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)

Microbial phytase hydrolyzes poorly degradable vegetable phytate P in the gastrointestinal tract of poultry; thereby increasing the availability of organic P to an extent that remains to be established. For this purpose, the P equivalency value of phytase in corn-soybean meal layer diets was assessed in three experiments (two short-term absorption studies and one performance trial lasting a complete production period). In the first absorption study, two basal diets containing 30 or 40 g Ca/kg diet were supplemented with either phytase [0, 250, or 500 phytase units (FTU)/kg diet] or with monocalcium phosphate (MCP; 0, 0.5, or 1.0 g P/kg diet) and fed to layers from 20 to 24 wk of age. The ileal absorption of Ca and P was measured during the last week. It was shown that 250 FTU/kg diet hydrolyzed an amount of phytate P that was equivalent to 1.3 g P from MCP. At the highest phytase inclusion level (500 FTU/ kg diet), a lower value of equivalency was observed, as P absorption was almost maximal at the lower level of phytase inclusion (250 FTU/kg diet). Phytase hydrolyzed phytate-bound P effectively at both Ca levels, although this degradation was significantly reduced by 12 percentage units at the higher dietary Ca level. The second absorption study, used 0, 250, and 500 FTU phytase/kg diet and 0 and 1.0 g P/kg diet of MCP. All diets were standardized at 35 g Ca/kg diet. The ileal absorption of Ca and P was determined at 24 and 36 wk of age. These values were significantly reduced in 36-wk-old hens compared to 24-wk-old hens. At 24 wk of age, phytic acid P degradation was significantly improved with increasing levels of phytase up to the maximum inclusion level of 500 FTU/kg diet (maximum phytic acid-P degradation at the end of the small intestine was 66%). In this experiment, the dose of 250 FTU/kg diet was equivalent to 0.8 g MCP-P. In Experiment 3, either phytase or MCP-P was added to a corn-soybean meal layer diet, containing 40 g Ca/kg diet and 3.6 g P/kg diet, at levels of 0, 100, 200, and 300 FTU/kg or levels of 0, 0.3, 0.6, and 0.9 g MCP-P/kg, respectively. Production performance was measured from 18 to 68 wk of age. Diets were consumed ad libitum. Growth, production performances (except kilograms of feed per kilogram of egg), and tibia parameters were significantly improved by dietary supplementation of the negative control diet with either phytase or MCP-P. Growth, egg production, and feed conversion ratio of the hens from the supplemented groups remained good throughout the experiment. No phytase dose effects on the production characteristics or tibia parameters were observed, indicating that the P requirements of the laying hens were met throughout the production period even at the lowest level of supplementation.
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PMID:The efficacy of phytase in corn-soybean meal-based diets for laying hens. 935 48

The characterization of the multiple inositol polyphosphate phosphatase (MIPP) is fundamental to our understanding of how cells control the signalling activities of 'higher' inositol polyphosphates. We now describe our isolation of a 2.3 kb cDNA clone of a rat hepatic form of MIPP. The predicted amino acid sequence of MIPP includes an 18 amino acid region that aligned with approximately 60% identity with the catalytic domain of a fungal inositol hexakisphosphate phosphatase (phytase A); the similarity encompassed conservation of the RHGXRXP signature of the histidine acid phosphatase family. A histidine-tagged, truncated form of MIPP was expressed in Escherichia coli and the enzymic specificity of the recombinant protein was characterized: Ins(1,3,4,5,6)P5 was hydrolysed, first to Ins(1,4,5,6)P4 and then to Ins(1,4,5)P3, by consecutive 3- and 6-phosphatase activities. Inositol hexakisphosphate was catabolized without specificity towards a particular phosphate group, but in contrast, MIPP only removed the beta-phosphate from the 5-diphosphate group of diphosphoinositol pentakisphosphate. These data, which are consistent with the substrate specificities of native (but not homogeneous) MIPP isolated from rat liver, provide the first demonstration that a single enzyme is responsible for this diverse range of specific catalytic activities. A 2.5 kb transcript of MIPP mRNA was present in all rat tissues that were examined, but was most highly expressed in kidney and liver. The predicted C-terminus of MIPP is comprised of the tetrapeptide SDEL, which is considered a signal for retaining soluble proteins in the lumen of the endoplasmic reticulum; the presence of this sequence provides a molecular explanation for our earlier biochemical demonstration that the endoplasmic reticulum contains substantial MIPP activity [Ali, Craxton and Shears (1993) J. Biol. Chem. 268, 6161-6167].
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PMID:Molecular cloning and expression of a rat hepatic multiple inositol polyphosphate phosphatase. 935 36

Two experiments using 413 crossbred growing-finishing pigs were conducted to assess the use of a commercial microbial phytase (Natuphos) in corn-soybean meal diets to improve phytate P bioavailability and thus reduce inorganic P supplementation and fecal P excretion. In Exp. 1 (n = 189), the following diets were used: 1) .50/.40% total P, respectively, for grower and finisher phases, and no phytase; 2) .40/.35% P and no phytase; 3) diet 2 plus 250 U phytase/kg; and 4) diet 2 plus 500 U phytase/ kg. The total Ca level was .58/.48% for diet 1 and .53/.43% Ca for diets 2, 3, and 4 in the grower and finisher phases, respectively. Feeding the low-P diet without supplemental phytase resulted in an overall 18% reduction in ADG (P < .05), 15% reduction in ADFI (P < .05), and 3% poorer feed efficiency (P < .08). Adding 250 to 500 U phytase/kg to the low-P diet restored ADG, ADFI, and feed conversion to levels not significantly different from and within 96% of that observed for pigs fed the adequate-P diet. The overall apparent digestibility of P was linearly (P < .01) improved with addition of 250 and 500 U phytase/kg to the low-P diet, but Ca and DM digestibilities were not affected by phytase or P level. In Exp. 2 (n = 224) the following diets were used: 1) .38/.33% total P, respectively, for grower and finisher phases, and no phytase; 2) .42/.37% P and no phytase; 3) .46/.41% P and no phytase; 4) diet 1 plus 167 U/kg phytase; 5) diet 1 plus 333 U/kg phytase; and 6) diet 1 plus 500 U/kg phytase. All diets contained .41/.36% Ca for grower and finisher phases, respectively. Pigs fed the low-P control diet grew slower (P < .01) and less efficiently (P < .10) than pigs fed diets with added P or phytase. With increasing levels of supplemental phytase or P there was a linear increase (P < .01) in ADG, digestibility of P, and digested P and a quadratic improvement (P < .05) in feed efficiency. Tenth rib mineralization based on shear force and ash were linearly increased (P < .08 to .001) as phytase or P was added to the low-P diet. There were generally no effects of P or phytase level on carcass quality. Using prediction equations derived from the response traits of ADG and P digestibility in Exp. 1 and ADG, P digestibility, and bone shear force in Exp. 2 to added phytase or P, we estimated that 500 U phytase released an amount of phytate P that was approximately equivalent to .87 to .96 g of P from dicalcium-monocalcium phosphate supplements. Fecal P excretion was estimated to be reduced 21.5%.
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PMID:Phytase supplementation of low-phosphorus growing-finishing pig diets improves performance, phosphorus digestibility, and bone mineralization and reduces phosphorus excretion. 941 91

The extracellular activity of Aspergillus niger phytase at the end of the growth phase was 132 nkat/mL in a laboratory bioreactor. The purified enzyme has molar mass approximately 100 kDa, pH optimum at 5.0, temperature optimum at 55 degrees C and high pH and temperature stability. The Km for dodecasodium phytate, calcium phytate and 4-nitrophenyl phosphate are 0.44, 0.45 and 1.38 mmol/L, respectively. The enzyme is noncompetively inhibited by inorganic monophosphate (Ki = 2.85 mmol/L) and by Cu2+, Zn2+, Hg2+, Sn2+, Cd2+ ions and strongly by F- ones; it is activated by Ca2+, Mg2+ and Mn2+ ions. The substrate specificity of phytase is broad with the highest affinity to calcium phytate.
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PMID:Characterization of phytase produced by Aspergillus niger. 944 82

Phytase hydrolyzes phytate to release inorganic phosphate, which would decrease the addition of phosphorus to feedstuffs for monogastric animals and thus reduce environmental pollution. The gene encoding phytase from Bacillus sp. DS11 was cloned in Escherichia coli and its sequence determined. A 560-bp DNA fragment was used as a probe to screen the genomic library. It was obtained through PCR of Bacillus sp. DS11 chromosomal DNA and two oligonucleotide primers based on N-terminal amino acid sequences of the purified protein and the cyanogen bromide-cleaved 21-kDa fragment. The phy cloned was encoded by a 2.2-kb fragment. This gene comprises 1152 nucleotides and encodes a polypeptide of 383 amino acids with a deduced molecular mass of 41,808 Da. Phytase was produced to 20% content of total soluble proteins in E. coli BL21 (DE3) using the pET22b(+) vector with the inducible T7 promoter. This is the first nucleic sequence report on phytase from a bacterial strain.
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PMID:Cloning of the thermostable phytase gene (phy) from Bacillus sp. DS11 and its overexpression in Escherichia coli. 959 81

Phytase catalyses the release of phosphate from phytate (myo-inositol hexakisphosphate), the predominant form of phosphorus in cereal grains, oilseeds and legumes. The presence of phytase activity was investigated in 334 strains of 22 species of obligately anaerobic ruminal bacteria. Measurable activities were demonstrated in strains of Selenomonas ruminantium, Megasphaera elsdenii, Prevotella ruminicola, Mitsuokella multiacidus and Treponema spp. Strains isolated from fermentations with cereal grains proved to have high activity, and activity was particularly prevalent in S. ruminantium, with over 96% of the tested strains being positive. The measured phytase activity was found exclusively associated with the bacterial cells and was produced in the presence of approximately 14 mM phosphate. The most highly active strains were all S. ruminantium, with the exception of the one Mitsuokella multiacidus strain examined. Phytase activity varied greatly among positive strains but activities as high as 703 nmol phosphate released (ml culture)-1 were measured for a S. ruminantium strain and 387 nmol phosphate released (ml culture)-1 for the Mitsuokella multiacidus strain.
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PMID:Phytase activity of anaerobic ruminal bacteria. 963 27

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

The phyA gene encoding an extracellular phytase from the thermophilic fungus Thermomyces lanuginosus was cloned and heterologously expressed, and the recombinant gene product was biochemically characterized. The phyA gene encodes a primary translation product (PhyA) of 475 amino acids (aa) which includes a putative signal peptide (23 aa) and propeptide (10 aa). The deduced amino acid sequence of PhyA has limited sequence identity (ca. 47%) with Aspergillus niger phytase. The phyA gene was inserted into an expression vector under transcriptional control of the Fusarium oxysporum trypsin gene promoter and used to transform a Fusarium venenatum recipient strain. The secreted recombinant phytase protein was enzymatically active between pHs 3 and 7.5, with a specific activity of 110 micromol of inorganic phosphate released per min per mg of protein at pH 6 and 37 degrees C. The Thermomyces phytase retained activity at assay temperatures up to 75 degrees C and demonstrated superior catalytic efficiency to any known fungal phytase at 65 degrees C (the temperature optimum). Comparison of this new Thermomyces catalyst with the well-known Aspergillus niger phytase reveals other favorable properties for the enzyme derived from the thermophilic gene donor, including catalytic activity over an expanded pH range.
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PMID:Molecular characterization and expression of a phytase gene from the thermophilic fungus Thermomyces lanuginosus. 979 1

Three experiments were conducted with 96 growing Landrace x Yorkshire x Duroc crossbreds to determine the collective effectiveness of cereal phytase from wheat middlings, microbial phytase, and citric acid in improving phytate-P bioavailability in corn-soy diets. In Exp. 1, 40 gilts (7 wk old) were fed five diets for 8 wk. Diets 1, 2, and 3 were low-P, corn-soybean meal diets (CSB) + 0, .1, or .2% inorganic P (Pi) as calcium phosphate, respectively. Diet 4 was a similar corn-soy diet that included 15% wheat middlings (461 cereal phytase U/kg). Diet 5 was the CSB + microbial phytase (1,200 U/kg; Natuphos, BASF, Mount Olive, NJ). In Exp. 2, 16 barrows (8 wk old) were fed two diets for 6 wk. Diet 1 was the same as Diet 3 of Exp. 1 (.2% Pi). Diet 2 was Diet 4 of Exp. 1 + microbial phytase (300 U/kg). In Exp. 3, 40 barrows and gilts (6 wk old) were fed four diets for 6 wk. Diets 1 and 2 were the same as those in Exp. 2. Diet 3 was Diet 2 of Exp. 2 + 1.5% citric acid. Diet 4 was similar to Diet 3 but contained 10 instead of 15% wheat middlings. In Exp. 1, pigs fed the low-P, CSB (Diet 1) had lower (P < .05) ADG, ADFI, plasma Pi concentration, bone strength, and mobility score than pigs of the other four treatments. Measurements for pigs fed the 15% wheat middlings diet were not significantly different from those of pigs fed the CSB + .1% Pi or microbial phytase. In Exp. 2, ADG (P=.06) during wk 1 to 3 and gain:feed ratio (P < .02) and plasma Pi concentration (P < .005) during all weeks favored pigs fed the CSB + .2% Pi compared with the other diet including 15% wheat middlings. In Exp. 3, identical ADG during all weeks and similar plasma Pi concentrations at wk 4 and 6 were observed between pigs fed the two citric acid diets (Diets 3 and 4) and the CSB + .2% Pi (Diet 1). Pigs fed Diet 4 (10% wheat middlings) had even higher (P < .02) gain:feed ratio during wk 1 to 3 than those fed Diet 1. It seems feasible to completely replace calcium phosphate with 10 to 15% wheat middlings, 300 U microbial phytase/ kg, and 1.5% citric acid in the corn-soy diets for growing pigs.
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PMID:Adding wheat middlings, microbial phytase, and citric acid to corn-soybean meal diets for growing pigs may replace inorganic phosphorus supplementation. 981 6

Phytases (myo-inositol hexakisphosphate phosphohydrolases) are found naturally in plants and microorganisms, particularly fungi. Interest in these enzymes has been stimulated by the fact that phytase supplements increase the availability of phosphorus in pig and poultry feed and thereby reduce environmental pollution due to excess phosphate excretion in areas where there is intensive livestock production. The wild-type phytases from six different fungi, Aspergillus niger, Aspergillus terreus, Aspergillus fumigatus, Emericella nidulans, Myceliophthora thermophila, and Talaromyces thermophilus, were overexpressed in either filamentous fungi or yeasts and purified, and their biophysical properties were compared with those of a phytase from Escherichia coli. All of the phytases examined are monomeric proteins. While E. coli phytase is a nonglycosylated enzyme, the glycosylation patterns of the fungal phytases proved to be highly variable, differing for individual phytases, for a given phytase produced in different expression systems, and for individual batches of a given phytase produced in a particular expression system. Whereas the extents of glycosylation were moderate when the fungal phytases were expressed in filamentous fungi, they were excessive when the phytases were expressed in yeasts. However, the different extents of glycosylation had no effect on the specific activity, the thermostability, or the refolding properties of individual phytases. When expressed in A. niger, several fungal phytases were susceptible to limited proteolysis by proteases present in the culture supernatant. N-terminal sequencing of the fragments revealed that cleavage invariably occurred at exposed loops on the surface of the molecule. Site-directed mutagenesis of A. fumigatus and E. nidulans phytases at the cleavage sites yielded mutants that were considerably more resistant to proteolytic attack. Therefore, engineering of exposed surface loops may be a strategy for improving phytase stability during feed processing and in the digestive tract.
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PMID:Biophysical characterization of fungal phytases (myo-inositol hexakisphosphate phosphohydrolases): molecular size, glycosylation pattern, and engineering of proteolytic resistance. 992 54


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