Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.1.3.8 (phytase)
 1,997 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Ninety-six crossbred young pigs (body weight 7.8 kg) were used in a 5-week trial to determine the effectiveness of microbial phytase (EC 3.1.3 26) in improving the bioavailabilities of P and other nutrients in maize-soyabean-meal diets and, thus, replacing inorganic P with phytase. A 2 x 5 factorial arrangement of treatments was employed with two available P (aP) levels (0.7 and 1.6 g/kg) and five phytase levels (0, 350, 700, 1050, 1400 U (the quantity of enzyme that liberates 1 mumol inorganic phosphate/min from 5.1 mm-sodium phytate at pH 5.5 and 37 degrees)/kg diet). In addition, two extra diets were formulated to supply the National Research Council (1988) recommended level of aP (3.2 g/kg) with 0 or 1400 U phytase. The addition of graded levels of phytase resulted in linear increases in average daily weight gain, average daily feed intake and weight gain:feed intake for pigs fed on diets containing 0.7 or 1.6 g aP/kg (P < 0.04). Also, the addition of phytase linearly increased apparent digestibilities of P and Ca (P < 0.01), whereas faecal P excretion was linearly decreased (P < 0.01). Linear increases in shear force, shear energy and ash content of both the metacarpal and tenth rib, and shear stress of the metacarpal were found to respond to added phytase (P < 0.01). These improvements in performance, apparent P absorption and bone measurements by phytase were also observed by increasing dietary aP levels for most measurements. Adding 1400 U phytase to the 3.2 g aP/kg diet further increased average daily weight gain, average daily feed intake, apparent absorption of P, Ca and N and metatarsal shear force and ash content (P < 0.01 to 0.05). Generally, maximum responses occurred at a phytase level of 1050 U/kg diet for the 0.7 g aP/kg diets and 700 U for the 1.6 g aP/kg diets. Based on non-linear and linear response equations generated for the phytase and aP levels, the average function of the equivalency of P (Y, g/kg) v. microbial phytase (X, U/kg) was developed across aP levels of 0.7 and 1.6 g/kg for average daily weight gain and apparent digestibility of P: Y = 2.622-2.559e 0.00185X. The replacement of 1 g inorganic P as defluorinated phosphate would require about 246 U microbial phytase. This represents 41% of released P from phytate.
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PMID:Replacement of inorganic phosphorus by microbial phytase for young pigs fed on a maize-soyabean-meal diet. 894 63

Two chick assays were conducted in an attempt to understand how 1alpha-hydroxylated cholecalciferol compounds [1,25-(OH)2 D3 and 1alpha-OH D3] function in chicks to improve utilization of phytate-bound phosphorus (P) and trace minerals. Mucosal tissue from chicks fed a P-deficient corn-soybean meal diet, with or without supplemental 1alpha-OH D3, was incubated with sodium phytate. Inorganic P (Pi) release from sodium phytate, a measure of mucosal phytase activity, was not influenced by 1alpha-OH D3 presence in the diet. Increasing doses of mucosal protein in tubes containing sodium phytate resulted in marked increases (P < 0.01) in Pi release, but 1alpha-OH D3 in the diet from which the duodenal mucosal tissue was obtained had no effect on Pi release. Similarly, addition of either 1alpha-OH D3 or 1,25-(OH)2 D3 directly to the incubation tubes had no effect on Pi production. Efficacy of supplemental 1alpha-OH D3 and phytase was also tested in cecectomized vs. sham-operated chicks that were fed P-deficient and cholecalciferol-adequate corn-soybean meal diets. Removal of the twin ceca was done in an attempt to remove much of the intestinal microbial activity, and in turn, much of the gut microbial phytase activity. Marked increases (P < 0.01) in bone ash occurred in response to phytase or 1alpha-OH D3 supplementation, and cecectomized birds responded to either addition in the same manner as sham-operated controls. The data suggest that the marked phytate-P releasing capacity of dietary 1alpha-OH D3 or 1, 25-(OH)2 D3 is not caused by an increased specific activity of intestinal phytase.
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PMID:1alpha-hydroxycholecalciferol does not increase the specific activity of intestinal phytase but does improve phosphorus utilization in both cecectomized and sham-operated chicks fed cholecalciferol-adequate diets. 931 64

A recombinant form of Escherichia coli phytase, which hydrolyzes phytic acid into phosphate and myo-inositol, has been expressed, purified and crystallized. Crystals have been obtained by the method of bulk crystallization in 10 mM sodium acetate buffer (pH 4.5) without using a conventional precipitant. The enzyme crystallized in space group P21, with unit-cell dimensions a = 74.9, b = 72.2, c = 82.4 A, and beta = 92.0 degrees. Crystals diffract to at least 2.2 A at a rotating-anode X-ray source and a 2.3 A resolution data set has been collected, giving completeness of 98.0% and an Rsym of 0.072. Assuming there are two phytase molecules in the asymmetric unit, the solvent content is calculated to be 42.1%. A self-rotation function shows a clear twofold non-crystallographic symmetry relating two molecules of E. coli phytase in the asymmetric unit.
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PMID:Purification, crystallization and preliminary X-ray analysis of the Escherichia coli phytase. 976 63

Bacterial strains were isolated from the pig colon to screen for phytase and acid phosphatase activities. Among 93 colonies, Colony 88 had the highest activities for both enzymes and was identified as an Escherichia coli strain. Using primers derived from the E. coli pH 2.5 acid phosphatase appA sequence (Dassa et al. (1990), J. Bacteriol. 172, 5497-5500), we cloned a 1482 bp DNA fragment from the isolate. In spite of 95% homology between the sequenced gene and the appA, 7 amino acids were different in their deduced polypeptides. To characterize the properties and functions of the encoded protein, we expressed the coding region of the isolated DNA fragment and appA in Pichia pastoris, respectively, as r-appA2 and r-appA. The recombinant protein r-appA2, like r-appA and the r-phyA phytase expressed in Aspergillus niger, was able to hydrolyze phosphorus from sodium phytate and p-nitrophenyl phosphate. However, there were distinct differences in their pH profiles, Km and Vmax for the substrates, specific activities of the purified enzymes, and abilities to release phytate phosphorus in soybean meal. In conclusion, the DNA fragment isolated from E. coli in pig colon seems to encode for a new acid phosphatase/phytase and is designated as E. coli appA2.
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PMID:Cloning, sequencing, and expression of an Escherichia coli acid phosphatase/phytase gene (appA2) isolated from pig colon. 1009 20

High iron consumption has been proposed to relate to an increase in the risk of colon cancer, whereas high levels of supplemental sodium phytate effectively reduce iron-induced oxidative injury and reverse iron-dependent augmentation of colorectal tumorigenesis. However, the protective role of intrinsic dietary phytate has not been determined. In this study, we examined the impact of removing phytate present in a corn-soy diet by supplemental microbial phytase on susceptibility of pigs to the oxidative stress caused by a moderately high dietary iron intake. Thirty-two weanling pigs were fed the corn-soy diets containing two levels of iron (as ferrous sulfate, 80 or 750 mg/kg diet) and microbial phytase (as Natuphos, BASF, Mt. Olive, NJ, 0 or 1200 units/kg). Pigs fed the phytase-supplemented diets did not receive any inorganic phosphorus to ensure adequate degradation of phytate. After 4 months of feeding, liver, colon, and colon mucosal scrapings were collected from four pigs in each of the four dietary groups. Colonic lipid peroxidation, measured as thiobarbituric acid reacting substances (TBARS), was increased by both the high iron (P< 0.0008) and phytase (P< 0.04) supplementation. Both TBARS and F2-isoprostanes, an in vivo marker of lipid peroxidation, in colonic mucosa were affected by dietary levels of iron (P< 0.03). Mean hepatic TBARS in pigs fed the phytase-supplemented, high iron diet was 43%-65% higher than that of other groups although the differences were nonsignificant. Moderately high dietary iron induced hepatic glutathione peroxidase activity (P= 0.06) and protein expression, but decreased catalase (P< 0.05) in the colonic mucosa. In conclusion, intrinsic phytate in corn and soy was protective against lipid peroxidation in the colon associated with a moderately high level of dietary iron.
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PMID:Dietary intrinsic phytate protects colon from lipid peroxidation in pigs with a moderately high dietary iron intake. 1032 Jun 35

Differential agar media for the detection of microbial phytase activity use the disappearance of precipitated calcium or sodium phytate as an indication of enzyme activity. When this technique was applied to the study of ruminal bacteria, it became apparent that the method was unable to differentiate between phytase activity and acid production. Strong positive reactions (zones of clearing around microbial colonies) observed for acid producing, anaerobic bacteria, such as Streptococcus bovis, were not corroborated by subsequent quantitative assays. Experimentation revealed that acidic solutions generated false positive results on the selected differential medium. Empirical studies undertaken to find a solution to this limitation determined the false positive results could be eliminated through a two step counterstaining treatment (cobalt chloride and ammonium molybdate/ammonium vanadate) which reprecipitates acid solubilized phytate. This report discusses the application of the developed two step counterstaining treatment for the screening of phytase producing ruminal bacteria as well as its use in phytase zymogram assays.
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PMID:A novel staining method for detecting phytase activity. 1057 3

A (31)P NMR method for quantitative determination of inositol phosphates in simple incubation samples of sodium phytate and Aspergillus niger phytase and in different types of complex samples, such as diets, digesta, and feces, is described. The inositol phosphates in complex samples were extracted with HCl, concentrated, and purified using freeze-drying and filtration and subsequently determined at pH 12.6 in aqueous solution using a (31)P NMR method. The (31)P NMR technique has as its main advantages over the HPLC techniques that it does not necessitate standards that may cause background matrix effects and that the spectra of inositol phosphates and orthophosphate appear in the same run without further sampling errors. The results of inositol hexaphosphate analysis with HPLC can be confirmed by this (31)P NMR method. Contents of inositol tetra-, tri-, di-, and monophosphate in the biological samples appear to be quantitatively not important. The (31)P NMR method can be applied for use in animal nutrition in general and studies of using phytase in diets for farm animals in particular, by measuring the content of inositol phosphates in feed ingredients, complete feeds, ileal contents, and feces of pigs and poultry.
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PMID:Quantification of inositol phosphates using (31)P nuclear magnetic resonance spectroscopy in animal nutrition. 1060 82

Five groups of individually housed albino rats (n = 7 each, initial average weight = 42 g) were fed diets based on corn starch and casein over a 4-week period. All diets were supplemented with 35 mg/kg of iron from FeSO4 x 7 H2O. Group I (control) was fed the basal diet free of phytic acid (PA) and phytase. By replacing corn starch by 7.5 g (groups II and IV) and 15 g phytic acid (groups III and V) from sodium phytate per kg diet, molar PA/iron ratios of 18 and 36 were obtained. In groups IV and V, 1000 U phytase from Aspergillus niger per kg diet were added. Food conversion efficiency ratio and growth rate as well as iron in plasma and spleen, hemoglobin, red blood cell count and erythrocyte zinc protoporphyrin were not influenced by the different dietary treatments. Dietary phytate reduced apparent iron absorption in groups II and III. Furthermore hematocrit, transferrin saturation and iron concentration in liver and femur were lowered in rats fed diets with PA, while total and latent iron-binding capacity of plasma increased. Microbial phytase supplementation (groups IV and V) partly counteracted the antinutritive effects of phytic acid on iron availability.
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PMID:Supplemental sodium phytate and microbial phytase influence iron availability in growing rats. 1061 76

Aspergillus fumigatus phytase is a heat-stable enzyme of great potential. Our objective was to determine if a high level of functional expression of the A. fumigatus phytase gene could be produced in Pichia pastoris and how the recombinant phytase reacted to different substrates, heating conditions, and proteases. A 1.4-kb DNA fragment containing the coding region of the gene was inserted into the expression vector pPICZalphaA and expressed in P. pastoris as an active, extracellular phytase (r-Afp). The yield was 729 mg of purified protein per liter of culture, with a specific activity of 43 units/mg of protein. The enzyme r-Afp shared similar pH and temperature optima, molecular size, glycosylation extent, and specificity for p-nitrophenyl phosphate and sodium phytate to those of the same enzyme expressed in A. niger. Given 20 min of exposure to 65 to 90 degrees C, the enzyme retained 20 to 39% higher residual activity in 10 and 200 mM sodium acetate than that in sodium citrate. The enzyme seemed to be resistant to pepsin digestion, but was degraded by high levels of trypsin. In conclusion, P. pastoris is a potential host to express high levels of A. fumigatus phytase and the thermostability of the recombinant enzyme is modulated by the specificity of buffer used in the heat treatment.
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PMID:Expression of the Aspergillus fumigatus phytase gene in Pichia pastoris and characterization of the recombinant enzyme. 1067 11

Several bioassays were conducted with young chicks and pigs fed phosphorus (P)-deficient corn-soybean meal diets. With diets for chicks containing .62% Ca and .42% P (.10% available P), graded doses of a citric acid + sodium citrate (1:1, wt:wt) mixture (0, 1, 2, 4, or 6% of diet) resulted in linear (P < .01) increases in both weight gain and tibia ash. Relative to chicks fed no citric acid, tibia ash (%) and weight gain (g/d) were increased by 43 and 22%, respectively, in chicks fed 6% citric acid. Additional chick trials showed that 6% citric acid alone or sodium citrate alone was as efficacious as the citric acid + sodium citrate mixture and that 1,450 U/kg of phytase produced a positive response in bone ash and weight gain in chicks fed a diet containing 6% citrate. Varying the Ca:available P ratio with and without citrate supplementation indicated that citric acid primarily affected phytate-P utilization, not Ca, in chicks. Moreover, chicks did not respond to citrate supplementation when fed a P-deficient (.13% available P), phytate-free casein-dextrose diet. Young pigs averaging 10 to 11 kg also were used to evaluate citric acid efficacy in two experiments. A P-deficient corn-soybean meal basal diet was used to construct five treatment diets that contained 1) no additive, 2) 3% citric acid, 3) 6% citric acid, 4) 1,450 U/kg phytase, and 5) 6% citric acid + 1,450 U/kg phytase. Phytase supplementation increased (P < .01) weight gain, gain:feed, and metatarsal ash, whereas citric acid addition increased only gain:feed (P < .05) and metatarsal ash (P < .08). A subsequent 22-d pig experiment was conducted to evaluate the effect of lower levels of citric acid (0, 1, 2, or 3%) or 1,450 U/kg phytase addition to a P-deficient corn-soybean meal diet. Phytase supplementation improved (P < .01) all criteria measured. Weight gain and gain:feed data suggested a response to citric acid addition, but this was not supported by fibula ash results (P > .10). The positive responses to phytase were much greater than those to citric acid in both pig experiments. Thus, dietary citric acid effectively improved phytate P utilization in chicks but had a much smaller effect in pigs.
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PMID:The effects of citric acid on phytate-phosphorus utilization in young chicks and pigs. 1076 76


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