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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 flour was fermented with Aspergillus usamii to improve the availabilities of dietary zinc and iron through the degradation of phytate. Three kinds of experimental diets that differed in protein sources were prepared: one consisting of 40% fermented soybean flour (RS diet), one consisting of 40% fermented soybean flour (FS diet), and one consisting of 20% regular soybean flour and 20% fermented soybean flour (RF diet). Zinc solubilities in the upper and the lower segments of the small intestine were higher in rats fed the FS diet than in rats fed the RS diet. The FS group showed higher solubility of iron in the lower small intestine than the RS group did. Zinc concentrations in the femur and plasma and iron concentrations in the liver and plasma were higher in the FS group than in the RS group. These results suggested that the fermentation of soybean flour improved the availabilities of dietary zinc and iron, which may be induced by increasing the solubilities of these minerals in the small intestine through the reduction of phytate content. Femoral and plasma zinc concentrations in the RF group were higher than in the RS group, but lower than in the FS group. No difference was noted in liver and plasma iron concentrations between the RF group and the FS group. Although phytase activity in FS degrades phytate in the RF diet, higher activity may be needed to degrade phytate completely.
J Nutr Sci Vitaminol (Tokyo) 1998 Dec
PMID:Fermentation of soybean flour with Aspergillus usamii improves availabilities of zinc and iron in rats. 1019 18

Differential agar media for the detection of microbial phytase activity use the disappearance of precipitated calcium or sodium phytate as an indication of enzyme activity. When this technique was applied to the study of ruminal bacteria, it became apparent that the method was unable to differentiate between phytase activity and acid production. Strong positive reactions (zones of clearing around microbial colonies) observed for acid producing, anaerobic bacteria, such as Streptococcus bovis, were not corroborated by subsequent quantitative assays. Experimentation revealed that acidic solutions generated false positive results on the selected differential medium. Empirical studies undertaken to find a solution to this limitation determined the false positive results could be eliminated through a two step counterstaining treatment (cobalt chloride and ammonium molybdate/ammonium vanadate) which reprecipitates acid solubilized phytate. This report discusses the application of the developed two step counterstaining treatment for the screening of phytase producing ruminal bacteria as well as its use in phytase zymogram assays.
J Microbiol Methods 1999 Dec
PMID:A novel staining method for detecting phytase activity. 1057 3

A (31)P NMR method for quantitative determination of inositol phosphates in simple incubation samples of sodium phytate and Aspergillus niger phytase and in different types of complex samples, such as diets, digesta, and feces, is described. The inositol phosphates in complex samples were extracted with HCl, concentrated, and purified using freeze-drying and filtration and subsequently determined at pH 12.6 in aqueous solution using a (31)P NMR method. The (31)P NMR technique has as its main advantages over the HPLC techniques that it does not necessitate standards that may cause background matrix effects and that the spectra of inositol phosphates and orthophosphate appear in the same run without further sampling errors. The results of inositol hexaphosphate analysis with HPLC can be confirmed by this (31)P NMR method. Contents of inositol tetra-, tri-, di-, and monophosphate in the biological samples appear to be quantitatively not important. The (31)P NMR method can be applied for use in animal nutrition in general and studies of using phytase in diets for farm animals in particular, by measuring the content of inositol phosphates in feed ingredients, complete feeds, ileal contents, and feces of pigs and poultry.
J Agric Food Chem 1999 Dec
PMID:Quantification of inositol phosphates using (31)P nuclear magnetic resonance spectroscopy in animal nutrition. 1060 82

The objective of this study was to investigate the influence of microbial phytase and calcium supplementation to diets for growing pigs on the retention of lead in the kidney, liver, muscle, brain, and bone (phalanx 1). The experiments were carried out with barrows over the body weight range from 17 to 50 kg. The average lead concentration of the diets was 1.45 mg/kg dry matter. Diets were prepared with or without a supplement of 800 units of microbial phytase. The calcium concentration in the diets was 6.53 or 13.4 g/kg dry matter. The addition of microbial phytase showed an increase of lead concentration in bone. By increasing the calcium concentration to 13.4 g/kg dry matter, it was possible to avoid the phytase-induced increase of lead retention in bone.
Biol Trace Elem Res 1999 Dec
PMID:Influence of microbial phytase and dietary calcium on the accumulation of lead in different organs of pigs. 1061 63

An experiment employing a factorial arrangement of two levels of Ca, two levels of available P (AP), and three levels of phytase enzyme was carried out with 360 ISA White layers from 18 to 67 wk of age. The Ca levels were maintained at 3.7 and 4.0% throughout the experiment. The AP levels were 0.2 and 0.4% for the high and low treatments until 55 wk of age and were reduced to 0.11 and 0.22% thereafter. Phytase enzyme levels were 0, 250, and 500 phytase units (FTU)/kg of feed. In the period before Week 55, either level of AP was likely adequate for maximum production. However, when lower levels of AP were fed after this time, low AP was associated with reduced BW and egg production, and enzyme supplementation was able to compensate for low AP. In this period, high AP and the highest level of phytase produced negative effects on BW, egg weight, and the feed conversion ratio. The ratio of Ca to AP was important; shell quality was best with high or low levels of both. With high levels of Ca, enzyme supplementation compensated for low levels of AP and overcompensated with a high level of AP. These effects were reduced or absent with low levels of Ca. It is clear from this study that phytase enzyme can compensate for low levels of AP in diets based on corn and soybean meal, but that the optimum level of supplementation depends as well on the Ca level.
Poult Sci 1999 Dec
PMID:The effect of phosphorus, phytase enzyme, and calcium on the performance of layers fed corn-based diets. 1062 50

1. Seven-day old male broilers (n=900) were fed on wheat-sorghum-soyabean meal-based diets containing 3 concentrations of phytic acid (10.4, 13.2 and 15.7 g/kg; equivalent to 2.9, 3.7 and 4.4 g/kg phytate phosphorus), 2 of non-phytate phosphorus (2.3 and 4.5 g/kg) and 3 of microbial phytase (Natuphos 5000 L; 0, 400 and 800 FTU/kg) in a 19-d trial. The dietary phytic acid contents were manipulated by the inclusion of rice pollard. 2. Each dietary treatment was fed to 5 pens (10 birds/pen) from 7 to 25 d of age. Records of body weight, food intake and mortality were maintained. On d 25, all surviving birds were killed and toe samples were obtained for toe ash measurements. 3. Increasing dietary phytic acid negatively influenced body weight gain, food intake and food/gain. These adverse effects were partially overcome by the addition of microbial phytase. 4. Supplemental phytase caused improvements in weight gain and food efficiency of broilers but the magnitude of the responses was greater in low non-phytate phosphorus diets, resulting in significant non-phytate phosphorus x phytase interactions. 5. Toe ash contents were improved by phytase addition but the response was greater at the highest concentration of phytic acid, resulting in a significant phytic acid x phytase interaction. Responses were also greater in low non-phytate phosphorus diets as indicated by significant non-phytate phosphorus x phytase interaction. 6. In general, there was very little difference in the responses to phytase additions at 400 and 800 FTU/kg. 7. The performance responses to added phytase in birds receiving adequate non-phytate phosphorus diets provide evidence for the influence of the enzyme on animal performance independent of its effect on phosphorus availability.
Br Poult Sci 1999 Dec
PMID:Response of broiler chickens to microbial phytase supplementation as influenced by dietary phytic acid and non-phytate phosphorus contents. I. Effects on bird performance and toe ash. 1067 Jun 79

A simple and inexpensive high-performance thin-layer chromatography (HPTLC) method for the analysis of inositol mono- to hexakisphosphates on cellulose precoated plates is described. Plates were developed in 1-propanol-25% ammonia solution-water (5:4:1) and substance quantities as low as 100-200 pmol were detected by molybdate staining. Chromatographic mobilities of nucleotides and phosphorylated carbohydrates were also characterized. Charcoal treatment was employed to separate nucleotides from inositol phosphates with similar R(F) values prior to HPTLC analysis. Practical application of the HPTLC system is demonstrated by analysis of grain extracts from wild type and low-phytate mutant barley as well as phytate degradation products resulting from barley phytase activity.
J Chromatogr B Biomed Sci Appl 1999 Dec 24
PMID:High-performance thin-layer chromatography method for inositol phosphate analysis. 1067 2

Phytic acid (myo-inositol hexakisphosphate, InsP(6)) hydrolysis by Bacillus phytase (PhyC) was studied. The enzyme hydrolyses only three phosphates from phytic acid. Moreover, the enzyme seems to prefer the hydrolysis of every second phosphate over that of adjacent ones. Furthermore, it is very likely that the enzyme has two alternative pathways for the hydrolysis of phytic acid, resulting in two different myo-inositol trisphosphate end products: Ins(2,4,6)P(3) and Ins(1,3,5)P(3). These results, together with inhibition studies with fluoride, vanadate, substrate and a substrate analogue, indicate a reaction mechanism different from that of other phytases. By combining the data presented in this study with (1) structural information obtained from the crystal structure of Bacillus amyloliquefaciens phytase [Ha, Oh, Shin, Kim, Oh, Kim, Choi and Oh (2000) Nat. Struct. Biol. 7, 147-153], and (2) computer-modelling analyses of enzyme-substrate complexes, a novel mode of phytic acid hydrolysis is proposed.
Biochem J 2000 Dec 15
PMID:Analysis of myo-inositol hexakisphosphate hydrolysis by Bacillus phytase: indication of a novel reaction mechanism. 1110 66

Previously, sequence comparisons between a mesophilic enzyme and a more thermostable homologue were shown to be a feasible approach to successfully predict thermostabilizing amino acid substitutions. The 'consensus approach' described in the present paper shows that even a set of amino acid sequences of homologous, mesophilic enzymes contains sufficient information to allow rapid design of a thermostabilized, fully functional variant of this family of enzymes. A sequence alignment of homologous fungal phytases was used to calculate a consensus phytase amino acid sequence. Upon construction of the synthetic gene, recombinant expression and purification, the first phytase obtained, termed consensus phytase-1, displayed an unfolding temperature (T(m)) of 78.0 degrees C which is 15-22 degrees C higher than the T(m) values of all parent phytases used in its design. Refinement of the approach, combined with site-directed mutagenesis experiments, yielded optimized consensus phytases with T(m) values of up to 90.4 degrees C. These increases in T(m) are due to the combination of multiple amino acid exchanges which are distributed over the entire sequence of the protein and mainly affect surface-exposed residues; each individual substitution has a rather small thermostabilizing effect only. Remarkably, in spite of the pronounced increase in thermostability, catalytic activity at 37 degrees C is not compromised. Thus, the design of consensus proteins is a potentially powerful and novel alternative to directed evolution and to a series of rational approaches for thermostability engineering of enzymes and other proteins.
Biochim Biophys Acta 2000 Dec 29
PMID:The consensus concept for thermostability engineering of proteins. 1115 Jun 16

The tools of molecular and cellular biology can be used to precisely describe traits in terms of a sequence of nucleic acids when their molecular and cellular bases are well understood. The entire genome of elite production birds, however, cannot be written as a series of A's, T's, C's, and G's because the interaction between alleles at the same and different loci is too large and there is likely to be many genotypes that encode the same production trait phenotype. A first draft of the genetic map of the chicken is anticipated within the next few years, but a complete molecular description of the genome of birds with elite production characteristics is not anticipated in the near future. Quantitative genetics will remain the cornerstone of breeding programs for production traits. Novel sequences encoding traits such as enhanced nutritional capability (e.g., expression of phytase) and resistance to specific diseases could be introduced into lines of chickens using the tools of molecular and cellular biology. Cloning could be used by the poultry industry to disperse highly desirable genotypes without the need for grandparent and parent flocks for multiplication.
Poult Sci 2001 Dec
PMID:From chicken coops to genome maps: generating phenotype from the molecular blueprint. 1177 77


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