to Positive Control diet. Ash, phosphorus, and calcium contents were higher in the Negative Control + phytase diet compared with those observed in the Negative Control diet without enzyme (p < 0.05). The Positive Control + phytase diet provided higher ash contents (p < 0.05), but calcium and phosphorus deposition was similar to those obtained with Positive Control diet. Phytase inclusion allows for a reduction in the diet cost per ton of produced feed. In conclusion, dietary supplementation with bacterial phytase produced from Escherichia coli for broiler chickens is recommended, as it provided increases in production performance, nutrient digestibility, and energy metabolizability and a reduction in the diet cost.
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Moradi, Soudabeh, Mohammad Reza Abdollahi, Arash Moradi, and Leili Jamshidi. "Effect of Bacterial Phytase on Growth Performance, Nutrient Utilization, and Bone Mineralization in Broilers Fed Pelleted Diets." Animals 13, no. 9 (April 24, 2023): 1450. http://dx.doi.org/10.3390/ani13091450.
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The influence of a bacterial 6-phytase on growth performance, coefficient of apparent ileal digestibility (CAID) of nutrients, blood parameters, and bone mineralization in broilers was evaluated. A total of 630 one-day-old male broilers were allocated to 7 dietary treatments, including positive control (PC) diet containing dicalcium phosphate, the PC marginally reduced in available P (avP) by 0.1% and calcium (Ca) by 0.2% vs. PC (NC1) or moderately reduced by 0.15 and 0.3% vs. PC (NC2), respectively, and four further diets comprising the NC1 and NC2 supplemented with 500 or 1000 FTU/kg of phytase in starter and finisher phases. A constant Ca to avP ratio was maintained across all diets. The body weight gain (BWG) and feed per unit gain (FCR) of birds fed NC1 and NC2 diets supplemented with phytase (500 and 1000 U/kg) was equivalent to that of birds fed the PC diet at 35 days. Phytase supplementation in the NC1 diet linearly increased the CAID of nitrogen (N) (p < 0.01), phosphorus (P) (p < 0.01), and Ca (p < 0.05). Additionally, phytase reduced (p < 0.01) excreta P concentration by approximately 27%, improved (p < 0.001) toe ash, and tended to increase tibia ash (p = 0.08), comparable with the PC. In conclusion, the addition of bacterial 6-phytase dosed in the range of 500–1000 FTU/kg was effective in replacing 1.5 g/kg avP and 3 g/kg Ca in broilers fed pelleted diets, using bone quality, BWG, and FCR as outcome measures.
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Bilyeu, Kristin D., Peiyu Zeng, Patricia Coello, Zhanyuan J. Zhang, Hari B. Krishnan, April Bailey, Paul R. Beuselinck, and Joe C. Polacco. "Quantitative Conversion of Phytate to Inorganic Phosphorus in Soybean Seeds Expressing a Bacterial Phytase." Plant Physiology 146, no. 2 (December 27, 2007): 468–77. http://dx.doi.org/10.1104/pp.107.113480.
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Cowieson, A. J., F. Fru-Nji, and O. Adeola. "Dietary phosphate equivalence of four forms of Pi contrasted with a novel microbial phytase from Citrobacter braakii in broiler chickens." Animal Production Science 55, no. 9 (2015): 1145. http://dx.doi.org/10.1071/an14489.
Full textAbstract:
Two 21-day studies were conducted with broilers to evaluate the efficacy of a bacterial 6-phytase from Citrobacter braakii and compare four Pi sources. The four sources were phosphates of monocalcium (MCP), dicalcium (DCP), tricalcium (TCP) with potassium phosphate (KH2PO4) acting as a ‘positive control’ reference. In each study, 336 4-day-old male birds (Ross 708) were blocked based on initial bodyweight (BW) and randomly allotted to one of seven diets with six replicate cages of eight birds each. Access to experimental diets and water from Days 4 to 25 post-hatching was ad libitum. In the first study, the seven diets were: (1) a low-P negative control (NC) corn-soybean meal basal diet formulated to contain crude protein (CP), Ca, total P, and non-phytate P (nPP) at 218, 9.0, 4.5, and 2.0 g/kg, respectively; (2) NC plus 0.75 g Pi from KH2PO4/kg; (3) NC plus 0.75 g Pi from MCP/kg; (4) NC plus 0.75 g Pi from DCP/kg; (5) NC plus 0.75 g Pi from TCP/kg; (6) NC plus phytase at 500 FYT/kg; (7) NC plus phytase at 1000 FYT/kg. Feeding the low-P NC diet reduced (P < 0.01) BW gain (BWG), feed intake (FI), and tibia ash. Supplementing the NC with Pi or phytase linearly improved (P < 0.01) BWG, FI, and tibia ash. Supplementing the NC with 0.75 g/kg Pi from MCP, DCP, or TCP were equipotent in improving BWG and FI; however, percent tibia ash was higher (P < 0.05) in birds fed MCP than either DCP or TCP. The second study was similar to the first study except that Ca, total P, and nPP in the NC were reduced to 7.0, 4.2, and 1.8 g/kg, respectively; and Pi from MCP, DCP, and TCP were reduced to 0.6 g/kg. Similar to observations in the first study, the low-P NC diet reduced (P < 0.01) BWG, FI, and tibia ash; and supplementing the NC with Pi or phytase linearly improved (P < 0.01) BWG, FI, P digestibility and tibia ash. Furthermore, supplementing the NC with 0.75 g Pi from MCP, DCP, or TCP per kg diet were equipotent in improving BWG, FI, and tibia ash though MCP resulted in superior (P < 0.05) retention of P compared with TCP. Results from both studies showed that the phytase was efficacious in releasing phytate-P for growth and bone mineralisation in chickens compensating, at least, the spared Pi, and that Pi sources commonly used in formulating diets of chickens may be different in their potential to supply digestible P.