Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

---

Query: EC:3.1.3.8 (phytase)
1,997 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Supplementation with phytase is an effective way to increase the availability of phosphorus in seed-based animal feed. The biochemical characteristics of an ideal phytase for this application are still largely unknown. To extend the biochemical characterization of wild-type phytases, the catalytic properties of a series of fungal phytases, as well as Escherichia coli phytase, were determined. The specific activities of the fungal phytases at 37 degreesC ranged from 23 to 196 U. (mg of protein)-1, and the pH optima ranged from 2.5 to 7.0. When excess phytase was used, all of the phytases were able to release five phosphate groups of phytic acid (myo-inositol hexakisphosphate), which left myo-inositol 2-monophosphate as the end product. A combination consisting of a phytase and Aspergillus niger pH 2.5 acid phosphatase was able to liberate all six phosphate groups. When substrate specificity was examined, the A. niger, Aspergillus terreus, and E. coli phytases were rather specific for phytic acid. On the other hand, the Aspergillus fumigatus, Emericella nidulans, and Myceliophthora thermophila phytases exhibited considerable activity with a broad range of phosphate compounds, including phenyl phosphate, p-nitrophenyl phosphate, sugar phosphates, alpha- and beta-glycerophosphates, phosphoenolpyruvate, 3-phosphoglycerate, ADP, and ATP. Both phosphate liberation kinetics and a time course experiment in which high-performance liquid chromatography separation of the degradation intermediates was used showed that all of the myo-inositol phosphates from the hexakisphosphate to the bisphosphate were efficiently cleaved by A. fumigatus phytase. In contrast, phosphate liberation by A. niger or A. terreus phytase decreased with incubation time, and the myo-inositol tris- and bisphosphates accumulated, suggesting that these compounds are worse substrates than phytic acid is. To test whether broad substrate specificity may be advantageous for feed application, phosphate liberation kinetics were studied in vitro by using feed suspensions supplemented with 250 or 500 U of either A. fumigatus phytase or A. niger phytase (Natuphos) per kg of feed. Initially, phosphate liberation was linear and identical for the two phytases, but considerably more phosphate was liberated by the A. fumigatus phytase than by the A. niger phytase at later stages of incubation.
...
PMID:Biochemical characterization of fungal phytases (myo-inositol hexakisphosphate phosphohydrolases): catalytic properties. 992 55

A 20-wk feeding trial (21 to 40 wk of age) was conducted to evaluate the effects of phytase supplementation on egg production, egg quality, nutrient retention, and P excretion of laying hens fed diets containing different levels of P. Nine hundred and sixty ISA Brown hens were randomly allocated to completely randomized block arrangement of four diets: corn-soybean diet (1.4% tricalcium phosphate, TCP) without (T1, control) and with phytase (T2); 0.7% TCP (T3) or 0% TCP (T4) diet with phytase. Dietary microbial phytase was added at a level of 500 U/kg. Both hen-day and hen-housed egg production of T2 were significantly (P < 0.05) higher than other treatments, which were not different among themselves. Egg weights were also significantly (P < 0.05) different among treatments ,with T2 being the highest. Feed consumption of T2 was significantly (P < 0.05) higher than other treatments but feed conversion ratio was not significantly different from others. Specific gravity and shell thickness of the eggs were highest in the control (T1) but eggshell strength and broken egg to total egg ratio were not different among treatments. Haugh units were not different among treatments. Retention of Ca, P, Mg, Fe, and Zn were greater (P < 0.05) in phytase-supplemented groups. There were significant (P < 0.05) differences in excretion of ash, P, and Zn. The excretion of these components were highest in the control, whereas P excretion was significantly lower in the T3 and T4 groups. In conclusion, supplementation of the microbial phytase to normal corn-soybean diet improved egg production and can reduce TCP level in the diet without affecting egg production and egg quality. Significant reduction of P excretion can be also achieved.
...
PMID:Effects of microbial phytase supplementation on egg production, eggshell quality, and mineral retention of laying hens fed different levels of phosphorus. 1002 51

Two experiments (Exp.) were conducted to determine the growth response of White Pekin ducks to inclusion of microbial phytase in finisher diet. In Exp. 1, 1-d-old male ducks (240 total) were reared in litter-floor pens and fed regular starter diet until 3 wk of age. At 3 wk of age, ducks were randomly divided into six groups of 10 ducks each and each group was fed one of four diets. Three finisher diets containing 16% CP and 0.18% available phosphorus (AP) without supplemental P were formulated with microbial phytase (Natuphos) added at 0, 750, or 1,500 phytase units/kg of diet. The fourth diet was a control finisher diet that was supplemented with dicalcium phosphate (DCP) to supply dietary AP of 0.41%. Group BW and feed intake were measured weekly to assess growth response. At 6 wk of age, leg bones (tibia, femur, metatarsus) from five randomly selected ducks were removed and analyzed for bone characteristics. In Exp. 2, a total of 120 ducks reared as in Exp. 1 were randomly divided into six groups of five ducks each and fed one of four diets. A basal finisher diet was formulated to contain 16% CP and 0.18% AP. Monosodium phosphate was added to the basal diet to give dietary AP levels of 0.18, 0.27, and 0.36%. The fourth diet was the basal diet supplemented with microbial phytase (750 phytase units/kg of diet). Ducks were fed these diets from 3 to 6 wk of age. At the end of the study, ducks were bled by cardiac puncture and blood plasma was analyzed for P concentration. Leg bones from all ducks were removed and analyzed for bone characteristics as in Exp. 1. Feed intake increased linearly with increased level of dietary phytase, whereas the weight gain response was quadratic only during the last week of Exp. 1. In Exp. 2, there was a quadratic response for weight gain due to dietary AP. Weight gain due to phytase (750 units) was not different from ducks fed diets at 0 or 0.18% AP. Plasma P concentration increased linearly as dietary AP increased. Plasma P levels of ducks fed phytase were similar to those of ducks fed 0.18% AP but lower than in ducks fed 0.27% AP. Estimates of AP resulting from the addition of 750 units of phytase to basal diet were 0.05 and 0.07% based on plasma P concentration and weight gain, respectively. Using regression analysis, the AP due to phytase effect in the diet was estimated to range from 0.06 to 0.08%. Results suggest that phytase can be used in finisher diets similar to the one used in this study for ducks from 3 to 6 wk of age to improve growth performance and leg bone development similar to ducks fed diets supplemented with P from inorganic sources.
...
PMID:Microbial phytase in finisher diets of White Pekin ducks: effects on growth performance, plasma phosphorus concentration, and leg bone characteristics. 1009 Feb 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.
...
PMID:Cloning, sequencing, and expression of an Escherichia coli acid phosphatase/phytase gene (appA2) isolated from pig colon. 1009 20

The effect of body weight on P digestibility and on efficacy of supplemental Aspergillus niger phytase was studied in two experiments with young growing pigs. Excreta were collected quantitatively. All diets contained 2.0 g digestible P per kg dry matter at a maximum and renal P excretion never exceeded 15 mg/d. When dietary P mainly originated from monocalcium-phosphate, both P digestibility and Ca net absorption linearly increased by 3.6 and 5.6 percentage units, respectively, when BW increased from 15 to 35 kg. With a similar range in BW, P digestibility and Ca net absorption were unaffected by BW when P mainly originated from maize, barley and soybean meal. In both types of diet, crude protein digestibility increased with increasing body weight, whereas organic matter digestibility was effected by BW only in the diet containing maize, barley and soybean meal. Phytase (400 U/kg) almost doubled P digestibility when supplemented to a diet with P mainly originating from maize, soybean meal and barley. This effect of phytase supplementation was equal in pigs at 15.7 kg BW (33 vs. 55%) and at 39.1 kg BW (32 vs. 56%). Digestibility of any organic fraction was unaffected by supplemental phytase. With regard to on-farm conditions, it appears eligible from this results to apply digestibility coefficients for P determined in growing-finishing pigs for piglets as well.
...
PMID:Effect of body weight on phosphorus digestibility and efficacy of a microbial phytase in young pigs. 1054 67

A semisynthetic peroxidase was designed by exploiting the structural similarity of the active sites of vanadium dependent haloperoxidases and acid phosphatases. Incorporation of vanadate ion into the active site of phytase (E.C. 3.1.3.8), which mediates in vivo the hydrolysis of phosphate esters, leads to the formation of a semisynthetic peroxidase, which catalyzes the enantioselective oxidation of prochiral sulfides with H(2)O(2) affording the S-sulfoxide, e.g. in 66% ee at 100% conversion for thioanisole. Under reaction conditions the semi-synthetic vanadium peroxidase is stable for over 3 days with only a slight decrease in turnover frequency. Polar water-miscible cosolvents, such as methanol, dioxane, and dimethoxyethane, can be used in concentrations of 30% (v/v) at a small penalty in activity and enantioselectivity. Among the transition metal oxoanions that are known to be potent inhibitors, only vanadate resulted in a semisynthetic peroxidase when incorporated into phytase. A number of other acid phosphatases and hydrolases were tested for peroxidase activity, when incorporated with vanadate ion. Phytases from Aspergillus ficuum, A. fumigatus, and A. nidulans, sulfatase from Helix pomatia, and phospholipase D from cabbage catalyzed enantioselective oxygen transfer reactions when incorporated with vanadium. However, phytase from A. ficuum was unique in also catalyzing the enantioselective sulfoxidation, albeit at a lower rate, in the absence of vanadate ion.
...
PMID:The rational design of semisynthetic peroxidases. 1058 39

Bacillus amyloliquefaciens DS11 phytase (DS11 phytase) and Aspergillus ficuum phytase (AF phytase) activities were investigated by measuring the release of phosphate from phytate in animal feedstuff such as wheat bran, corn meal, soybean meal and rice flour at pH 5 and 7. In all the tested feedstuff, the enzymatic activity of DS11 phytase was more active at pH 7, but that of AF phytase was more active at pH 5. From these results, the phytate in the gastrointestinal tract could be degraded in the small intestine or stomach by DS11 or AF phytase, respectively. In conclusion, the results presented in this paper indicated that different combination ratios of DS11 and AF phytase, depending on the kind of feedstuff, might effectively induce more enzymatic activity both in the stomach and small intestine in terms of the pH of the gastrointestinal tract.
...
PMID:Comparative enzymatic hydrolysis of phytate in various animal feedstuff with two different phytases. 1059 87

Phytases hydrolyze phytic acid to less phosphorylated myo-inositol derivatives and inorganic phosphate. A thermostable phytase is of great value in applications for improving phosphate and metal ion availability in animal feed, and thereby reducing phosphate pollution to the environment. Here, we report a new folding architecture of a six-bladed propeller for phosphatase activity revealed by the 2.1 A crystal structures of a novel, thermostable phytase determined in both the partially and fully Ca2+-loaded states. Binding of two calcium ions to high-affinity calcium binding sites results in a dramatic increase in thermostability (by as much as approximately 30 degrees C in melting temperature) by joining loop segments remote in the amino acid sequence. Binding of three additional calcium ions to low-affinity calcium binding sites at the top of the molecule turns on the catalytic activity of the enzyme by converting the highly negatively charged cleft into a favorable environment for the binding of phytate.
...
PMID:Crystal structures of a novel, thermostable phytase in partially and fully calcium-loaded states. 1065 18

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.
...
PMID:Expression of the Aspergillus fumigatus phytase gene in Pichia pastoris and characterization of the recombinant enzyme. 1067 11

The localization of phytase (myo-inositol-hexaphosphate phosphohydrolase) in the ruminal bacteria, Selenomonas ruminantium JY35 and Mitsuokella multiacidus 46/5(2), was determined with transmission electron microscopy. Phosphate produced from the enzymatic dephosphorylation of the calcium salt of phytic acid is precipitated as calcium phosphate. The calcium is then replaced with lead to produce electron-dense lead phosphate. This deposition of lead phosphate localized phytase in S. ruminantium JY35 and M. multiacidus 46/5(2) to the outer membrane, and confirmed intracellular expression of the enzyme in Escherichia coli pSrP.2, the recombinant clone which possesses the gene (phyA) encoding phytase (phyA) in S. ruminantium.
...
PMID:Localization of phytase in Selenomonas ruminantium and Mitsuokella multiacidus by transmission electron microscopy. 1077 78


<< Previous 1 2 3 4 5 6 7 8 9 10 Next >>

---