• Korean Journal of Organic Agriculture
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• v.9 no.1
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• pp.51-60
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• 2001

An experiment was carried out to evaluate the effects of adding phytase on nutrient digestibility in growing pigs. Three barrow pigs averaging 28.5$\pm$3.1kg of body weight were allotted to three treatments by 3$\times$3 latin square. Treatments included 1) com-soybean meal based-control diet and 2) and 3) control diet with phytase 500 and 1,000 unit. There were no effects of treatments on dry matter and nitrogen digestibility (P〉0.05). Ash, Ca and P digestibility in pigs fed diet with phytase were greater than those in pigs fed control diet(P〈0.05). An increased phosphorus intake was observed from pigs fed diet with treatments compared to that from pigs fed control diet. Pigs fed diet with adding phytase had improved P absorption compared to pigs fed control diet(P〈0.05). There was no significantly effect on fecal and urine excretion(P〈0.05). Ca intake was increased for pigs fed adding phytase compared to pigs fed control diet. No evident effect was observed on Ca fecal excretion among dietary treatments, however, urine excretion of pigs fed diet with treatments was decreased compared to pigs fed control diet. Cystine, isoleucine, tyrosine and phenylalanine digestibility of amino acid were greater for pigs fed diet with treatments than those for pigs fed control diet(P〈0.05). Asparagine, threonine, serine, methionine and leucine digestibility were improved for pigs fed with phytase 500 unit compared to pigs fed control diet. Lysine, histidine, arginine and other animo acids digestibility were higher for pigs fed diet with phytase than control diet, but there were no significant effects by the treatments(P〉0.05). In conclusion, the results of the experiments suggest that phytase supplementation in the diets could be a very beneficial strategy in environmental aspects.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.11
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• pp.1561-1567
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• 2000

An experiment was conducted to investigate the effect of dietary protein levels and supplementation of phytase and pentosanase in wheat-soybean meal diet on the performance and output of N and P in broilers. Addition of phytase alone or in combination with pentosanase to reduced or control protein diets did not affect average final body weight of mixed sexes. However, addition of phytase and pentosanase in combination to reduced protein diets in male broilers significantly depressed body weights. Intestinal viscosity of 21d broilers was significantly decreased by addition of phytase and pentosanase alone or in combination. Tibia ash content was significantly increased by phytase supplementation. Supplementation of phytase alone and in combination with pentosanase to reduced protein diets significantly decreased P in manure and daily output of P. Daily N output was lowest in the reduced protein diet supplemented with phytase and pentosanase combination. The retention of DM, N and P was highest in the reduced protein diet supplemented with phytase and pentosanase combination. In conclusion, supplementation of phytase alone or in combination with pentosanase to reduced protein diets can decrease output of N and P. But the combination of the enzymes has no beneficial effects on the performance of broilers, especially those on wheat-soybean meal diet with reduced protein level.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.5
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• pp.673-680
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• 2000

An experiment was conducted with day-old 300 commercial male broiler chicks (Arbor Acres$^{(R)}$) to evaluate the efficacy of crude phytase preparerations produced from a culture of Aspergillus ficcum. The experiment consisted of five dietary treatments; T1, com-soy control diet with 0.45% non-phytate phosphorus (NPP) for starter period and 0.35% NPP for grower period; T2, control - 0.1% NPP; T3, control 0.2% NPP; T4, T3+600 U of crude phytase (broth+cell); and T5, T3+600 U of crude phytase (broth). The body weight gain, feed intake, and feed/gain of chickens fed T1 diet was highest (p<0.01) among treatments. BW gain and feed intake of T4 and T5 were greater than those of T3 but were less than those of T1 and T2. T3 was highest in mortality among treatments. Decreasing the NPP level lowered availability of DM, crude ash, ether extract, crude fiber, Zn, and Fe but supplementation of crude phytase preparations improved the availability of these nutrients as well as those of Ca, P and Cu. Excretion of P and Cu significantly decreased as the NPP level in the diet decreased. Further reduction of P and Cu excretion and reduction of Ca, Mg and Fe excretion were achieved by supplementation of crude phytase preparations. The serum concentrations of Ca, P, Mg, Zn, Fe, and Cu were significantly increased by crude phytase supplementation. The weight and length of tibia, and contents of crude ash, Ca, P, Mg, and Zn were adversely affected by lowering NPP level but partially recovered by supplementation of crude phytase preparations. In conclusion, lowering NPP level in the broiler diet significantly depressed the performance. Supplementation of crude phytase preparations produced from Aspergillus ficuum could partially recover the depression.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.8
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• pp.1158-1164
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• 2003

In two feeding experiments male and mixed-sex broiler chicks were offered diets based on sorghum and a wheatsorghum blend with two tiers of nutrient specifications, without and with microbial phytase (600 and 800 FTU/kg), from 7-25 and 1-42 days post-hatch, respectively. The nutrient specifications for protein, amino acids, energy density and phosphorus (P) of standard diets were reduced to formulate the modified diets on a least-cost basis. Calculated differences in nutrient specifications between standard and modified diets ranged from 14.3 to 17.1 g/kg crude protein, 0.24 to 0.40 MJ/kg apparent metabolisable energy (AME) and 1.06 to 1.20 g/kg available P. In both experiments, reduced nutrient specifications had a negative impact on growth rates and feed efficiency and phytase supplementation had a positive influence on growth performance and protein efficiency ratios (PER). Phytase addition to the less expensive, modified diets either partially or entirely compensated for reduced growth performance and, consequently, feed costs per kg of live weight gain were reduced. In Experiment 1, phytase increased (p<0.001) nitrogen-corrected AME (AMEn) from 15.39 to 15.89 MJ/kg dry matter. For nitrogen (N) retention there was an interaction (p<0.05) between diet type and phytase as the effects of phytase on N retention were more pronounced in the modified diets, with an increase from 0.512 to 0.561. These results demonstrate the positive effects of phytase on protein and energy utilisation, in addition to its established liberation of phytate-bound P and illustrate the feasibility of assigning nutrient replacement values to the feed enzyme for consideration in least-cost ration formulations. Further work is, however, required to define the most appropriate reductions in nutrient specifications in association with phytase supplementation.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.11
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• pp.1642-1649
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• 2003

An experiment was conducted to evaluate the effects of phytase supplementation on the growth performance, nutrients utilization and bone mineralization in broiler chickens. Day-old broiler chicks (n=480) were equally devided into eight treatment groups and fed maize or wheat based isocaloric, isonitrogenous and isocalcium diets having two non phytate phosphorus (NPP) concentrations (0.50% and 0.30%) and two phytase levels (0 and 500 phytase units/kg diet) in a 42 days growth trial. Maize based dietary treatments were MC (NPP 0.50%, MN (NPP 0.30%), MNP (MN+500 units of phytase) and MCP (MC+500 units of phytase), whereas wheat based experimental diets were WC (NPP 0.50%), WN (NPP 0.30%), WNP (WN+500 units of phytase) and WCP (WC+500 units of phytase). The NPP levels were maintained by dicalcium phosphate. Reduction in dietary NPP depressed live weight gain and feed intake and increased feed conversion ratio (FCR). Phytase supplementation to low NPP (0.30%) diets significantly (p<0.05) improved the growth performances of broilers. The supplementation to low NPP diets allowed complete, safe and economic replacement of dietary inorganic P (dicalcium phosphate) to reduce feed cost per kg live weight gain of broilers. Reduction in dietary NPP did not affect retention of nutrients except phosphorus (P) but had a significant (p<0.05) depression in tibia ash and minerals (Ca, P) concentration in serum and tibia ash. Phytase supplementation at low NPP level was effective (p<0.05) in improving the retention of dry matter, Ca and P and Ca and P concentration in serum and tibia ash. However, the supplementation was not effective at high level of NPP (0.50%). There were no significant (p>0.05) differences in carcass quality among dietary treatments. The response of phytase was greater in low NPP and maize based diets as compared with high NPP and wheat based diets, respectively. The results show that phytase supplementation to low NPP (0.30%) diets improved the growth performance, relative retention of nutrients (N, Ca and P) and minerals (Ca, P) status of blood and bone in broiler chickens, with a better efficacy in maize based diets.

• Journal of Microbiology and Biotechnology
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• v.11 no.6
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• pp.915-921
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• 2001

Phytase improves the bioavailability of phytate phosphorus in plant foods to humans and animals, and reduces the phosphorus pollution of animal waste. In order to express a high level of fungal phytase in Saccharomyces cerevisiae, various expression vectors were constructed with different combinations of promoters, translation enhancers, signal peptides, and terminator. Three different promoters fused to the phytase gene (phyA) from Aspergillus niger were tested: a galactokinase (GAL1) promoter, glyceraldehyde-3-phosphate dehydrogenase (GPD) promoter, and yeast hybrid ADH2-GPD promoter consisting of alcohol dehydrogenase II (ADH2) and a GPD promoter. The signal peptides of phytase, glucose oxidase (GO), and rice amylase 1A(RAmy1A) were included. Plus, the translation enhancers of the ${\Omega}$ sequence and UTR70 from the tobacco mosaic virus (TMV) and spinach, respectively, were also tested. Among the recombinant vectors, pGphyA06 containing the GPD promoter, the ${\Omega}$ sequence, RAmy1A, and GAL7 terminator expressed the highest phytase activity in a culture filtrate, which was estimated at 20 IU/ml. An intracellular localization of the expressed phytase activity in a culture filtrate, which was estimated at 20 IU/ml. An intracellular localization of the expressed phytase was also performed by inserting an endoplasmic reticulum (ER) retention signal, KDEL sequence, into the C-terminus of the phytase within the vector pHphyA-6. It appeared that the KDEL sequence directed most of the early expression of phytase into the intracellular compartment yet more than $60\%$ of the total phytase activity was still retained within the cell even after the prolonged (>3 days) incubation of the transformant. However, the intracellular enzyme activity of the transformant without a KDEL sequence was as high as that of the extracellular one, thereby strongly suggesting that the secretion of phytase in S. cerevisiae appeared to be the rate-limiting step for the expression of a large amount of extracellular recombinant phytase, when compared with other yeasts.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.3
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• pp.394-402
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• 2003

Individual and combined supplementation of phosphorus-adequate, wheat-based broiler diets with exogenous phytase and xylanase was evaluated in three experiments. The effects of the enzyme combination in lysine-eficient diets containing wheat and sorghum were more pronounced than those of the individual feed enzymes. The inclusion of phytase plus xylanase improved (p<0.05) weight gains (7.3%) and feed efficiency (7.0%) of broilers (7-28 days post-hatch) and apparent metabolisable energy (AME) by 0.76 MJ/kg DM. Phytase plus xylanase increased (p<0.05) the overall, apparent ileal digestibility of amino acids by 4.5% (0.781 to 0.816); this was greater than the responses to either phytase (3.6%; 0.781 to 0.809) or xylanase (0.7%; 0.781 to 0.784). Absolute increases in amino acid digestibility with the combination exceeded the sum of the individual increases generated by phytase and xylanase for alanine, aspartic acid, glutamic acid, glycine, histidine, isoleucine, phenylalanine, threonine, tyrosine and valine. These synergistic responses may have resulted from phytase and xylanase having complementary modes of action for enhancing amino acid digestibilities and/or facilitating substrate access. The two remaining experiments were almost identical except wheat used in Experiment 2 had a higher phytate concentration and a lower estimated AME content than wheat used in Experiment 3. Individually, phytase and xylanase were generally more effective in Experiment 2, which probably reflects the higher dietary substrate levels present. Phytase plus xylanase increased (p<0.05) gains (15.4%) and feed efficiency (7.0%) of broiler chicks from 4-24 days post-hatch in Experiment 2; whereas, in Experiment 3, the combination increased (p<0.05) growth to a lesser extent (5.6%) and had no effect on feed efficiency. This difference in performance responses appeared to be 'rotein driven'as the combination increased (p<0.05) nitrogen retention in Experiment 2 but not in Experiment 3; whereas phytase plus xylanase significantly increased AME in both experiments. In Experiments 2 and 3 the combined inclusion levels of phytase and xylanase were lower that the individual additions, which demonstrates the benefits of simultaneously including phytase and xylanase in wheat-based poultry diets.

• Microbiology and Biotechnology Letters
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• v.30 no.1
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• pp.1-7
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• 2002

Phytase from Escherichia coli WC7 was purified from cell extracts and its molecular mass was estimated to be 45 kDa by SDS-PAGE. Its optimum temperature and pH for phytate hydrolysis was 6$0^{\circ}C$ and pH 5.0, respectively. The enzyme was stable up to 6$0^{\circ}C$ and over broad pH range (pH 2-12). The enzyme had higher affinity for sodium phytate than p-nitrophenylphosphate (pNPP). That is, the apparent Km value for sodium phytate and pNPP were $0.15\pm$0.02 mM and 2.82$\pm$0.05 mM, respectively. The gene encoding the phytase was cloned in E. coli XL1-Blue. Sequence analysis showed an open reading frame of 1241 Up encoding a signal peptide (22 aa) and a mature enzyme (410 aa). WC7 phytase was expressed up to 17.5 U/ml in the transformed E. coli XL1-Blue/pUEP, which was 23-fold higher than the activity from wild strain.

• Animal Bioscience
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• v.34 no.6
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• pp.1049-1060
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• 2021

Objective: This study assessed the effect of different levels of xylanase, β-glucanase and phytase on intestinal enzyme activities and tibia bone development in broiler chickens fed wheat-based diets. Methods: Twelve experimental diets were formulated using a 3×2×2 factorial design (three doses of phytase and two doses of both xylanase and β-glucanase) and offered to 648 day-old Ross 308 male chicks having 6 replicates groups with 9 birds per replicate and lasted for 35 days. Results: An interaction between the enzymes products improved (p<0.01) the activity of chymotrypsin. Protein content at d 10 was highest (p<0.001) with addition of phytase while general proteolytic activity (GPA) (p<0.02) and lipase activity (p<0.001) were decreased. At d 24, there were improvements in protein content (p<0.01) and lipase (p<0.04) with supplementation of superdose phytase. Addition of superdose phytase decreased in chymotrypsin (p<0.02), trypsin (p<0.01) and GPA (p<0.001). The optimum dose of xylanase decreased the chymotrypsin activity (p = 0.05), while the GPA (p<0.001) was increased with the optimum level of β-glucanase. Superdose phytase supplementation at d 10 improved maltase (p = 0.05), sucrase (p<0.001) and alkaline phosphatase (p<0.001) activities in the jejunum while aminopeptidase activity was highest (p<0.005) with the low level of phytase. Protein content of jejunum mucosa was bigger (p<0.001) in birds fed superdose phytase while maltase activity (p<0.001) at d 24 was reduced by this treatment. Sucrase (p<0.04) and aminopeptidase activities (p<0.001) improved when diets supplemented with low levels of phytase. Tibia bone breaking strength was highest (p<0.04) with addition of low level of superdose phytase or optimum level of β-glucanase. Bone dry matter content decreased (p<0.04) when diets supplemented with phytase. Conclusion: From the results obtained in this study, supplementation of superdose phytase was the most effective, however, the cost-benefit analysis of the use of such a dose needs to be evaluated.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.7
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• pp.957-961
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• 2000

A series of experiments was conducted to evaluate phytin phosphohydrolysis actlVlty in the rumen and to isolate phytase positive rumen bacteria. Endogenous phytase activity of wheat bran was estimated and compared with that of bacterial phytin phosphohydrolysis. Substantial phytase activity was detected in wheat bran during in vitro rumen incubation. Bacterial phytase activity was suggested not to be high. Only two facultative anaerobes, Klebsiella sp. and Corynebacterium sp. were isolated as phytase producing organisms. These belonged to a minor microbial group in the rumen population. Protozoal fraction showed an initial velocity of phytin phosphohydrolysis 7 times higher than the bacterial fraction.