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

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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)

Soybean phytase (myo-inositol-hexakisphosphate phosphohydrolase; EC 3.1.3.8) was purified from 10-day-old germinating cotyledons using a four-step purification scheme. Phytase was separable from the major acid phosphatase present, and stained as a minor band of the three acid phosphatases detectable by activity staining after gel electrophoresis. The purified enzyme exhibited two closely migrating bands on sodium dodecyl sulfate-polyacrylamide gel electrophoresis of approximately 59 and 60 KDa. The molar extinction coefficient of the enzyme at 280 nm was estimated to be 7.5 X 10(4) M-1 cm-1. The isoelectric point of phytase, as judged by the elution profile on chromatofocusing, was about 5.5. The enzyme was totally absorbed to a Procion Red HE3B column and eluted as a single protein component at a salt concentration of 250-300 mM. The enzyme possessed a high affinity for phytic acid (apparent Km = 48 microM), and was strongly inhibited by phosphate (apparent Ki = 18 microM), vanadate, and fluoride. Characteristic of other plant phytases, the pH and temperature optima were 4.5-4.8 and 55 degrees C, respectively.
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PMID:Purification and characterization of phytase from cotyledons of germinating soybean seeds. 282 33

Soybean acid phosphatase (orthophosphoric-monoester phosphohydrolase, EC 3.1.3.2) was completely separated from phytase (EC 3.1.3.8) isolated from cotyledons of germinating seeds and purified to homogeneity. A four-step purification regimen consisting of ammonium sulfate fractionation, and ion-exchange, affinity, and chromatofocusing gel chromatographies was employed to achieve a homogeneous preparation. Acid phosphatase activity appeared as a major band of the three forms of acid phosphatase identified on native gels. The purified enzyme had a molecular weight of 53,000 when electrophoresed on 8% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and a molecular weight of 53,000 from its mobility in a Fracto-gel TSK HW-50F gel permeation column. The molar extinction coefficient of the enzyme at 278 nm was estimated to be 4.2 X 10(4) M-1 cm-1. The isoelectric point of the protein, as revealed by chromatofocusing, was about 6.7. The optimal pH for activity, like other plant acid phosphatases, was 5.0. While the enzyme failed to accommodate phytate as a substrate, the enzyme did exhibit a broad substrate selectivity. The affinity of the enzyme for p-nitrophenyl phosphate was high (Km = 70 microM), and activity was competitively inhibited by orthophosphate (Ki = 280 microM). The estimated catalytic turnover number (Kcat) of the enzyme for p-nitrophenyl phosphate was about 430 per second. Although the purified enzyme was stable at 0 degrees C and exhibited maximum catalytic activity at 60 degrees C, thermal inactivation studies indicated that the enzyme lost 100% activity after treatment at 68 degrees C for 10 min.
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PMID:Purification and characterization of acid phosphatase from cotyledons of germinating soybean seeds. 282 34

Low-phytate wheat bran was produced by enzymatic hydrolysis and extraction. Rat bioassay methods were utilized to determine bioavailability of iron and zinc in the low-phytate brans and to study the effect of dietary phytate/zinc molar ratio on zinc bioavailability when the phytate source was bran. Endogenous phytase activity hydrolyzed 80-100% of the phytate when wheat bran was incubated in water overnight. The relative biological values of the iron in raw bran and phytate-free bran were 98 and 113, respectively, compared to 100 for ferrous ammonium sulfate in a hemoglobin repletion assay. Low-phytate brans with phytate/zinc molar ratios of 8 or less were equivalent to zinc sulfate as dietary sources of zinc for growth of rats. Rats fed diets that contained wheat bran with zinc sulfate added to reduce the dietary phytate/zinc molar ratio from 40 or 50 to 20 grew at the same rate as rats fed a phytate-free diet, but femur zinc values were lower than those in the reference group. Gel filtration chromatography of extracts of raw and low-phytate brans suggested that zinc might be associated with phytate in wheat bran.
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PMID:Bioavailability to rats of iron and zinc in wheat bran: response to low-phytate bran and effect of the phytate/zinc molar ratio. 625 2

Iron bioavailability from an infant cereal made of wheat flour with a low extraction rate (70%) and cow milk was measured in infants by using a stable-isotope technique. A dephytinized infant cereal was prepared by adding commercial phytase during manufacture, resulting in degradation of 88% of the native phytic acid. Paired comparisons were made to evaluate the effect of phytic acid on iron bioavailability. Both infant cereals contained identical amounts of ascorbic acid and had a molar ratio of ascorbic acid to iron of 2:1. Iron was added as ferrous sulfate. No difference in iron bioavailability was observed in this study; the geometric mean was 8.7% (range: 3.8-16.9%) and 8.5% (range: 3.4-21.4%) from the cereal with native phytic acid (0.08% phytic acid) and the dephytinized cereal (0.01% phytic acid), respectively. Dephytinization of infant cereals containing a relatively low native phytic acid content and high amounts of ascorbic acid is thus unnecessary to ensure adequate bioavailability of iron.
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PMID:Bioavailability in infants of iron from infant cereals: effect of dephytinization. 909 72

An Fe depletion-repletion chick bioassay was conducted to determine whether supplemental microbial phytase or 1 alpha-hydroxycholecalciferol (1 alpha-OH D3) would improve the bioavailability of Fe in soybean meal (SBM). Weight gain, hemoglobin, and hematocrit were markedly improved when increasing levels (0, 10, 20, and 80 mg/kg) of Fe from analytical grade ferrous sulfate (FeSO4.7H2O) were added to the Fe-deficient casein-dextrose basal diet containing 20 mg Fe/kg. Addition of 19 mg Fe/kg from SBM to the basal diet improved (P < 0.05) hemoglobin and hematocrit, but the response was less than that obtained from 10 mg Fe/kg from FeSO4.7H2O. Phytase (1,430 units/kg), 1 alpha-OHD3 (10 micrograms/kg), or the combination, added to the SBM-fortified basal diet did not further improve hematocrit or hemoglobin, indicating that Fe bioavailability of SBM was not increased by either of these feed additives. Based on standard-curve methodology, and using hemoglobin as a criterion, the relative bioavailability of Fe was 38.5% for SBM, 21.0% for SBM+phytase, 23.2% for SBM+1 alpha-OHD3, and 29.2% for SBM+phytase+ 1 alpha-OHD3.
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PMID:Iron bioavailability in soybean meal as affected by supplemental phytase and 1 alpha-hydroxycholecalciferol. 931 19

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

Dietary phytase supplementation improves bioavailabilities of phytate-bound minerals such as P, Ca, and Zn to pigs, but its effect on Fe utilization is not clear. The efficacy of phytase in releasing phytate-bound Fe and P from soybean meal in vitro and in improving dietary Fe bioavailability for hemoglobin repletion in young, anemic pigs was examined. In Exp. 1, soybean meal was incubated at 37 degrees C for 4 h with either 0, 400, 800, or 1,200 units (U) of phytase/kg, and the released Fe and P concentrations were determined. In Exp. 2, 12 anemic, 21-d-old pigs were fed either a strict vegetarian, high-phytate (1.34%) basal diet alone, or the diet supplemented with 50 mg Fe/kg diet (ferrous sulfate) or phytase at 1,200 U/kg diet (Natuphos, BASF, Mt. Olive, NJ) for 4 wk. In Exp. 3, 20 anemic, 28-d-old pigs were fed either a basal diet with a moderately high phytate concentration (1.18%) and some animal protein or the diet supplemented with 70 mg Fe/kg diet, or with one of two types of phytase (Natuphos or a new phytase developed in our laboratory, 1,200 U/kg diet) for 5 wk. In Exp. 2 and 3, diets supplemented with phytase contained no inorganic P. In Exp. 1, free P concentrations in the supernatant increased in a phytase dose-dependent fashion (P<.05), whereas free Fe concentrations only increased at the dose of 1,200 U/kg (P<.10). In Exp. 2 and 3, dietary phytase increased hemoglobin concentrations and packed cell volumes over the unsupplemented group; these two measures, including growth performance, were not significantly different than those obtained with dietary supplemental Fe. In conclusion, both sources of phytase effectively degraded phytate in corn-soy diets and subsequently released phytate-bound Fe from the diets for hemoglobin repletion in young, anemic pigs.
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PMID:Phytase improves iron bioavailability for hemoglobin synthesis in young pigs. 1046 92

Phytases produced by numerous microorganisms and plants degrade phytic acid that has chelated with metal ions in food and feed. It is important to study phytase for the role of metal ions in nutrition of animals and humans as well as in the reduction of organic phosphate content of aqueous environment. This article reports on solid-state fermentation of phytase from a new substrate of cassava dregs. Large quantities of cassava dregs are produced in tropical areas as a byproduct of cassava starch processing. Protein and inorganic salts were found to be low in cassava dregs. Cassava dregs could be employed for phytase synthesis after the addition of a nitrogen source and mineral salts. Ammonium nitrate was the best nitrogen source among the nitrogen sources investigated, including beef extract, yeast extract, urea, ammonium nitrate, sodium nitrate, and ammonium sulfate. Sodium dodecyl sulfate promoted phytase production from cassava dregs. A maximum phytase yield of 6.73 U/g of dry mass was obtained. The obtained phytase was stable at feed-processing temperature, since 70% of initial enzyme activity was maintained after 30 min of treatment at 75 degrees C.
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PMID:Solid-state fermentation of phytase from cassava dregs. 1196 5

A phytase (EC 3.1.3.8) from Pseudomonas syringae MOK1 was purified to apparent homogeneity in two steps employing cation and an anion exchange chromatography. The molecular weight of the purified enzyme was estimated to be 45 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. The optimal activity occurred at pH 5.5 and 40 degrees C. The Michaelis constant (Km) and maximum reaction rate (Vmax) for sodium phytate were 0.38 mM and 769 U/mg of protein, respectively. The enzyme was strongly inhibited by Cu2+, Cd2+, Mn2+, and ethylenediaminetetraacetic acid (EDTA). It showed a high substrate specificity for sodium phytate with little or no activity on other phosphate conjugates. The enzyme efficiently released orthophosphate from wheat bran and soybean meal.
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PMID:Purification and characterization of a phytase from Pseudomonas syringae MOK1. 1462 9

Copper is often added to broiler diets at prophylactic concentrations as an antimicrobial despite purported chelation with and reduced utilization of phytin phosphorus. Therefore, male chicks were fed 0, 62.5, 125, 250, or 375 ppm Cu from Cu sulfate in combination with 600 phytase units (FTU)/kg phytase from 9 to 22 d of age (6 cages/diet, 8 birds/cage). Nonphytate phosphorus (NPP) and Ca were formulated to 0.2 and 0.7% of the diet, respectively. Three additional control diets were formulated to contain 0.27, 0.34, and 0.40% NPP, each with 0.7% Ca. Birds fed increasing concentrations of Cu with 600 FTU phytase/kg had linear reductions in performance characteristics (P < or = 0.05). Birds fed increasing concentrations of Cu with 600 FTU phytase/kg had linear increases in toe ash percentage (P < or = 0.027), but tibia ash percentage was not affected (P > 0.05). Birds fed increasing Cu concentrations with 600 FTU phytase/kg had linear reductions in apparent P retention as a percentage of total P (P < or = 0.0005). Supplementation with increasing concentrations of Cu to a diet containing 600 FTU phytase/kg resulted in decreases in 21 d BW, BW gain, feed consumption, feed conversion, tibia and toe ash weights, and apparent P retention as a percentage of total P. In this experiment, Cu supplementation did not reduce the efficacy of phytase (i.e., improvement in apparent P retention with phytase supplementation) but did decrease apparent P retention, BW gain, feed consumption, feed conversion, and tibia ash and toe ash weights.
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PMID:The effects of copper on the efficacy of phytase, growth, and phosphorus retention in broiler chicks. 1533 8


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