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1.
Phytate Hydrolysis in Rat Gastrointestinal Tracts, as Observed by 31P Fourier Transform Nuclear Magnetic Resonance Spectroscopy
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Phytate hydrolysis was followed through rat gastrointestinal tracts by 31P nuclear magnetic resonance spectroscopy. No phytate hydrolysis products were detected in the diet, stomach, or small intestine. It was concluded that cecal bacteria were responsible for phytate hydrolysis, which continued in the colon and fecal pellet. 相似文献
2.
《Archives of animal nutrition》2013,67(4):255-280
The present study gives an overview on the whole mechanism of phytate degradation in the gut and the enzymes involved. Based on the similarity of the human and pigs gut, the study was carried out in pigs as model for humans. To differentiate between intrinsic feed phytases and endogenous phytases hydrolysing phytate in the gut, two diets, one high (control diet) and the other one very low in intrinsic feed phytases (phytase inactivated diet) were applied. In the chyme of stomach, small intestine and colon inositol phosphate isomers and activities of phytases and alkaline phosphatases were determined. In parallel total tract phytate degradation and apparent phosphorus digestibility were assessed. In the stomach chyme of pigs fed the control diet, comparable high phytase activity and strong phytate degradation were observed. The predominant phytate hydrolysis products were inositol phosphates, typically formed by plant phytases. For the phytase inactivated diet, comparable very low phytase activity and almost no phytate degradation in the stomach were determined. In the small intestine and colon, high activity of alkaline phosphatases and low activity of phytases were observed, irrespective of the diet fed. In the colon, stronger phytate degradation for the phytase inactivated diet than for the control diet was detected. Phytate degradation throughout the whole gut was nearly complete and very similar for both diets while the apparent availability of total phosphorus was significantly higher for the pigs fed the control diet than the phytase inactivated diet. The pathway of inositol phosphate hydrolysis in the gut has been elucidated. 相似文献
3.
Phytic acid is naturally occurring compound that reduces intestinal absorption of many metals. Early work suggests that some dietary phytate may be hydrolyzed in the large intestines by bacteria, but more recently nutritionists have suggested that a mucosal enzyme is responsible. This paper reports a study intended to resolve this controversy. The hydrolysis of dietary phytic acid was measured in germfree and conventional rats fed either of two diets that differed in their calcium content. Negligible phytate hydrolysis occurred in the germfree rats, whereas 22 and 56% of the phytic acid was hydrolyzed by conventional rats fed high- and low-calcium diets, respectively. We concluded that bacteria were responsible for the hydrolysis of phytate in these diets and that any activity of endogenous enzyme was negligible. 相似文献
4.
Greiner R 《The protein journal》2004,23(8):567-576
A periplasmatic phytate-degrading enzyme from Pantoea agglomerans isolated from soil was purified about 470-fold to apparent homogeneity with a recovery of 16% referred to the phytate-degrading activity in the crude extract. It behaved as a monomeric protein with a molecular mass of about 42 kDa. The purified enzyme exhibited a single pH optimum at 4.5. Optimum temperature for the degradation of phytate was 60°C. The kinetic parameters for the hydrolysis of sodium phytate were determined to be KM = 0.34 mmol/l and kcat = 21 s-1 at pH 4.5 and 37°C. The enzyme exhibited a narrow substrate selectivity. Only phytate and glucose-1-phosphate were identified as good substrates. Since this Pantoea enzyme has a strong preference for glucose-1-phosphate over phytate, under physiological conditions glucose-1-phosphate is its most likely substrate. The maximum amount of phosphate released from phytate by the purified enzyme suggests myo-inositol pentakisphosphate as the final product of enzymatic phytate degradation. 相似文献
5.
Junichi Inagawa Isao Kiyosawa Taro Nagasawa 《Bioscience, biotechnology, and biochemistry》2013,77(11):3027-3032
The effects of phytate on the hydrolysis of lactose with β-galactosidases from bovine liver and Escherichia coli were investigated. The activities of both β-galactosidases were decreased to the same extent by increased concentrations of phytate. The rates of inhibition of β-galactosidase activity from E. coli in a reaction mixture containing 10 mm of phytate were 78.9% and 64.4%, respectively, in the absence of and with 4 mm of Mg2 +. Therefore, it was found that the stimulatory effect of Mg2+ was hardly affected by the presence of phytate in the range from 2 to 10 mm. The β-galactosidase activity was also not influenced by preincubating β-galactosidase or lactose with phytate. Kinetic studies showed that the inhibition of β-galactosidase activity by phytate was of an uncompetitive type with a Ki value of 3.46 mm. Therefore, it is considered that phytate may interact with a complex of ß-galactosidase and lactose. 相似文献
6.
Lassen SF Breinholt J Østergaard PR Brugger R Bischoff A Wyss M Fuglsang CC 《Applied and environmental microbiology》2001,67(10):4701-4707
Phytases catalyze the hydrolysis of phosphomonoester bonds of phytate (myo-inositol hexakisphosphate), thereby creating lower forms of myo-inositol phosphates and inorganic phosphate. In this study, cDNA expression libraries were constructed from four basidiomycete fungi (Peniophora lycii, Agrocybe pediades, a Ceriporia sp., and Trametes pubescens) and screened for phytase activity in yeast. One full-length phytase-encoding cDNA was isolated from each library, except for the Ceriporia sp. library where two different phytase-encoding cDNAs were found. All five phytases were expressed in Aspergillus oryzae, purified, and characterized. The phytases revealed temperature optima between 40 and 60 degrees C and pH optima at 5.0 to 6.0, except for the P. lycii phytase, which has a pH optimum at 4.0 to 5.0. They exhibited specific activities in the range of 400 to 1,200 U. mg, of protein(-1) and were capable of hydrolyzing phytate down to myo-inositol monophosphate. Surprisingly, (1)H nuclear magnetic resonance analysis of the hydrolysis of phytate by all five basidiomycete phytases showed a preference for initial attack at the 6-phosphate group of phytic acid, a characteristic that was believed so far not to be seen with fungal phytases. Accordingly, the basidiomycete phytases described here should be grouped as 6-phytases (EC 3.1.3.26). 相似文献
7.
Kinetics of phytate hydrolysis by Aspergillus niger phytase and correlation between the amount of released phosphate and creation of lower myo-inositol phosphates were investigated. Phytase was able to hydrolyze myo-inositol hexakis-, pentakis-, tetrakis-, and trisphosphates. Finally, about 56% of total phosphate were released and myo-inositol bisphosphate was detected as the end-product. 相似文献
8.
Saccharomyces cerevisiae CY phytase-producing cells were immobilized in calcium alginate beads and used for the degradation of phylate. The maximum
activity and immobilization yield of the immobilized phytase reached 280 mU/g-bead and 43%, respectively. The optimal pH of
the immobilized cell phytase was not different from that of the free cells. However, the optimum temperature for the immobilized
phytase was 50°C, which was 10°C higher than that of the free cells; pH and thermal stability were enhanced as a consequence
of immobilization. Using the immobilized phytase, phytate was degraded in a stirred tank bioreactor. Phytate degradation,
both in a buffer solution and in soybean-curd whey mixture, showed very similar trends. At an enzyme dosage of 93.9 mU/g-phytate,
half of the phytate was degraded after 1 h of hydrolysis. The operational stability of the immobilized beads was examined
with repeated batchwise operations. Based on 50% conversion of the phytate and five times of reuse of the immobilized beads,
the specific degradation (g phytate/g dry cell weight) for the immobilized phytase increased 170% compared to that of the
free phytase. 相似文献
9.
Purification and properties of phytate-specific phosphatase from Bacillus subtilis. 总被引:6,自引:0,他引:6
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An enzyme which liberates Pi from myo-inositol hexaphosphate (phytic acid) was shown to be present in culture filtrates of Bacillus subtilis. It was purified until it was homogeneous by ultracentrifugation, but it still showed two isozymes on polyacrylamide gel electrophoresis. The enzyme differed from other previously known phytases in its metal requirement and in its specificity for phytate. It had a specific requirement for Ca2+ for its activity. The enzyme hydrolyzed only phytate and had no action on other phosphate esters tested. This B. subtilis phytase is the only known phytate-specific phosphatase. The products of hydrolysis of phytate by this enzyme were Pi and myo-inositol monophosphate. The enzyme showed optimum activity at pH 7.5. It was inhibited by Ba2+, Sr2+, Hg2+, Cd2+, and borate. Its activity was unaffected by urea, diisopropylfluorophosphate, arsenate, fluoride, mercaptoethanol, trypsin, papain, and elastase. 相似文献
10.
Presence of and phosphate release from polyphosphates or phytate phosphate in lake sediments 总被引:2,自引:0,他引:2
NaOH is often used as an extractant for thefractionation of sediment-bound phosphates. Besidesorthophosphate, a certain quantity
of phosphate whichis called ‘non-reactive NaOH-extractable phosphate’is also extracted. In recent literature it has beensuggested
that this fraction consists of bacterialpolyphosphates and might be responsible for the phosphate release from aquatic sediments
under anoxicconditions.
In a previous publication we have already shown thatNaOH is not an accurate extractant as both theconcentration of NaOH and
the duration of theextraction have an influence on the quantity ofphosphate extracted, due to the hydrolysis of organicphosphates.
In this article we show that cold NaOH does not onlyextract iron-bound phosphate but phytate phosphate aswell. Non-reactive
phosphate in this extract was notrelated to the presence of polyphosphate, but mainlyto phytate and humic phosphates. As it
has been shownthat phytate may disappear from sediments when theybecome anoxic, we suggest that phytate mineralizationmay
be an important mechanism for anoxic phosphaterelease from sediments.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
11.
Greiner R 《The protein journal》2004,23(8):577-585
High-pressure liquid chromatography (HPLC) analysis established myo-inositol pentakisphosphate as the final product of phytate dephosphorylation by the phytate-degrading enzyme from Pantoea agglomerans. Neither product inhibition by phosphate nor inactivation of the Pantoea enzyme during the incubation period were responsible for the limited phytate hydrolysis as shown by addition of phytate-degrading enzyme and phytate, respectively, after the observed stop of enzymatic phytate degradation. In additon, the Pantoea enzyme did not possess activity toward the purified myo-inositol pentakisphosphate. Using a combination of High-Performance Ion Chromatography (HPIC) analysis and kinetic studies, the nature of the generated myo-inositol pentakisphosphate was established. The data demonstrate that the phytate-degrading enzyme from Pantoea agglomerans dephosphorylates myo-inositol hexakisphosphate in a stereospecific way to finally D-myo-inositol(1,2,4,5,6)pentakisphosphate. 相似文献
12.
Effect of phytate-removal and deamidation of soybean proteins on calcium absorption in the in situ rats 总被引:1,自引:0,他引:1
Kumagai H Koizumi A Sato N Ishikawa Y Suda A Sakurai H Kumagai H 《BioFactors (Oxford, England)》2004,22(1-4):21-24
Soybean proteins were deamidated by cation-exchange resins after phytate, the inhibitor for calcium absorption from the small intestine, was removed in order to provide the enhancement function of calcium absorption to soybean proteins. About 92% of the phosphorus was removed from the soybean proteins by anion-exchange-resin treatment, indicating that most of the phytate was removed. About 70% of the acid amide was deamidated by cation-exchange-resin treatment, and phytate-removed and deamidated soybean proteins (PrDS) having high calcium binding properties were obtained. PrDS were hydrolyzed by digestive enzymes and their calcium-binding properties and the enhancement function of the calcium absorption from the small intestine of rats were examined. As a result, PrDS retained their high calcium binding properties even after hydrolysis by digestive enzymes. In situ experiments showed that PrDS and their hydrolysates enhanced the calcium absorption from the intestine. 相似文献
13.
Urbano G López-Jurado M Aranda P Vidal-Valverde C Tenorio E Porres J 《Journal of physiology and biochemistry》2000,56(3):283-294
This review describes the present state of knowledge about phytic acid (phytate), which is often present in legume seeds. The antinutritional effects of phytic acid primarily relate to the strong chelating associated with its six reactive phosphate groups. Its ability to complex with proteins and particularly with minerals has been a subject of investigation from chemical and nutritional viewpoints. The hydrolysis of phytate into inositol and phosphates or phosphoric acid occurs as a result of phytase or nonenzymatic cleavage. Enzymes capable of hydrolysing phytates are widely distributed in micro-organisms, plants and animals. Phytases act in a stepwise manner to catalyse the hydrolysis of phytic acid. To reduce or eliminate the chelating ability of phytate, dephosphorylation of hexa- and penta-phosphate forms is essential since a high degree of phosphorylation is necessary to bind minerals. There are several methods of decreasing the inhibitory effect of phytic acid on mineral absorption (cooking, germination, fermentation, soaking, autolysis). Nevertheless, inositol hexaphosphate is receiving increased attention owing to its role in cancer prevention and/or therapy and its hypocholesterolaemic effect. 相似文献
14.
Phytase (myo-inositol hexaphosphate phosphohydrolase) belongs to phosphatases. It catalyzes the hydrolysis of phytate to less-phosphorylated inorganic phosphates and phytate. Phytase is used primarily for the feeding of simple hermit animals in order to increase the usability of amino acids, minerals, phosphorus and energy. In the present study, phytase isolation from the Lactobacillus coryniformis strain, isolated from Lor cheese sources, phytase purification and characterization were studied. The phytase was purified in simple three steps. The enzyme was obtained with 2.60% recovery and a specific activity of 202.25 (EU/mg protein). The molecular mass of the enzyme was determined to be 43.25 kDa with the sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) method. The optimum temperature and pH for the enzyme were found as 60 °C and 5.0 and respectively. To defined the substrate specificity of the phytase, the hydrolysis of several phosphorylated compounds by the purified enzyme was studied and sodium phytate showed high specificity. Furthermore, the effects of Ca2+, Ag+, Mg2+, Cu2+, Co2+, Pb2+, Zn2+ and Ni2+ metal ions on the enzyme were studied. 相似文献
15.
Sajidan A Farouk A Greiner R Jungblut P Müller EC Borriss R 《Applied microbiology and biotechnology》2004,65(1):110-118
Klebsiella sp. strain ASR1 isolated from an Indonesian rice field is able to hydrolyse myo-inositol hexakis phosphate (phytate). The phytase protein was purified and characterised as a 42 kDa protein accepting phytate, NADP and sugar phosphates as substrates. The corresponding gene (phyK) was cloned from chromosomal DNA using a combined approach of protein and genome analysis, and expressed in Escherichia coli. The recombinant enzyme was identified as a 3-phytase yielding myo-inositol monophosphate, Ins(2)P, as the final product of enzymatic phytate hydrolysis. Based on its amino acid sequence, PhyK appears to be a member of a hitherto unknown subfamily of histidine acid phytate-degrading enzymes with the active site RHGXRXP and HD sequence motifs, and is different from other general phosphatases and phytases. Due to its ability to degrade sodium phytate to the mono phosphate ester, the phyK gene product is an interesting candidate for industrial and agricultural applications to make phytate phosphorous available for plant and animal nutrition.Electronic Supplementary Material Supplementary material is available in the online version of this article at 相似文献
16.
Preparation of inositol phosphates from sodium phytate by enzymatic and nonenzymatic hydrolysis 总被引:6,自引:0,他引:6
Procedures for preparing myo-inositol bis-, tris-, tetrakis-, and pentakisphosphates from sodium phytate were established. Hydrolysis was achieved by autoclaving or enzymatic treatment; the inositol phosphates were separated by anion-exchange chromatography and were identified by fast atom bombardment-mass spectrometry. Enzymatic hydrolysis was more specific than autoclaving for isomer formation, whereas autoclaving was more efficient for producing the bis- and trisphosphates, which did not accumulate in significant amounts under the conditions of enzymatic hydrolysis. Sodium salts of the inositol phosphates were more powdery and less hygroscopic than the potassium salts. The procedures were satisfactory for producing gram quantities of each inositol phosphate, amounts adequate for animal studies of effects on mineral bioavailability. 相似文献
17.
A periplasmatic phytase from a bacterium isolated from Malaysian waste water was purified about 173-fold to apparent homogeneity
with a recovery of 10% referred to the phytase activity in the crude extract. It behaved as a monomeric protein with a molecular
mass of about 42 kDa. The purified enzyme exhibited a single pH optimum at 4.5. Optimum temperature for the degradation of
phytate was 65°C. The kinetic parameters for the hydrolysis of sodium phytate were determined to be K
M = 0.15 mmol/l and k
cat = 1164 s−1 at pH 4.5 and 37°C. The purified enzyme was shown to be highly specific. Among the phosphorylated compounds tested, phytate
was the only one which was significantly hydrolysed. Some properties such as considerable activity below pH 3.0, thermal stability
and resistance to pepsin make the enzyme attractive for an application as a feed supplement. 相似文献
18.
Biochemical properties and substrate specificities of alkaline and histidine acid phytases 总被引:8,自引:0,他引:8
Phytases are a special class of phosphatase that catalyze the sequential hydrolysis of phytate to less-phosphorylated myo-inositol derivatives and inorganic phosphate. Phytases are added to animal feedstuff to reduce phosphate pollution in the environment, since monogastric animals such as pigs, poultry, and fish are unable to metabolize phytate. Based on biochemical properties and amino acid sequence alignment, phytases can be categorized into two major classes, the histidine acid phytases and the alkaline phytases. The histidine acid phosphatase class shows broad substrate specificity and hydrolyzes metal-free phytate at the acidic pH range and produces myo-inositol monophosphate as the final product. In contrast, the alkaline phytase class exhibits strict substrate specificity for the calcium–phytate complex and produces myo-inositol trisphosphate as the final product. This review describes recent findings that present novel viewpoints concerning the molecular basis of phytase classification. 相似文献
19.
Kumagai H Koizumi A Suda A Sato N Sakurai H Kumagai H 《Bioscience, biotechnology, and biochemistry》2004,68(7):1598-1600
Soybean globulins were deamidated after removing phytate using ion-exchange resins, and then hydrolyzed by digestive enzymes. The phytate-removed deamidated soybean globulins (PrDS) retained high calcium-binding ability even after the hydrolysis by digestive enzymes. PrDS and its hydrolysates enhanced calcium absorption from the small intestine when injected into the small intestine together with a calcium solution. 相似文献
20.
《Bioscience, biotechnology, and biochemistry》2013,77(7):1598-1600
Soybean globulins were deamidated after removing phytate using ion-exchange resins, and then hydrolyzed by digestive enzymes. The phytate-removed deamidated soybean globulins (PrDS) retained high calcium-binding ability even after the hydrolysis by digestive enzymes. PrDS and its hydrolysates enhanced calcium absorption from the small intestine when injected into the small intestine together with a calcium solution. 相似文献