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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)

The thermostable phytase from Bacillus amyloliquefaciens DS11 hydrolyzes phytate (myo-inositol hexakisphosphate, IP6) to less phosphorylated myo-inositol phosphates in the presence of Ca2+. In this report, we discuss the unique Ca2+-dependent catalytic properties of the phytase and its specific substrate requirement. Initial rate kinetic studies of the phytase indicate that the enzyme activity follows a rapid equilibrium ordered mechanism in which binding of Ca2+ to the active site is necessary for the essential activation of the enzyme. Ca2+ turned out to be also required for the substrate because the phytase is only able to hydrolyze the calcium-phytate complex. In fact, both an excess amount of free Ca2+ and an excess of free phytate, which is not complexed with each other, can act as competitive inhibitors. The Ca2+-dependent catalytic activity of the enzyme was further confirmed, and the critical amino acid residues for the binding of Ca2+ and substrate were identified by site-specific mutagenesis studies. Isothermal titration calorimetry (ITC) was used to understand if the decreased enzymatic activity was related to poor Ca2+ binding. The pH dependence of the Vmax and Vmax/Km consistently supported these observations by demonstrating that the enzyme activity is dependent on the ionization of amino acid residues that are important for the binding of Ca2+ and the substrate. The Ca2+-dependent activation of enzyme and substrate was found to be different from other histidine acid phytases that hydrolyze metal-free phytate.
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PMID:Calcium-dependent catalytic activity of a novel phytase from Bacillus amyloliquefaciens DS11. 1158 67

The possible use of phytase as a breadmaking improver has been tested in whole wheat breads by adding different amounts of fungal phytase. The effect of phytase addition on the fermentation stage and the final bread quality was analyzed. The phytase addition shortened the fermentation period, without affecting the bread dough pH. Regarding the whole wheat bread, a considerable increase of the specific bread volume, an improvement of the crumb texture, and the width/height ratio of the bread slice were obtained. An in vitro assay revealed that the improving effect of phytase on breadmaking might be associated with the activation of alpha-amylase, due to the release of calcium ions from calcium-phytate complexes promoted by phytase activity. As a conclusion, phytase offers excellent possibilities as a breadmaking improver, with two main advantages: first, the nutritional improvement produced by decreasing phytate content, and second, all the benefits produced by alpha-amylase addition can be obtained by adding phytase, which promotes the activation of endogenous alpha-amylase.
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PMID:Use of fungal phytase to improve breadmaking performance of whole wheat bread. 1171 42

A total of 72 barrows (initial body weight 16.7 kg) was used, to evaluate the influence of microbial phytase supplementation alone or in combination with calcium to barley soybean meal diets on the accumulation of cadmium (Cd) in kidney, liver, muscle, brain and bone. The control group received the basal diet with 6 g Ca and a low native Cd concentration of 0.03 mg/kg dry matter (DM). In the experimental groups 2, 3, 4 and 5 dietary cadmium concentration was elevated to 0.78 mg/kg DM. The diet of group 3 was supplemented with 800 U microbial phytase/kg, the diet of group 4 with 6 g Ca/kg. The diet of group 5 contained both supplements. The addition of microbial phytase caused an increase of Cd retention in kidney and liver at 30 and 50 kg body weight. This effect was counteracted by the contemporary addition of calcium. A supplementation of Ca alone showed no effect on the Cd accumulation in kidney and liver. In muscle, brain and bone no effects of phytase and calcium on the accumuLation of Cd could be found.
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PMID:The influence of dietary microbial phytase and calcium on the accumulation of cadmium in different organs of pigs. 1178 74

A novel phytase gene ( phyL) was cloned from Bacillus licheniformis by multiple steps of degenerate and inverse PCR. The coding region of the phyL gene was 1,146 bp in size and a promoter region of approximately 300 bp was identified at the upstream sequence. This gene, together with a phytase gene ( 168phyA) identified in the B. subtilis strain 168 genome by a homology search, was cloned and over-expressed in B. subtilis using a phi105MU331 prophage vector system. Up to 35 units of phytase/ml were secreted into the culture media; and mature enzymes of around 44-47 kDa were purified for characterization. Both phytases exhibited broad temperature and pH optima and showed high thermostability. Of the two, the phytase encoded by phyL exhibited higher thermostability, even at a lower calcium concentration, as it was able to recover 80% of its original activity after denaturation at 95 degrees C for 10 min. With their neutral pH optima and good temperature stabilities, these Bacillus phytases are good candidates for animal feed applications and transgenic studies.
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PMID:Molecular cloning and the biochemical characterization of two novel phytases from B. subtilis 168 and B. licheniformis. 1211 Nov 45

The aim of this study was to investigate the effect of various calcium:total phosphorus (Ca:tP) ratios due to addition of microbial phytase and 1,25-dihydroxycholecalciferol [1,25-(OH)2D3] on the absorption levels of minerals. In a 42-day experiment repeated three times, 144 day-old male broiler chicks (ISA 220) were divided into six groups of eight chicks each. Diets containing two different (1:1 and 2:1) Ca levels were prepared. Groups 1, 2 and 3 received a diet of 1:1 Ca:tP ratio while Groups 4, 5 and 6 a diet of 1:2 Ca:tP ratio. These diets contained 1,25-(OH)2D3 and phytase in levels of 5 micrograms/kg and 600 FYT/kg, respectively. The faeces was collected to analyse the absorption of minerals. At the end of the study, the absorption levels of Ca, P, Zn, Mn and Cu were increased by the addition of phytase enzyme (p < 0.05). This effect was obvious in the 3rd week. On the other hand, in the 6th week only Ca and P absorption levels were influenced positively by the addition of phytase enzyme. The results proved the positive effect of phytase, an enzyme which is used for increasing the utilisation rate of phytate P, Ca and other minerals in broilers.
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PMID:Effects of microbial phytase and 1,25-dihydroxycholecalciferol on the absorption of minerals from broiler chicken diets containing different levels of calcium. 1223 72

1. Large White male turkeys from a heavy commercial male-line were fed on 16 diets containing 4 concentrations of calcium (6, 10, 14 and 18 g/kg) and available phosphorus (3, 5, 7, 9 g/kg) in a 4 x 4 factorial experiment with three replicates (pens). Turkeys were weighed and food intakes recorded from 4 to 7, 8 to 10 and 11 to 13 weeks of age. 2. The diets containing 6 g/kg calcium and 5, 7 or 9 g/kg available phosphorus concentrations were associated with lower body weighs at 10 and 13 weeks of age. 3. Fewer than 6% of the turkeys had an abnormal gait at 13 weeks of age. 4. The optimum dietary concentrations were 10 g/kg of calcium and 3 g/kg of available phosphorus. It was concluded that these concentrations should be fed at least to 13 weeks of age. 5. Retentions of dietary calcium and phosphorus averaged 300 g/kg of intake. 6. Utilisation of dietary phytate ranged from 5 to 11 g/kg and it is recommended that organic phosphorus should be ignored in the formulation of diets for growing turkeys unless they are supplemented with a phytase enzyme.
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PMID:Effects of dietary calcium and phosphorus on mineral retention, growth, feed efficiency and walking ability in growing turkeys. 1236 18

Extracellular phytase produced by Aspergillus niger ATCC 9142 was purified to homogeneity by employing an initial ultrafiltration step, followed by chromatography using ion exchange, gel filtration and chromatofocusing steps. The purified enzyme was an 84 kDa, monomeric protein. It possessed a temperature optimum of 65 degrees C, and a pH optimum of 5.0. Km and Vmax values of 100 microM and 7 nmol/s, respectively, were recorded and these values fall well within the range of those previously reported for microbial phytases. Substrate specificity studies indicated that, while the enzyme could hydrolyse a range of non-phytate-based phosphorylated substrates, its preferred substrate was phytate. Phytase activity was moderately stimulated in the presence of Mg2+, Mn2+, Cu2+, Cd2+, Hg2+, Zn2+ and F- ions. Activity was not significantly affected by Fe2- or Fe3- and was moderately inhibited by Ca2+. The enzyme displayed higher thermostability at 80 degrees C than did two commercial phytase products. Initial characterisation of the purified enzyme suggested that it could be a potential candidate for use as an animal feed supplement.
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PMID:Purification and characterization of extracellular phytase from Aspergillus niger ATCC 9142. 1265 85

An experiment was conducted to compare two common methods of estimating bone ash from growing broiler chicks (A = autoclaving; B = boiling/extracting). Ross x Ross 1-d-old broiler chicks were fed a corn-soy, phosphorus-deficient diet (22.7% CP, 1% calcium, 0.22% non-phytate phosphorus), with 0, 750, 1,500, 3,000, or 6,000 units of phytase (FTU) to produce bones with a range of ash. The methods were compared with bones from chicks at 7, 14, and 21 d of age. Left legs were used for method B, and right legs were used for method A. Data was analyzed by the general linear models procedure of SAS software, and differences between means were detected using the Duncan's new multiple range test at the 0.05 level. Variance estimates were the mean square errors (from SAS outputs). Sample sizes needed to detect a 2% difference in bone ash were calculated using the method of Zar. The addition of 6,000 FTU/kg increased tibia ash from 26 to 37%, 29 to 41%, and 33 to 43% on Days 7, 14 and 21, respectively (method B). With bones from 7-d-old chicks, the same number of samples was necessary to detect a 2% difference using methods A and B. With bones from 14- and 21-d-old chicks, approximately 50 and 150% more samples were necessary, respectively, using method A. The autoclaving method is less labor-intensive and requires no toxic solvents, but for older birds, many more samples or replications are needed to detect the same treatment differences.
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PMID:Power of two methods for the estimation of bone ash of broilers. 1270 2

Information is needed on organic polyphosphates such as myo-inositol 1,2,3,5/4,6-hexakis dihydrogenphosphate or phytate (IP6) contribution to the sources and sinks of dissolved phosphorus (PO4-P) in the soil-manure-water system. Effects of Na+, Ca2+, Al3+, and Fe3+ and cation to IP6-P mole ratios on the enzymatic dephosphorylation of IP6 were studied to determine controlling mechanisms of dephosphorylation and persistence in manure. Phytate- and PO4-P were analyzed by high-performance liquid chromatography. Phytate dephosphorylation by Aspergillus ficuum (Reichardt) Henn. phytase EC 3.1.3.8 decreases by 50 +/- 3.6 and 40 +/- 4% at pH 4.5 and 6, respectively, as Ca2+ concentrations increase and cation to IP6-P mole ratios reach 6:6. Polyanionic IP6 has a high affinity for Al3+ and Fe3+ and reductions in dephosphorylation average 27 and 32% at a cation to IP6-P mole ratio of 1:6 for Al3+ and Fe3+, respectively, while reaching more than 99% at a mole ratio of 6:6. A phytase-hydrolyzable phosphorus (PHP) fraction is native to ruminant animal manure and is proportional to total solids (TS) concentration in 1 to 100 g L(-1) suspensions. Added phytase, in effect, increases water-extractable P content of manure and the risk of environmental P dispersion. As the bioavailability and ecological effect of IP6-P appear to be regulated not only by pH-controlled enzyme activity but also by the associated counterions, the differential protective effects of cations influence the accuracy of manure PHP fraction estimates and increase phytate resistance to enzymatic dephosphorylation that may lead to its persistence in manure.
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PMID:Polyvalent cation effects on myo-inositol hexakis dihydrogenphosphate enzymatic dephosphorylation in dairy wastewater. 1270 95

Absorption of selenium and copper is much lower in ruminants than in nonruminants. The low absorption of these minerals in ruminants is due to modifications that occur in the rumen environment. Selenium bioavailability is reduced by high dietary sulfur and the presence of cyanogenetic glycosides in certain legumes. Feeding organic selenium from selenomethionine or selenized yeast results in much higher tissue and milk selenium concentrations than are obtained with selenite. High dietary molybdenum in combination with moderate to high dietary sulfur results in formation of thiomolybdates in the rumen. Thiomolybdates greatly reduce copper absorption, and certain thiomolybdate species can be absorbed and interfere systemically with copper metabolism. Independent of molybdenum, high dietary sulfur reduces copper absorption perhaps via formation of copper sulfide. High dietary iron also reduces copper bioavailability. Dietary factors that affect bioavailability of zinc in ruminants are not well defined. Phytate does not affect zinc absorption in ruminants because microbial phytase in the rumen degrades phytate. Manganese is very poorly absorbed in ruminants, and limited research suggests that high dietary calcium and phosphorus may reduce manganese absorption.
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PMID:Trace mineral bioavailability in ruminants. 1273 Apr 54


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