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Sample records for beta-propeller phytase genes

  1. The diversity and abundance of phytase genes (beta-propeller phytases) in bacterial communities of the maize rhizosphere

    NARCIS (Netherlands)

    Cotta, S.R.; Cavalcante Franco Dias, A.; Seldin, L.; Andreote, F. D.; van Elsas, J. D.

    2016-01-01

    The ecology of microbial communities associated with organic phosphorus (P) mineralization in soils is still understudied. Here, we assessed the abundance and diversity of bacteria harbouring genes encoding beta-propeller phytases (BPP) in the rhizosphere of traditional and transgenic maize cultivat

  2. Optimization of five environmental factors to increase beta-propeller phytase production in Pichia pastoris and impact on the physiological response of the host.

    Science.gov (United States)

    Viader-Salvadó, José M; Castillo-Galván, Miguel; Fuentes-Garibay, José A; Iracheta-Cárdenas, María M; Guerrero-Olazarán, Martha

    2013-01-01

    Recently, we engineered Pichia pastoris Mut(s) strains to produce several beta-propeller phytases, one from Bacillus subtilis and the others designed by a structure-guided consensus approach. Furthermore, we demonstrated the ability of P. pastoris to produce and secrete these phytases in an active form in shake-flask cultures. In the present work, we used a design of experiments strategy (Simplex optimization method) to optimize five environmental factors that define the culture conditions in the induction step to increase beta-propeller phytase production in P. pastoris bioreactor cultures. With the optimization process, up to 347,682 U (82,814 U/L or 6.4 g/L culture medium) of phytase at 68 h of induction was achieved. In addition, the impact of the optimization process on the physiological response of the host was evaluated. The results indicate that the increase in extracellular phytase production through the optimization process was correlated with an increase in metabolic activity of P. pastoris, shown by an increase in oxygen demand and methanol consumption, that increase the specific growth rate. The increase in extracellular phytase production also occurred with a decrease in extracellular protease activity. Moreover, the optimized culture conditions increased the recombinant protein secretion by up to 88%, along with the extracellular phytase production efficiency per cell.

  3. Identification of a phytase gene in barley (Hordeum vulgare L..

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    Fei Dai

    Full Text Available BACKGROUND: Endogenous phytase plays a crucial role in phytate degradation and is thus closely related to nutrient efficiency in barley products. The understanding of genetic information of phytase in barley can provide a useful tool for breeding new barley varieties with high phytase activity. METHODOLOGY/PRINCIPAL FINDINGS: Quantitative trait loci (QTL analysis for phytase activity was conducted using a doubled haploid population. Phytase protein was purified and identified by the LC-ESI MS/MS Shotgun method. Purple acid phosphatase (PAP gene was sequenced and the position was compared with the QTL controlling phytase activity. A major QTL for phytase activity was mapped to chromosome 5 H in barley. The gene controlling phytase activity in the region was named as mqPhy. The gene HvPAP a was mapped to the same position as mqPhy, supporting the colinearity between HvPAP a and mqPhy. CONCLUSIONS/SIGNIFICANCE: It is the first report on QTLs for phytase activity and the results showed that HvPAP a, which shares a same position with the QTL, is a major phytase gene in barley grains.

  4. Phytase.

    Science.gov (United States)

    Wodzinski, R J; Ullah, A H

    1996-01-01

    Of all the sources of phytase that have been studied (plant, animal, and microorganisms), the highest yields are produced by a wild-type strain A. niger NRRL 3135 (12.7 mg P/hr/ml = 6.8 microns P/ml/min = 113.9 nKat/ml) in a mineral salt medium in which total phosphate (4 mg %) is limiting for growth and cornstarch and glucose are the carbon sources. Synthesis of the enzyme is repressed by phosphate in the wild-type strain. Aspergillus niger NRRL 3135 produces two phytases one with pH optima at 2.5 and 5.5 (phyA) and one with an optimum at pH 2.0 (phyB). It also produces a pH 6.0 optimum phosphatase that has no phytase activity. These three glycoproteins have been purified to homogeneity, characterized, sequenced, and cloned. The sequences have been compared to each other, other phytases, and to known phosphatases. Their homology has been determined. The active sites of phytases show remarkable homology to the active site residues of the members of a particular class of acid phosphatase (histidine phosphatase). The most conserved sequence is RHGXRXP. Phytase has been covalently immobilized on Fractogel TSK HW-75 F and glutaraldehyde-activated silicate. It has been immobilized on agarose. Losses of activity have been noted on immobilization but these may be minimized by future research. It should be possible to commercially produce and recover penta-, tetra-, tri-, di-, and monoinositol phosphates using immobilized phytase if markets develop for those products. Phytase (phyA) from A. niger NRRL 3135 has been cloned into an A. niger glucoamylase producing strain CBS 513.88 using a construct that has a glucoamylae promoter and an A. niger NRRL 3135 leader sequence, and that is devoid of phosphate repression. The yield of the secreted enzyme was increased 52-fold above that of wild-type A. niger NRRL 3135. The bioengineered organism produces 270 microns P/ml/min (4500 nKat/ml) which is approximately 7.9 g/liter in the medium. The yield of the secreted enzyme was

  5. Identification of novel phytase genes from an agricultural soil-derived metagenome.

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    Tan, Hao; Mooij, Marlies J; Barret, Matthieu; Hegarty, Pardraig M; Harington, Catriona; Dobson, Alan D W; O'Gara, Fergal

    2014-01-01

    Environmental microorganisms are emerging as an important source of new enzymes for wide-scale industrial application. In this study, novel phytase genes were identified from a soil microbial community. For this, a function-based screening approach was utilized for the identification of phytase activity in a metagenomic library derived from an agricultural soil. Two novel phytases were identified. Interestingly, one of these phytases is an unusual histidine acid phosphatase family phytase, as the conserved motif of the active site of PhyX possesses an additional amino acid residue. The second phytase belongs to a new type, which is encoded by multiple open reading frames (ORFs) and is different to all phytases known to date, which are encoded by a single ORF.

  6. Expression of Bacillus Species Isolated Phytase Gene from Soil by PCR Method

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    Nazemi, A. (PhD

    2014-06-01

    Full Text Available Background and Objective: Recognizing and using of isolated phytase in the soil microorganisms are paramount importance to produce the Phytase enzyme utilized commercially in different industries. This study was conducted to recognize different bacillus species which are Phytase producers and detection of the gene that can produce this enzyme. Material and Methods: Soil samples were gathered through different parts of mountainous areas. The early isolation of bacillus was carried out in Bacillus Medium Agar. After isolating the bacteria and genome extraction, the responsible gene of enzyme producer recognized and amplified by PCR method. The size of this protein and the optimal production situation in supplemental exploitation such as SDS-PAGE and the enzymatic activity of its size were evaluated. Results: Of 40 samples, one bacterium secreting Phytase enzyme was isolated. This bacterium was sequenced and recognized Bacillus Sobtlis species that is classified in STR Genus. The size of protein phytase produced by this gene was about 45 KD and the enzyme activity at 55 degrees was measured about 5.65 in wavelength of 415 NM. The phytase gene with the size of 1200 bp was propagated. Conclusion: the microorganisms, in natural conditions, produce Phytase enzyme in limited amount and with the quality appropriate to microorganisms. Thus, isolating the bacilli producing Phytase enzyme and purifying this protein are highly significant. Key words: Bacillus Subtilis; Phytase; SDS-PAGE; Enzymatic Activity; Polymerization Chain Reaction

  7. Heterologous Expression of Pantoea Agglomerans Phytase Gene Optimized for Plant-Host Expression

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    N.N. Khabipova

    2016-04-01

    Full Text Available Here we report expression and characterization of recombinant bacterial phytase PaPhyC from Pantoea sp. Codon-optimized phytase gene was expressed E.coli BL21 pLysS and protein expression was confirmed by Western blotting. Recombinant protein expressed in E.coli has high phytase activity. We show that PaPhyC recombinant phytase has different molecular masses when expressed in bacteria and plants, suggesting that possible protein glycosylation in plants may influence its overall size.

  8. Recombinant HAP Phytase of the Thermophilic Mold Sporotrichum thermophile: Expression of the Codon-Optimized Phytase Gene in Pichia pastoris and Applications.

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    Ranjan, Bibhuti; Satyanarayana, T

    2016-02-01

    The codon-optimized phytase gene of the thermophilic mold Sporotrichum thermophile (St-Phy) was expressed in Pichia pastoris. The recombinant P. pastoris harboring the phytase gene (rSt-Phy) yielded a high titer of extracellular phytase (480 ± 23 U/mL) on induction with methanol. The recombinant phytase production was ~40-fold higher than that of the native fungal strain. The purified recombinant phytase (rSt-Phy) has the molecular mass of 70 kDa on SDS-PAGE, with K m and V max (calcium phytate), k cat and k cat/K m values of 0.147 mM and 183 nmol/mg s, 1.3 × 10(3)/s and 8.84 × 10(6)/M s, respectively. Mg(2+) and Ba(2+) display a slight stimulatory effect, while other cations tested exert inhibitory action on phytase. The enzyme is inhibited by chaotropic agents (guanidinium hydrochloride, potassium iodide, and urea), Woodward's reagent K and 2,3-bunatedione, but resistant to both pepsin and trypsin. The rSt-Phy is useful in the dephytinization of broiler feeds efficiently in simulated gut conditions of chick leading to the liberation of soluble inorganic phosphate with concomitant mitigation in antinutrient effects of phytates. The addition of vanadate makes it a potential candidate for generating haloperoxidase, which has several applications.

  9. Expression of an Aspergillus niger Phytase Gene (phyA) in Saccharomyces cerevisiae

    OpenAIRE

    Han, Yanming; Wilson, David B.; Lei, Xin Gen

    1999-01-01

    Phytase improves the bioavailability of phytate phosphorus in plant foods to humans and animals and reduces phosphorus pollution of animal waste. Our objectives were to express an Aspergillus niger phytase gene (phyA) in Saccharomyces cerevisiae and to determine the effects of glycosylation on the phytase’s activity and thermostability. A 1.4-kb DNA fragment containing the coding region of the phyA gene was inserted into the expression vector pYES2 and was expressed in S. cerevisiae as an act...

  10. Characterization and expression of codon optimized soybean phytase gene in E. coli.

    Science.gov (United States)

    Singh, Pritee; Punjabi, Mansi; Jolly, Monica; Rai, R D; Sachdev, Archana

    2013-12-01

    Phytic acid, the major storage form of phosphorus in plant seeds is degraded by the phytases to yield inositol and free phosphate, contributing thereby to the improved bioavailability of phytate phosphorus and essential minerals in plant foods and simultaneous reduction in phosphorus pollution of the terrestrial and aquatic ecosystems. As a possible strategy for altering seed phytate levels, the approach involving reduction of phytate content by ectopically expressing endogenous phytase gene during seed development of soybean (Glycine max L. cv. Pusa-20) was attempted in the present study. Semi-quantitative RT-PCR revealed the maximum expression of phytase gene transcripts in germinating cotyledons (approximately 10 days after germinations), compared to other vegetative tissues. A full-length phytase cDNA was amplified from the germinating seedlings by splicing by overlap extension (SOE)-PCR and its sequence analysis revealed an open-reading-frame of 1644 bp, including an N terminal signal peptide of 28 amino acids. Predicted amino acid sequence (547-aa) of molecular mass 62 kDa on alignment with related purple acid phosphatases in other plants shared five conserved domains and seven invariant amino acids involved in coordination of the metals in the binuclear center of purple acid phosphatases. Owing to a large number of E. coli low-usage codons in soybean phytase gene, the modified gene was cloned into a prokaryotic expression vector pET-28a (+) and its expression in E. coli was confirmed by SDS-PAGE and Western blot analysis. Bioassay of the crude expression product in E. coli revealed a functional phytase gene, showing a great potential for developing low phytate transgenic soybean through its seed-specific overexpression in the early stages of seed development.

  11. Gene cloning and soluble expression of Aspergillus niger phytase in E. coli cytosol via chaperone co-expression.

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    Ushasree, Mrudula Vasudevan; Vidya, Jalaja; Pandey, Ashok

    2014-01-01

    A phytase gene from Aspergillus niger was isolated and two Escherichia coli expression systems, based on T7 RNA polymerase promoter and tac promoter, were used for its recombinant expression. Co-expression of molecular chaperone, GroES/EL, aided functional cytosolic expression of the phytase in E. coli BL21 (DE3). Untagged and maltose-binding protein-tagged recombinant phytase showed an activity band of ~49 and 92 kDa, respectively, on a zymogram. Heterologously-expressed phytase was fractionated from endogenous E. coli phytase by (NH4)2SO4 precipitation. The enzyme had optimum activity at 50 °C and pH 6.5.

  12. Isolation and characterization of a novel phytase gene (Sphyl) from soybean (Glycine max (L.) Merr.)

    Institute of Scientific and Technical Information of China (English)

    Li GUO; Jiaojiao WANG; Kai XIAO

    2008-01-01

    A novel phytase gene Sphyl was isolated based on screening a cDNA library which was constructed from germinated soybean (Glycine max (L.) Merr. cv. Kefeng 6) cotyledon. The full-length cDNA of Sphyl was 1644 bp predicated to encode 547 amino acids including an N terminal signal peptide of 27 amino acids. Phylogenetic analysis indi-cated that Sphyl had high similarities with the phytase genes from Medicago truncatula and rice, and acid phosphatase genes from M. truncatula and Arabidopsis. Prokaryotic expression of Sphyl in Escherichia coli BL21 (DE3) showed that the induced protein had high phytase activities. The tran-scripts of Sphyl could be detected in various tissues, such as cotyledons, leaves, stems and roots of seedlings. The phytase activities in the above tissues were in accordance with their corresponding Sphyl transcripts. The transcripts of Sphyl in cotyledons showed an increasing trend from 5 to 30 days after germination, suggesting that Sphyl had involved the hydrolyses of the organic phosphorus compounds in seeds from the stages of early seed germination to young seedlings in soybean. Therefore, it was speculated that Sphyl played an important role during the seed germination and the growth of the seedlings by releasing inorganic phosphorus (P) from phosphorus reserve in seeds.

  13. Ectopic Expression of a Phytase Gene from Medicago truncatula Barrel Medic Enhances Phosphorus Absorption in Plants

    Institute of Scientific and Technical Information of China (English)

    Kai Xiao; Jian-Heng Zhang; Maria Harrison; Zeng-Yu Wang

    2006-01-01

    In the present study, the phosphorus-absorption capacity of transgenic Arabidopsis plants ectopically expressing a novel phytase gene from Medicago truncatula Barrel Medic was evaluated. A full-length cDNA encoding an extracellular form of phytase was isolated from the model legume M. truncatula. The phytase gene (MtPHY1) has an open reading frame of 1 632 bp predicted to encode 543 amino acids, including an Nterminal signal peptide of 27 amino acids. The genomic sequence of the MtPHY1 gene is 5 151 bp, containing seven exons interrupted by six introns. Under high-Pi (2 mmol/L) growth conditions, higher levels of MtPHY1transcripts accumulated in the leaf and stem than in the root. The transcript level was reduced in the stem and increased in the root, with no obvious changes in the hybridization signal detected in the leaf under lowPi (10 μmol/L) conditions. Chimeric transgenes were constructed by placing MtPHY1 under the control of the constitutive CaMV35S promoter and the root-specific MtPT1 promoter. Phytase activities in root apoplast of transgenic Arabidopsis were 12.3- to 16.2-fold of that in control plants. The phytase expressed was secreted into the rhizosphere, as demonstrated by HPLC analysis of phytate degradation by root exudates. Ectopic expression of MtPHY1 in Arabidopsis, leading to significant improvement in organic phosphorus absorption and plant growth, indicated that MtPHY1 has great potential for improving plant phosphorus absorption and phytoremediation.

  14. Evidence for the negative regulation of phytase gene expression in Streptomyces lividans and Streptomyces coelicolor.

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    Boukhris, Ines; Dulermo, Thierry; Chouayekh, Hichem; Virolle, Marie-Joëlle

    2016-01-01

    Sco7697, a gene encoding a phytase, enzyme able to degrade phytate (myo-inositol 1,2,3,4,5,6-hexakis phosphate), the most abundant phosphorus storing compound in plants is present in the genome of S. coelicolor, a soil born bacteria with a saprophytic lifestyle. The expression of this gene was previously shown to be induced in conditions of Pi limitation by the response regulator PhoP binding to an operator sequence, the PHO box, located upstream of the -35 promoter sequence. A close examination of the promoter region of sco7697 revealed the presence of another putative operator site, a Direct Repeat (DR), located downstream of the -10 promoter sequence. In order to determine whether this DR played a role in regulation of sco7697 expression, different variants of the phytase gene promoter region were transcriptionally fused to the ß-glucuronidase reporter gene (GUS). As expected, deletion of the PHO box led to abolition of sco7697 induction in conditions of Pi limitation. Interestingly, alteration of the DR correlated with a dramatic increase of GUS expression but only when PhoP was present. These results demonstrated that this DR is the site of strong negative regulation by an unknown repressor. The latter would impede the necessary activation of phytase expression by PhoP.

  15. Molecular Cloning of Phytase Gene from ASUIA279 and Its Expression in Pichia pastoris System

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    M. H. Mahamad Maifiah

    2011-12-01

    Full Text Available Phytases catalyze the hydrolysis of phytate (myo-inositol hexakisphosphate, one of the major storage form of phosphate in plants, with subsequent release of myo-inositol, phosphate and phytate-bound minerals. Non-ruminant animals such as chicken, swine and fish can't use the organic phosphorus and minerals from their diet because there is no phytase activity in their digestive tract. Phytate degrading enzyme is added to the animal feed diet to improve phosphorus availability from the dietary phytate and at the same time this lessen the phosphate pollution level in areas of intensive animal production as the phosphate would not be excreted out to the environment. ASUIA279, a bacterial strain isolated from Malaysian soil has potentially shown good phytase activity. In the present work, the gene encoding for phytase has been amplified from the plasmid DNA of recombinant ASUIA279(5 obtained from a previous study (unpublished data by a polymerase chain reaction (PCR methodology. The amplified phytase gene was extracted, purified then cloned into the pPICZA plasmid and transformed into Pichia pastorisX-33 strain for enzyme production.ABSTRAK: Fitase pemangkinan hidrolisis fitat (myo-inositol hexakisphosphate, merupakan salah satu cara penyimpanan utama fosfat dalam tumbuhan, dengan pelepasan berturut myo-inositol , fosfat dan galian terikat fitat. Haiwan bukan ruminan seperti ayam, khinzir dan ikan tidak dapat memanfaatkan fosforus organik serta galian yang diperolehi daripada makanan kerana tidak mempunyai aktiviti fitase di dalam saluran pencernaan mereka. Enzim pengecilan fitase dicampurkan ke dalam pemakanan haiwan untuk mempertingkatkan keperolehan fosforus dari fitat diet. Pada masa yang sama ia dapat mengurangkan tahap pencemaran fosfat di kawasan yang terdapat penternakan haiwan secara intensif agar fosfat tidak dikumuhkan ke persekitaran. ASUIA279, satu strain bakteria yang diasingkan daripada tanih di Malaysiamenunjukkan aktiviti fitase

  16. Expression of Aspergillus nidulans phy Gene in Nicotiana benthamiana Produces Active Phytase with Broad Specificities

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    Tae-Kyun Oh

    2014-09-01

    Full Text Available A full-length phytase gene (phy of Aspergillus nidulans was amplified from the cDNA library by polymerase chain reaction (PCR, and it was introduced into a bacterial expression vector, pET-28a. The recombinant protein (rPhy-E, 56 kDa was overexpressed in the insoluble fraction of Escherichia coli culture, purified by Ni-NTA resin under denaturing conditions and injected into rats as an immunogen. To express A. nidulans phytase in a plant, the full-length of phy was cloned into a plant expression binary vector, pPZP212. The resultant construct was tested for its transient expression by Agrobacterium-infiltration into Nicotiana benthamiana leaves. Compared with a control, the agro-infiltrated leaf tissues showed the presence of phy mRNA and its high expression level in N. benthamiana. The recombinant phytase (rPhy-P, 62 kDa was strongly reacted with the polyclonal antibody against the nonglycosylated rPhy-E. The rPhy-P showed glycosylation, two pH optima (pH 4.5 and pH 5.5, an optimum temperature at 45~55 °C, thermostability and broad substrate specificities. After deglycosylation by peptide-N-glycosidase F (PNGase-F, the rPhy-P significantly lost the phytase activity and retained 1/9 of the original activity after 10 min of incubation at 45 °C. Therefore, the deglycosylation caused a significant reduction in enzyme thermostability. In animal experiments, oral administration of the rPhy-P at 1500 U/kg body weight/day for seven days caused a significant reduction of phosphorus excretion by 16% in rat feces. Besides, the rPhy-P did not result in any toxicological changes and clinical signs.

  17. Expression of Aspergillus nidulans phy gene in Nicotiana benthamiana produces active phytase with broad specificities.

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    Oh, Tae-Kyun; Oh, Sung; Kim, Seongdae; Park, Jae Sung; Vinod, Nagarajan; Jang, Kyung Min; Kim, Sei Chang; Choi, Chang Won; Ko, Suk-Min; Jeong, Dong Kee; Udayakumar, Rajangam

    2014-09-03

    A full-length phytase gene (phy) of Aspergillus nidulans was amplified from the cDNA library by polymerase chain reaction (PCR), and it was introduced into a bacterial expression vector, pET-28a. The recombinant protein (rPhy-E, 56 kDa) was overexpressed in the insoluble fraction of Escherichia coli culture, purified by Ni-NTA resin under denaturing conditions and injected into rats as an immunogen. To express A. nidulans phytase in a plant, the full-length of phy was cloned into a plant expression binary vector, pPZP212. The resultant construct was tested for its transient expression by Agrobacterium-infiltration into Nicotiana benthamiana leaves. Compared with a control, the agro-infiltrated leaf tissues showed the presence of phy mRNA and its high expression level in N. benthamiana. The recombinant phytase (rPhy-P, 62 kDa) was strongly reacted with the polyclonal antibody against the nonglycosylated rPhy-E. The rPhy-P showed glycosylation, two pH optima (pH 4.5 and pH 5.5), an optimum temperature at 45~55 °C, thermostability and broad substrate specificities. After deglycosylation by peptide-N-glycosidase F (PNGase-F), the rPhy-P significantly lost the phytase activity and retained 1/9 of the original activity after 10 min of incubation at 45 °C. Therefore, the deglycosylation caused a significant reduction in enzyme thermostability. In animal experiments, oral administration of the rPhy-P at 1500 U/kg body weight/day for seven days caused a significant reduction of phosphorus excretion by 16% in rat feces. Besides, the rPhy-P did not result in any toxicological changes and clinical signs.

  18. A phytase gene is overexpressed in root nodules cortex of Phaseolus vulgaris-rhizobia symbiosis under phosphorus deficiency.

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    Lazali, Mohamed; Zaman-Allah, Mainassara; Amenc, Laurie; Ounane, Ghania; Abadie, Josiane; Drevon, Jean-Jacques

    2013-08-01

    Phosphorus is an essential nutrient for rhizobial symbioses to convert N2 into NH4 usable for N nutrition in legumes and N cycle in ecosystems. This N2 fixation process occurs in nodules with a high energy cost. Phytate is the major storage form of P and accounts for more than 50 % of the total P in seeds of cereals and legumes. The phytases, a group of enzymes widely distributed in plant and microorganisms, are able to hydrolyze a variety of inositol phosphates. Recently, phytase activity was discovered in nodules. However, the gene expression localization and its role in N2-fixing nodules are still unknown. In this work, two recombinant inbred lines (RILs) of common bean (Phaseolus vulgaris L.), selected as contrasting for N2 fixation under P deficiency, namely RILs 115 (P-efficient) and 147 (P-inefficient) were inoculated with Rhizobium tropici CIAT 899, and grown under hydroaeroponic conditions with sufficient versus deficient P supply. With in situ RT-PCR methodology, we found that phytase transcripts were particularly abundant in the nodule cortex and infected zone of both RILs. Under P deficiency, phytase transcripts were significantly more abundant for RIL115 than for RIL147, and more in the outer cortex than in the infected zone. Additionally, the high expression of phytase among nodule tissues for the P-deficient RIL115 was associated with an increase in phytase (33 %) and phosphatase (49 %) activities and efficiency in use of the rhizobial symbiosis (34 %). It is argued that phytase activity in nodules would contribute to the adaptation of the rhizobia-legume symbiosis to low-P environments.

  19. High mature grain phytase activity in the Triticeae has evolved by duplication followed by neofunctionalization of the purple acid phosphatase phytase (PAPhy) gene

    DEFF Research Database (Denmark)

    Madsen, Claus Krogh; Dionisio, Giuseppe; Holme, Inger

    2013-01-01

    The phytase activity in food and feedstuffs is an important nutritional parameter. Members of the Triticeae tribe accumulate purple acid phosphatase phytases (PAPhy) during grain filling. This accumulation elevates mature grain phytase activities (MGPA) up to levels between ~650 FTU/kg for barley...

  20. Mutations in the beta propeller WDR72 cause autosomal-recessive hypomaturation amelogenesis imperfecta.

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    El-Sayed, Walid; Parry, David A; Shore, Roger C; Ahmed, Mushtaq; Jafri, Hussain; Rashid, Yasmin; Al-Bahlani, Suhaila; Al Harasi, Sharifa; Kirkham, Jennifer; Inglehearn, Chris F; Mighell, Alan J

    2009-11-01

    Healthy dental enamel is the hardest and most highly mineralized human tissue. Though acellular, nonvital, and without capacity for turnover or repair, it can nevertheless last a lifetime. Amelogenesis imperfecta (AI) is a collective term for failure of normal enamel development, covering diverse clinical phenotypes that typically show Mendelian inheritance patterns. One subset, known as hypomaturation AI, is characterised by near-normal volumes of organic enamel matrix but with weak, creamy-brown opaque enamel that fails prematurely after tooth eruption. Mutations in genes critical to enamel matrix formation have been documented, but current understanding of other key events in enamel biomineralization is limited. We investigated autosomal-recessive hypomaturation AI in a consanguineous Pakistani family. A whole-genome SNP autozygosity screen identified a locus on chromosome 15q21.3. Sequencing candidate genes revealed a point mutation in the poorly characterized WDR72 gene. Screening of WDR72 in a panel of nine additional hypomaturation AI families revealed the same mutation in a second, apparently unrelated, Pakistani family and two further nonsense mutations in Omani families. Immunohistochemistry confirmed intracellular localization in maturation-stage ameloblasts. WDR72 function is unknown, but as a putative beta propeller is expected to be a scaffold for protein-protein interactions. The nearest homolog, WDR7, is involved in vesicle mobilization and Ca2+-dependent exocytosis at synapses. Vesicle trafficking is important in maturation-stage ameloblasts with respect to secretion into immature enamel and removal of cleaved enamel matrix proteins via endocytosis. This raises the intriguing possibility that WDR72 is critical to ameloblast vesicle turnover during enamel maturation.

  1. Histomorphology and small intestinal sodium-dependent glucose transporter 1 gene expression in piglets fed phytic acid and phytase-supplemented diets.

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    Woyengo, T A; Rodriguez-Lecompte, J C; Adeola, O; Nyachoti, C M

    2011-08-01

    An experiment was conducted to determine the effect of dietary phytic acid (PA) and phytase supplementation on small intestinal histomorphology and Na-dependent glucose transporter 1 (SGLT1) gene expression in piglets. Twenty-four piglets with an average initial BW of 7.60 ± 0.73 kg were randomly assigned to 3 experimental diets, to give 8 piglets per diet. The diets were a casein-cornstarch-based diet that was supplemented with 0 or 2% PA, or 2% PA (as Na phytate) plus an Escherichia coli-derived phytase at 500 phytase units/kg. The basal diet was formulated to meet the 1998 NRC energy, digestible AA, mineral, and vitamin requirements for piglets. After 10 d of feeding, the piglets were killed to determine small intestinal histomorphology and small intestinal SGLT1 gene expression. Phytic acid supplementation did not affect (P > 0.1) villus height (VH) and the VH-to-crypt depth (CD) ratio, but did decrease (P 0.1) VH, CD, and the VH-to-CD ratio. Phytic acid supplementation reduced SGLT1 gene expression in the duodenum, jejunum, and ileum by 1.1-, 5.4-, and 2.4-fold, respectively. Phytase supplementation increased SGLT1 gene expression in the jejunum by 2.6-fold, but reduced SGLT1 gene expression in the duodenum and ileum by 2.0- and 4.0-fold, respectively. In conclusion, PA reduced CD in the jejunum and SGLT1 gene expression in the duodenum, jejunum, and ileum, whereas phytase supplementation increased the expression of SGLT1 in the jejunum. The reduced SGLT1 gene expression by PA implies that PA reduces nutrient utilization in pigs partly through reduced expression of SGLT1, which is involved in glucose and Na absorption. The increased expression of SGLT1 in the jejunum by phytase supplementation implies that phytase alleviated the negative effects of PA partly through increased expression of SGLT1.

  2. Characterization, gene cloning, and sequencing of a fungal phytase, PhyA, from Penicillium oxalicum PJ3.

    Science.gov (United States)

    Lee, Seung Ho; Cho, Jaiesoon; Bok, Jinduck; Kang, Seungha; Choi, Yunjaie; Lee, Peter C W

    2015-01-01

    A phytase from Penicillium oxalicum PJ3, PhyA, was purified near to homogeneity with 427-fold increase in specific phytase activity by ammonium sulfate precipitation, gel filtration, and ion-exchange chromatographies. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and zymogram analysis of the purified enzyme indicated an estimated molecular mass of 65 kD. The optimal pH and temperature of the purified enzyme were pH 4.5 and 55°C, respectively. The enzyme activity was strongly inhibited by Ca(2+), Cu(2+), Zn(2+), and phenylmethylsulfonyl fluoride (PMSF). The Km value for sodium phytate was 0.545 mM with a Vmax of 600 U/mg of protein. The phyA gene was cloned, and it contains an open reading frame of 1,383 with a single intron (118 bp), and encodes a protein of 461 amino acids.

  3. Improvement of organic phosphate acquisition in transgenic tobacco plants by overexpression of a soybean phytase gene Sphyl

    Institute of Scientific and Technical Information of China (English)

    Li GUO; Yuxin ZHAO; Shuhua ZHANG; Haina ZHANG; Kai XIAO

    2009-01-01

    Due to the huge amount in the soil, phytate is an important potential source for providing the plants with available phosphorus (Pi) by the involved catalytic reaction of phytase. In this study, a construct fusing the open reading frame (ORF) of Sphyl into corresponding positions in the fragment of binary expression vector pBI121 was created and used to transform tobacco. Molecular identification by PCR and RT-PCR indicated the target gene Sphyl in the transgenic tobacco plants was transcribed under the regulation of an upstream promoter. Compared with the control plants, the phytase activities in all the transgenic plants were increased, with the increased range consistent with the expression levels in the transgenic plants. Under the growth conditions with phytate as the sole phosphorus source, the transgenic line 1 plants displayed a high expression level of Sphyl and shows notable improved growth performance, such as higher fresh weight and dry weight, as well as higher total P content and more accumulative P amount per plant than CK. This clearly indicated that overexpression of Sphyl could improve the phosphorus acquisition by the extruded Sphyl phytase in the rhizosphere, where this enzyme could catalyze the degradation of the phytate and release the available Pi for plants. The Sphyl gene seemed to have a potential value in the creation of new crop cultivars with high phosphorus use efficiency.

  4. Design and Expression of a Synthetic phyC Gene Encoding the Neutral Phytase in Pichia pastoris

    Institute of Scientific and Technical Information of China (English)

    Li-Kou ZOU; Hong-Ning WANG; Xin PAN; Tao XIE; Qi WU; Zi-Wen XIE; Wan-Rong ZHOU

    2006-01-01

    The 1074-bp phyCs gene (optimized phyC gene) encoding neutral phytase was designed and synthesized according to the methylotrophic yeast Pichia pastoris codon usage bias without altering the protein sequence. The expression vector, pP9K-phyCs, was linearized and transformed in P. pastoris. The yield of total extracellular phytase activity was 17.6 U/ml induced in Buffered Methanol-complex Medium (BMMY) and 18.5 U/ml in Wheat Bran Extract Induction (WBEI) medium at the flask scale, respectively,improving over 90 folds compared with the wild-type isolate. Purified enzyme showed temperature optimum of 70℃ and pH optimum of 7.5. The enzyme activity retained 97% of the relative activity after incubation at 80℃ for 5 min. Because of the heavy glycosylation the expressed phytase had a molecular size of approximately 51 kDa. After deglycosylation by endoglycosylase H (EndoHf), the enzyme had an apparent molecular size of 42 kDa. Its property and thermostability was affected by the glycosylation.

  5. Gene expression profiling in hepatic tissue of newly weaned pigs fed pharmacological zinc and phytase supplemented diets

    Directory of Open Access Journals (Sweden)

    Link Jane E

    2008-09-01

    Full Text Available Abstract Background Zinc (Zn is an essential trace element. However, Zn bioavailability from commonly consumed plants may be reduced due to phytic acid. Zn supplementation has been used to treat diarrheal disease in children, and in the U.S. swine industry at pharmacological levels to promote growth and fecal consistency, but underlying mechanisms explaining these beneficial effects remain unknown. Moreover, adding supplemental phytase improves Zn bioavailability. Thus, we hypothesized that benefits of pharmacological Zn supplementation result from changes in gene expression that could be further affected by supplemental phytase. The goal of this study was to investigate the effects of feeding newly weaned pigs dietary Zn (150, 1,000, or 2,000 mg Zn/kg as Zn oxide with or without phytase [500 phytase units (FTU/kg] for 14 d on hepatic gene expression. Liver RNA from pigs fed 150, 1,000, or 2,000 mg Zn/kg, or 1,000 mg Zn/kg with phytase (n = 4 per treatment was reverse transcribed and examined using the differential display reverse transcription polymerase chain reaction technique. Liver RNA from pigs fed 150 or 2,000 mg Zn/kg (n = 4 per treatment was also evaluated using a 70-mer oligonucleotide microarray. Results Expressed sequence tags for 61 putatively differentially expressed transcripts were cloned and sequenced. In addition, interrogation of a 13,297 element oligonucleotide microarray revealed 650 annotated transcripts (FDR ≤ 0.05 affected by pharmacological Zn supplementation. Seven transcripts exhibiting differential expression in pigs fed pharmacological Zn with sequence similarities to genes encoding GLO1, PRDX4, ACY1, ORM1, CPB2, GSTM4, and HSP70.2 were selected for confirmation. Relative hepatic GLO1 (P PRDX4 (P ACY1 (P HSP70.2 (P Conclusion Results suggest that feeding pharmacological Zn (1,000 or 2,000 mg Zn/kg affects genes involved in reducing oxidative stress and in amino acid metabolism, which are essential for cell

  6. Phytase-producing capacity of yeasts isolated from traditional African fermented food products and PHYPk gene expression of Pichia kudriavzevii strains.

    Science.gov (United States)

    Greppi, Anna; Krych, Łukasz; Costantini, Antonella; Rantsiou, Kalliopi; Hounhouigan, D Joseph; Arneborg, Nils; Cocolin, Luca; Jespersen, Lene

    2015-07-16

    Phytate is known as a strong chelate of minerals causing their reduced uptake by the human intestine. Ninety-three yeast isolates from traditional African fermented food products, belonging to nine species (Pichia kudriavzevii, Saccharomyces cerevisiae, Clavispora lusitaniae, Kluyveromyces marxianus, Millerozyma farinosa, Candida glabrata, Wickerhamomyces anomalus, Hanseniaspora guilliermondii and Debaryomyces nepalensis) were screened for phytase production on solid and liquid media. 95% were able to grow in the presence of phytate as sole phosphate source, P. kudriavzevii being the best growing species. A phytase coding gene of P. kudriavzevii (PHYPk) was identified and its expression was studied during growth by RT-qPCR. The expression level of PHYPk was significantly higher in phytate-medium, compared to phosphate-medium. In phytate-medium expression was seen in the lag phase. Significant differences in gene expression were detected among the strains as well as between the media. A correlation was found between the PHYPk expression and phytase extracellular activity.

  7. Enhanced secretion of natto phytase by Bacillus subtilis.

    Science.gov (United States)

    Tsuji, Shogo; Tanaka, Kosei; Takenaka, Shinji; Yoshida, Ken-ichi

    2015-01-01

    Phytases comprise a group of phosphatases that trim inorganic phosphates from phytic acid (IP6). In this study, we aimed to achieve the efficient secretion of phytase by Bacillus subtilis. B. subtilis laboratory standard strain 168 and its derivatives exhibit no phytase activity, whereas a natto starter secretes phytase actively. The natto phytase gene was cloned into strain RIK1285, a protease-defective derivative of 168, to construct a random library of its N-terminal fusions with 173 different signal peptides (SPs) identified in the 168 genome. The library was screened to assess the efficiency of phytase secretion based on clear zones around colonies on plates, which appeared when IP6 was hydrolyzed. The pbp SP enhanced the secretion of the natto phytase most efficiently, i.e. twice that of the original SP. Thus, the secreted natto phytase was purified and found to remove up to 3 phosphates from IP6.

  8. Microbial diversity of mangrove sediment in Shenzhen Bay and gene cloning, characterization of an isolated phytase-producing strain of SPC09 B. cereus.

    Science.gov (United States)

    Zhang, Shengpeng; Liao, Shao-An; Yu, Xiaoyuan; Lu, Hongwu; Xian, Jian-An; Guo, Hui; Wang, Anli; Xie, Jian

    2015-06-01

    Phytases hydrolyze phytate to release inorganic phosphate, which decreases the requirement for phosphorus in fertilizers for crops and thus reduces environmental pollutants. This study analyzed microbial communities in rhizosphere sediment, collected in September 2012 from Shenzhen Bay, Guangdong, China, using high-throughput pyrosequencing; the results showed that the dominant taxonomic phyla were Chloroflexi, Firmicutes, and Proteobacteria, and the proportion of the beneficial bacteria, Bacillus, was 4.95 %. Twenty-nine culturable, phytase-producing bacteria were isolated, their phosphorus solubilization capacity was analyzed, and they were taxonomically characterized. Their phylogenetic placement was determined using 16S ribosomal RNA (rRNA) gene sequence analysis. The result shows that most of the isolates are members of the order Bacillales, although seven strains of Enterobacteriales, two strains of Pseudomonadales, and one strain of Oceanospirillales were also identified. The phytase gene was cloned from SPC09, Bacillus cereus, which showed the highest phosphorus solubilizing ability among the isolated strains. The gene encoded a primary translation product of 335 amino acids. A construct including the 1005-nt ORF fragment, Bc-phy, was transformed into Escherichia coli. The recombinant phytase was produced and purified, which revealed the temperature optima at 60 °C and pH optima at 6.5. The assessment by quantitative PCR (qPCR) showed an abundance of bacteria containing the Bc-phy gene; the level was generally higher in the mangrove forest than in the tidal flats and in surface soil compared to bottom soil, and the highest value was obtained in June. Herein, we report on the cloning, characterization, and activity of a novel phytase isolated from a mangrove system.

  9. Ensifer meliloti overexpressing Escherichia coli phytase gene ( appA) improves phosphorus (P) acquisition in maize plants

    Science.gov (United States)

    Sharma, Vikas; Kumar, Ajit; Archana, G.; Kumar, G. Naresh

    2016-10-01

    The Escherichia coli phytase gene appA encoding enzyme AppA was cloned in a broad host range plasmid pBBR1MCS2 ( lac promoter), termed pVA1, and transformed into the Ensifer meliloti 1020. Transformation of pVA1 in Ensifer meliloti { E. m (pVA1)} increased its phosphatase and phytase activity by ˜9- and ˜50-fold, respectively, compared to the transformants containing empty plasmid as control { E. m (pBBR1MCS2)}. The western blot experiments using rabbit anti-AppA antibody showed that AppA is translocated into the periplasm of the host after its expression. Ensifer meliloti harboring AppA protein { E. m (pVA1)} and { E. m (pBBR1MCS2)} could acidify the unbuffered phytate minimal media (pH 8.0) containing Ca-phytate or Na-phytate as sole organic P (Po) source to below pH 5.0 and released P. However, both { E. m (pVA1)} and { E. m (pBBR1MCS2)} neither dropped pH of the medium nor released P when the medium was buffered at pH 8.0 using Tris-Cl, indicating that acidification of medium was important for the enzymatic hydrolysis of phytate. Further experiments proved that maize plants inoculated with { E. m. (pVA1)} showed increase in growth under sterile semi solid agar (SSA) medium containing Na-phytate as sole P source. The present study could be helpful in generating better transgenic bioinoculants harboring phosphate mineralization properties that ultimately promote plant growth.

  10. Ensifer meliloti overexpressing Escherichia coli phytase gene (appA) improves phosphorus (P) acquisition in maize plants.

    Science.gov (United States)

    Sharma, Vikas; Kumar, Ajit; Archana, G; Kumar, G Naresh

    2016-10-01

    The Escherichia coli phytase gene appA encoding enzyme AppA was cloned in a broad host range plasmid pBBR1MCS2 (lac promoter), termed pVA1, and transformed into the Ensifer meliloti 1020. Transformation of pVA1 in Ensifer meliloti {E. m (pVA1)} increased its phosphatase and phytase activity by ∼9- and ∼50-fold, respectively, compared to the transformants containing empty plasmid as control {E. m (pBBR1MCS2)}. The western blot experiments using rabbit anti-AppA antibody showed that AppA is translocated into the periplasm of the host after its expression. Ensifer meliloti harboring AppA protein {E. m (pVA1)} and {E. m (pBBR1MCS2)} could acidify the unbuffered phytate minimal media (pH 8.0) containing Ca-phytate or Na-phytate as sole organic P (Po) source to below pH 5.0 and released P. However, both {E. m (pVA1)} and {E. m (pBBR1MCS2)} neither dropped pH of the medium nor released P when the medium was buffered at pH 8.0 using Tris-Cl, indicating that acidification of medium was important for the enzymatic hydrolysis of phytate. Further experiments proved that maize plants inoculated with {E. m. (pVA1)} showed increase in growth under sterile semi solid agar (SSA) medium containing Na-phytate as sole P source. The present study could be helpful in generating better transgenic bioinoculants harboring phosphate mineralization properties that ultimately promote plant growth.

  11. Animal feed compositions containing phytase derived from transgenic alfalfa and methods of use thereof

    Science.gov (United States)

    Austin-Phillips, Sandra; Koegel, Richard G.; Straub, Richard J.; Cook, Mark

    1999-01-01

    A value-added composition of matter containing plant matter from transgenic alfalfa which expresses exogenous phytase activity is disclosed. The phytase activity is a gene product of an exogenous gene encoding for phytase which has been stably incorporated into the genome of alfalfa plants. The transgenic alfalfa expresses phytase activity in nutritionally-significant amounts, thereby enabling its use in animal feeds to eliminate the need for phosphorous supplementation of livestock, poultry, and fish feed rations.

  12. Animal feed compositions containing phytase derived from transgenic alfalfa and methods of use thereof

    Energy Technology Data Exchange (ETDEWEB)

    Austin-Phillips, Sandra (Madison, WI); Koegel, Richard G. (Madison, WI); Straub, Richard J. (Brooklyn, WI); Cook, Mark (Madison, WI)

    2001-01-01

    A value-added composition of matter containing plant matter from transgenic alfalfa which expresses exogenous phytase activity is disclosed. The phytase activity is a gene product of an exogenous gene encoding for phytase which has been stably incorporated into the genome of alfalfa plants. The transgenic alfalfa expresses phytase activity in nutritionally-significant amounts, thereby enabling its use in animal feeds to eliminate the need for phosphorous supplementation of livestock, poultry, and fish feed rations.

  13. Overexpression of phyA and appA genes improves soil organic phosphorus utilisation and seed phytase activity in Brassica napus.

    Directory of Open Access Journals (Sweden)

    Yi Wang

    Full Text Available Phytate is the major storage form of organic phosphorus in soils and plant seeds, and phosphorus (P in this form is unavailable to plants or monogastric animals. In the present study, the phytase genes phyA and appA were introduced into Brassica napus cv Westar with a signal peptide sequence and CaMV 35S promoter, respectively. Three independent transgenic lines, P3 and P11 from phyA and a18 from appA, were selected. The three transgenic lines exhibited significantly higher exuded phytase activity when compared to wild-type (WT controls. A quartz sand culture experiment demonstrated that transgenic Brassica napus had significantly improved P uptake and plant biomass. A soil culture experiment revealed that seed yields of transgenic lines P11 and a18 increased by 20.9% and 59.9%, respectively, when compared to WT. When phytate was used as the sole P source, P accumulation in seeds increased by 20.6% and 46.9% with respect to WT in P11 and a18, respectively. The P3 line accumulated markedly more P in seeds than WT, while no significant difference was observed in seed yields when phytate was used as the sole P source. Phytase activities in transgenic canola seeds ranged from 1,138 to 1,605 U kg(-1 seeds, while no phytase activity was detected in WT seeds. Moreover, phytic acid content in P11 and a18 seeds was significantly lower than in WT. These results introduce an opportunity for improvement of soil and seed phytate-P bioavailability through genetic manipulation of oilseed rape, thereby increasing plant production and P nutrition for monogastric animals.

  14. Cloning and Expression of Phytase appA Gene from Shigella sp. CD2 in Pichia pastoris and Comparison of Properties with Recombinant Enzyme Expressed in E. coli.

    Directory of Open Access Journals (Sweden)

    Moushree Pal Roy

    Full Text Available The phytase gene appAS was isolated from Shigella sp. CD2 genomic library. The 3.8 kb DNA fragment contained 1299 bp open reading frame encoding 432 amino acid protein (AppAS with 22 amino acid signal peptide at N-terminal and three sites of N-glycosylation. AppAS contained the active site RHGXRXP and HDTN sequence motifs, which are conserved among histidine acid phosphatases. It showed maximum identity with phytase AppA of Escherichia coli and Citrobacter braakii. The appAS was expressed in Pichia pastoris and E. coli to produce recombinant phytase rAppAP and rAppAE, respectively. Purified glycosylated rAppAP and nonglycosylated rAppAE had specific activity of 967 and 2982 U mg(-1, respectively. Both had pH optima of 5.5 and temperature optima of 60°C. Compared with rAppAE, rAppAP was 13 and 17% less active at pH 3.5 and 7.5 and 11 and 18% less active at temperature 37 and 50°C, respectively; however, it was more active at higher incubation temperatures. Thermotolerance of rAppAP was 33% greater at 60°C and 24% greater at 70°C, when compared with rAppAE. Both the recombinant enzymes showed high specificity to phytate and resistance to trypsin. To our knowledge, this is the first report on cloning and expression of phytase from Shigella sp.

  15. Cloning and Expression of Phytase appA Gene from Shigella sp. CD2 in Pichia pastoris and Comparison of Properties with Recombinant Enzyme Expressed in E. coli.

    Science.gov (United States)

    Pal Roy, Moushree; Mazumdar, Deepika; Dutta, Subhabrata; Saha, Shyama Prasad; Ghosh, Shilpi

    2016-01-01

    The phytase gene appAS was isolated from Shigella sp. CD2 genomic library. The 3.8 kb DNA fragment contained 1299 bp open reading frame encoding 432 amino acid protein (AppAS) with 22 amino acid signal peptide at N-terminal and three sites of N-glycosylation. AppAS contained the active site RHGXRXP and HDTN sequence motifs, which are conserved among histidine acid phosphatases. It showed maximum identity with phytase AppA of Escherichia coli and Citrobacter braakii. The appAS was expressed in Pichia pastoris and E. coli to produce recombinant phytase rAppAP and rAppAE, respectively. Purified glycosylated rAppAP and nonglycosylated rAppAE had specific activity of 967 and 2982 U mg(-1), respectively. Both had pH optima of 5.5 and temperature optima of 60°C. Compared with rAppAE, rAppAP was 13 and 17% less active at pH 3.5 and 7.5 and 11 and 18% less active at temperature 37 and 50°C, respectively; however, it was more active at higher incubation temperatures. Thermotolerance of rAppAP was 33% greater at 60°C and 24% greater at 70°C, when compared with rAppAE. Both the recombinant enzymes showed high specificity to phytate and resistance to trypsin. To our knowledge, this is the first report on cloning and expression of phytase from Shigella sp.

  16. Phytase activity in lichens.

    Science.gov (United States)

    Higgins, Niall F; Crittenden, Peter D

    2015-10-01

    Phytase activity was investigated in 13 lichen species using a novel assay method. The work tested the hypothesis that phytase is a component of the suite of surface-bound lichen enzymes that hydrolyse simple organic forms of phosphorus (P) and nitrogen (N) deposited onto the thallus surface. Hydrolysis of inositol hexaphosphate (InsP6 , the substrate for phytase) and appearance of lower-order inositol phosphates (InsP5 -InsP1 ), the hydrolysis products, were measured by ion chromatography. Phytase activity in Evernia prunastri was compared among locations with contrasting rates of N deposition. Phytase activity was readily measurable in epiphytic lichens (e.g. 11.3 μmol InsP6 hydrolysed g(-1)  h(-1) in Bryoria fuscescens) but low in two terricolous species tested (Cladonia portentosa and Peltigera membranacea). Phytase and phosphomonoesterase activities were positively correlated amongst species. In E. prunastri both enzyme activities were promoted by N enrichment and phytase activity was readily released into thallus washings. InsP6 was not detected in tree canopy throughfall but was present in pollen leachate. Capacity to hydrolyse InsP6 appears widespread amongst lichens potentially promoting P capture from atmospheric deposits and plant leachates, and P cycling in forest canopies. The enzyme assay used here might find wider application in studies on plant root-fungal-soil systems.

  17. Cisgenic barley with improved phytase activity

    DEFF Research Database (Denmark)

    Holme, Inger; Dionisio, Giuseppe; Brinch-Pedersen, Henrik

    2012-01-01

    The cisgenesis concept implies that plants are transformed only with their own genetic materials or genetic materials from closely related species capable of sexual hybridization. Furthermore, foreign sequences such as selection genes and vector-backbone sequences should be absent. We used a barley...... phytase gene (HvPAPhy_a) expressed during grain filling to evaluate the cisgenesis concept in barley. The marker gene elimination method was used to obtain marker-free plant lines. Here, the gene of interest and the selection gene are flanked by their own T-DNA borders to allow unlinked integration...... of the two genes. We analysed the transformants for co-transformation efficiency, increased phytase activities in the grain, integration of the kanamycin resistance gene of the vector-backbone and segregation between the HvPAPhy_a insert and the hygromycin resistance gene. The frequencies of the four...

  18. Cloning, Overexpression, and Characterization of a Metagenome-Derived Phytase with Optimal Activity at Low pH.

    Science.gov (United States)

    Tan, Hao; Wu, Xiang; Xie, Liyuan; Huang, Zhongqian; Gan, Bingcheng; Peng, Weihong

    2015-06-01

    A phytase gene was identified in a publicly available metagenome derived from subsurface groundwater, which was deduced to encode for a protein of the histidine acid phosphatase (HAP) family. The nucleotide sequence of the phytase gene was chemically synthesized and cloned, in order to further overexpress the phytase in Escherichia coli. Purified protein of the recombinant phytase demonstrated an activity for phytic acid of 298 ± 17 μmol P/min/mg, at the pH optimum of 2.0 with the temperature of 37 °C. Interestingly, the pH optimum of this phytase is much lower in comparison with most HAP phytases known to date. It suggests that the phytase could possess improved adaptability to the low pH condition caused by the gastric acid in livestock and poultry stomachs.

  19. Expression, purification and characterization of a phyAm-phyCs fusion phytase

    Institute of Scientific and Technical Information of China (English)

    Li-kou ZOU; Hong-ning WANG; Xin PAN; Guo-bao TIAN; Zi-wen XIE; Qi WU; Hui CHEN; Tao XIE; Zhi-rong YANG

    2008-01-01

    The phyAm gene encoding acid phytase and optimized neutral phytase phyCs gene were inserted into expression vector pPIC9K in correct orientation and transformed into Pichiapastoris in order to expand the pH profile of phytase and decrease the cost of production. The fusion phytase phyAm-phyCs gene was successfully overexpressed in P. pastoris as an active and ex-tracellular phytase. The yield of total extracellular fusion phytase activity is (25.4±0.53) U/ml at the flask scale and (159.1±2.92) U/ml for high cell-density fermentation, respectively. Purified fusion phytase exhibits an optimal temperature at 55 ℃ and an optimal pH at 5.5~6.0 and its relative activity remains at a relatively high level of above 70% in the range of pH 2.0 to 7.0. About 51% to 63% of its original activity remains after incubation at 75 ℃ to 95 ℃ for 10 min. Due to heavy glycosylation, the expressed fusion phytase shows a broad and diffuse band in SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). After deglycosylation by endoglycosidase H (EndoHf), the enzyme has an apparent molecular size of 95 kDa. The characterization of the fusion phytase was compared with those of phyCs and phyAm.

  20. Citrobacter amalonaticus Phytase on the Cell Surface of Pichia pastoris Exhibits High pH Stability as a Promising Potential Feed Supplement

    OpenAIRE

    Cheng Li; Ying Lin; Yuanyuan Huang; Xiaoxiao Liu; Shuli Liang

    2014-01-01

    Phytase expressed and anchored on the cell surface of Pichia pastoris avoids the expensive and time-consuming steps of protein purification and separation. Furthermore, yeast cells with anchored phytase can be used as a whole-cell biocatalyst. In this study, the phytase gene of Citrobacter amalonaticus was fused with the Pichia pastoris glycosylphosphatidylinositol (GPI)-anchored glycoprotein homologue GCW61. Phytase exposed on the cell surface exhibits a high activity of 6413.5 U/g, with an ...

  1. Phytases for Improved Iron Absorption

    DEFF Research Database (Denmark)

    Nielsen, Anne Veller Friis; Nyffenegger, Christian; Meyer, Anne S.

    2014-01-01

    Microbial phytases (EC 3.1.3.8) catalyse dephosphorylation of phytic acid, which is the primary storage compound for phosphorous in cereal kernels. The negatively charged phosphates in phytic acid chelate iron (Fe3+) and thus retards iron bioavailability in humans 1. Supplementation of microbial...... phytase can improve iron absorption from cereal-based diets 2. In order for phytase to catalyse iron release in vivo the phytase must be robust to low pH and proteolysis in the gastric ventricle. Our work has compared the robustness of five different microbial phytases, evaluating thermal stability......, activity retention, and extent of dephosphorylation of phytic acid in a simulated low pH/pepsin gastric environment. The five phytases responded differently to the robustness parameters: The Peniophora lycii phytase (Ronozyme NP) was the most thermostable, but the least robust enzyme at low pH, whereas...

  2. Regulation of Soluble Phosphate on the Ability of Phytate Mineralization and β-Propeller Phytase Gene Expression of Pseudomonas fluorescens JZ-DZ1, a Phytate-Mineralizing Rhizobacterium.

    Science.gov (United States)

    Shen, Lan; Wu, Xiao-Qin; Zeng, Qing-Wei; Liu, Hong-Bin

    2016-12-01

    Phytate-mineralizing rhizobacteria (PMR) play an important role in providing phosphorus for the sustainable plant growth. It is important to investigate the ability of PMR to produce phytase under different phosphate levels for its application. The effects of different concentrations of soluble phosphate on the ability of phytate mineralization of Pseudomonas fluorescens JZ-DZ1, a phytate-mineralizing rhizobacterium, were investigated in both solid and liquid media. The results on solid media showed that halo zone width gradually reduced with concentrations of soluble phosphate increasing from 0.05 to 20 mM, indicating the reduction of the ability of phytate mineralization. The results were consistent with the quantitative detection of phytase activity from the overall trend. An 1866-bp β-propeller phytase (BPP) gene (phyPf) was cloned from the strain, and the deduced amino acid sequence of phyPf shared 98 % of identity with a known BPP from Pseudomonas sp. BS10-3 (AJF36073.1). The results of relative real-time quantitative PCR assay showed that the expression of phyPf was induced by a low concentration (0.1 mM) of soluble phosphate, suggesting that BPP secretion was regulated by gene phyPf. The BPP-harboring bacterium P. fluorescens JZ-DZ1 with low phosphate-inducible ability of phytate mineralization could be potentially applied to promote phosphorus uptake for plants in the future.

  3. [Occupational allergies to phytase].

    Science.gov (United States)

    van Kampen, V; Merget, R; Brüning, T

    2008-12-01

    Phytases are phosphatases that can break down the undigestible phytic acid (phytate). They are frequently used as an animal feed supplement - often in poultry and swine - to enhance the nutritive value of plant material by liberation of inorganic phosphate from phytate. Exposure to enzyme dusts has long been known to cause occupational allergies. The present paper reviews the results of the evaluation of literature data concerning occupational airway sensitization due to phytases. Cases of specific airway sensitization caused by phytases could be shown clearly by the presented studies. As symptoms, results of skin prick tests, detection of specific IgE-antibodies and results of specific challenge tests are consistent, an immunologic mechanism can be assumed.

  4. Cloning of a novel phytase from an anaerobic rumen bacterium, Mitsuokella jalaludinii, and its expression in Escherichia coli

    Institute of Scientific and Technical Information of China (English)

    TAN Wan-qin; Phang Chiun Yee; Sieo Chin Chin; Yiap Beow Chin; Clemente Michael Wong Vui Ling; Norhani Abdullah; Son Radu; Ho Yin Wan

    2015-01-01

    The ful length phytase gene of Mitsuokel a jalaludini was successful y cloned and was found to be 1 047 bp in length, with 348 amino acids, and was designated as PHY7 phytase gene. A comparison of the sequence of PHY7 phytase gene of M. jalaludini with various microbial phytase gene sequences showed that it was not similar to those from other bacteria except Selenomonas ruminatium, thus suggesting that they may both express a new class of phytase. The PHY7 phytase gene was subsequently subcloned into bacterial expression vector, pET32a, for expression in Escherichia coli strain Ro-setta-gami. Expression of the recombinant phytase gene was optimised and characterised. The recombinant phytase was estimated to be approximately 55 kDa by SDS-PAGE analysis. The recombinant phytase exhibited optimum activity at 55°C, pH 4.5 and showed good pH stability from pH 3.5 to 5.5 (>78%relative activity). Metal ions such as Ca2+, Mg2+, and K+were found to exert signiifcant stimulatory effect on the recombinant phytase activity while Cu2+, Fe3+, and Zn2+greatly inhibited the enzyme activity. The recombinant phytase showed moderate resistance to trypsin proteolysis, but susceptible to pepsin proteolysis. The results of the study showed that several characteristics of recombinant phytase were slightly different from the native enzyme. Unfavourable characteristics such as reduced pH stability and metal ion effects should be taken into consideration during feed enzyme formulation.

  5. Cloning of a novel phytase from an anaerobic rumen bacterium, Mitsuokella jalaludinii, and its expression in Escherichia coli

    Institute of Scientific and Technical Information of China (English)

    TAN Wan-qin[1; Phang Chiun Yee[1; Sieo Chin Chin[1,2; Yiap Beow Chin[3; Clemente Michael Wong VuiLing[4; Norhani Abdullah[1,5,6; Son Radu[7; Ho Yin Wan[1

    2015-01-01

    The full length phytase gene of Mitsuokellajalaludiniiwas successfully cloned and was found to be 1 047 bp in length, with 348 amino acids, and was designated as PHY7 phytase gene. A comparison of the sequence of PHY7 phytase gene of M. jalaludinii with various microbial phytase gene sequences showed that it was not similar to those from other bacteria except Selenomonas ruminafium, thus suggesting that they may both express a new class of phytase. The PHY7 phytase gene was subsequently subcloned into bacterial expression vector, pET32a, for expression in Escherichia coil strain Ro- setta-gami. Expression of the recombinant phytase gene was optimised and characterised. The recombinant phytase was estimated to be approximately 55 kDa by SDS-PAGE analysis. The recombinant phytase exhibited optimum activity at 55~C, pH 4.5 and showed good pH stability from pH 3.5 to 5.5 (>78% relative activity). Metal ions such as Caz+, Mg2+, and K+ were found to exert significant stimulatory effect on the recombinant phytase activity while Cu2+, Fe3+, and Zn2+ greatly inhibited the enzyme activity. The recombinant phytase showed moderate resistance to trypsin proteolysis, but susceptible to pepsin proteolysis. The results of the study showed that several characteristics of recombinant phytase were slightly different from the native enzyme. Unfavourable characteristics such as reduced pH stability and metal ion effects should be taken into consideration during feed enzyme formulation.

  6. Transgenic Expression of the Recombinant Phytase in Rice (Oryza sativa)

    Institute of Scientific and Technical Information of China (English)

    LIU Qiao-quan; LI Qian-feng; JIANG Li; ZHANG Da-jiang; WANG Hong-mei; GU Ming-hong; YAO Quan-hong

    2006-01-01

    In most of the cereal crop, phytic acid is the main storage form of phosphorus, which can decrease the bioavailability of phosphate. Transgenic expression of phytase is regarded as an efficient way to release phosphate from phytate in transgenic plants.In this study, a plant expression vector, containing the recombinant phytase gene driven by the maize ubiquitin (Ubi) promoter was constructed and introduced into an elite rice variety via Agrobacterium-mediated transformation. During the experiment, a total of 15 independent transgenic rice lines were regenerated. The results of PCR and Southern blot indicated that the target gene was integrated into the genome of transgenic rice plants. Moreover, the RT-PCR analysis of total RNAs extracted from the immature seeds of several transgenic lines showed that the recombinant phytase gene could be normally expressed. The inorganic phosphorus content, both in the mature seeds and the leaf was significantly higher in the transgenic plants than in the untransformed wild type.

  7. A novel phytase with sequence similarity to purple acid phosphatases is expressed in cotyledons of germinating soybean seedlings.

    Science.gov (United States)

    Hegeman, C E; Grabau, E A

    2001-08-01

    Phytic acid (myo-inositol hexakisphosphate) is the major storage form of phosphorus in plant seeds. During germination, stored reserves are used as a source of nutrients by the plant seedling. Phytic acid is degraded by the activity of phytases to yield inositol and free phosphate. Due to the lack of phytases in the non-ruminant digestive tract, monogastric animals cannot utilize dietary phytic acid and it is excreted into manure. High phytic acid content in manure results in elevated phosphorus levels in soil and water and accompanying environmental concerns. The use of phytases to degrade seed phytic acid has potential for reducing the negative environmental impact of livestock production. A phytase was purified to electrophoretic homogeneity from cotyledons of germinated soybeans (Glycine max L. Merr.). Peptide sequence data generated from the purified enzyme facilitated the cloning of the phytase sequence (GmPhy) employing a polymerase chain reaction strategy. The introduction of GmPhy into soybean tissue culture resulted in increased phytase activity in transformed cells, which confirmed the identity of the phytase gene. It is surprising that the soybean phytase was unrelated to previously characterized microbial or maize (Zea mays) phytases, which were classified as histidine acid phosphatases. The soybean phytase sequence exhibited a high degree of similarity to purple acid phosphatases, a class of metallophosphoesterases.

  8. Assessment of a bacterial 6-phytase in the diets of broiler chickens.

    Science.gov (United States)

    Olukosi, O A; Kong, C; Fru-Nji, F; Ajuwon, K M; Adeola, O

    2013-08-01

    Two 21-d broiler experiments were conducted to assess the efficacy of a bacterial 6-phytase expressed in Aspergillus oryzae on growth performance, nutrient utilization, and intestinal molecular markers. Two hundred forty birds in 5 treatments (experiment 1) or 256 birds in 4 treatments (experiment 2) were used. The treatments included a negative control diet that was marginally deficient in P (NC) or NC plus tricalcium phosphate, 500, 1,000, or 2,000 phytase units/kg (experiment 1), and NC or NC plus monocalcium phosphate, 500 or 1,000 phytase units/kg (experiment 2). In both experiments, excreta were collected on d 19 to 21, whereas birds and feed were weighed and ileal digesta collected on d 21. For experiment 1, mucosa scraping was collected from the duodenum, jejunum, and ileum from all birds for quantification of expression level of gut level inflammatory cytokines, Toll-like receptors, and phosphate transporter (NaPi-IIb). In both experiments, tricalcium phosphate, monocalcium phosphate, and phytase supplementation improved (P phytase supplementation (experiment 1) and there was an increase (P phytase supplementation (experiment 2). Diets did not affect the expression of gut level cytokines, Toll-like receptors, or the mucin gene. Phytase supplementation tended to decrease (P phytase was effective in enhancing growth of broilers receiving low-P diets as well as in increasing efficiency of P utilization and phytic acid degradation.

  9. Expression of phytase gene in prokaryotic cell and application progress of phytase%植酸酶基因在原核细胞中表达及植酸酶应用进展

    Institute of Scientific and Technical Information of China (English)

    卢珊; 张守纯; 朱航; 梁志刚

    2007-01-01

    @@ 植酸酶(Phytase)是催化植酸及其盐类物质水解成肌醇和磷酸的一类酶的总称,广泛存在于微生物、麦芽、种子等中[1].植酸酶可使单胃动物对饲料中磷的利用率提高60%,粪便中磷的排出量减少40%,另外还可降低植酸的抗营养作用.因此植酸酶在饲料工业中的应用对提高养殖业效益和降低环境污染有重要意义.生产植酸酶的表达系统包括真核表达系统和原核表达系统.真核表达系统主要有丝状真菌表达系统和酵母表达系统;原核表达系统主要是大肠杆菌表达系统.

  10. Cisgenic Barley with Improved Phytase Activity

    DEFF Research Database (Denmark)

    Holme, Inger; Dionisio, Giuseppe; Brinch-Pedersen, Henrik

    2010-01-01

    based on the cisgenesis concept. In contrast to transgenesis, cisgenesis implies that the plant is transformed only with its own or very closely related genetic material. Furthermore, all “helper” genes and gene sequences of foreign nature are removed from the transformed plant lines. Cisgenic crops...... are accordingly very similar to those generated by conventional breeding. The cisgenesis concept allows for the introduction of extra gene copies of a particular gene to accentuate the trait. We are using a barley purple acid phosphatase expressed during grain filling as candidate gene for cisgenesis. A genomic...... barley lambda library has been used to isolate the genomic clone of this phytase including 2.3 kb of the promoter region and 600 bp of the terminator region. The clone has been inserted into a cisgenic Agrobacterium vector where both the gene of interest and the selection gene are flanked by their own T...

  11. Cisgenic barley with improved phytase activity

    DEFF Research Database (Denmark)

    Holme, Inger; Dionisio, Giuseppe; Brinch-Pedersen, Henrik

    2010-01-01

    based on the cisgenesis concept. In contrast to transgenesis, cisgenesis implies that the plant is transformed only with its own or very closely related genetic material. Furthermore, all “helper” genes and gene sequences of foreign nature are removed from the transformed plant lines. Cisgenic crops...... are accordingly very similar to those generated by conventional breeding. The cisgenesis concept allows for the introduction of extra gene copies of a particular gene to accentuate the trait. We are using a barley purple acid phosphatase expressed during grain filling as candidate gene for cisgenesis. A genomic...... barley lambda library has been used to isolate the genomic clone of this phytase including 2.3 kb of the promoter region and 600 bp of the terminator region. The clone has been inserted into a cisgenic Agrobacterium vector where both the gene of interest and the selection gene are flanked by their own T...

  12. Isolation and molecular characterization of thermostable phytase from Bacillus subtilis (BSPhyARRMK33).

    Science.gov (United States)

    Reddy, Chinreddy Subramanyam; Achary, V Mohan Murali; Manna, Mrinalini; Singh, Jitender; Kaul, Tanushri; Reddy, Malireddy K

    2015-03-01

    The thermostable phytase gene was isolated from Bacillus subtilis ARRMK33 (BsPhyARRMK33). The gene has an ORF of 1152 bp and that encodes a protein of 383 amino acids. Sequence analysis showed high homology with Bacillus sp. phytase proteins, but no similarity was found with other phytases. SDS-PAGE analysis exhibited a predicted molecular mass of 42 kDa. Homology modeling of BsPhyARRMK33 protein based on Bacillus amyloliquefaciens crystal structure disclosed its β-propeller structure. BsPhyARRMK33 recombinant plasmid in pET-28a(+) was expressed in Rosetta gami B DE3 cells and the maximum phytase activity 15.3 U mg(-1) obtained. The enzyme exhibits high thermostability at various temperatures and broad pH ranges. The recombinant protein retained 74% of its original activity after incubation at 95 °C for 10 min. In the presence of Ca(2+), the recombinant phytase activity was maximal where as it was inhibited by EDTA. The optimal pH and temperature for the recombinant phytase activity is achieved at 7.0 and 55 °C, respectively. Thermostable nature and wide range of pH are promising features of recombinant BsPhyARRMK33 protein that may be employed as an efficient alternative to commercially known phytases and thereby alleviate environmental eutrophication.

  13. Microbial phytases in phosphorus acquisition and plant growth promotion.

    Science.gov (United States)

    Singh, Bijender; Satyanarayana, T

    2011-04-01

    Phosphorus (P) is one of the major constituents in energy metabolism and biosynthesis of nucleic acids and cell membranes with an important role in regulation of a number of enzymes. Soil phosphorous is an important macronutrient for plant growth. Phosphorus deficiency in soil is a major problem for agricultural production. Total soil P occurs in either organic or in organic form. Phytic acid as phytate (salts of phytic acid) is the major form of organic phosphorus in soil and it is not readily available to plants as a source of phosphorus because it either forms a complex with cations or adsorbs to various soil components. Phosphate solubilizing microorganisms are ubiquitous in soils and could play an important role in supplying P to plants. Microorganisms utilizing phytate are found in cultivated soils as well as in wetland, grassland and forest soils. Various fungi and bacteria (including plant growth promoting rhizobacteria) hydrolyze this organic form of phosphorus secreting phosphatases such as phytases and acidic/alkaline phosphatases. A large number of transgenic plants have been developed which were able to utilize sodium phytate as sole source of phosphorus. However, the recombinant phytases were similar to their wild type counterparts in terms of their properties. Increased phytase/phosphatase activity in transgenic plants may be an effective approach to promote their phytate-phosphorus utilization. The extracellular phytase activity of transgenic plant roots is a significant factor in the utilization of phosphorus from phytate. Furthermore, this indicated that an opportunity exists for using gene technology to improve the ability of plants to utilize accumulated forms of soil organic phosphorus. This review is focused on the role of phytases and phytase producing microbes in promoting the growth of different plants.

  14. 芽孢杆菌中性植酸酶基因的原核表达及酶学性质分析%Prokaryotic Expression and Enzymological Property Analysis of Neutral Phytase Gene from Bacillus sp.

    Institute of Scientific and Technical Information of China (English)

    李镇刚; 赵爱春; 王茜龄; 金筱耘; 李军; 余茂德

    2012-01-01

    When used as feedstuff additive, phytase can effectively improve the utilization of phosphorus in feedstuff, reduce environmental pollution caused by fecal phosphorus excretion, and reduce anti-nutritional effect of phytic acid. In order to obtain highly active phytase with stable performance, the mature peptide coding sequence of neutral phytase gene phyC (GenBank accession number; FJ986327) was amplified from Bacillus sp. By PCR and cloned into prokaryot-ic expression vector pET-28a( +). The recombinant plasmid was subsequently transformed into E. Coli BL21 (DE3) for expression. Large amount of inclusion body protein was obtained after induction with 0.5 mmol/L IPTG for 4 h at 37 ℃, and the soluble protein could be easily obtained by induction with 0. 5 mmol/L IPTG for 6 h at 25 ℃. After being purified with Ni-NTA affinity chromatography, enzymological properties of the recombinant phytase were assayed. The results showed that the obtained phytase had favorable heat tolerance. The optimum reaction temperature was 55 ℃, and more than 20% of enzyme activity was maintained after incubation at 70 ℃ for 10 min. The enzyme also had high resistance to acid and alkali. The optimum pH value was 6. 0 to 7. 0, and more than 80%, 70% and 40% of enzyme activity was maintained when incubated at pH 5. 5~9. 0, pH 5. 0 ~10. 0 and pH 2. 0~4. 0 for 60 min, respectively. Using the con-structed prokaryotic expression vector containing coding sequence of Bacillus neutral phytase gene phyC of which signal peptide sequence had been removed, neutral phytase with stable performance could be highly expressed in E. Coli under the optimized induced expression condition.%植酸酶作为饲料添加剂能够有效提高动物对饲料中磷的利用率及减少粪便中磷排放对环境的污染,并降低植酸的抗营养作用.为了获得性能稳定的高活性植酸酶,采用PCR扩增芽孢杆菌(Bacillus sp.)中性植酸酶基因phyC(GenBank登录号:FJ986327)的成熟肽

  15. Purification, biochemical characterization, and genetic cloning of the phytase produced by Burkholderia sp. strain a13.

    Science.gov (United States)

    Graminho, Eduardo Rezende; Takaya, Naoki; Nakamura, Akira; Hoshino, Takayuki

    2015-01-01

    A phytase-producing bacterium, Burkholderia sp. a13 (JCM 30421), was isolated from Lake Kasumigaura by enrichment cultivation using minimum medium containing phytic acid as the sole phosphorus source. The phytase production by strain a13 was induced by the presence of phytic acid and repressed by the addition of glucose. The purified enzyme had a molecular weight of 44 kDa and a phytase activity of 174 μmol min(-1) mg(-1). The enzyme showed broad substrate specificity, but the highest activity was observed with phytic acid. The enzyme activity was strongly inhibited by Cu(2+), Zn(2+), Hg(2+), and iodoacetic acid, indicating the requirement of a thiol group for the activity. Genetic cloning reveals that the mature portion of this enzyme consists of 428 amino acids with a calculated molecular weight of 46 kDa. The amino acid sequence showed the highest similarity to the phytase produced by Hafnia alvei with 48% identity; it also contained histidine acid phosphatase (HAP) motifs (RHGXRXP and HD), indicating the classification of this enzyme in the HAP phytase family. We have successfully expressed the cloned gene in Escherichia coli from its putative initiation codon, showing that the gene actually encodes the phytase.

  16. Phytase in non-ruminant animal nutrition: a critical review on phytase activities in the gastrointestinal tract and influencing factors

    OpenAIRE

    Dersjant-Li, Yueming; Awati, Ajay; Schulze, Hagen; Partridge, Gary

    2014-01-01

    This review focuses on phytase functionality in the digestive tract of farmed non-ruminant animals and the factors influencing in vivo phytase enzyme activity. In pigs, feed phytase is mainly active in the stomach and upper part of the small intestine, and added phytase activity is not recovered in the ileum. In poultry, feed phytase activities are mainly found in the upper part of the digestive tract, including the crop, proventriculus and gizzard. For fish with a stomach, phytase activities...

  17. Identification and characterization of a mesophilic phytase highly resilient to high-temperatures from a fungus-garden associated metagenome.

    Science.gov (United States)

    Tan, Hao; Wu, Xiang; Xie, Liyuan; Huang, Zhongqian; Peng, Weihong; Gan, Bingcheng

    2016-03-01

    Phytases are enzymes degrading phytic acid and thereby releasing inorganic phosphate. While the phytases reported to date are majorly from culturable microorganisms, the fast-growing quantity of publicly available metagenomic data generated in the last decade has enabled bioinformatic mining of phytases in numerous data mines derived from a variety of ecosystems throughout the world. In this study, we are interested in the histidine acid phosphatase (HAP) family phytases present in insect-cultivated fungus gardens. Using bioinformatic approaches, 11 putative HAP phytase genes were initially screened from 18 publicly available metagenomes of fungus gardens and were further overexpressed in Escherichia coli. One phytase from a south pine beetle fungus garden showed the highest activity and was then chosen for further study. Biochemical characterization showed that the phytase is mesophilic but possesses strong ability to withstand high temperatures. To our knowledge, it has the longest half-life time at 100 °C (27 min) and at 80 °C (2.1 h) as compared to all the thermostable phytases publicly reported to date. After 100 °C incubation for 15 min, more than 93 % of the activity was retained. The activity was 3102 μmol P/min/mg at 37 °C and 4135 μmol P/min/mg at 52.5 °C, which is higher than all the known thermostable phytases. For the high activity level demonstrated at mesophilic temperatures as well as the high resilience to high temperatures, the phytase might be promising for potential application as an additive enzyme in animal feed.

  18. Citrobacter amalonaticus phytase on the cell surface of Pichia pastoris exhibits high pH stability as a promising potential feed supplement.

    Science.gov (United States)

    Li, Cheng; Lin, Ying; Huang, Yuanyuan; Liu, Xiaoxiao; Liang, Shuli

    2014-01-01

    Phytase expressed and anchored on the cell surface of Pichia pastoris avoids the expensive and time-consuming steps of protein purification and separation. Furthermore, yeast cells with anchored phytase can be used as a whole-cell biocatalyst. In this study, the phytase gene of Citrobacter amalonaticus was fused with the Pichia pastoris glycosylphosphatidylinositol (GPI)-anchored glycoprotein homologue GCW61. Phytase exposed on the cell surface exhibits a high activity of 6413.5 U/g, with an optimal temperature of 60°C. In contrast to secreted phytase, which has an optimal pH of 5.0, phytase presented on the cell surface is characterized by an optimal pH of 3.0. Moreover, our data demonstrate that phytase anchored on the cell surface exhibits higher pH stability than its secreted counterpart. Interestingly, our in vitro digestion experiments demonstrate that phytase attached to the cell surface is a more efficient enzyme than secreted phytase.

  19. Ectopic expression of a soybean phytase in developing seeds of Glycine max to improve phosphorus availability.

    Science.gov (United States)

    Chiera, Joseph M; Finer, John J; Grabau, Elizabeth A

    2004-12-01

    A transgenic approach was used to alter soybean seed phytate content by expressing a soybean phytase gene (GmPhy) during seed development to degrade accumulating phytic acid (IP6). An expression vector containing the soybean phytase cDNA controlled by the seed-specific beta-conglycinin promoter (alpha'-subunit) was used to transform embryogenic soybean cultures. Plants from four independent transgenic lines were analyzed for transgene integration and seed IP6 levels. The reduction in IP6 levels in transgenic seeds compared to control 'Jack' soybeans ranged from 12.6 to 24.8 as determined by HPLC. A low copy transformant was propagated to the T4 generation and examined in more detail for phytase expression and enzyme activity during seed development. Expression of phytase mRNA and phytase activity increased during seed development, consistent with the use of an embryo-specific promoter. Ectopic phytase expression during seed development offers potential as an effective strategy for reducing phytate content in soybean seed.

  20. Phytases for improved iron absorption

    DEFF Research Database (Denmark)

    Nielsen, Anne Veller Friis; Meyer, Anne S.

    2016-01-01

    Phytase enzymes present an alternative to iron supplements, because they have been shown to improve iron absorption by means of catalysing the degradation of a potent iron absorption inhibitor: phytic acid. Phytic acid is a hexaphosphate of inositol and is particularly prevalent in cereal grains,...

  1. 一株产植酸酶青霉的分类鉴定及其植酸酶基因克隆和在毕赤酵母中的表达%Identification of a Penicillium isolate, gene cloning of phytase and its expression in Pichia pastoris

    Institute of Scientific and Technical Information of China (English)

    包佳源; 马忠友; 戚明玲; 黄勃; 樊美珍

    2012-01-01

    Phytases are a group of enzymes capable of releasing phosphate from phytate, and have been applied mainly to animal (swine and poultry) diets in order to improve phytate P digestibility and mineral bioavail-ability. RCEF4908, a strain with high phytase yield, was screened from a lot of phytase-producted fungal isolates. According to morphological characters and internal transcribed spacer sequences of rDNA, and it was identified as Penicillium oxalicum Currie & Thorn. The fragment of the mature phytase was cloned directly from DNA of P. oxalicum RCEF4908 with the special primers designed from phytase gene in GenBank No. AY071824. The deduced amino acid sequence of this phytase with the conserved motifs RHGXRXP and HD belongs to a typical histidine acid phosphatases. The phytase was overexpressed in the recombinant P49, and its specific activity could reach 8 000 U·mL-1 after induced 144h at optimum pH 5.5, and showed high activity with phytic acid sodium salt at a pH range of 2.5 to 7.0. This recombinant phytase studied here can retain 53.8%, 52.5% and 45.2% of its activity after denaturation at 90 ℃ for 10, 20 and 30 min, respectively. The broad pH optima and high thermostability of the phytase makes it a better candidate for feed-pelleting applications.%植酸酶是催化植酸盐水解成磷酸盐的一类酶的总称,添加到猪和家禽动物的饲料中可以提高植酸磷的利用率和促进对矿质元素的吸收利用.根据形态特征和ITS分子序列,作者对该实验室筛选得到的高产植酸酶菌株青霉RCEF4908进行了鉴定,确定该菌株在系统分类上属于草酸青霉Penicillium oxalicum Currie & Thom.利用特异性引物扩增出植酸酶成熟肽基因片段,蛋白质序列含有保守区域RHGXRXP和HD,属于组氨酸酸性磷酸酶家族.通过基因操作成功实现了植酸酶在Pichia pastoris GS 115的表达.转化子P49在诱导培养144 h后酶活性可以达到约8 000U·mL-1.重组植酸酶在pH 2.5~7.0

  2. Efficient production of mutant phytase (phyA-7) derived from Selenomonas ruminantium using recombinant Escherichia coli in pilot scale.

    Science.gov (United States)

    Chi-Wei Lan, John; Chang, Chih-Kai; Wu, Ho-Shing

    2014-09-01

    A mutant gene of rumen phytase (phyA-7) was cloned into pET23b(+) vector and expressed in the Escherichia coli BL21 under the control of the T7 promoter. The study of fermentation conditions includes the temperature impacts of mutant phytase expression, the effect of carbon supplements over induction stage, the inferences of acetic acid accumulation upon enzyme expression and the comparison of one-stage and two-stage operations in batch mode. The maximum value of phytase activity was reached 107.0 U mL(-1) at induction temperature of 30°C. Yeast extract supplement demonstrated a significant increase on both protein concentration and phytase activity. The acetic acid (2 g L(-1)) presented in the modified synthetic medium demonstrated a significant decrease on expressed phytase activity. A two-stage batch operation enhanced the level of phytase activity from 306 to 1204 U mL(-1) in the 20 L of fermentation scale. An overall 3.7-fold improvement in phytase yield (35,375.72-1,31,617.50 U g(-1) DCW) was achieved in the two-stage operation.

  3. Overexpression and biochemical characterization of a thermostable phytase from Bacillus subtilis US417 in Pichia pastoris.

    Science.gov (United States)

    Hmida-Sayari, Aïda; Elgharbi, Fatma; Farhat, Ameny; Rekik, Hatem; Blondeau, Karine; Bejar, Samir

    2014-09-01

    The overexpression of the native gene encoding the thermostable Bacillus subtilis US417 phytase using Pichia pastoris system is described. The phytase gene, in which the sequence encoding the signal peptide was replaced by that of the α-factor of Saccharomyces cerevisiae, was placed under the control of the methanol-inducible promoter of the alcohol oxidase 1 gene and expressed in Pichia pastoris. Small-scale expression experiments and activity assays were used to screen positive colonies. A recombinant strain was selected and produces 43 and 227 U/mL of phytase activity in shake flasks and in high-cell-density fermentation, respectively. The purified phytase was glycosylated protein and varied in size (50-65 kDa). It has a molecular mass of 43 kDa when it was deglycosylated. The purified r-PHY maintains 100% of its activity after 10 min incubation at 75 °C and pH 7.5. This thermostable phytase, which is also active over broad pH ranges, may be useful as feed additives, since it can resist the temperature used in the feed-pelleting process.

  4. Soybean phytase and nucleic acid encoding the same

    OpenAIRE

    1999-01-01

    Isolated soybean phytase polypeptides and isolated nucleic acids encoding soybean phytases are provided. The invention is also directed to nucleic acid expression constructs, vectors, and host cells comprising the isolated soybean phytase nucleic acids, as well as methods for producing recombinant and non-recombinant purified soybean phytase. The invention also relates to transgenic plants expressing the soybean phytase, particularly expression under seed-specific expression control elements.

  5. Trace element inhibition of phytase activity.

    Science.gov (United States)

    Santos, T; Connolly, C; Murphy, R

    2015-02-01

    Nowadays, 70 % of global monogastric feeds contains an exogenous phytase. Phytase supplementation has enabled a more efficient utilisation of phytate phosphorous (P) and reduction of P pollution. Trace minerals, such as iron (Fe), zinc (Zn), copper (Cu) and manganese (Mn) are essential for maintaining health and immunity as well as being involved in animal growth, production and reproduction. Exogenous sources of phytase and trace elements are regularly supplemented to monogastric diets and usually combined in a premix. However, the possibility for negative interaction between individual components within the premix is high and is often overlooked. Therefore, this initial study focused on assessing the potential in vitro interaction between inorganic and organic chelated sources of Fe, Zn, Cu and Mn with three commercially available phytase preparations. Additionally, this study has investigated if the degree of enzyme inhibition was dependent of the type of chelated sources. A highly significant relationship between phytase inhibition, trace mineral type as well as mineral source and concentration, p phytases for Fe and Zn, as well as for Cu with E. coli and Aspergillus niger phytases. Different chelate trace mineral sources demonstrated diversifying abilities to inhibit exogenous phytase activity.

  6. Cloning and characterization of the first actinomycete β-propeller phytase from Streptomyces sp. US42.

    Science.gov (United States)

    Boukhris, Ines; Farhat-Khemakhem, Ameny; Bouchaala, Kameleddine; Virolle, Marie-Joëlle; Chouayekh, Hichem

    2016-10-01

    A gene encoding an extracellular phytase was cloned for the first time from an Actinomycete, Streptomyces sp. US42 and sequenced. The sequence of this gene revealed an encoded polypeptide (PHY US42) exhibiting one and six residues difference with the putative phytases of Streptomyces lividans TK24 and Streptomyces coelicolor A3(2), respectively. The molecular modeling of PHY US42 indicated that this phytase belongs to the group of β-propeller phytases that are usually calcium-dependent. PHY US42 was purified and characterized. Its activity was calcium-dependent and maximal at pH 7 and 65 °C. The enzyme was perfectly stable at pH ranging from 5 to 10 and its thermostability was greatly enhanced in the presence of calcium. Indeed, PHY US42 maintained 80% of activity after 10 min of incubation at 75 °C in the presence of 5 mM CaCl2 . PHY US42 was also found to exhibit high stability after incubation at 37 °C for 1 h in the presence of bovine bile and digestive proteases like of pepsin, trypsin, and chymotrypsin. Considering its biochemical properties, PHY US42 could be used as feed additive in combination with an acid phytase for monogastric animals.

  7. Bacillus phytases: Current status and future prospects.

    Science.gov (United States)

    Borgi, Mohamed Ali; Boudebbouze, Samira; Mkaouar, Héla; Maguin, Emmanuelle; Rhimi, Moez

    2015-01-01

    Phytases catalyze the hydrolysis of phytic acid in a stepwise manner to lower inositol phosphates, myo-inositol (having important role in metabolism and signal transduction pathways), and inorganic phosphate. These enzymes have been widely used in animal feed in order to improve phosphorus nutrition and to decrease pollution in animal waste. Compared to previously described phytases, the phytase (PhyL) from Bacillus licheniformis ATCC 14580 has attractive biochemical properties which can increase the profitability of several biotechnological procedures (animal nutrition, humain health…etc). Due to its amino acid sequence with critical substitutions, the PhyL could be a model to enhance other phytases features, in terms of thermal stability and high activity. Otherwise, an engineered PhyL, with low pH optimum, will represent a challenge within the class of β- propeller phytases.

  8. Phytase in non-ruminant animal nutrition: a critical review on phytase activities in the gastrointestinal tract and influencing factors.

    Science.gov (United States)

    Dersjant-Li, Yueming; Awati, Ajay; Schulze, Hagen; Partridge, Gary

    2015-03-30

    This review focuses on phytase functionality in the digestive tract of farmed non-ruminant animals and the factors influencing in vivo phytase enzyme activity. In pigs, feed phytase is mainly active in the stomach and upper part of the small intestine, and added phytase activity is not recovered in the ileum. In poultry, feed phytase activities are mainly found in the upper part of the digestive tract, including the crop, proventriculus and gizzard. For fish with a stomach, phytase activities are mainly in the stomach. Many factors can influence the efficiency of feed phytase in the gastrointestinal tract, and they can be divided into three main groups: (i) phytase related; (ii) dietary related and (iii) animal related. Phytase-related factors include type of phytase (e.g. 3- or 6-phytase; bacterial or fungal phytase origin), the pH optimum and the resistance of phytase to endogenous protease. Dietary-related factors are mainly associated with dietary phytate content, feed ingredient composition and feed processing, and total P, Ca and Na content. Animal-related factors include species, gender and age of animals. To eliminate the antinutritional effects of phytate (IP6), it needs to be hydrolyzed as quickly as possible by phytase in the upper part of the digestive tract. A phytase that works over a wide range of pH values and is active in the stomach and upper intestine (along with several other characteristics and in addition to being refractory to endogenous enzymes) would be ideal.

  9. Phytase-Producing Bacteria from Extreme Regions in Indonesia

    Directory of Open Access Journals (Sweden)

    Sajidan

    2015-10-01

    Full Text Available ABSTRACTIn this study, 154 isolates capable of producing extracellular phytate-degrading activity were isolated from four soil samples from volcanic areas in Central Java, Indonesia. Six strains with high phytate-degrading activity were selected for strain identification and characterization of the corresponding phytate-degrading enzyme. Blast analysis of 16S rRNA gene sequences revealed high similarities for all the six isolates to reference sequences belonging to the genusBacillus. Isolates MS5, MC6, D10 and D16 showed 99% sequence identity toB. cereus, while isolate MC8 exhibited 99% sequence identity toB. aryabhatti and D6 99% sequence identity toB. psychrotolerans. The crude extracellular phytase preparations from the isolates showed following optimal conditions for phytate dephosphorylation: pH 4.0 and 50°C (isolate D10, pH 5.0 and 60°C (isolate MC6, and isolate MS5, pH 6.0 and 50°C (isolate D16 and pH 6.0 and 60°C (isolate D6 and pH 6.0 and 40°C (isolate MC8. Zn2+ and Fe3+ strongly inhibited phytate dephosphorylation with all phytase preparations studied. In the presence of Ca2+, an increase in phytase activity of 10-15% was obtained.

  10. Quantitative conversion of phytate to inorganic phosphorus in soybean seeds expressing a bacterial phytase.

    Science.gov (United States)

    Bilyeu, Kristin D; Zeng, Peiyu; Coello, Patricia; Zhang, Zhanyuan J; Krishnan, Hari B; Bailey, April; Beuselinck, Paul R; Polacco, Joe C

    2008-02-01

    Phytic acid (PA) contains the major portion of the phosphorus in the soybean (Glycine max) seed and chelates divalent cations. During germination, both minerals and phosphate are released upon phytase-catalyzed degradation of PA. We generated a soybean line (CAPPA) in which an Escherichia coli periplasmic phytase, the product of the appA gene, was expressed in the cytoplasm of developing cotyledons. CAPPA exhibited high levels of phytase expression, >or=90% reduction in seed PA, and concomitant increases in total free phosphate. These traits were stable, and, although resulted in a trend for reduced emergence and a statistically significant reduction in germination rates, had no effect on the number of seeds per plant or seed weight. Because phytate is not digested by monogastric animals, untreated soymeal does not provide monogastrics with sufficient phosphorus and minerals, and PA in the waste stream leads to phosphorus runoff. The expression of a cytoplasmic phytase in the CAPPA line therefore improves phosphorus availability and surpasses gains achieved by other reported transgenic and mutational strategies by combining in seeds both high phytase expression and significant increases in available phosphorus. Thus, in addition to its value as a high-phosphate meal source, soymeal from CAPPA could be used to convert PA of admixed meals, such as cornmeal, directly to utilizable inorganic phosphorus.

  11. Unraveling the major triticeae phytases – towards cereals with high grain phytase activity

    DEFF Research Database (Denmark)

    Madsen, Claus Krogh

    Phytic acid is the main storage form of phosphate in plant seeds. Phosphorus in this form is largely indigestible by monogastric animals due to insufficient activity of the hydrolytic enzyme phytase in their digestive tract and in most plant seeds. Moreover as a strong chelator, undigested phytic...... acid constitutes the most important anti nutritional factor for the bioavailability of minerals. However, the mature grain phytase activity varies considerably between plant species and between cultivars. Cereals like rye, wheat and barley possess significant phytase activity with the potential...... to contribute to the hydrolysis of phytic acid in food and feed. In contrast rice and maize possess insufficient phytase activity in the mature grain to have a notable effect on the phytic acid. The aim of the present study was to (1) uncover the genetic basis for the major phytases in cereal grains (2) explain...

  12. Transgenic Soybean(Glycine max·L)Plants with phyA Gene ImproVed Phytase ActiVity%植酸酶基因phyA转化大豆品种研究

    Institute of Scientific and Technical Information of China (English)

    闫瑞叶; 李喜焕; 李桂兰; 常文锁; 张彩英

    2011-01-01

    organic phosphorus(P)is the main form of phosphorus in soil,of which the phytate accounted for more than 50%.The available way that improves Plant P nutrition is to hydrolyze phytate into inorganic phosphorus(Pi)to increase the soil P utilization efficiency.ln this study,a phytase gene(phyA),driven by the root-specific pyk10 promoter and directed by a carrot extracellular targeting peptide,was introduced into different soybean varieties(Jidou 12,Jidou 16,wuxing 1 and Jilin 35)by Agrobacterium-mediated and pollen tube pathway transformation method.PCR analysis results showed that the phyA was successfully integrated into the soybean genome.Total 114,101 and 28 PCR positive plants were obtained from T0,T1 and T2 generation,respectively.The results also showed that the T4 generation plants could grow better than wild-type when cultured under the condjtion in which phytate was the sole source of phosphorus.And the secreted phytase activities of roots also indicated that three transgenic lines were increased by 5%,13%and 24%compared to the wild-type plants,respectively.%土壤磷多以有机态形式存在,而植酸磷占一半以上,分解利用植酸态磷是提高土壤磷利用效率、改善植物磷素营养的新途径.利用大豆农杆菌介导子叶节转化与花粉管通道转化技术,将含有根特异启动子pyk10、信号肽S和植酸酶phyA的嵌和基因(KSA)转入冀豆12、冀豆16、五星1号和吉林35中.经PCR检测,共获得T0阳性植株114个,T1阳性植株101个,T2阳性材料28个.通过将T4代转基因株系和野生型对照在仅含植酸磷的营养液中进行培养发现,转基因植株在植酸磷条件下生长状况优于对照,并且3个转基因株系根系分泌型植酸酶活性分别比野生型提高5%、13%和24%.

  13. Phytase activity in brown rice during steeping and sprouting

    NARCIS (Netherlands)

    Ou, K.; Cheng, Y.; Xing, Y.; Lin, L.; Nout, M.J.R.; Liang, J.

    2011-01-01

    Phytase in brown rice will be activated and accumulated during seed germination. Changes of phytase activity in brown rice during two stages of germination (steeping and sprouting) affected by process conditions were studied. It was shown that steeping led to significant decrease of phytase activity

  14. 小麦植酸酶基因TaPHY1的克隆、分子特征和表达特性研究%Cloning, molecular characterization, and expression pattern of TaPHY1, a novel phytase gene in wheat(Triticum aestivum L.)

    Institute of Scientific and Technical Information of China (English)

    刘天龙; 许振龙; 郭程谨; 路文静; 李小娟; 谷俊涛; 肖凯

    2011-01-01

    Using a wheat purple acid phosphatase (PAP)-type phytase gene referred to PAPhya1 as a query, a novel phytase gene to be homologous to PAPhya1 that has not been characterized and functionally analyzed was identified and designated as TaPHYl in this study. The full length of TaPHY1 is 1771 bp, encoding a 548-aa polypeptide. TaPHY1 contains three conserved signature domains generally identified in PAP-type phytases, including metal-lophos, purple acid phosphatase and metallo-dependent phosphatases. Phylogenetic analysis revealed that TaPHY1 shares high similarities to the previously reported phyase genes such as PAPhya1~PAPPhya3 and PAPhyb1 ~PAPhyb2, as well as high similarities to PAP-type phytase genes in other plant species including the barley (H. Vulgare) and rye (S. Cereale).TaPHY1 is abundantly expressed in germinating seed and little expressed in the primary roots and leaves. The expressed TaPHY1 based on the prokaryotic expression system exhibited strong capability to degrade the phytate, a major component of organic phosphate compound in the mature seed. Taken together, as a novel PAP-type phytase gene in wheat, TaPHY1 plays crucial roles on degradation of the seed phytic acid and its derivatives, and provides the Pi for the novel organs of the young seedlings.%利用小麦植酸酶基因PAPhya1进行同源查找,获得1个与该基因高度同源、尚未开展分子特征和生物学功能研究的小麦新型植酸酶基因TaPHY1.TaPHY1的全长cDNA序列为1771 bp,编码548个氨基酸残基,TaPHY1含有紫色酸性磷酸酶(PAP)类植酸酶通常含有的保守域Metallophos、Purple acid phosphatase和Metallo-dependent phosphatases.系统进化分析表明,TaPHY1除与已报道的小麦PAP类植酸酶基因PAPhya1~PAPPhya3和PAPh yb1~PAPh yb2高度同源外,还与部分大麦和黑麦的PAP型植酸酶基因在序列上高度相似.TaPHY1在萌发种子中呈高丰度表达,而在根叶中的表达水平很低.对原核

  15. Effects of inositol, inositol-generating phytase B applied alone, and in combination with 6-phytase A to phosphorus-deficient diets on laying performance, eggshell quality, yolk cholesterol, and fatty acid deposition in laying hens.

    Science.gov (United States)

    Zyla, K; Mika, M; Duliński, R; Swiatkiewicz, S; Koreleski, J; Pustkowiak, H; Piironen, J

    2012-08-01

    Phytase B, a product of Aspergillus niger phyB gene expressed in Trichoderma reesei, which increased myo-inositol concentrations in 20 mM sodium phytate solution 7.5-fold during 120-min incubation, a combination of phytase B with 6-phytase A, and pure myo-inositol were tested as feed supplements in Bovans Brown laying hens. In the 2-factorial experiment (2×5), birds from wk 50 to 62 were fed 2 basal diets, corn-soybean (CSM) or wheat-soybean (WSM), using 12 one-hen cages per treatment. For both basal diets, the dietary treatments included negative control (0.08% nonphytate P in CSM, 0.13% nonphytate P in WSM; NC); internal control groups, NC+0.04% nonphytate P from monocalcium phosphate, MCP (IC); NC+0.1% of myo-inositol (Inos), NC+phytase B at 1,300 units of phytase B-acid phosphatase activity (AcPU)/kg (PhyB), NC+phytase B at 1,300 AcPU/kg+6-phytase A at 300 FTU/kg (PhyA+B). Feed intake, laying performance, and eggshell quality were determined. The total lipid and cholesterol contents as well as fatty acid profile were assessed in egg yolks collected from hens fed CSM diets, as was fatty acid profile. The hens fed the WSM diet consumed significantly more feed, laid a higher mass of eggs daily with higher mean weights, and had a higher hen-day egg production than the birds receiving the CSM diets. Similarly, higher values for yolk weights, shell weights, shell thickness, shell density, and breaking strengths were determined in the eggs laid by the hens fed the WSM diets. In hens fed either the CSM diets with phytase B alone, or in combination with 6-phytase A, enhanced feed intakes, egg mass, and hen-day egg production were recorded. Phytases also enhanced the eggshell quality parameters in the hens fed both variants of the diets. Phytase B alone, or in combination with 6-phytase A, reduced the total lipid and cholesterol concentrations in egg yolks collected from the hens fed the CSM diets, whereas the combination of both phytases improved the n-6:n-3

  16. Phytase application in chewing gum - A technical assessment

    DEFF Research Database (Denmark)

    Nielsen, Anne Veller Friis; Meyer, Anne S.

    2016-01-01

    Phytase catalysis has been shown to improve iron absorption by dephosphorylation of the potent iron chelator, phytic acid, found in high amounts in cereals. Recently, the World Health Organization evaluated the phytase from Aspergillus niger as safe for use in human food. The phytase may work...... either prior to ingestion, i.e. in the food, or post ingestion, i.e. in the human gastrointestinal tract. We have assessed the technical aspects of formulation and release of phytase added to chewing gum as a delivery vehicle. Phytases from Aspergillus niger and Escherichia coli incorporated into chewing...... gum were released quantitatively upon chewing and retained phytase activity (50-80% of the enzyme activity added was released within 10 minutes). Initial evaluations of phytase chewing gum shelf life showed good stability after 48 days of storage of the chewing gum at ambient conditions....

  17. Phytase-active lactic acid bacteria from sourdoughs

    DEFF Research Database (Denmark)

    Nuobariene, Lina; Cizeikiene, Dalia; Gradzeviciute, Egle

    2015-01-01

    at conditions optimal for leavening of bread dough (pH 5.5 and 30°C). The phytase active isolates belonged to the species Lactobacillus panis, Lactobacillus reuteri, Lactobacillus fermentum, and Pediococcus pentosaceus. Phytase activities of the tested LAB isolates were both extra- and intra...... bacteria (LAB) which could be used as a starter to increase mineral bioavailability in whole-meal bread. Hence, LAB isolates were isolated from Lithuanian sourdoughs, tested for phytase activity, and phytase active isolates were identified. Studies of phytase activity of the isolates were carried out......-cellular. The highest extracellular phytase production was found in L.panis with a volumetric phytase activity of 140U/mL. Phytate degradation in whole-wheat dough fermented with L.panis or L.fermentum was 90% and 70%, respectively....

  18. A type IV translocated Legionella cysteine phytase counteracts intracellular growth restriction by phytate.

    Science.gov (United States)

    Weber, Stephen; Stirnimann, Christian U; Wieser, Mara; Frey, Daniel; Meier, Roger; Engelhardt, Sabrina; Li, Xiaodan; Capitani, Guido; Kammerer, Richard A; Hilbi, Hubert

    2014-12-05

    The causative agent of Legionnaires' pneumonia, Legionella pneumophila, colonizes diverse environmental niches, including biofilms, plant material, and protozoa. In these habitats, myo-inositol hexakisphosphate (phytate) is prevalent and used as a phosphate storage compound or as a siderophore. L. pneumophila replicates in protozoa and mammalian phagocytes within a unique "Legionella-containing vacuole." The bacteria govern host cell interactions through the Icm/Dot type IV secretion system (T4SS) and ∼300 different "effector" proteins. Here we characterize a hitherto unrecognized Icm/Dot substrate, LppA, as a phytate phosphatase (phytase). Phytase activity of recombinant LppA required catalytically essential cysteine (Cys(231)) and arginine (Arg(237)) residues. The structure of LppA at 1.4 Å resolution revealed a mainly α-helical globular protein stabilized by four antiparallel β-sheets that binds two phosphate moieties. The phosphates localize to a P-loop active site characteristic of dual specificity phosphatases or to a non-catalytic site, respectively. Phytate reversibly abolished growth of L. pneumophila in broth, and growth inhibition was relieved by overproduction of LppA or by metal ion titration. L. pneumophila lacking lppA replicated less efficiently in phytate-loaded Acanthamoeba castellanii or Dictyostelium discoideum, and the intracellular growth defect was complemented by the phytase gene. These findings identify the chelator phytate as an intracellular bacteriostatic component of cell-autonomous host immunity and reveal a T4SS-translocated L. pneumophila phytase that counteracts intracellular bacterial growth restriction by phytate. Thus, bacterial phytases might represent therapeutic targets to combat intracellular pathogens.

  19. Introducing Phytase Gene phyA into Different Soybean Varieties from Transgenic Line Utilizing Sexual Hybridization%杂交转育植酸酶phyA大豆阳性材料筛选研究

    Institute of Scientific and Technical Information of China (English)

    李喜焕; 刘渊; 闫瑞叶; 孔佑宾; 李桂兰; 常文锁; 张彩英

    2013-01-01

    Phosphorus ( P) is one of the most important inorganic nutrients that can significantly affect plant growth and metabolism. However,30 to 40 percent of the unavailable P in agricultural soils exists as phytate,which can not directly be absorbed by plant except for resolved by phytase. So selecting or developing new varieties that can resolve phytate-P through the modern transgenic breeding approach provided a new opportunity to improve the efficiency of phosphorus by plants. In this paper,a phytase gene(phyA) , isolated from Aspergillus ficuum, was introduced into soybean by Agrobacterium-mediated transformation, and then thirty-eight soybean crosses had been made between transgenic lines(JL35-phyA,male parent)and other non-transgenic varieties(female parent). PCR and RT-PCR results showed that the phyA was successfullly incorporated into soybean genome and expressed in transgenic line JL35-phyA. Futhermore,the pods,seeds,and weights per plant and weight 100 seeds of JL35-phyA were higher than wildtype JL35 significantly. The results also showed that 427 PCR positive transgenic F1 plants from the thirty-eight cross combinations above and 377 PCR positive transgenic F2 plants from the progenies of the cross combination JD17 × JL35-phyA were obtained by the sexual hybridization transfer method. Analysis results of these progenies showed that the variation of agronomic characters were abundant,and some excellent transgenic lines were selected. So it provides many important genetic resources to transgenic breeding of phyA in soybean.%植酸及其盐类占土壤非有效态磷30%~40%,利用转基因技术结合常规育种手段培育能够分解利用植酸磷的作物新品种是解决这一问题的最新途径.本研究以农杆菌转化子叶节所获得的JL35-phyA为试材,采用PCR与RT-PCR进行目的基因检测,获得转基因阳性材料;随后将这些阳性材料与38个常规大豆杂交,实现phyA向不同大豆品种的转育.结果表明,利

  20. Improvement of megaprimer method for site-directed mutagenesis and its application to phytase

    Institute of Scientific and Technical Information of China (English)

    Haiqiang LU; Hongwei YU; Runfang GUO; Yingmin JIA

    2009-01-01

    Site-directed mutagenesis is used extensively for probing gene function. In this paper we describe an improved megaprimer method to the site-directed muta-genesis of phytase from Aspergillus niger, which allowed the mutations to be performed more efficiently in less time than other traditional methods. Three rounds of PCR and two pairs of primers were required in this method, and additionally, the restriction enzyme Dpn I was used for the elimination of template instead of the gel purification in this process. The entire procedure was performed in one tube. Moreover, this method was easier for obtaining large mutant genes than other methods. We successfully carried out the site-directed mutagenesis of phytase by adopting this method.

  1. Phytase-mediated mineral solubilization from cereals underin vitrogastric conditions

    DEFF Research Database (Denmark)

    Nielsen, Anne V. F.; Meyer, Anne S.

    2016-01-01

    cereal phytic acid at similar rates and to similar extents. Microbial phytase-catalysed phytate dephosphorylation was accompanied by increased iron and zinc release from the cereal substrates. For wheat bran at pH 5, the endogenous wheat phytase activity produced mineral release equal to or better than...... that of the microbial phytases. No increases in soluble cadmium, lead or arsenic were observed with microbial phytase-catalyzed phytate dephosphorylation. CONCLUSION Microbial phytase treatment abated phytate chelation hence enhanced the release of iron and zinc from the phytate-rich cereals at the simulated gastric...... conditions. The data infer that acid stable microbial phytases can help improve iron bioavailability from phytate-rich cereal substrates via post-ingestion activity....

  2. The degradation of phytate by microbial and wheat phytases is dependent on the phytate matrix and the phytase origin

    DEFF Research Database (Denmark)

    Brejnholt, Sarah M; Dionisio, Giuseppe; Glitsoe, Vibe

    2011-01-01

    BACKGROUND: Phytases increase utilization of phytate phosphorus in feed. Since wheat is rich in endogenous phytase activity it was examined whether wheat phytases could improve phytate degradation compared to microbial phytases. Moreover, it was investigated whether enzymatic degradation of phytate...... is influenced by the matrix surrounding it. Phytate degradation was defined as the decrease in the sum of InsP6 + InsP5. RESULTS: Endogenous wheat phytase effectively degraded wheat InsP6 + InsP5 at pH 4 and pH 5, while this was not true for a recombinant wheat phytase or phytase extracted from wheat bran. Only...... microbial phytases were able to degrade InsP6 + InsP5 in the entire pH range from 3 to 5, which is relevant for feed applications. A microbial phytase was efficient towards InsP6 + InsP5 in different phytate samples, whereas the ability to degrade InsP6 + InsP5 in the different phytate samples ranged from...

  3. Cloning, recombinant expression and characterization of a new phytase from Penicillium chrysogenum.

    Science.gov (United States)

    Ribeiro Corrêa, Thamy Lívia; de Queiroz, Marisa Vieira; de Araújo, Elza Fernandes

    2015-01-01

    The phy gene, which encodes a phytase in Penicillium chrysogenum CCT 1273, was cloned into the vector pAN-52-1-phy and the resulting plasmid was used for the cotransformation of Penicillium griseoroseum PG63 protoplasts. Among the 91 transformants obtained, 23 were cotransformants. From there, the phytase activity of these 23 transformants was evaluated and P. griseoroseum T73 showed the highest. The recombinant strain P. griseoroseum T73 contained the phy gene integrated in at least three sites of the genome and showed a 5.1-fold increase in phytase activity in comparison to the host strain (from 0.56 ± 0.2 to 2.86 ± 0.4 U μg protein(-1)). The deduced PHY protein has 483 amino acids; an isoelectric point (pI) higher than that reported for phytases from filamentous fungi (7.6); higher activity at pH 2.0 (73%), pH 5.0 (100%) and 50 °C; and is stable at pH values 3.0-8.0 and temperatures 70-80 °C. PHY produced by the recombinant strain P. griseoroseum T73 was stable after four weeks of storage at -20, 8 and 25 °C and was effective in releasing Pi, especially from soybeans. The data presented here show that P. griseoroseum is a successful host for expression of heterologous protein and suggest the potential use of PHY in the animal nutrition industry.

  4. Performance of microbial phytases for gastric inositol phosphate degradation

    DEFF Research Database (Denmark)

    Nielsen, Anne Veller Friis; Nyffenegger, Christian; Meyer, Anne S.

    2015-01-01

    Microbial phytases catalyze dephosphorylation of phytic acid, thereby potentially releasing chelated iron and improving human iron absorption from cereal-based diets. For this catalysis to take place in vivo, the phytase must be robust to low pH and proteolysis in the gastric ventricle. This stud...

  5. Phytase, a new life for an “old” enzyme

    Science.gov (United States)

    Phytase represents a group of phosphohydrolytic enzymes that initiate stepwise removals of phosphate from phytate. Simple-stomached species such as swine, poultry, and fish require extrinsic phytase to digest phytate: the major form of phosphorus in plant feeds. Consequently, this enzyme is suppleme...

  6. [Effects of phytase transgenic corn planting on soil nematode community].

    Science.gov (United States)

    Zhao, Zong-Chao; Su, Ying; Mou, Wen-Ya; Liu, Man-Qiang; Chen, Xiao-Yun; Chen, Fa-Jun

    2014-04-01

    A healthy soil ecosystem is essential for nutrient cycling and energy conversion, and the impact of exogenous genes from genetically modified crops had aroused wide concerns. Phytase transgenic corn (i. e., the inbred line BVLA430101) was issued a bio-safety certificate on 27 September 2009 in China, which could improve the efficiency of feed utilization, reduce environmental pollution caused by animal manure. In this study, the abundance of trophic groups, community structure and ecological indices of soil nematodes were studied over the growing cycle of phytase transgenic corn (ab. transgenic corn) and control conventional parental corn (ab. control corn) in the field. Totally 29 and 26 nematode genera were isolated from transgenic corn and control corn fields, respectively. The abundances of bacterivores and omnivores-predators, the total number of soil nematodes, and the Shannon index (H) were significantly greater under transgenic corn than under control corn, while the opposite trend was found for the relative abundance of herbivores and the maturity index (Sigma MI) of soil nematodes. Repeated-measures analysis of variance (ANOVA) did not detect any significant effects of transgenic corn on the composition and abundance of nematode trophic groups and ecological indices of soil nematodes. Furthermore, the Student-T test showed that the abundances of bacterivores and omnivores-predators and the total number of soil nematodes during the milk-ripe stage were significant higher in the transgenic corn field than in the control corn field. The effects of transgenic corn planting on soil nematodes might be related to the increase in the nitrogen content of field soil under transgenic corn compared to control corn.

  7. Secretive expression of Aspergillus fumigatus phytase in tobacco improves phosphorus nutrition in plant

    Institute of Scientific and Technical Information of China (English)

    Wang Yan; Gao Xiaorong; Su Qiao; An Lijia

    2007-01-01

    To generate transgenic plants capable of utilizing exogenous phytate,an Aspersgillus fumigatus phytase gene(fphyA) was constitutively expressed in tobacco and recombinant enzyme was secreted from plant roots into the rhizosphere using the signal sequence from tobacco calretieulin.After 40 days of plant growth in hydroponic media,phytase activities in leaves,stems,roots and growth media of transgenic plants were 8.6-fold,7.4-fold,12.6-fold and 14.3-fold higher than those of wild-type plants.Signifi-cant improvements in plant growth and phosphoms(P)utilization were observed in the transgenic plants.When phytate was supplied as the sole P source.45-day-old transgenic tobaccos accumulated 1.0-fold and 0.5-fold more shoot and root biomass than wild-type tobaccos.with a concomitant of 1.7-fold increase in total P concentration.These results indicate that secretive expression of the A.fumigatus phytase improves acquisition and use of P from phytate in plants.

  8. Characterization of an extremely salt-tolerant and thermostable phytase from Bacillus amyloliquefaciens US573.

    Science.gov (United States)

    Boukhris, Ines; Farhat-Khemakhem, Ameny; Blibech, Monia; Bouchaala, Kameleddine; Chouayekh, Hichem

    2015-09-01

    The extracellular phytase produced by the Bacillus amyloliquefaciens US573 strain, isolated from geothermal soil located in Southern Tunisia was purified and characterized. This calcium-dependent and bile-stable enzyme (PHY US573) was optimally active at pH 7.5 and 70 °C. It showed a good stability at pH ranging from 4 to 10, and especially, an exceptional thermostability as it recovered 50 and 62% of activity after heating for 10 min at 100 and 90 °C, respectively. In addition, PHY US573 was found to be extremely salt-tolerant since it preserved 80 and 95% of activity in the presence of 20 g/l of NaCl and LiCl, respectively. The gene corresponding to PHY US573 was cloned. It encodes a 383 amino acids polypeptide exhibiting 99% identity with the highly thermostable phytases from Bacillus sp. MD2 and B. amyloliquefaciens DS11 (3 and 5 residues difference, respectively), suggesting the existence of common molecular determinants responsible for their remarkable heat stability. Overall, our findings illustrated that in addition to its high potential for application in feed industry, the salt tolerance of the PHY US573 phytase, may represent an exciting new avenue for improvement of phosphorus-use efficiency of salt-tolerant plants in soils with high salt and phytate content.

  9. A Phytase-Based Reporter System for Identification of Functional Secretion Signals in Bifidobacteria.

    Directory of Open Access Journals (Sweden)

    Annika Osswald

    Full Text Available Health-promoting effects have been attributed to a number of Bifidobacterium sp. strains. These effects as well as the ability to colonise the host depend on secreted proteins. Moreover, rational design of protein secretion systems bears the potential for the generation of novel probiotic bifidobacteria with improved health-promoting or therapeutic properties. To date, there is only very limited data on secretion signals of bifidobacteria available. Using in silico analysis, we demonstrate that all bifidobacteria encode the major components of Sec-dependent secretion machineries but only B. longum strains harbour Tat protein translocation systems. A reporter plasmid for secretion signals in bifidobacteria was established by fusing the coding sequence of the signal peptide of a sialidase of Bifidobacterium bifidum S17 to the phytase gene appA of E. coli. The recombinant strain showed increased phytase activity in spent culture supernatants and reduced phytase levels in crude extracts compared to the control indicating efficient phytase secretion. The reporter plasmid was used to screen seven predicted signal peptides in B. bifidum S17 and B. longum E18. The tested signal peptides differed substantially in their efficacy to mediate protein secretion in different host strains. An efficient signal peptide was used for expression and secretion of a therapeutically relevant protein in B. bifidum S17. Expression of a secreted cytosine deaminase led to a 100-fold reduced sensitivity of B. bifidum S17 to 5-fluorocytosine compared to the non-secreted cytosine deaminase suggesting efficient conversion of 5-fluorocytosine to the cytotoxic cancer drug 5-fluorouracil by cytosine deaminase occurred outside the bacterial cell. Selection of appropriate signal peptides for defined protein secretion might improve therapeutic efficacy as well as probiotic properties of bifidobacteria.

  10. Analysis of myo-inositol hexakisphosphate hydrolysis by Bacillus phytase

    DEFF Research Database (Denmark)

    Kerovuo, J.; Rouvinen, J.; Hatzack, Frank-Andreas

    2000-01-01

    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...... 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...

  11. Heat-resistant bacterial phytase in broiler pelleted diets

    Directory of Open Access Journals (Sweden)

    TC de F Carlos

    2015-03-01

    Full Text Available The objective of the study was to evaluate the effects of a heat-resistant bacterial phytase added to pelleted diets on mineral digestibility, live performance, carcass traits, and bone quality of broilers. Three treatments were evaluated: Positive control; negative control, with 0.10 points reduction in calcium level and 0.15 points reduction in available phosphorus level; and negative control + phytase at 500 FTU/kg. Mineral digestibility and bone quality results demonstrated that the evaluated phytase resisted pelleting as it increased the utilization of the minerals present in the diet.

  12. Phytase transgenic corn in nutrition of laying hens: residual phytase activity and phytate phosphorus content in the gastrointestinal tract.

    Science.gov (United States)

    Gao, C Q; Ji, C; Zhao, L H; Zhang, J Y; Ma, Q G

    2013-11-01

    The residual activities of transgenic corn-derived and 2 commercial microbial phytases (PA and PB) along the gastrointestinal tract (GIT) of laying hens were compared to evaluate their relative resistance to hydrolysis in the GIT when added to P-deficient diets. The treatments consisted of a negative control (NC) diet containing 0.10 nonphytate P and an NC diet supplemented with transgenic corn-derived phytase (TCDP), PA, and PB at 500 to 5,000 FTU/kg of diet, respectively. Seven diets were fed to Hy-Line Brown laying hens (n = 504; 8 replicates of 9 hens per treatment) for 21 d. At the end of the experiment, the hens were killed and digesta samples from the crop, proventriculus and gizzard, jejunum, and ileum were collected and analyzed for residual phytase activities and phytate P content. Phytase activity in the transgenic corn was determined to be 8,980 FTU/kg of DM. The residual phytase activities along the GIT had increased (P corn is as efficacious as the commercial microbial phytases (PA and PB) in P-deficient diets for the improvement of phytate P digestibility, which would eliminate the need for supplemental phytase and corn separately in laying hen diets.

  13. Effect of dietary phytase transgenic corn on physiological characteristics and the fate of recombinant plant DNA in laying hens.

    Science.gov (United States)

    Gao, Chunqi; Ma, Qiugang; Zhao, Lihong; Zhang, Jianyun; Ji, Cheng

    2014-01-01

    The study aimed to evaluate the potential effects of feeding with phytase transgenic corn (PTC) on organ weight, serum biochemical parameters and nutrient digestibility, and to determine the fate of the transgenic DNA in laying hens. A total of 144 50-week-old laying hens were grouped randomly into 2 treatments, with 8 replicates per treatment and 9 hens per replicate. Each treatment group of hens was fed with diets containing 62.4% non-transgenic conventional corn (CC) or PTC for 16 weeks. The phytase activity for CC was 37 FTU/kg of DM, whereas the phytase activity for PTC was 8,980 FTU/kg of DM. We observed that feeding PTC to laying hens had no adverse effect on organ weight or serum biochemical parameters (p>0.05). A fragment of a poultry-specific ovalbumin gene (ov) was amplified from all tissues of hens showing that the DNA preparations were amenable to PCR amplification. Neither the corn-specific invertase gene (ivr) nor the transgenic phyA2 gene was detected in the breast muscle, leg muscle, ovary, oviduct and eggs. The digestibility data revealed no significant differences between the hens that received the CC- and PTC-based diets in the digestibility of DM, energy, nitrogen and calcium (p>0.05). Phosphorus digestibility of hens fed the PTC-based diet was greater than that of hens fed the CC-based diet (58.03% vs 47.42%, phens. No recombinant phyA2 gene was detected in muscle tissues and reproductive organs of laying hens. The novel plant phytase was efficacious in improving the phosphorus digestibility of laying hens.

  14. Construction of a transformation system for the stable expression of foreign genes in Chlorella sp.

    Institute of Scientific and Technical Information of China (English)

    Wang Yiyun; Gao Xiaorong; Wang Changhai

    2007-01-01

    A stable transformation system for the expression of foreign genes in the unicellular green marine alga (Chlorella sp. MACC/C95) was established. Using electroporation, the alga was transformed with a plasmid containing the phytase gene under the control of CaMV35S promoter and the neomycin phosphotransferase(npt)as a seleetable marker gene. The integration of the phytase gene into the Chlorella genome was revealed by PCR and Southern blotting analysis. RT-PCR analysis revealed the expression of phytasegene at the transcript level. The enhanced activity of phytase enzyme in the transformants confirmed the integration and successful expression of phytase gene. The introduced phytase gene and its protein expression were stably maintained for at least 30 generations in media devoid of selectable antibiotics G418. This is an important step toward the production of useful foreign proteins in Chlorella sp. MACC/C95.

  15. Hydrolysis of precipitated phytate by three distinct families of phytases

    OpenAIRE

    Lim, BL; Tang, J.; Leung, A.; Leung, C

    2006-01-01

    While genetically modified plants that secrete histidine acid phosphatases (HAPs), β-propeller phytases (BPPs) and purple acid phosphatases (PAPs) have been shown to assimilate soluble phytate, little is known about whether these plants have the ability to hydrolyze precipitated phytate. In this study, the ability of representative members of these three classes of phytases to hydrolyze metal-phytate salts and to hydrolyze phytate adsorbed to aluminum precipitates was compared. All three phyt...

  16. Phytase Production and Development of an Ideal Dephytinization Process for Amelioration of Food Nutrition Using Microbial Phytases.

    Science.gov (United States)

    Jain, Jinender; Singh, Bijender

    2016-10-29

    Development of an ideal process for reduction of food phytates using microbial phytases is a demanding task by all food and feed industries all over the world. Phytase production by Bacillus subtilis subsp. subtilis JJBS250 isolated from soil sample was optimized in submerged fermentation using statistical tools. Among all the culture variables tested, sucrose, sodium phytate and Tween-80 were identified as the most significant variables using the Placket-Burman design. Further optimization of these variables resulted in a 6.79-fold improvement in phytase production (7170 U/L) as compared to unoptimized medium. Supplementation of microbial phytases (fungal and bacterial) resulted in improved bioavailability of nutritional components with the concomitant liberation of inorganic phosphorus, reducing sugar, soluble protein and amino acids, thus mitigating anti-nutritional properties of phytic acid.

  17. Molecular and biochemical characteristics of recombinant β-propeller phytase from Bacillus licheniformis strain PB-13 with potential application in aquafeed.

    Science.gov (United States)

    Kumar, Vinod; Sangwan, Punesh; Verma, A K; Agrawal, Sanjeev

    2014-05-01

    Phytic acid is the major storage form of organic phosphorus in nature- and plant-based animal feed. It forms insoluble complexes with nutritionally important metals and proteins that are unavailable for monogastric or agastric animals. Phytases initiate the stepwise hydrolysis of phytic acid and release inorganic orthophosphate. In the present investigation, the phytase gene from a phytase producing Bacillus licheniformis strain PB-13 was successfully expressed in Escherichia coli BL21. Recombinant phytase 'rPhyPB13' was found to be catalytically active, with an activity of 0.97 U/mL and specific activity of 0.77 U/mg. The rPhyPB13 was purified to 14.10-fold using affinity chromatography. Similar to other β-propeller phytases, purified rPhyPB13 exhibited maximal activity at pH 6.0-6.5 and 60 °C in the presence of 1 mM Ca(2+) and was highly active over a wider pH range (pH 4.0-8.0) and high temperature (80 °C). It has shown maximum activity towards Na-phytate as substrate. The observed K m , V max and k cat of purified rPhyPB13 were 1.064 mM, 1.32 μmol/min/mg and 27.46 s(-1), respectively. PhyPB13 was resistant to trypsin inactivation, activated in presence of Ca(2+) and inhibited in presence of EDTA. Crude rPhyPB13 has good digestion efficiency for commercial feed and soybean meal. These results indicate that PhyPB13 is a β-propeller phytase that has application potential in aquaculture feed.

  18. Chromobacterium sp. from the tropics: detection and diversity of phytase activity

    Directory of Open Access Journals (Sweden)

    Patrícia S Costa

    2011-03-01

    Full Text Available Phytases are a group of enzymes that catalyze phytic acid hydrolysis with release of phosphorus (P. The ability of Chromobacterium sp. to produce phytase was detected in 115 out of 118 candidate bacteria isolated from different Brazilian biomas. This is the first report revealing the genus Chromobacterium as phytase producer.

  19. Enzymatic Comparisons of Aspergillus niger PhyA and Escherichia coli AppA2 Phytases

    Science.gov (United States)

    This study was to compare three phytase activity assays and kinetics of Aspergillus niger PhyA and Escherichia coli AppA2 phytases expressed in Pichia pastoris at the observed stomach pH of 3.5. In Experiment 1, equivalent phytase activities in the crude preparations of PhyA and AppA2 were tested ...

  20. Research Progress and Safety Evaluation of Phytase Transgenic Maize%转植酸酶基因玉米的研究与安全评价

    Institute of Scientific and Technical Information of China (English)

    熊建文; 彭端; 覃晓娟; 王继瑞

    2011-01-01

    Phytase is one of most widespread feed additives, which exists widely in nature. It can enhance the phosphorus and mineral absorption in monogastric animals, and it is significant for reducing the level of phosphorus output in nature. Maize is the major ingredient of animal feed, and it may enhance cost or reduce efficiency to maize by adding phytase. Recently, with the development of gene engineering technique, many phytase genes from fungus have been cloned and successfully expressed in the microorganisms and plants. In this review, we mainly introduced the microorganism resource of phytase gene and the relative investigation achievements, expounded the methods of transgenic technology and the research progress of the transgenic maize plants expressing phytase gene. Further, we also evaluated the safety of phytase transgenic maize. According to this article, we hope that it would improve the quality of maize, reduce the cost of production and promote the application of phytase in feed industry in the future.%植酸酶是应用最广泛的饲料添加剂之一,它在自然界中广泛存在,能提高饲料中磷的利用率,对减轻动物高磷粪便导致的环境水域污染有着重要意义.玉米是动物饲料的主要原料,在玉米饲料中添加植酸酶易造成饲料成本升高和生产效率降低等问题.近年来随着基因工程技术的发展,已克隆出多个真菌植酸酶基因,并成功在微生物、植物系统中表达.本文主要介绍了植酸酶基因的微生物来源与相关研究成果,阐述了转基因技术的常用方法和转植酸酶基因玉米的研究进展,并详细评价了转植酸酶基因玉米的安全性.通过对转植酸酶基因玉米的深入探讨,对改善玉米的品质,降低成本及推广植酸酶在饲料工业中的应用具有重要意义.

  1. Effect of nonphytate phosphorus and phytase levels on broiler femur.

    Directory of Open Access Journals (Sweden)

    Luana Martins Schaly

    2009-03-01

    Full Text Available This experiment was carried out to evaluate the effect of nonphytate phosphorus (NPP and phytase levels on the weight, morphometry and weight/length index (WLI of broiler femurs at 21 and 42 days of age. One thousand, two hundred chicks were allocated in a completely randomized design and 4 x 3 factorial arrangement (NPP x phytase levels, with four replicates. The NPP levels, at each phase were of 0.45, 0.37, 0.29 and 0.21% in the initial phase, 0.41, 0.33, 0.25 and 0.17% in the growth phase, and of 0.37, 0.29, 0.21 and 0.13% in the final phase. The phytase levels used were 0, 500 and 1000U/kg of diet. At 21 and 42 days of age, 48 birds were sacrificed for femur collection. At 21 days, there was no effect (P > 0.05 of NPP x phytase interaction on bone parameters, but the NPP reduction decreased (P < 0.05 the weight, length and WLI of the femurs, and the inclusion of 500U/kg of phytase improved (P < 0.05 the weight and WLI of the bones. At 42 days of age, NPP x phytase interaction was significant (P < 0.05 for the weight and length, and birds fed diets with no phytase had femurs that were lighter and shorter when the lowest NPP levels were evaluated. However, the inclusion of 500 or 1000U/kg of phytase produced weights and lengths similar to those produced by treatment with recommended NPP levels, and the lower NPP levels evaluated caused a reduction (P < 0.05 in the diameter and WLI of femurs. It was concluded that diets with 0.29, 0.25 or 0.21% of NPP, with 500 U/kg of phytase, could be used with no negative effect on the femur quality in broilers from one to 42 days of age.

  2. Microbial Phytase and Phosphorus Utilization by Broiler Chickens

    Directory of Open Access Journals (Sweden)

    Martin Kliment

    2012-05-01

    Full Text Available The aim of study was to investigate the mathematical and statistical assesment of the micorbial 6-phytase efficacy on phosphorus utilization at broiler chickens Cobb 500. Broiler chickens fed commercial feed mixtures based on soyabean-maize meal. Each feed mixture was fed ad libitum to chickens in boxes in commercial poultry farm. The trial consited of three groups of broiler chickens, one control group (CG and two trial groups, in which were broiler chickens fed by feed mixtures with decreased phosphorus content (TG1 and with microbial 6-phytase (TG2. A body weight of chickens at the end of the trial (42 day was 1900.0 g compared with 1883,0 g (TG1 and 1827.0 g (CG with not statistically significant differences (P≥0.05. Phosphorus, calcium and magnesium content in blood serum of broiler chickens in every group was not staticstically significant (P≥0.05. Phosphorus content in broiler chickens excreta was most higher in in control group (4.2556 g/kg in comparison with trial group (2.0911 g/kg were was microbial 6-phytase added and in trial group (3.1851 g/kg were was phosphorus content in feed mixtures decreased. In addition we concluded that microbial 6-phytase. Phytase addition into feed mixtures has not negative effect on broiler chickens growth ability and health, and helped to better utilization of phytate phosphorus from feed mixtures in relation to excreted phosphorus.

  3. Characteristics of Recombinant Phytase (rSt-Phy) of the Thermophilic mold Sporotrichum thermophile and its applicability in dephytinizing foods.

    Science.gov (United States)

    Ranjan, Bibhuti; Singh, Bijender; Satyanarayana, T

    2015-12-01

    Sporotrichum thermophile produces very low titres of phytase (St-Phy) extracellularly, which is acidstable, thermostable, and protease insensitive with broad substrate specificity, and therefore, the gene encoding phytase (St-Phy) has been cloned and expressed in E. coli. The purified recombinant phytase (rSt-Phy) has the molecular mass of 55 kDa with Km and Vmax (calcium phytate), kcat and kcat/Km of 0.143 mM, 185.05 nmoles mg(-1)  s(-1), 5.1 × 10(3) s(-1), and 3.5 × 10(7) M(-1) s(-1), respectively. Mg(2+) and Ba(2+) display slight stimulatory effect on the enzyme, while it is inhibited by other ions to a varied extent. The enzyme is also inhibited by chaotropic agents (guanidinium hydrochloride, potassium iodide, and urea), Woodward's reagent K, and 2,3-butanedione but resistant to both pepsin and trypsin. The rSt-Phy is useful in dephytinization of tandoori and naan (unleavened flat Indian breads), and bread, liberating soluble inorganic phosphate that mitigates anti-nutrient effects of phytic acid.

  4. Phytate phosphorus utilization and intestinal phytase activity in laying hens

    Directory of Open Access Journals (Sweden)

    Alaeldein M. Abudabos

    2012-01-01

    Full Text Available The first aim of this study was to examine the effects of strain and age on phytate phosphorus (PP utilization and intestinal phytase activity (IPA in two strains of laying hens, Hy-line Brown (HB and Hy-line White W-36 (HW at 32, 52 and 72 weeks of age. A digestion trial was conducted using the indicator method and birds were sampled to measure IPA. The second aim was to examine the effect of feed grade exogenous phytase enzyme on total (TP, water soluble (WSP and phytate phosphorus (PP excretion in the HW strain fed varying levels of phosphorus. Hens were fed three concentrations of available P (AP: 0.2, 0.3 and 0.4%. Each level of P was supplemented with three levels of commercial feed grade exogenous phytase enzyme (0, 300, and 600 FTU/kg and the amount of TP, WSP and PP in excreta per 100 g of feed consumed was calculated. The HB retained more PP as compared to HW. Intestinal phytase activity showed a significant (P<0.01 age effect with the highest activity occurring at 32 weeks. There were significant differences in the amount of TP and SP excreted between birds receiving the 3 levels of phosphorus with 300 units phytase (P<0.01. The results of this study showed that layers are capable of utilizing PP, and that utilization is regulated by IPA and varies with age. Exogenous phytase improved PP utilization but it increased the amount of TP and WSP in excreta.

  5. Cloning and characterization of purple acid phosphatase phytases from wheat, barley, maize, and rice

    DEFF Research Database (Denmark)

    Dionisio, G.; Madsen, C. K.; Holm, P. B.;

    2011-01-01

    Barley (Hordeum vulgare) and wheat (Triticum aestivum) possess significant phytase activity in the mature grains. Maize (Zea mays) and rice (Oryza sativa) possess little or virtually no preformed phytase activity in the mature grain and depend fully on de novo synthesis during germination. Here...... type (OsPAPhy_b and ZmPAPhy_b, respectively) were identified. HvPAPhy_a and HvPAPhy_b1/b2 share 86% and TaPAPhya1/a2 and TaPAPhyb1/b2 share up to 90% (TaPAPhy_a2 and TaPAPhy_b2) identical amino acid sequences. despite of this, PAPhy_a and PAPhy_b isogenes are differentially expressed during grain...... development and germination. In wheat, it was demonstrated that a and b isogene expression is driven by different promoters (approximately 31% identity). TaPAPhy_a/b promoter reporter gene expression in transgenic grains and peptide mapping of TaPAPhy purified from wheat bran and germinating grains confirmed...

  6. Comparative Proteomics of Leaves from Phytase-transgenic Maize and the Non-transgenic Isogenic Variety

    Directory of Open Access Journals (Sweden)

    Yanhua Tan

    2016-08-01

    Full Text Available To investigate unintended effects in genetically modified crops (GMCs, a comparative proteomics analysis between the leaves of the phytase-transgenic maize and those of non-transgenic plants was performed by using two-dimensional gel electrophoresis and mass spectrometry. A total of 57 differentially expressed protein spots (DEPs were successfully identified, which represented 44 unique proteins. Functional classification of the identified unique proteins showed that these proteins were predominantly involved in carbohydrate transport and metabolism, followed by post-translational modification. KEGG pathway analysis revealed that most of the DEPs participated in carbon fixation in photosynthesis. Comparison of the changes in the protein and gene transcript levels of the identified unique proteins showed that most proteins had a similar pattern of changes between proteins and transcripts. Our results suggested that although some significant differences were observed, the proteomic patterns were not substantially altered between the leaves of phytase-transgenic maize and its non-transgenic isogenic type. Moreover, none of the DEPs was identified as a new toxic protein or an allergenic protein. The differences of proteome between the two kinds of maize leaves might be attributed to both genetic modification and hybrid influence.

  7. Contribution of intestinal- and cereal-derived phytase activity on phytate degradation in young broilers.

    Science.gov (United States)

    Morgan, N K; Walk, C L; Bedford, M R; Burton, E J

    2015-07-01

    There is little consensus as to the capability of poultry to utilize dietary phytate without supplemental phytase. Therefore, an experiment was conducted to examine the extent to which endogenous phytase of intestinal and cereal origin contributes to phytate degradation in birds aged 0 to 14 d posthatch. Ross 308 broilers (n = 720) were fed one of 4 experimental diets with differing dietary ingredient combinations and approximate total phytate levels of 10 g/kg, dietary phytase activity analyzed at 460 U/kg, dietary calcium (Ca) levels of 11 g/kg, and nonphytate-phosphorus (P) levels of 4 g/kg. Broiler performance, gizzard, duodenum, jejunum and ileum pH, Ca and P digestibility and solubility, amount of dietary phytate hydrolyzed in the gizzard, jejunum, and ileal digesta phytase activity were analyzed at d 4, 6, 8, 10, 12, and 14 posthatch. Intestinal endogenous phytase activity increased significantly (P Phytase activity and phytate hydrolysis continued to increase with age, with a greater phytase activity and associated increase in phytate hydrolysis and mineral utilization between d 10 and 12. Gizzard and jejunum Ca and P solubility and ileal Ca and P digestibility increased significantly (P phytase activity recovered in the ileum was approximately 45 U/kg. There were strong correlations between phytase activity measured in the ileum and phytate hydrolyzed in the gizzard (r = 0.905, P phytase activity is responsible for phytate-P hydrolysis in broilers.

  8. Yeast Acid Phosphatases and Phytases: Production, Characterization and Commercial Prospects

    Science.gov (United States)

    Kaur, Parvinder; Satyanarayana, T.

    The element phosphorus is critical to all life forms as it forms the basic component of nucleic acids and ATP and has a number of indispensable biochemical roles. Unlike C or N, the biogeochemical cycling of phosphorus is very slow, and thus making it the growth-limiting element in most soils and aquatic systems. Phosphohydrolases (e.g. acid phosphatases and phytases) are enzymes that break the C-O-P ester bonds and provide available inorganic phosphorus from various inassimilable organic forms of phosphorus like phytates. These enzymes are of significant value in effectively combating phosphorus pollution. Although phytases and acid phosphatases are produced by various plants, animals and micro organisms, microbial sources are more promising for the production on a commercial scale. Yeasts being the simplest eukaryotes are ideal candidates for phytase and phos-phatase research due to their mostly non-pathogenic and GRAS status. They have not, however, been utilized to their full potential. This chapter focuses attention on the present state of knowledge on the production, characterization and potential commercial prospects of yeast phytases and acid phosphatases.

  9. Interrelationships of glycosylation and aggregation kinetics for Peniophora lycii phytase

    DEFF Research Database (Denmark)

    Høiberg-Nielsen, R.; Fuglsang, C.C.; Arleth, L.

    2006-01-01

    The kinetics of thermally induced aggregation of the glycoprotein Peniophora lycii phytase (Phy) and a deglycosylated form (dgPhy) was studied by dynamic (DLS) and static (SLS) light scattering. This provided a detailed insight into the time course of the formation of small aggregates (similar...

  10. Regulation of Raoultella terrigena comb.nov. phytase expression.

    Science.gov (United States)

    Zamudio, Marcela; González, Aracely; Bastarrachea, Fernando

    2002-01-01

    Phytases catalyze the release of phosphate from phytate (myo-inositol hexakisphosphate) to inositol polyphosphates. Raoultella terrigena comb.nov. phytase activity is known to increase markedly after cells reach the stationary phase. In this study, phytase activity measurements made on single batch cultures indicated that specific enzyme activity was subject to catabolite repression. Cyclic AMP (cAMP) showed a positive effect in expression during exponential growth and a negative effect during stationary phase. RpoS exhibited the opposite effect during both growth phases; the induction to stationary phase decreased twofold in the rpoS::Tn10 mutant, but the effect of RpoS was not clearly determined. Two phy::MudI1734 mutants, MW49 and MW52, were isolated. These formed small colonies in comparison with the MW25 parent strain when plated on Luria-Bertani (LB) or LB supplemented with glucose. They did not grow in minimal media or under anaerobiosis, but did grow aerobically on LB and LB glucose at a lower rate than did MW25. The beta-galactosidase activity level in these mutants increased three to four fold during stationary growth in LB glucose and during anaerobiosis. Addition of cAMP during the exponential growth of MW52 on LB glucose provoked a decrease in beta-galactosidase activity during the stationary phase, confirming its negative effect on phytase expression during stationary growth.

  11. Stereospecificity of inositol hexakisphosphate dephosphorylation by Paramecium phytase

    NARCIS (Netherlands)

    Kaay, Jeroen van der; Haastert, Peter J.M. van

    1995-01-01

    InsP(6) is an abundant compound in many micro-organisms, plants and animal cells. Its function and route of synthesis are still largely unknown. Degradation of InsP(6) is mediated by phytase, which in most organisms dephosphorylates InsP(6) in a relatively non-specific way. In the micro-organism Par

  12. Ethanol tolerance in Aspergillus niger and Escherichia coli phytase

    Science.gov (United States)

    The expanded use of corn and other grain for biofuels have created an increased supply of dried grains with soluble (DDGS) and other byproducts of ethanol fermentation. Elevated levels of phytic acid in this DDGS indicate that ethanol is denaturing the native phytase produced by the yeast, Saccharo...

  13. Effect of exogenous phytase on degradation of inositol phosphate in dairy cows

    DEFF Research Database (Denmark)

    Brask-Pedersen, Dorte Buus; Glitsø, Lene Vibe; Skov, L.K.

    2013-01-01

    The effect of exogenous phytase on inositol phosphate degradation in the rumen of dairy cows was investigated in a 4 × 4 Latin square design. Four lactating Danish Holstein cows fitted with ruminal, duodenal, and ileal cannulas were offered a total mixed ration (TMR) with a high content of inositol...... phosphate and supplemented with 1 of 4 concentrations of phytase [none, low, medium, or high, corresponding to 23, 2,023, 3,982, and 6,015 phytase units/kg of dry matter (DM)]. Exogenous phytase lead to a higher rumen pool of phytase. Inositol phosphate content in digesta samples from rumen, duodenum, ileum......, and feces was almost entirely composed of myo-inositol hexakisphosphate (InsP6), indicating that degradation of this compound is the rate-limiting step in inositol phosphate degradation in the digestive tract. Ruminal and total-tract degradations of InsP6 were higher when exogenous phytase was added...

  14. Variation in activity of root extracellular phytase between genotypes of barley

    DEFF Research Database (Denmark)

    Asmar, Mohammad Farouq

    1997-01-01

    Barley genotypes grown in nutrient solution under P nutrient stress and sterile conditions were compared in activity of root-associated and root-released extracellular phytase. The activity of root-associated phytase of all genotypes was about 10 times higher than that of root-released phytase...... and the genotypes performed differently with regard to the activity of the enzymes. The winter barley genotype, Marinka had the highest activity of root-associated extracellular phytase which differed significantly from Alexis and Senate, but not from Regatta. Alexis showed the lowest activity of root......-released extracellular phytase which differed significantly from those of Marinka and Regatta, but not from Senate. Generally, there was a significant correlation between the activity of root-associated and released extracellular phytase....

  15. SCREENING OF THERMOPHYLIC MICROORGANISM FROM IJEN CRATER BANYUWANGI AS PHYTASE ENZYME PRODUCER

    OpenAIRE

    Kusumadjaja, Aline Puspita; Budiati, Tutuk; Puspaningsih, Ni Nyoman Tri; Sajidan, Sajidan

    2010-01-01

    Phytase is enzyme which hydrolysis phytic acid to anorganic phosphate and myo-inositol pentakis-, tetrakis-, tris-, bis-, and monophosphate. The use of phytase in feed industry can overcome environment and nutrition problems which were arisen from unmetabolism phytic acid or its salt by poultry, swine and fish. The feed industry needs a thermostable enzyme due to the need of high temperature in pelleting process, i.e. 81 °C. By using thermostabile phytase, the pelleting process will not affec...

  16. ADOPTING SELECTED HYDROGEN BONDING AND IONIC INTERACTIONS FROM ASPERGILLUS FUMIGATUS PHYTASE STRUCTURE IMPROVES THE THERMOSTABILITY OF ASPERGILLUS NIGER PHYA PHYTASE

    Science.gov (United States)

    Although it has been widely used as a feed supplement to reduce manure phosphorus pollution of swine and poultry, Aspergillus niger PhyA phytase is unable to withstand heat inactivation during feed pelleting. Crystal structure comparisons with its close homolog, the thermostable Aspergillus fumigatu...

  17. Extracellular expression of alkaline phytase in Pichia pastoris: Influence of signal peptides, promoters and growth medium

    Directory of Open Access Journals (Sweden)

    Mimi Yang

    2015-06-01

    Full Text Available Alkaline phytase isolated from pollen grains of Lilium longiflorum (LlALP possesses unique catalytic and thermal stability properties that suggest it has the potential to be used as a feed supplement. However, substantial amounts of active enzymes are needed for animal feed studies and endogenous levels of LlALP in lily pollen are too low to provide the required amounts. Active rLlALP2 (coded by LlAlp2, one of two isoforms of alkaline phytase cDNA identified in lily pollen has been successfully expressed in intracellular compartments of Pichia pastoris, however enzyme yields have been modest (25–30 mg/L and purification of the enzyme has been challenging. Expression of foreign proteins to the extracellular medium of P. pastoris greatly simplifies protein purification because low levels of endogenous proteins are secreted by the yeast. In this paper, we first describe the generation of P. pastoris strains that will secrete rLlALP2 to the extracellular medium. Data presented here indicates that deletion of native signal peptides at the N- and C-termini of rLlALP2 enhanced α-mating factor (α-MF-driven secretion by four-fold; chicken egg white lysozyme signal peptide was ineffective in the extracellular secretion of rLlALP2. Second, we describe our efforts to increase expression levels by employing a constitutive promoter from the glyceraldehyde-3-phosphate dehydrogenase gene (PGAP in place of the strong, tightly controlled promoter of alcohol oxidase 1 gene (PAOX1. PGAP enhanced the extracellular expression levels of rLlALP2 compared to PAOX1. Finally, we report on the optimization of the culture medium to enhance yields of rLlALP2. The strength of PGAP varies depending on the carbon source available for cell growth; secreted expression of rLlALP2 was highest when glycerol was the carbon source. The addition of histidine and Triton X-100 also enhanced extracellular expression. Taken together, the employment of PGAP under optimized culture

  18. 转植酸酶玉米生育期及秸秆还田期根际微生物数量和细菌菌群多样性%Quantity of Microorganism and Diversity of Bacterial Community in Rhizosphere of Transgenic Maize with Phytase Gene in Growth and Straw-returning Periods

    Institute of Scientific and Technical Information of China (English)

    张林森; 杨正友; 孙红炜; 李凡; 杨淑珂; 段丽君; 路兴波

    2013-01-01

    在大田栽培条件下,以转植酸酶玉米10TPY005、非转植酸酶亲本玉米为试材,于2011年在玉米的不同生育期以及秸秆还田期对玉米根际土壤细菌、放线菌和真菌数量的变化情况进行了测定.结果表明:在同一时期同一种微生物数量在转植酸酶玉米、非转基因玉米及空白对照中基本相同;不同细菌菌群在不同时期内数量变化情况不同:氨化细菌的数量在生育期内明显高于秸秆还田期,好气性纤维素分解菌及好气性固氮菌的数量在生育期内明显低于秸秆还田期;整个生育期和秸秆还田期转植酸酶玉米3种群落特征参数(优势集中性、群落多样性、均匀度指数)均高于非转植酸酶玉米,同时生育期内3种生物群落特征参数也明显高于秸秆还田期.%The transgenic maize 10TPY005 with phytase gene and its non - transgenic parent were grown in field to study the quantitative changes of bacteria, actinomycetes and fungi in maize rhizosphere at different growth stages and straw - returning stage in 2011. The diversity of the same microorganism at the same stage of transgenic maize was similar to that of non - transgenic maize and blank control, but the diversity of different communities varied at different stages. The quantity of ammonifying bacteria was significantly higher in growth period than that in straw - returning period, however, the quantities of aerobic cellulose - decomposing bacteria and aerobic nitrogen fixing bacteria were lower in growth period. In the whole growth period and straw - returning period, the characteristic parameters (Simpson index, evenness index and the Shannon - Wiener index) of the three populations were higher in transgenic maize than those in non - transgenic maize. And the three parameters in growth period were also obviously higher than those in straw - returning period.

  19. Effect of phytase supplementation on growing pigs performance

    Directory of Open Access Journals (Sweden)

    Šefer D.

    2012-01-01

    Full Text Available Dietary concentrations of phytate are crucial for its anti-nutritive properties and its negative impact on P availability. The increase of dietary phytate level is shown to increase endogenous losses of amino acids and minerals in pigs. The partial availability of the P component of phytate to simple-stomached species attains importance as the world's rock phosphate reserves are not renewable, which could lead to a P supply crisis in the future. Supplementing phytase is becoming increasingly common as a method to improve the availability of P in plant ingredients containing high levels of phytate P. Fourty-eight pigs (Swedish Landrace boars × Dutch landrace sows weaned at day 35 with an initial BW of 8.72±0.28 kg were used for a 40-day weaner performance study. The study was structured as a complete randomized design to evaluate the response of weaner pigs to four concentrations of microbial phytase produced by Aspergillus niger: (T1 basal diet; (T2 basal diet + 1000 FTU/kg; (T3 diet with decreased dicalcium phosphate + 1000 FTU/kg; and (T4 diet with no dicalcium phosphate + 1000 FTU/kg. Control group of piglets (T1 has achieved standard body mass while addition of phytase in meal increased body mass by 6.59% in T2 and 7.52% in T3. Phytase supplementation prevented decreased body weight gain diets where available phosphorous level was reduced by 50. The amount of consumed feed per day was not significantly different. Feed efficiency of T2 and T3 groups was by 3.23% better and of T4 for 11.29% lower compared to the control group of piglets (T1. Lower production results achieved by the use of low phosphorous diets can be avoided to a certain level by the use of microbial phytase. The use of phytase in pig diet significantly improved phosphorous availability, as well as of other mineral substances from the phytate complex. [Projekat Ministarstva nauke Republike Srbije, br. III46002

  20. Characterization of phytase activity from cultivated edible mushrooms and their production substrates.

    Science.gov (United States)

    Collopy, Patrick D; Royse, Daniel J

    2004-12-15

    Phytase is used commercially to maximize phytic acid degradation and to decrease phosphorus levels in poultry and swine manure. To determine phytase content in edible mushrooms, basidiomata of Agaricus bisporus and three specialty mushrooms (Grifola frondosa, Lentinula edodes, and Pleurotus cornucopiae) and spent mushroom substrate (SMS) were surveyed. Enzyme activity ranged from 0.046 to 0.074 unit/g of tissue for four A. bisporus types (closed and open whites and closed and open browns) grown at The Pennsylvania State University's Mushroom Test Demonstration Facility (MTDF). The addition of various nutrient supplements to phase II mushroom production substrate did not alter phytase activity in A. bisporus. Portabella mushrooms (open brown) obtained from a commercial farm had significantly higher levels of phytase activity (0.211 unit/g of tissue) compared to A. bisporus grown at the MTDF. Of the specialty mushrooms surveyed, maitake (G. frondosa) had 20% higher phytase activity (0.287 unit/g of tissue) than commercial portabella mushrooms. The yellow oyster mushroom (P. cornucopiae) ranked second in level of phytase activity (0.213 unit/g of tissue). Shiitake (L. edodes) contained the least amount of phytase in basidiomata (0.107 unit/g of tissue). Post-crop steam treatment (60 degrees C, 24 h) of SMS reduced phytase activity from 0.074 to 0.018 unit/g. Phytase was partially purified from commercially grown portabella basidiomata 314-fold with an estimated molecular mass of 531 kDa by gel filtration chromatography. The optimum pH for activity was 5.5, but appreciable phytase activity was observed over the range of pH 5.0-8.0. Partially purified A. bisporus phytase was inactivated following a 10-min incubation at > or =60 degrees C.

  1. Enhanced submerged Aspergillus ficuum phytase production by implementation of fed-batch fermentation.

    Science.gov (United States)

    Coban, Hasan B; Demirci, Ali

    2014-12-01

    Phytase is an important feed and food additive, which is both used in animal and human diets. Phytase has been used to increase the absorption of several divalent ions, amino acids, and proteins in the bodies and to decrease the excessive phosphorus release in the manure to prevent negative effects on the environment. To date, microbial phytase has been mostly produced in solid-state fermentations with insignificant production volumes. There are only a few studies in the literature that phytase productions were performed in submerged bench-top reactor scale. In our previous studies, growth parameters (temperature, pH, and aeration) and important fermentation medium ingredients (glucose, Na-phytate, and CaSO4) were optimized. This study was undertaken for further enhancement of phytase production with Aspergillus ficuum in bench-top bioreactors by conducting fed-batch fermentations. The results showed that addition of 60 g of glucose and 10 g of Na-phytate at 96 h of fermentation increased phytase activity to 3.84 and 4.82 U/ml, respectively. Therefore, the maximum phytase activity was further enhanced with addition of glucose and Na-phytate by 11 and 40 %, respectively, as compared to batch phytase fermentations. It was also reported that phytase activity increased higher in early log stage additions than late log stage additions because of higher microbial activity. In addition, the phytase activity in fed-batch fermentation did not drop significantly as compared to the batch fermentation. Overall, this study shows that fungal phytase can be successfully produced in submerged fed-batch fermentations.

  2. Molecular and biochemical characteristics of β-propeller phytase from marine Pseudomonas sp. BS10-3 and its potential application for animal feed additives.

    Science.gov (United States)

    Nam, Seung-Jeung; Kim, Young-Ok; Ko, Tae-Kyung; Kang, Jin-Ku; Chun, Kwang-Hoon; Auh, Joong-Hyuck; Lee, Chul-Soon; Lee, In-Kyu; Park, Sunghoon; Oh, Byung-Chul

    2014-10-01

    Phytate is an antinutritional factor that impacts the bioavailability of essential minerals such as Ca(2+), Mg(2+), Mn(2+), Zn(2+), and Fe(2+) by forming insoluble mineral-phytate salts. These insoluble mineral-phytate salts are hydrolyzed rarely by monogastric animals, because they lack the hydrolyzing phytases and thus excrete the majority of them. The β-propeller phytases (BPPs) hydrolyze these insoluble mineral-phytate salts efficiently. In this study, we cloned a novel BPP gene from a marine Pseudomonas sp. This Pseudomonas BPP gene (PsBPP) had low sequence identity with other known phytases and contained an extra internal repeat domain (residues 24-279) and a typical BPP domain (residues 280-634) at the C-terminus. Structurebased sequence alignment suggested that the N-terminal repeat domain did not possess the active-site residues, whereas the C-terminal BPP domain contained multiple calcium-binding sites, which provide a favorable electrostatic environment for substrate binding and catalytic activity. Thus, we overexpressed the BPP domain from Pseudomonas sp. to potentially hydrolyze insoluble mineral-phytate salts. Purified recombinant PsBPP required Ca(2+) or Fe(2+) for phytase activity, indicating that PsBPP hydrolyzes insoluble Fe(2+)-phytate or Ca2+-phytate salts. The optimal temperature and pH for the hydrolysis of Ca(2+)-phytate by PsBPP were 50°C and 6.0, respectively. Biochemical and kinetic studies clearly showed that PsBPP efficiently hydrolyzed Ca(2+)-phytate salts and yielded myo-inositol 2,4,6-trisphosphate and three phosphate groups as final products. Finally, we showed that PsBPP was highly effective for hydrolyzing rice bran with high phytate content. Taken together, our results suggest that PsBPP has great potential in the animal feed industry for reducing phytates.

  3. 农杆菌介导植酸酶基因转化大豆的研究%Transformation of Phytase Gene into Soybean via Agrobacterium-mediated Method

    Institute of Scientific and Technical Information of China (English)

    刘琦; 夏善勇; 张军; 段忠卫; 刘鑫磊; 李希臣

    2011-01-01

    以大豆胚尖为受体,利用农杆菌介导法将来源于泡盛曲霉的植酸酶基因phy转入黑农37和吉林小粒豆中.此法取材方便,操作简单,产生嵌合体的可能件低.在筛选培养基中经草胺膦(1.0 mg·L-1)筛选,获得抗性植侏.对获得的草胺磷抗性植株采用除草剂(10%Basta)筛查表型鉴定与目的基因、筛选标记基因PCR分子检测相结合的方法,检测结果直观、可信度高.最终获得5株阳性植株,转化率为0.5%.%The Phy gene cloned from Aspergillus awamori was transformed into soybean varieties Heinong37 and Jilinxiaoli soybean through Agrobacterium-mediated method with soybean embryonic tips as acceptors. The method has advantage of easy operation and lower chimera rate. The regenerated plants were screened by glufosinate-ammonium( 1.0 mg · L-1 )of medium.The regenerated plantlets resistant to glufosinate-ammonium were detected by combining the PCR analysis of the target gene as well as selective marker gene and the herbicide screening. The result was intuitive and credible. Finally, 5 positive plants were obtained, and the transformation efficiency reached 0. 5%.

  4. Effect of ph and temperature on the activity of phytase products used in broiler nutrition

    Directory of Open Access Journals (Sweden)

    L de P Naves

    2012-09-01

    Full Text Available The activity of three commercial microbial phytase (Aspergillus oryzae, A. niger, and Saccharomyces cerevisae products used in broiler nutrition was determined at different pH (2.0 to 9.0 and temperature (20 to 90°C values. Enzymatic activity was determined according to the reaction of the phytase with its substrate (sodium phytate, in four replicates, and was expressed in units of phytase activity (FTU. A. oryzae phytase exhibited optimal activity at pH 4.0 and 40°C, but its absolute activity was the lowest of the three phytases evaluated. A. niger phytase exhibited maximal activity close to pH 5.0 and 45ºC, whereas S. cerevisae phytase presented its highest activity at pH close to 4.5 and temperatures ranging between 50 and 60°C. It was concluded that A. niger and S. cerevisae phytase products exhibited the highest absolute activities in vitro at pH and temperature values (pH lower than 5.0 and 41ºC corresponding to the ideal physiological conditions of broilers, which would theoretically allow high hydrolysis rate of the phytate contained in the feed.

  5. The efficacy of phytase in corn-soybean meal-based diets for laying hens

    NARCIS (Netherlands)

    Klis, van der J.D.; Versteegh, H.A.J.; Simons, P.C.M.; Kies, A.K.

    1997-01-01

    Microbial phytase hydrolyzes poorly degradable vegetable phytate P in the gastrointestinal tract of poultry; thereby increasing the availability of organic P to an extent that remains to be established. For this purpose, the P equivalency value of phytase in cornsoybean meal layer diets was assessed

  6. Kinetic Characterization of O-Phospho-L-Tyrosine Phosphohydrolase Activity of Two Fungal Phytases.

    Science.gov (United States)

    Fungal phytases belonging to 'Histidine Acid Phosphatase' or HAP class of phosphomonoesterase that catalyzes the hydrolysis of phytic acid could also hydrolyze O-phospho-tyrosine. Two phytases from Aspergillus niger and Aspergillus awamori with pH optima 2.5 were tested for phospho-tyrosine hydrola...

  7. Improvement of phosphorus availability by microbial phytase in broilers and pigs

    NARCIS (Netherlands)

    Simons, P.C.M.; Versteegh, H.A.J.; Jongbloed, A.W.; Kemme, P.A.; Slump, P.; Bos, K.D.; Wolters, M.G.E.; Beudeker, R.F.; Verschoor, G.J.

    2005-01-01

    Techniques have been developed to produce microbial phytase for addition to diets for simple-stomached animals, with the aim to improve phosphorus availability from phytate-P in plant sources. The activityof the crude microbial phytase showed pH optima at pH 5-5 and 2·5. The enzyme was able to degra

  8. Phytase studies in pigs and poultry: effect on protein digestion and energy utilization

    NARCIS (Netherlands)

    Kies, A.K.

    2005-01-01

    Phytase is applied for improving digestibility of phosphorus in pig and poultry diets. Indepen-dently, phytase also improves animal performance. The mechanisms to explain this effect were investigated and quantified. Protein can be complexed with phytate, especially under the acid conditions that oc

  9. Hydrolysis of organic phosphates by commercially available phytases: Biocatalytic potentials and ion on their enzymatic activities

    Science.gov (United States)

    Commercially available phytases from wheat and the fungus Aspergillus ficuum have been used in environmental and agricultural phosphorus research. In order to better understand the biochemical properties of these two phytases, in vitro experiments were conducted to study their catalytic potentials...

  10. Effects of phytase supplementation on phosphorus retention in broilers and layers: A meta-analysis

    NARCIS (Netherlands)

    Bougouin, A.; Appuhamy, J.A.D.R.N.; Kebreab, E.; Dijkstra, J.; Kwakkel, R.P.; France, J.

    2014-01-01

    Phytase, a widely used feed additive in poultry diets, increases P availability and subsequently reduces inorganic-P supplementation and P-excretion. Phytase supplementation effect on P-retention in poultry has been investigated, but the effect sizes were highly variable. The present study’s objecti

  11. Different site-specific N-glycan types in wheat (Triticum aestivum L.) PAP phytase

    DEFF Research Database (Denmark)

    Dionisio, G.; Brinch-Pedersen, H.; Welinder, Karen Gjesing

    2011-01-01

    Phytase activity in grain is essential to make phosphate available to cell metabolism, and in food and feed. Cereals contain the purple acid phosphatase type of phytases (PAPhy). Mature wheat grain is dominated by TaPAPhy_a which, in the present work, has been characterized by extensive peptide...

  12. Improving the neutral phytase activity from Bacillus amyloliquefaciens DSM 1061 by site-directed mutagenesis.

    Science.gov (United States)

    Xu, Wei; Shao, Rong; Wang, Zupeng; Yan, Xiuhua

    2015-03-01

    Neutral phytase is used as a feed additive for degradation of anti-nutritional phytate in aquatic feed industry. Site-directed mutagenesis of Bacillus amyloliquefaciens DSM 1061 phytase was performed with an aim to increase its activity. Mutation residues were chosen based on multiple sequence alignments and structure analysis of neutral phytsaes from different microorganisms. The mutation sites on surface (D148E, S197E and N156E) and around the active site (D52E) of phytase were selected. Analysis of the phytase variants showed that the specific activities of mutants D148E and S197E remarkably increased by about 35 and 13% over a temperature range of 40-75 °C at pH 7.0, respectively. The k cat of mutants D148E and S197E were 1.50 and 1.25 times than that of the wild-type phytase, respectively. Both D148E and S197E showed much higher thermostability than that of the wild-type phytase. However, mutants N156E and D52E led to significant loss of specific activity of the enzyme. Structural analysis revealed that these mutations may affect conformation of the active site of phytase. The present mutant phytases D148E and S197E with increased activities and thermostabilities have application potential as additives in aquaculture feed.

  13. Microparticle-enhanced Aspergillus ficuum phytase production and evaluation of fungal morphology in submerged fermentation.

    Science.gov (United States)

    Coban, Hasan B; Demirci, Ali; Turhan, Irfan

    2015-06-01

    Phytase can be used in animal's diets to increase the absorption of several divalent ions, amino acids and proteins and to decrease the excessive phosphorus release in manure to prevent negative effects on the environment. This study aimed to enhance the current submerged fungal phytase productions with a novel fermentation technique by evaluating the effect of the various microparticles on Aspergillus ficuum phytase production. It was observed that microparticles prevented bulk fungal pellet growth, decreased average fungal pellet size and significantly increased phytase activity in the submerged fermentation. Microbial structure imaging results showed that the average fungal pellet radius decreased from 800 to 500 and 200 µm by addition of 15 g/L aluminum oxide and talcum, respectively, in shake-flask fermentation. Also, addition of 15 g/L of talcum and aluminum oxide increased phytase activity to 2.01 and 2.93 U/ml, respectively, compared to control (1.02 U/ml) in shake-flask fermentation. Additionally, phytase activity reached 6.49 U/ml within 96 h of fermentation with the addition of 15 g/L of talcum, whereas the maximum phytase activity was only 3.45 U/ml at 120 h of fermentation for the control in the 1-L working volume bioreactors. In conclusion, microparticles significantly increased fungal phytase activity and production yield compared to control fermentation.

  14. Characteristics Of Phytases From Soybeans And Microorganisms Involved In The Tempe Production

    Directory of Open Access Journals (Sweden)

    K A Buckle2

    2004-12-01

    Full Text Available Tempe was prepared from Forest variety of soybeans inoculated with pure culture of the mould Rhizopus oligosporus strain CT11K2. Partially purified phytases from soybeans, mould of the R. oligosporus strain CT11K2, yeasts (Endomycopsis burtonii, Candida diddensiae and Candida tropicalis and bacteria (Streptococcus faecium and Streptococcus dysgalactiae involved during tempe production were prepared according to procedure developed by Sutardi (1988. The crude phytases were characterized and phytic acid, inorganic and total phosphorous content of soybeans, intermediate products and tempe, were also determined.Results show that phytases, produced by microorganisms involved in tempe production, and by endogenous soybean phytase, had significantly different characteristics especially on optimum pH, temperature and substrate, Km and Vmax values, inactivation and activation energy and also temperature coefficient. All phytases contribute to the reduction in phytic acid content; and mould phytase, especially the extra-cellular enzyme, and the endogenous soybean phytase, showed the dominant effect on phytic acid degradation compared with other microbial phytases and physical treatments such as soaking, boiling and steaming.Phytic acid content of soybeans was reduced by more than 50% from about 10.4 to about 4.9 - 5.0 mg/g (DWB in fresh tempe (after 40 h fermentation at 30oC

  15. Isolation of a phytase with distinctive characteristics from an edible mushroom, Pleurotus eryngii.

    Science.gov (United States)

    Li, Miao; Wang, Hexiang; Ng, Tzi Bun

    2013-04-01

    From the fresh fruiting bodies of the mushroom Pleurotus eryngii a phytase with a molecular mass of 14 kDa was isolated. The isolation protocol entailed ion exchange chromatography on DEAE-cellulose and CM-cellulose, affinity chromatography on Affi-gel blue gel, and ion exchange chromatography on Q-Sepharose. The phytase was unadsorbed on DEAE-cellulose, CM-cellulose and Affi-gel blue gel, and adsorbed on Q-Sepharose. It appeared as a single band in SDSPAGE. It exhibited maximal activity at around 37°C. Its activity underwent little changes over the range of pH 3.0 to 9.0. The aforementioned characteristics are different from those of animal, plant and bacterial phytases. The low molecular mass and pH stability of P. eryngii phytase also distinguish it from mushroom phytases and other fungal phytases reported earlier. The purified enzyme exhibited a broad substrate specificity on a range of phosphorylated compounds, and the phytase demonstrated the N-terminal sequence ADNVYRHDNN which shows little homology to known phytases. It inhibited proliferation of human nasopharyngeal carcinoma CNE2 cells, hepatoma HepG2 cells and breast cancer MCF7 cells with an IC(50) of 1.9 μM, 2.9 μM, and 1.0 μM, respectively.

  16. Tethered phytic acid as a probe for measuring phytase activity.

    Science.gov (United States)

    Berry, Duane F; Berry, David A

    2005-06-15

    A novel approach for measuring phytase activity is presented. We have developed a new chromophoric substrate analog of phytic acid, 5-O-[6-(benzoylamino)hexyl]-d-myo-inositol-1,2,3,4,6-pentakisphosphate that permits direct measurement of the phosphate ester bond-cleavage reaction using HPLC. This compound, along with its dephosphorylated T-phosphatidylinositol intermediates, are quantified using reversed phase chromatography with UV detection.

  17. Identification and characterization of a phytase of potential commercial interest.

    Science.gov (United States)

    Casey, Anne; Walsh, Gary

    2004-06-10

    Phytases catalyse the hydrolytic degradation of phytic acid and its salts and are added to monogastric animal feed to ameliorate the negative environmental and nutritional consequences of dietary phytate. Screening of 58 microbial strains identified a phytase produced by Rhizopus oligosporus ATCC 22959 that displayed physicochemical characteristics likely to render it of potential industrial interest. The 124 kDa enzyme was purified to homogeneity by anion exchange chromatography, gel filtration and chromatofocusing. The monomeric glycosylated enzyme (30.5% total carbohydrate) displayed maximum activity at 65 degrees C and pH 5.0. It displayed a Km of 10.4 microM, a Vmax of 1.32 nmols(-1) and a Kcat of 51 s(-1). It is acid tolerant, retaining full activity after incubation at pH 2.0 for 6h. HPLC analysis indicated the enzyme's ability to almost completely degrade phytate. Substrate specificity studies showed its ability to dephosphorylate several additional phosphorylated molecules. Activity was unaffected or moderately stimulated by a range of metal ions with only Ca2+ exerting a modest (13%) inhibitory effect. The enzyme is significantly more thermostable at 80 degrees C and retains a significantly greater proportion of maximal activity at physiological temperatures than do two commercial phytases tested for comparative purposes. This may render it of industrial interest.

  18. Phosphorus digestibility response of growing pigs to phytase supplementation of triticale distillers' dried grains with solubles.

    Science.gov (United States)

    Xue, P C; Adeola, O

    2015-02-01

    An experiment was conducted in growing pigs to determine the true total-tract digestibility (TTTD) of P in triticale distillers' dried grains with solubles (DDGS) with or without phytase using the regression method. Six diets were formulated in a 3 × 2 factorial arrangement, including 3 levels of triticale DDGS (300, 400, or 500 g/kg) and phytase (0 or 500 phytase units [FTU]/kg of diet). A total of 48 barrows (initial BW 22.2 ± 1.3 kg) were assigned to the 6 diets in a randomized complete block design. There was a 5-d adjustment period followed by a 5-d total collection of feces. The results show that P intake, fecal P output, and digested P increased linearly ( phytase on apparent total-tract digestibility (ATTD) of P. In diets without added phytase, the ATTD of P in triticale DDGS was 65.0, 67.7, and 63.2% for the diets with 300, 400, and 500 g/kg triticale DDGS, respectively; the corresponding values for diets with added phytase were 77.3, 76.3, and 75.7%. By regressing daily digested P against daily P intake, the TTTD of P was estimated at 75.4% for triticale DDGS or 81.1% with added phytase, respectively. In conclusion, the TTTD of P in triticale DDGS without supplemental phytase was 75.4%, and it was 81.1% in the presence of phytase at 500 FTU/kg of the diet, but the difference was not statistically significant. For triticale DDGS, the supplementation of 500 FTU/kg phytase in diet could increase the ATTD of P ( < 0.001) but not the TTTD of P.

  19. Novel Glucose-1-Phosphatase with High Phytase Activity and Unusual Metal Ion Activation from Soil Bacterium Pantoea sp. Strain 3.5.1.

    Science.gov (United States)

    Suleimanova, Aliya D; Beinhauer, Astrid; Valeeva, Liia R; Chastukhina, Inna B; Balaban, Nelly P; Shakirov, Eugene V; Greiner, Ralf; Sharipova, Margarita R

    2015-10-01

    Phosphorus is an important macronutrient, but its availability in soil is limited. Many soil microorganisms improve the bioavailability of phosphate by releasing it from various organic compounds, including phytate. To investigate the diversity of phytate-hydrolyzing bacteria in soil, we sampled soils of various ecological habitats, including forest, private homesteads, large agricultural complexes, and urban landscapes. Bacterial isolate Pantoea sp. strain 3.5.1 with the highest level of phytase activity was isolated from forest soil and investigated further. The Pantoea sp. 3.5.1 agpP gene encoding a novel glucose-1-phosphatase with high phytase activity was identified, and the corresponding protein was purified to apparent homogeneity, sequenced by mass spectroscopy, and biochemically characterized. The AgpP enzyme exhibits maximum activity and stability at pH 4.5 and at 37°C. The enzyme belongs to a group of histidine acid phosphatases and has the lowest Km values toward phytate, glucose-6-phosphate, and glucose-1-phosphate. Unexpectedly, stimulation of enzymatic activity by several divalent metal ions was observed for the AgpP enzyme. High-performance liquid chromatography (HPLC) and high-performance ion chromatography (HPIC) analyses of phytate hydrolysis products identify dl-myo-inositol 1,2,4,5,6-pentakisphosphate as the final product of the reaction, indicating that the Pantoea sp. AgpP glucose-1-phosphatase can be classified as a 3-phytase. The identification of the Pantoea sp. AgpP phytase and its unusual regulation by metal ions highlight the remarkable diversity of phosphorus metabolism regulation in soil bacteria. Furthermore, our data indicate that natural forest soils harbor rich reservoirs of novel phytate-hydrolyzing enzymes with unique biochemical features.

  20. Effect of citrate on Aspergillus niger phytase adsorption and catalytic activity in soil

    Science.gov (United States)

    Mezeli, Malika; Menezes-Blackburn, Daniel; Zhang, Hao; Giles, Courtney; George, Timothy; Shand, Charlie; Lumsdon, David; Cooper, Patricia; Wendler, Renate; Brown, Lawrie; Stutter, Marc; Blackwell, Martin; Darch, Tegan; Wearing, Catherine; Haygarth, Philip

    2015-04-01

    Current developments in cropping systems that promote mobilisation of phytate in agricultural soils, by exploiting plant-root exudation of phytase and organic acids, offer potential for developments in sustainable phosphorus use. However, phytase adsorption to soil particles and phytate complexion has been shown to inhibit phytate dephosphorylation, thereby inhibiting plant P uptake, increasing the risk of this pool contributing to diffuse pollution and reducing the potential benefits of biotechnologies and management strategies aimed to utilise this abundant reserve of 'legacy' phosphorus. Citrate has been seen to increase phytase catalytic efficiency towards complexed forms of phytate, but the mechanisms by which citrate promotes phytase remains poorly understood. In this study, we evaluated phytase (from Aspergillus niger) inactivation, and change in catalytic properties upon addition to soil and the effect citrate had on adsorption of phytase and hydrolysis towards free, precipitated and adsorbed phytate. A Langmuir model was fitted to phytase adsorption isotherms showing a maximum adsorption of 0.23 nKat g-1 (19 mg protein g-1) and affinity constant of 435 nKat gˉ1 (8.5 mg protein g-1 ), demonstrating that phytase from A.niger showed a relatively low affinity for our test soil (Tayport). Phytases were partially inhibited upon adsorption and the specific activity was of 40.44 nKat mgˉ1 protein for the free enzyme and 25.35 nKat mgˉ1 protein when immobilised. The kinetics of adsorption detailed that most of the adsorption occurred within the first 20 min upon addition to soil. Citrate had no effect on the rate or total amount of phytase adsorption or loss of activity, within the studied citrate concentrations (0-4mM). Free phytases in soil solution and phytase immobilised on soil particles showed optimum activity (>80%) at pH 4.5-5.5. Immobilised phytase showed greater loss of activity at pH levels over 5.5 and lower activities at the secondary peak at pH 2

  1. The effect of dietary phytase supplementation on the N-balance of growing pigs

    Directory of Open Access Journals (Sweden)

    Veronika Halas

    2010-01-01

    Full Text Available Some studies suggest that dietary phytase enhance the growth rate of pigs fed P-adequate diets. This may be due to an increased N digestibility and/or improved protein gain. The aim was to study the effect of dietary phytase supplementation on the N-balance of growing pigs upon protein limiting condition. A total of 24 hybrid individually kept barrows (25kg BW were assigned into 4 treatments. Diet in AP0 (AP: adequate protein contained 190g/kg crude protein and no phytase supplementation, diets in RP0, RP500 and RP1000 (RP: reduced protein contained 160g/kg crude protein and 0, 500 and 1000 FTU/kg phytase supplementation, respectively. The balance trial consisted of 7 days adaptation and 5 days collection, during which the feces and urine were collected quantitatively. Additional phytase to low protein diets increased the N-retention of the pigs (P0.05. Dietary treatments did not affect the digestibility of protein, however, 500 FTU/kg phytase supplementation increased the efficiency of N-retention. Our results show that the protein content of the feed for pigs of 20-30kg can be reduced from 190 to 160 g/kg if the diet is supplemented with 500 FTU/kg phytase without weakening the N-balance of pigs.

  2. The effect of dietary phytase on broiler performance and digestive, bone, and blood biochemistry characteristics

    Directory of Open Access Journals (Sweden)

    JPL de Sousa

    2015-03-01

    Full Text Available The dietary inclusion of phytase increases nutrient and energy bioavailability for broilers. The effect of phytase increases nutrients and energy bioavailability for either the objective of this experiment was to evaluate the influence of nutrient and energy reduction in diets supplemented with phytase on the performance, gastrointestinal pH, organ and bone composition, and blood biochemistry of broilers between eight and 21 days of age. In the study, 1.120 male Cobb 500(r broilers, with 161±1g average weight, were used. At eight days of age, birds were distributed according to a completely randomized experimental design with seven treatments in a 3x2+1 factorial arrangement with eight replicates of 20 broiler each. Treatments corresponded to reduction of calcium (Ca and phosphorus (P, amino acids and energy, or reduction of Ca, P, amino acids and energy; supplementation or not of phytase; and a positive control treatment. Broiler fed the diet with reduced Ca and P levels and phytase supplementation presented the best performance of all groups. The diet with reduced amino acid and energy levels and phytase addition reduced gizzard and proventriculus pH. Dietary Ca and P reduction increased relative liver and heart weights, as well as albumin blood levels. The bones of broilers fed phytase-supplemented diets presented higher ash content.

  3. Studies towards the stabilisation of a mushroom phytase produced by submerged cultivation.

    Science.gov (United States)

    Spier, Michele Rigon; Behsnilian, Diana; Zielinski, Acácio; Konietzny, Ursula; Greiner, Ralf

    2015-10-01

    A novel phytase from Ganoderma australe G24 was produced by submerged cultivation and recovery. Liquid and solid forms of phytase were developed; both types of product were formulated using different additives. Ganoderma australe G24 phytase was very stable in liquid form with NaCl and sodium acetate buffer. Solid form products were obtained by spray-drying using different polymers to encapsulate the phytase and the capsules obtained were analyzed by electron microscopy. Micrographs confirmed micro and nanoparticles formed with maltodextrin (300 nm to 7-8 µm) without the presence of agglomerates. The use of maltodextrin for solid formulation of G. australe G24 phytase is recommended, and resulted in good stability after the drying process and during storage (shelf life). Kinetic models of phytase inactivation in the microencapsulated powders over time were proposed for the different stabilizing additives. Inactivation rate constants, half-lives and D values (decimal reduction time) were obtained. Phytase encapsulated with maltodextrin remained stable after 90 days, with k 0.0019 day(-1) and a half-life (t1/2) of 367.91 days(-1).

  4. Evaluation of the compositional and nutritional values of phytase transgenic corn to conventional corn in roosters.

    Science.gov (United States)

    Gao, C Q; Ma, Q G; Ji, C; Luo, X G; Tang, H F; Wei, Y M

    2012-05-01

    Three experiments were conducted to evaluate the compositional and nutritional values of corn grains [phytase transgenic corn (PTC) and isogenic conventional corn (CC)] and compare the efficacy of corn-based phytase and extraneous microbial phytase for enhancing the utilization of phytate phosphorus (P) in single corn or corn-soybean mixed meals (corn:soybean = 2.5:1, wt:wt) fed to roosters. Following a 48-h fasting period, 16 roosters were given 50 g of each sample via crop intubation and excreta were collected for 48 h. Nitrogen-free and phosphorus-free diets were used to evaluate endogenous amino acid and endogenous P losses, respectively. Chemical composition was not different between PTC and CC, whereas the phytase content for PTC was greater than CC (8,047 vs. 37 FTU/kg of corn, DM basis; P 0.05) between roosters fed PTC and extraneous microbial phytase in equivalent FTU/kg of diets. The results of this study indicated that the chemical composition, TME, and true amino acid availability in PTC are essentially equivalent to that in CC, and the true P utilization for roosters is higher in PTC than in CC. Corn expressing phytase is as efficacious as equivalent microbial phytase when supplemented in corn-soybean diets for chickens.

  5. Super High Dosing with a Novel Buttiauxella Phytase Continuously Improves Growth Performance, Nutrient Digestibility, and Mineral Status of Weaned Pigs.

    Science.gov (United States)

    Zeng, Zhikai; Li, Qingyun; Tian, Qiyu; Zhao, Panfeng; Xu, Xiao; Yu, Shukun; Piao, Xiangshu

    2015-11-01

    This study was conducted to evaluate the efficacy of a novel Buttiauxella phytase to pigs fed P-deficient, corn-soybean meal diets. One hundred and twenty crossbred piglets (9.53 ± 0.84 kg) were allocated to one of five treatments which consisted of four low P diets (0.61 % Ca and 0.46 % total P) supplemented with 0, 500, 1,000, or 20,000 FTU/kg phytase as well as a positive control diet (0.77 % Ca and 0.62 % total P). Each treatment had six replicated pens with four pigs per pen. Pigs were fed the experimental diets for 28 days. Phytase supplementation linearly improved (P phytase (20,000 FTU/kg) further increased (P phytase inclusion group, as well as ATTD of Ca and P. Metacarpal bone characteristics and several trace mineral concentration in bone, plasma, or organ tissues were linearly (P phytase. Super high dosing with phytase (20,000 FTU/kg) supplementation improved (P phytase supplementation (500 or 1,000 FTU/kg). In conclusion, supplementation of 500 FTU of Buttiauxella phytase/kg and above effectively hydrolyzed phytate in a low-P corn-soybean diet for pigs. In addition, a super high dosing with phytase (20,000 FTU/kg) improved macro- or micro mineral availability and growth performance.

  6. Optimization of phytase production from escherichia coli by altering solids-state fermentation conditions

    OpenAIRE

    McKinney, K.; Combs, J.; Becker, P.; Humphries, A.; K. Filer; Vriesekoop, F.

    2015-01-01

    Cultivation of Escherichia coli on wheat-bran substrate under various Solid-State Fermentation (SSF) conditions was evaluated for phytase yield along with the enzyme activity profile as a potential, low-cost alternative to submerged-liquid fermentation. The maximum phytase activity achieved by E. coli was 350 ± 50 SPU of phytase activity per gram of bran, incubated for 96 h with a substrate bed moisture content of 70% (w/v) at 37 °C with a relative air humidity of 90%, and supplemented with 1...

  7. Cultivation Conditions for Phytase Production from Recombinant Escherichia coli DH5α

    OpenAIRE

    Rafidah Mohd Ariff; Anwar Fitrianto; Mohd Yazid Abd. Manap; Aini Ideris; Azhar Kassim; Afinah Suhairin; Anis Shobirin Meor Hussin

    2013-01-01

    Response surface methodology (RSM) was used to optimize the cultivation conditions for the production of phytase by recombinant Escherichia coli DH5α. The optimum predicted cultivation conditions for phytase production were at 3 hours seed age, a 2.5% inoculum level, an L-arabinose concentration of 0.20%, a cell concentration of 0.3 (as measured at 600 nm) and 17 hours post-induction time with a predicted phytase activity of 4194.45 U/mL. The model was validated and the results showed no sign...

  8. Effect of sodium fluoride on phytase activity during germination of Pisum sativum L. seeds

    Energy Technology Data Exchange (ETDEWEB)

    Hauskrecht, I.

    1972-01-01

    The effects of fluoride ion, in concentrations varying from 0 to 4 mM F/sup -/, on phytase activity in the course of germination of Pisum sativum L. seeds and in the course of incubation in vitro are reported. The fluoride ion in 1 mM concentration cause a significant 80% decrease of the phytase activity in the incubation medium and a 24% decrease in the course of germination. The inhibition of phytase, as well as its synthesis, by fluoride ion during germination may influence the metabolism of phosphorus at the initial stage of ontogenesis.

  9. Growth performance, diet nutrient digestibility, and bone mineralization in weaned pigs fed pelleted diets containing thermostable phytase.

    Science.gov (United States)

    Yáñez, J L; Landero, J L; Owusu-Asiedu, A; Cervantes, M; Zijlstra, R T

    2013-02-01

    Traditional supplemental dietary phytase loses activity during steam pelleting. The thermal tolerance and bioefficacy of a phytase product with a thermoprotective coating [coated phytase (C-phytase)] was compared in mash and pelleted diets to a traditional, uncoated phytase (U-phytase) added to a negative control (NC) diet, formulated with reduced dietary Ca and P, and compared with a corn-soybean meal based positive control (POC) diet. Growth performance, nutrient digestibility, and third metacarpal bone characteristics were response variables. Weaned pigs (n = 56; 8.20 ± 0.5 kg initial BW; 28 d of age) were individually housed and randomly allotted to 1 of 7 diets for 21 d. The diets were 1) POC mash, 2) NC mash, 3) NC pelleted at 90°C, 4) NC mash + 500 U/kg U-phytase, 5) NC mash + 500 U/kg C-phytase, 6) NC + 500 U/kg C-phytase pelleted at 80°C, and 7) NC + 500 U/kg C-phytase pelleted at 90°C. The POC and NC diets were formulated to be isoenergetic and isolysinic. The content of Ca and available P was 1.01 and 0.40% and 0.83 and 0.22% in the POC and NC diets, respectively. Pig BW and feed intake were measured on d 7, 14, and 21, and feces were collected for 2 d. On d 21, pigs were killed and ileal digesta and the third metacarpal bone collected. Pigs fed POC had greater (P pigs fed NC. Pelleting the NC diet did not improve performance, nutrient digestibility, or P use. Adding the U-phytase to NC mash diet increased (P pigs fed NC. Pigs fed C-phytase in NC mash diets had increased (P pigs fed NC but not different than pigs fed U-phytase NC mash diets. Pigs fed pelleted NC diet with C-phytase had a greater (P pigs fed mash NC diet with C-phytase but had similar growth performance, AID of CP and AA, and bone mineralization to pigs fed U-phytase. In conclusion, release and bioefficacy of phytase after pelleting was not affected by the thermal protective coating.

  10. Effect of exogenous phytase on feed inositol phosphate hydrolysis in an in vitro rumen fluid buffer system

    DEFF Research Database (Denmark)

    Brask-Pedersen, Dorte Niss; Glitsø, Lene Vibe; Skov, L.K.

    2011-01-01

    Three in vitro experiments using a rumen fluid buffer system were performed to investigate the effect of addition of 4 experimental phytases (Phy1, Phy2, Phy3, and Phy4) compared with no addition of phytase on feed inositol phosphate hydrolysis in wheat and rapeseed cake to determine which of the 4...... phytases was most suitable under rumen-like conditions. The feedstuffs were incubated with a mixture of physiological buffer, ruminal fluid, and exogenous phytase at pH 6.2, after which the samples were incubated for different periods. Incubations were stopped using HCl, and the samples were analyzed...... for inositol phosphates via high performance ion chromatography. Addition of phytase (Phy1) resulted in enhanced degradation of myo-inositol hexakisphosphate (InsP6) in rapeseed cake, whereas addition of exogenous phytase did not improve the degradation of InsP6 in wheat. Only rapeseed cake was therefore used...

  11. Meta-analysis of effects of microbial phytase on digestibility and bioavailability of copper and zinc in growing pigs.

    Science.gov (United States)

    Bikker, P; Jongbloed, A W; Thissen, J T N M

    2012-12-01

    A meta-analysis was conducted to determine the effect of microbial phytase in pig diets on digestibility and bioavailability of Cu and Zn. Studies (n = 22) into effects of microbial phytase on digestibility and plasma levels of Cu and Zn were included in a dataset and regression analysis was performed to quantify the effect of Aspergillus niger derived 3-phytase in studies (n = 14) with a maximum dietary Zn content of 100 mg/kg and a maximum Cu addition of 20 mg/kg. Phytase inclusion increased digestibility of Zn (P = 0.003) and plasma Zn content (P < 0.001) without affecting digestibility and plasma level of Cu. Based on a comparison with dose-response studies with Zn, we estimated 500 phytase units (FTU) of microbial phytase is equivalent to 27 mg of Zn from ZnSO(4).

  12. Differences in phytase activity and phytic acid content between cultivated and Tibetan annual wild barleys.

    Science.gov (United States)

    Dai, Fei; Qiu, Long; Xu, Yang; Cai, Shengguan; Qiu, Boyin; Zhang, Guoping

    2010-11-24

    The Qinghai-Tibetan Plateau in China is considered to be one of the original centers of cultivated barley. At present, little is known about the phytase activity (Phy) or phytic acid content (PA) in grains of Tibetan annual wild barley. Phy and PA were determined in grains of 135 wild and 72 cultivated barleys. Phy ranged from 171.3 to 1299.2 U kg(-1) and from 219.9 to 998.2 U kg(-1) for wild and cultivated barleys, respectively. PA and protein contents were much higher in wild barley than in cultivated barley. Tibetan annual wild barley showed a larger genetic diversity in phytase activity and phytic acid and protein contents and is of value for barley breeding. There is no significant correlation between phytase activity and phytic acid or protein content in barley grains, indicating that endogenous phytase activity had little effect on the accumulation of phytic acid.

  13. Establishment of Phytase-Specific Qualitative PCR Detection Method and Construction of a Positive Plasmid Molecule%植酸酶基因定性PCR检测方法及阳性质粒分子的构建

    Institute of Scientific and Technical Information of China (English)

    李俊; 刘信; 曹应龙; 武玉花; 厉建萌; 吴刚; 张丽; 卢长明

    2012-01-01

    Phytase gene is valuably applicated in agricultural production, especially in genetic engineering of crops. In order to meet the requirements of safety regulation of transgenic crops, the gene-specific qualitative PCR detection method targeting the fungal-originated phytase gene was developed. A primer pair Phytase-F5/R5 yielding a 389 bp amplicon was selected from 11 primer pairs, then the PCR reaction system was optimized by improving Mg2+ concentration, primer concentration and primer anneal temperature. Twenty transgenic and nontransgenic lines from different crops were used as templates in PCR, showing that the PCR method had good amplification specificity. The results of sensitivity testing indicated that the phytase amplicon was still observed when the template concentration was down to 0.05%, which reaches the national standards for GMO (Genetically Modified Organism) detection method. In addition, we cointegrated the phytase gene and the endogenous reference genes from six major crops of wheat, rice, cotton, soybeans, corn, and rapeseed into a vector, yielding a positive plasmid molecule pBS Endoge-nous-phytase. The positive plasmid molecule was suitable for screening phytase gene in the six crops about wheat, soybeans, corn, cotton, rice, and rape. This study provides positive materials and detection method for safety regulation of genetically modified crops carrying phytase gene.%植酸酶基因在农业生产中具有很高的应用价值,正被广泛用于农作物的基因工程研究.为了满足转植酸酶基因作物安全监管的需要,通过优化PCR检测条件,建立了植酸酶基因特异性定性PCR检测方法.该方法具有良好的扩增特异性和检测灵敏度,可以满足我国植酸酶转基因作物监管需要.此外,我们将6种主要农作物(小麦、水稻、棉花、大豆、玉米和油菜)的内标准基因和植酸酶基因克隆到同一载体上,构建成了阳性质粒分子pBS Endogenousphytase

  14. Pharmacological zinc and phytase supplementation enhance metallothionein mRNA abundance and protein concentration in newly weaned pigs.

    Science.gov (United States)

    Martínez, Michelle M; Hill, Gretchen M; Link, Jane E; Raney, Nancy E; Tempelman, Robert J; Ernst, Catherine W

    2004-03-01

    The swine industry feeds pharmacological zinc (Zn) to newly weaned pigs to improve health. Because most swine diets are plant-based with a high phytic acid content, we hypothesized that adding phytase to diets could reduce the amount of Zn required to obtain beneficial responses. The role of metallothionein (MT) in Zn homeostasis could be important in this positive response. Thus, the goal of this study was to investigate the effect of dietary Zn and phytase on relative MT mRNA abundance and protein concentration in newly weaned pigs. Diets containing adequate (150 mg Zn/kg) or pharmacological concentrations of Zn (1000 or 2000 mg Zn/kg), as zinc oxide, with or without phytase [0, 500 phytase units (FTU)/kg, Natuphos, BASF] were fed in a 3 x 2 factorial design. Plasma and tissue minerals were measured in pigs killed after 14 d of dietary intervention. Hepatic and renal relative MT mRNA abundance and protein were greater (P pigs fed 1000 mg Zn/kg with phytase, or 2000 mg Zn/kg with or without phytase vs. the remaining treatments. Intestinal mucosa MT mRNA abundance and protein were greater (P pigs fed 2000 mg Zn/kg with phytase than in pigs fed 2000 mg Zn/kg alone or 1000 mg Zn/kg with phytase. Pigs fed 1000 mg Zn/kg plus phytase or 2000 mg Zn/kg with or without phytase had higher plasma, hepatic, and renal Zn than those fed the adequate Zn diets or 1000 mg Zn/kg. We conclude that feeding 1000 mg Zn/kg with phytase enhances MT mRNA abundance and protein and Zn absorption to the same degree as 2000 mg Zn/kg with and without phytase.

  15. Effect of exogenous phytase on feed inositol phosphate hydrolysis in an in vitro rumen fluid buffer system.

    Science.gov (United States)

    Brask-Pedersen, D N; Glitsø, L V; Skov, L K; Lund, P; Sehested, J

    2011-02-01

    Three in vitro experiments using a rumen fluid buffer system were performed to investigate the effect of addition of 4 experimental phytases (Phy1, Phy2, Phy3, and Phy4) compared with no addition of phytase on feed inositol phosphate hydrolysis in wheat and rapeseed cake to determine which of the 4 phytases was most suitable under rumen-like conditions. The feedstuffs were incubated with a mixture of physiological buffer, ruminal fluid, and exogenous phytase at pH 6.2, after which the samples were incubated for different periods. Incubations were stopped using HCl, and the samples were analyzed for inositol phosphates via high performance ion chromatography. Addition of phytase (Phy1) resulted in enhanced degradation of myo-inositol hexakisphosphate (InsP(6)) in rapeseed cake, whereas addition of exogenous phytase did not improve the degradation of InsP(6) in wheat. Only rapeseed cake was therefore used subsequently. All 4 phytases increased degradation of InsP(6) in rapeseed cake in the in vitro system, and degradability of InsP(6) increased with higher incubation time and higher phytase dosages, independent of phytase. Addition of 2 units of phytase per gram of substrate of the phytases Phy1, Phy2, Phy3, and Phy4 led to an undegraded InsP(6) content of 56, 49, 70, and 18%, respectively, when incubated with rapeseed cake for 6h, indicating that Phy2 and Phy4 were the most effective phytases. However, Phy2 had a higher specific activity than Phy4, as 60% of the original InsP(6) content was remaining after 3h when 5mg of enzyme protein per gram of substrate of Phy2 was added to rapeseed cake, whereas 150 mg of enzyme protein per gram of substrate of Phy4 was necessary to achieve a similar result. Therefore, Phy2 appeared to be most applicable under rumen-like conditions.

  16. Linking ileal digestible phosphorus and bone mineralization in broiler chickens fed diets supplemented with phytase and highly soluble calcium.

    Science.gov (United States)

    Adeola, O; Walk, C L

    2013-08-01

    The objectives of this study were to determine the ileal digestibility of P in potassium phosphate, phytase-related ileal digestible P release, bone-mineralization-based ileal digestible P equivalency of phytase, and phytase-related efficiency of ileal digestible P utilization for bone mineralization in broiler chickens at 2 dietary concentrations of highly soluble Ca (HSC). Birds were sorted by BW at d 15 posthatch and assigned to 8 cages per diet with 8 birds per cage. Twelve diets were arranged in a 2 × 6 factorial of HSC at 5 or 6 g/kg and P supply treatment at 6 levels consisting of 4 added P levels (P from KH2PO4 added at 0, 0.7, 1.4, or 2.1 g/kg of diet) or 2 added phytase levels (500 or 1,000 phytase units). On d 24 posthatch, ileal digesta were collected for ileal P digestibility (IPD) determination and the left tibia was collected from the 4 heaviest birds in each cage for bone ash determination. Weight gain, G:F, and tibia ash were higher (P phytase linearly increased (P phytase showed a reduction (P phytase intake indicated that 1,000 units of added phytase released 1.701 or 1.561 g of digestible P in diets containing 5 or 6 g of added HSC/kg, respectively. Polynomial regressions of tibia ash on digestible P or phytase intake in diets containing 5 or 6 g of added HSC/kg at 1,000 phytase units gave digestible P equivalency of 1.487 or 1.448 g, respectively. Thus, phytase-related efficiency of ileal digestible P utilization for bone mineralization was 87.4 and 92.8% in diets containing 5 or 6 g of added HSC/kg, respectively.

  17. Supplementation of laying japanese quail with amylase, phytase and protease

    Directory of Open Access Journals (Sweden)

    Josimar Santana Ribeiro

    2015-07-01

    Full Text Available The objective of this study was to evaluate the effects of supplementation of diets for laying Japanese quail with amylase, phytase and protease alone or in combination. Three-hundred quail were assigned to a completely randomized design consisting of five treatments and six repetitions, with 10 animals per experimental unit. The treatments were: a control diet and diets supplemented with 300 ppm amylase, 300 ppm protease and 500 phytase units (FTU/kg and the combination of these enzymes. In the diets containing the enzymes, the nutritional requirements of one or more of the following components were reduced: protein, digestible amino acids, energy, calcium and phosphorus, giving priority to the use of enzymes. The evaluations were performed over four periods of 21 days each. Performance (mean egg production, feed intake, mean egg weight, and feed conversion, egg quality (proportion of egg constituents and specific egg weight, and dietary nutrient digestibility (apparent digestibility coefficient of dry matter and crude protein were evaluated. There was no significant effect of the treatments on the variables analyzed (P>0.05, indicating that the enzymes, alone or in combination, have beneficial effects, maintaining performance and egg quality of Japanese quail.

  18. Modulation of small intestinal phosphate transporter by dietary supplements of mineral phosphorus and phytase in broilers.

    Science.gov (United States)

    Huber, Korinna; Zeller, Ellen; Rodehutscord, Markus

    2015-05-01

    Dietary phosphorus (P) is known as a main modulator of phosphate (Pi) transporter expression. The effect of supplemented mineral P with or without phytase on protein expression of two sodium-dependent Pi (NaPi) transporters and a calcium channel was studied in the small intestine of broilers. Thirty-six broilers were randomly assigned to six different diets at 15 days of age. Two levels of total P (tP, adjusted by monocalcium phosphate (MCP) supplementation), 0.39% (BD-) and 0.47% (BD+) were fed until day 25; and at each tP level, three levels of phytase were used with 0, 500, and 12,500 FTU/kg of an E. coli phytase. Mucosa samples from jejunum and ileum were taken and apical membranes were isolated by MgCl2 precipitation. Protein expression of NaPi IIb, NaPi type III (PiT1) and the calcium channel TRPV6 were semiquantitatively measured by Western blotting and jejunal mucosal phytase activity by measurement of Pi release. The jejunal NaPi IIb transporter was expressed with two distinct bands, which were modulated differently by diet. NaPi IIb Band1 increased (P phytase supplementation but was not affected by MCP supplementation. This inverse modulation of Band1 and Band2 was significantly related to the amount of net absorbed P with higher expression of Band1 at higher amounts of net absorbed P. In addition, a second Pi transporter, PiT1, was detected in which ileal expression decreased (P phytase supplementation. The expression of the calcium channel TRPV6 was increased in BD+ groups. A trend for an interaction between MCP and phytase supplementation on mucosal phytase activity was observed (P = 0.079) with a decrease in activity when BD+ with 12,500 FTU/kg phytase was fed. Chicken intestinal epithelial cells responded to dietary supplemented phytase and MCP by changing the Pi transporter expression in apical membranes. In conclusion, availability of Pi is most likely the key modulator of transporter protein expression. However, a contribution of lower inositol

  19. Characteristics Of Phytases From Soybeans And Microorganisms Involved In The Tempe Production

    OpenAIRE

    K A Buckle2); Sutardi*

    2004-01-01

    Tempe was prepared from Forest variety of soybeans inoculated with pure culture of the mould Rhizopus oligosporus strain CT11K2. Partially purified phytases from soybeans, mould of the R. oligosporus strain CT11K2, yeasts (Endomycopsis burtonii, Candida diddensiae and Candida tropicalis) and bacteria (Streptococcus faecium and Streptococcus dysgalactiae) involved during tempe production were prepared according to procedure developed by Sutardi (1988). The crude phytases were characterized and...

  20. Overexpression and functional characterization of an Aspergillus niger phytase in the fat body of transgenic silkworm, Bombyx mori.

    Science.gov (United States)

    Xu, Hanfu; Liu, Yaowen; Wang, Feng; Yuan, Lin; Wang, Yuancheng; Ma, Sanyuan; Beneš, Helen; Xia, QingYou

    2014-08-01

    In a previous study, we isolated 1,119 bp of upstream promoter sequence from Bmlp3, a gene encoding a member of the silkworm 30 K storage protein family, and demonstrated that it was sufficient to direct fat body-specific expression of a reporter gene in a transgenic silkworm, thus highlighting the potential use of this promoter for both functional genomics research and biotechnology applications. To test whether the Bmlp3 promoter can be used to produce recombinant proteins in the fat body of silkworm pupae, we generated a transgenic line of Bombyx mori which harbors a codon-optimized Aspergillus niger phytase gene (phyA) under the control of the Bmlp3 promoter. Here we show that the Bmlp3 promoter drives high levels of phyA expression in the fat body, and that the recombinant phyA protein is highly active (99.05 and 54.80 U/g in fat body extracts and fresh pupa, respectively). We also show that the recombinant phyA has two optimum pH ranges (1.5-2.0 and 5.5-6.0), and two optimum temperatures (55 and 37 °C). The activity of recombinant phyA was lost after high-temperature drying, but treating with boiling water was less harmful, its residual activity was approximately 84% of the level observed in untreated samples. These results offer an opportunity not only for better utilization of large amounts of silkworm pupae generated during silk production, but also provide a novel method for mass production of low-cost recombinant phytase using transgenic silkworms.

  1. Bacterial phytase: potential application, in vivo function and regulation of its synthesis

    Directory of Open Access Journals (Sweden)

    Ursula Konietzny

    2004-06-01

    Full Text Available The stepwise release of phosphate from phytate, the major storage form of phosphate in plant seeds and pollen, is initiated by a class of enzymes that have been collectively called phytases. The classification is solely due to the in vitro capability of these enzymes to accept phytate as a substrate. Phytases have been studied intensively in recent years because of the great interest in such enzymes for reducing phytate content in animal feed and food for human consumption. They have a wide distribution in plants, microorganisms, and in some animal tissues. Due to several biological characteristics, such as substrate specificity, resistance to proteolysis and catalytic efficiency, bacterial phytases have considerable potential in commercial applications. In bacteria, phytase is an inducible enzyme and its expression is subjected to a complex regulation, but phytase formation is not controlled uniformly among different bacteria. It was suggested that phytase is not required for balanced growth of bacterial cells, but may be synthesised in response to a nutrient or energy limitation.

  2. Production of partially phosphorylated myo-inositol phosphates using phytases immobilised on magnetic nanoparticles.

    Science.gov (United States)

    Greiner, Ralf; Konietzny, Ursula; Blackburn, Daniel Menezes; Jorquera, Milko A

    2013-08-01

    Phytases of different origin were covalently bound onto Fe3O4 magnetic nanoparticles (12 nm). Binding efficiencies of all three phytases were well above 70% relative to the number of aldehyde groups available on the surface of the magnetic nanoparticles. Temperature stability for all three phytases was enhanced as a consequence of immobilisation, whereas pH dependence of enzyme activity was not affected. Maximum catalytic activity of the immobilised phytases was found at 60°C (rye), 65°C (Aspergillus niger) and 70°C (Escherichia albertii). The immobilised enzymes exhibited the same excellent substrate specificities and unique myo-inositol phosphate phosphatase activities as their soluble counterparts. However, the catalytic turnover number dropped drastically for the immobilised phytases. The amount of the desired partially phosphorylated myo-inositol phosphate isomer could be easily controlled by the contact time of substrate solution and immobilised enzymes. The immobilised phytases showed a high operational stability by retaining almost full activity even after fifty uses.

  3. Production and characterization of phytase from Bacillus spp. as feed additive in aquaculture

    Directory of Open Access Journals (Sweden)

    Rande B. Dechavez

    2011-07-01

    Full Text Available Phytases are phosphohydrolases that catalyze the release of phosphate from phytate (myo inositol hexakisphosphate, the major phosphorus (P form mostly occurring in animal feeds of plant origin. These enzymes can be supplemented in animal diets to reduce inorganic phosphorus supplementation and fecal phosphorus excretion. Four species of Bacillus namely, B. pumilus , B.megaterium , B. coagulans , and B. licheniformis were used to study the biochemical characteristics of their phytases. All the strains investigated were able to hydrolyze extracellular phytate. The activity of phytase increased markedly at the late stationary phase in all the species tested. Highest enzyme activity was found in phytase from B. megaterium after the 4th day of culture. The crude phytases from the different Bacillus strains were optimally active at pH values ranging 5.5 to 7.0 at 37 0 C and retained their activity at temperatures up to 80 0 C. The enzymes exhibited thermostability, retaining ~50 %activity at 70 0 C and were fairly stable up to pH 10. These properties indicate that the Bacillus phytases appear to be suitable for animal feed supplementation in aquaculture to improve the bioavailability of phosphorus.

  4. Formulated products containing a new phytase from Schyzophyllum sp. phytase for application in feed and food processing

    Directory of Open Access Journals (Sweden)

    Denise Naomi Salmon

    2011-12-01

    Full Text Available A new formulated product containing high yield of phytase from Schizophyllum sp., an important mushroom used for medicinal studies, was developed for application in feed industries and for future use in food processing. The enzyme presented a high activity yield 55.5 U/mL and 6240 U/gds in liquid and solid formulated product, respectively. It showed a good shelf-life in concentrated product, retaining 67.8% of its activity after 60 days of storage at room temperature and 90% of the activity was maintained in the liquid formulation after the same period. Powder bioformulated product maintained 77% of its activity after two months of storage, without the addition of chemical additives, which was named as a new bioformulated product containing high quantities of phytase. After separation and concentration steps, enzyme stability was monitored in two forms: liquid and solid. The liquid product was stable with the presence of manitol and polyethylene glycol at 1% (w/v, while solid product was the most stable product without the presence of chemical additives.

  5. The supplementation of phytase RONOZYME P on the growth and the utilisation of phosphorus by sex-reversed red tilapia (Oreochromis niloticus Linn.)

    OpenAIRE

    Jirawat Tudkaew,; Jacques Gabaudan; Wutiporn Phromkunthong

    2008-01-01

    The effect of phytase supplementation to a low fish meal based diet on growth performance and phosphorus utilisation was investigated in sex-reversed red tilapia. Diets were prepared without phytase or inorganic phosphorus supplementation, with phytase, with supplemented inorganic phosphorus and with both phytase and supplemental inorganic phosphorus. Available phosphorus was set below requirement and the total phosphorus set to meet requirement for tilapia. After 8 weeks, there were signific...

  6. Phosphorus utilization and characterization of ileal digesta and excreta from broiler chickens fed diets varying in cereal grain, phosphorus level, and phytase addition

    Science.gov (United States)

    Both endogenous and exogenous phytase present in poultry feeds can alter phytate utilization as well as the solubility of phosphorus (P) excreted, which can be a concern from an environmental perspective. To address the effects of endogenous phytase present in cereal grains and exogenous phytase ad...

  7. The interplay of dietary nutrient specification and varying calcium to total phosphorus ratio on efficacy of a bacterial phytase: 1. Growth performance and tibia mineralization.

    Science.gov (United States)

    Olukosi, O A; Fru-Nji, F

    2014-12-01

    A 14-d experiment was conducted to study the effects of 2 dietary variables on efficacy of a 6-phytase from Citrobacter braakii on broiler growth performance and tibia mineralization. Diets were formulated with or without nutrient matrix values for phytase as negative or positive control (NC or PC, respectively) and with 2 Ca:total P (tP; 2:1 or 2.5:1). The diets were supplemented with 0, 1,000, or 2,000 phytase units (FYT)/kg, thus producing a 2 × 2 × 3 factorial arrangement. Birds and feed were weighed on d 7 and 21, and tibia bones were collected from all the birds on d 21. The main effects of nutrient matrix, Ca:tP, and phytase supplementation were significant (P phytase and matrix × phytase interactions were significant (P phytase increased weight gain (P phytase increased (P Phytase supplementation of diets with 2:1 Ca:tP increased (P phytase supplementation increased (P phytase supplementation (1,000 FYT/kg) was in NC diets with narrow Ca:tP, whereas the best response to higher level of phytase supplementation (2,000 FYT/kg) was achieved in diets in PC diets with wide Ca:tP.

  8. Elimination of a disulfide bridge in Aspergillus niger NRRL 3135 Phytase (PhyA) enhances heat tolerance and optimizes its temperature versus activity profile

    Science.gov (United States)

    The utilization of microbial phytases in animal feed, rich in phytate, and intended for animals with simple stomachs is now widely accepted. The commercial phytases currently available are all histidine acid phosphatases (HAP) and have been termed histidine acid phytases (HAPhy). The HAPhy enables ...

  9. Impact of Assay conditions on activity estimate and kinetics comparison of Aspergillus niger PhyA and Escherichia coli AppA2 phytases

    Science.gov (United States)

    This study was to compare three phytase activity assays and kinetics of Aspergillus niger PhyA and Escherichia coli AppA2 phytases expressed in Pichia pastoris at the observed stomach pH of 3.5. In Experiment 1, equivalent phytase activities in the crude preparations of PhyA and AppA2 were tested ...

  10. Barley HvPAPhy_a as transgene provides high and stable phytase activities in mature barley straw and in grains

    DEFF Research Database (Denmark)

    Holme, Inger; Dionisio, Giuseppe; Madsen, Claus Krogh

    2017-01-01

    , the green leaves and in the dry mature vegetative plant parts left after harvest of the grains. The T2-generation of HvPAPhy_a transformed barley showed phytase activity increases up to 19-fold (29 000 phytase units (FTU) per kg in mature grains). Moreover, also in green leaves and mature dry straw, phytase...

  11. Phytase modulates ileal microbiota and enhances growth performance of the broiler chickens.

    Science.gov (United States)

    Ptak, Anna; Bedford, Michael R; Świątkiewicz, Sylwester; Żyła, Krzysztof; Józefiak, Damian

    2015-01-01

    Phytase is well studied and explored, however, little is known about its effects on the microbial ecology of the gastrointestinal tract. In total, 400 one-day-old female Ross 308 chicks were randomly distributed to four experimental groups. The dietary treatments were arranged as a 2 × 2 complete factorial design, with the factors being adequate (PC) or insufficient calcium (Ca) and digestible phosphor (dP)(NC) and with or without 5000 phytase units (FTU)/kg of Escherichia coli 6-phytase. The gastrointestinal tract pH values, ileal microbial communities and short-chain fatty acid concentrations in the digesta were determined. The reduction in Ca and dP concentration significantly affected pH in the crop and caeca, and addition of phytase to the NC resulted in a pH increase in the ileum. The reduction in Ca and dP concentration significantly lowered, while phytase supplementation increased ileal total bacterial counts. Additionally, the deficient diet reduced butyrate- but increased lactate-producing bacteria. The addition of phytase increased Lactobacillus sp./Enterococcus sp. whereas in case of Clostridium leptum subgroup, Clostridium coccoides-Eubacterium rectale cluster, Bifidobacterium sp. and Streptococcus/Lactococcus counts, a significant Ca and dP level x phytase interaction was found. However, the recorded interactions indicated that the effects of phytase and Ca and dP levels were not consistent. Furthermore, the reduction of Ca and dP level lowered Clostridium perfringens and Enterobacteriaceae counts. The analysis of fermentation products showed that reducing the Ca and dP content in the diet reduced total SCFA, DL-lactate, and acetic acid in the ileum whereas phytase increased concentrations of these acids in the NC group. This suggests that P is a factor which limits fermentation in the ileum. It may be concluded that phytase plays a role in modulating the gut microbiota of chicken, however, this is clearly linked with the levels of P and Ca in a diet.

  12. Impact of response criteria (tibia ash weight vs. percent) on phytase relative non phytate phosphorus equivalance.

    Science.gov (United States)

    Li, W; Angel, R; Kim, S-W; Jiménez-Moreno, E; Proszkowiec-Weglarz, M; Plumstead, P W

    2015-09-01

    The current study was conducted to evaluate the impacts of using tibia ash percentage or ash weight as the response criteria on estimated phytase relative equivalence. Straight run broilers were fed treatment (Trt) diets from 7 to 21 d age (6 birds/pen, 8 pens/Trt). The corn-soy based Trt were formulated to contain 0.80% Ca and 4 non-phytate phosphorus (nPP) concentrations (0.20, 0.27, 0.34, and 0.40%). Monocalcium phosphate was the inorganic phosphate source added to achieve 4 different dietary nPP concentrations and against which the nPP relative equivalence of phytase was determined. A 6-phytase (Danisco Animal Nutrition, DuPont Industrial Biosciences, Marlborough, UK) was added at 500 or 1,000 phytase unit ( FTU: )/kg to the 0.20% nPP diet resulting 6 total Trts. Tibia ash was determined at 21 d age. Phytase fed at 500 or 1,000 FTU/kg increased tibia ash weight and ash percentage compared to that of birds fed 0.20% nPP diet without phytase (Pphytase nPP relative equivalence. The R2 obtained from pen value regressions were 0.81 and 0.84, for tibia ash weight and percentage, respectively. Ash percentage from birds fed 500 and 1,000 FTU phytase/kg fell within the range obtained with the MCP additions. Ash weight (842 mg/tibia) from birds fed 1,000 FTU phytase/kg exceeded (Pphytase/kg Trt. The nPP relative equivalence in birds fed 500 FTU phytase/kg were 0.117 and 0.168% based on ash percentage and weight, respectively (Pphytase/kg was 0.166% for ash percentage. Results suggested that ash weight better reflects the amount of bone mineralization as compared to ash percentage and using ash percentage may lead to an underestimation of phytase efficacy.

  13. Used of microbial phytase to replace inorganic phosphorus in sex-reversed red tilapia: 1 dose response

    Directory of Open Access Journals (Sweden)

    Wutiporn Phromkunthong

    2006-07-01

    Full Text Available Sex-reversed red tilapia of average initial body weight 5.5 g were fed seven practical diets containing 0, 500, 1,000, 2,000 and 4,000 units of microbial phytase/kg and two diets containing 0.2 and 0.3% feed grade dicalcium phosphate (DCP (but no microbial phytase, respectively. The experiment was carried out in 235- l glass aquaria filled with 180 l water and attached with a closed-recirculating water system with 0.8 l/min flow rate. The experimental period was 10 weeks. All experimental diets were formulated with plant-based protein of 30% and 6% fat. Results indicated an improvement in apparent digestibility coefficient of phosphorus (ADCP in fish given phytase supplemented feed. There was no difference in ADCP when 1,000 unit phytase/kg diet or higher phytase levels (2,000 and 4,000 unit phytase/kg diet or 0.2 and 0.3% DCP were supplemented. A significant increase was noted for hemoglobin in tilapia that received 1,000 unit phytase/kg diet or higher levels compared to the control. Serum phosphorus ma kedly increased when the fish were given feeds with 1,000 unit phytase/kg diet and over, while the supplementation of 500 unit phytase/kg diet and over increased serum zinc level. Higher levels of phosphorus were retained in bone whereas lower levels of phosphorus presented in the feces of tilapia fed feeds supplemented with phytase. Growth performance was markedly influenced when the fish were given feed with 4,000 unit phytase/kg diet.

  14. Purification, sequencing and evaluation of a divergent phytase from Penicillium oxalicum KCTC6440.

    Science.gov (United States)

    Kim, Bong-Hyun; Lee, Ji Yeon; Lee, Peter C W

    2015-01-01

    A fungal strain producing high levels of phytase was purified to homogeneity from Penicillium oxalicum KCTC6440 (PhyA). The molecular mass of the purified PhyA was 65 kDa and optimal activity occurred at 55°C. The enzyme was stable in a pH range of 4.5-6.5, with an optimum performance at pH 5.5. The Km value for the substrate sodium phytate was 0.48 mM with a Vmax of 672 U/mg. The enzyme was inhibited by Ca(2+), Cu(2+), and Zn(2+), and slightly enhanced by EDTA. The PhyA efficiently released phosphate from feedstuffs such as soybean, rich bran and corn meal. The PhyA gene was cloned in two steps of degenerate PCR and inverse PCR and found to comprise 1501 bp and encode 461 amino acid residues. The enzyme was found to have only 13 amino acids differing to the known PhyA from other Penicillium sp., but has distinct enzyme characteristics. Computational analysis showed that PhyA possessed more positively charged residues in the active sites compared to other PhyA molecules, which may explain the broader pH spectrum.

  15. A novel phosphorus biofertilizer based on cattle manure and phytases-nanoclay complexes

    Science.gov (United States)

    Blackburn, Daniel; Jorquera, Milko; Greiner, Ralf; Velasquez, Gabriela; Mora, María de la Luz

    2013-04-01

    Phytate and other phytase labile organic phosphorus (P) are abundant in both soils and manures. These recalcitrant forms of P accumulate in soils by their interaction with mineral particles. The aim of this work was to evaluate the potential of treating cattle manure with phytases stabilized in allophanic nanoclays, as a novel P biofertilization technology for crops grown in volcanic soils (Andisol). Two Andisols and two manures with contrasting inorganic Pcontent were used: Low P soil from Piedras Negras series (SPN-LP); High P soil from Freire Series (SF-HP); Low P Waste (WPN-LP); High P Waste (WF-HP). The used Andisols and manures were incubated with phytase-nanoclay complexes and the inorganic P was determined in the NaOH-EDTA and bicarbonate extracts. The WPN-LP was also inoculated with an alkaline β-propeller phytase (BPP) producing bacterium. The incubated SPN-LP and SPN-LP-WPN-LP mixture were evaluated for their P supplying capacity to wheat plants under greenhouse conditions. Our resultsindicated that the treatment of cattle manure with phytase stabilized in nanoclays resulted in a significant (P≤0.0.5) increase in the inorganic P. The use of phytase treated cattle manure increased 10% plant dry weight and 39% P concentration in wheat plants under greenhouse conditions, being equivalent to a P fertilizer dose of about 150 kg of P ha-1. In the case of low P cattle manure inoculated with BPP producing bacterium, inorganic P increased 10% in soil extracts (NaOH EDTA and Bicarbonate). However, the application of this treated manure did not result in a significant response to wheat growth and P acquisition. Our results suggest that this novel approach of incubating cattle manure with phytase stabilized in nanoclays enhances organic P cycling and P nutrition of plants grown under P-deficient soils.

  16. Expression of bifidobacterial phytases in Lactobacillus casei and their application in a food model of whole-grain sourdough bread.

    Science.gov (United States)

    García-Mantrana, Izaskun; Yebra, María J; Haros, Monika; Monedero, Vicente

    2016-01-04

    Phytases are enzymes capable of sequentially dephosphorylating phytic acid to products of lower chelating capacity and higher solubility, abolishing its inhibitory effect on intestinal mineral absorption. Genetic constructions were made for expressing two phytases from bifidobacteria in Lactobacillus casei under the control of a nisin-inducible promoter. L. casei was able of producing, exporting and anchoring to the cell wall the phytase of Bifidobacterium pseudocatenulatum. The phytase from Bifidobacterium longum spp. infantis was also produced, although at low levels. L. casei expressing any of these phytases completely degraded phytic acid (2mM) to lower myo-inositol phosphates when grown in MRS medium. Owing to the general absence of phytase activity in lactobacilli and to the high phytate content of whole grains, the constructed L. casei strains were applied as starter in a bread making process using whole-grain flour. L. casei developed in sourdoughs by fermenting the existing carbohydrates giving place to an acidification. In this food model system the contribution of L. casei strains expressing phytases to phytate hydrolysis was low, and the phytate degradation was mainly produced by activation of the cereal endogenous phytase as a consequence of the drop in pH. This work shows the capacity of lactobacilli to be modified in order to produce enzymes with relevance in food technology processes. The ability of these strains in reducing the phytate content in fermented food products must be evaluated in further models.

  17. Evaluation of different microbial phytases on phosphorus digestibility in pigs fed a wheat and barley based diet

    DEFF Research Database (Denmark)

    Poulsen, Hanne Damgaard; Blaabjerg, Karoline; Strathe, Anders Bjerring

    2010-01-01

    Phytase increases the availability of phytate phosphorus (P) in plant feedstuffs resulting in a reduced need for inorganic P addition and therefore minimized P excretion. Thus, the majority of pig feed is supplemented with microbial phytases. The present study aimed to examine three commercial...

  18. Evolution and diversity of PAPhy_a phytase in the genepool of wheat (Triticum aestivum L., Poaceae)

    DEFF Research Database (Denmark)

    Madsen, Claus Krogh; Petersen, Gitte; Seberg, Ole

    2017-01-01

    Higher phytase activity in food and feedstuffs is desirable in order to counter the antinutritional effects of phytate. The most promising platform where this might be achieved through plant breeding is wheat and its Triticeae relatives. They already accumulate notable amounts of phytase...

  19. The effects of supplementation of a novel bacterial 6-phytase on mineral digestibility and plasma minerals in lactating sows.

    Science.gov (United States)

    Nasir, Z; Broz, J; Zijlstra, R T

    2012-12-01

    Pigs digest P in plant feedstuffs poorly because pigs do not produce sufficient endogenous phytase to hydrolyze P from phytate (inositol hexaphosphate). Supplementation of phytase to diets of piglets and grower-finisher pigs increased digestibility of minerals including P and Ca; however, data on phytase efficacy in lactating sows are scarce. Therefore, effects of adding a bacterial 6-phytase expressed in a strain of Aspergillus oryzae (Ronozyme HiPhos; DSM Nutritional Products, Basel, Switzerland) on apparent total tract digestibility (ATTD) of P and Ca was assessed in 45 lactating sows. Three diets were prepared: (i) positive control (PC; 0.52% available P), a regular sow diet containing inorganic P, (ii) negative control (NC; 0.20% available P) without inorganic P, and (iii) NC + 500 units of phytase/kg diet. Each diet was fed randomly to 15 sows for 21 d (from 5 d prior to farrowing to 15 d after farrowing). At day 15 after farrowing, ATTD of P did not differ between PC and NC. Phytase supplementation to NC increased (P 0.05) ATTD of CP and Ca. On day 1 after farrowing, plasma P was 0.66 mmol/L lower (P Phytase supplementation to NC increased (P phytase supplementation increased P bioavailability in lactating sows.

  20. Meta-analysis of effects of microbial phytase on digestibility and bioavailability of copper and zinc in growing pigs

    NARCIS (Netherlands)

    Bikker, P.; Jongbloed, A.W.; Thissen, J.T.N.M.

    2012-01-01

    A meta-analysis was conducted to determine the effect of microbial phytase in pig diets on digestibility and bioavailability of Cu and Zn. Studies (n = 22) into effects of microbial phytase on digestibility and plasma levels of Cu and Zn were included in a dataset and regression analysis was perform

  1. Mineral absorption and excretion as affected by microbial phytase and their effect on energy metabolism in young piglets

    NARCIS (Netherlands)

    Kies, A.K.; Gerrits, W.J.J.; Schrama, J.W.; Heetkamp, M.J.W.; Linden, van der K.L.; Zandstra, T.; Verstegen, M.W.A.

    2005-01-01

    Positive effects of dietary phytase supplementation on pig performance are observed not only when phosphorus is limiting. Improved energy utilization might be one explanation. Using indirect calorimetry, phytase-induced changes in energy metabolism were evaluated in young piglets with adequate phosp

  2. Influence of dietary phytic acid and source of microbial phytase on ileal endogenous amino acid flows in broiler chickens.

    Science.gov (United States)

    Cowieson, A J; Ravindran, V; Selle, P H

    2008-11-01

    The effects of phytic acid and 2 sources of exogenous phytase (bacterial vs. fungal) on the flow of endogenous amino acids at the terminal ileum of broilers were assessed using the enzyme-hydrolyzed casein method. Phytic acid (as the sodium salt) was included in a purified diet at 8.5 and 14.5 g/kg, and each diet was fed without or with a fungal (Aspergillus niger-derived) or a bacterial (Escherichia coli-derived) microbial phytase at 500 phytase units/kg of diet. Increasing the concentration of phytic acid in the diet from 8.5 to 14.5 g/kg increased (P acids by an average of 68%, with a range from 17% for proline to 145% for phenylalanine. The flow of endogenous aspartic acid, serine, glutamic acid, glycine, leucine, tyrosine, phenylalanine, and histidine were increased by more than the mean, indicating changes in the composition of endogenous protein in response to the presence of higher concentrations of phytic acid. Supplementation of both phytases reduced (P acids, but the reduction (P = 0.06) was greater for the bacterial phytase compared with the fungal phytase. These data suggest that a substantial part of the amino acid and energy responses observed following phytase supplementation in broiler chickens stems from reduced endogenous amino acid flows and that the capacity of different phytases to counteract the antinutritive properties of phytic acid vary.

  3. Dephytinisation with Intrinsic Wheat Phytase and Iron Dephytinisation with Intrinsic Wheat Phytase and Iron Fonio (Digitaria exilis) Meals in West African Women

    NARCIS (Netherlands)

    Koreissi, Y.; Fanou-Fogny, N.M.L.; Moretti, D.; Schuth, S.; Dossa, R.A.M.; Egli, I.; Zimmerman, M.B.; Brouwer, I.D.

    2013-01-01

    Low iron and high phytic acid content make fonio based meals a poor source of bioavailable iron. Phytic acid degradation in fonio porridge using whole grain cereals as phytase source and effect on iron bioavailability when added to iron fortified fonio meals were investigated. Grains, nuts and seeds

  4. Enhancement of thermostability and kinetic efficiency of Aspergillus niger PhyA phytase by site-directed mutagenesis.

    Science.gov (United States)

    Hesampour, Ardeshir; Siadat, Seyed Ehsan Ranaei; Malboobi, Mohammad Ali; Mohandesi, Nooshin; Arab, Seyed Shahriar; Ghahremanpour, Mohammad Mehdi

    2015-03-01

    Phytase efficiently catalyzes the hydrolysis of phytate to phosphate; it can be utilized as an animal supplement to provide animals their nutrient requirements for phosphate and to mitigate environmental pollution caused by unutilized feed phosphate. Owing to animal feed being commonly pelleted at 70 to 90 °C, phytase with a sufficiently high thermal stability is desirable. Based on the crystal structure of PhyA and bioinformatics analysis at variant heat treatments, 12 single and multiple mutants were introduced by site-directed mutagenesis in order to improve phytase thermostability. Mutated constructs were expressed in Pichia pastoris. The manipulated phytases were purified; their biochemical and kinetic investigation revealed that while the thermostability of six mutants was improved, P9 (T314S Q315R V62N) and P12 (S205N S206A T151A T314S Q315R) showed the highest heat stability (P phytase to be used as an animal feed supplement.

  5. Response of broiler chickens to different levels of calcium, non-phytate phosphorus and phytase.

    Science.gov (United States)

    Akter, M; Graham, H; Iji, P A

    2016-12-01

    1. Five hundred and seventy six-d old Ross 308 broiler chicks (6 cages per diet, 8 birds per cage in 3 × 2 × 2 factorial arrangement) were fed on maize-soybean meal-based diets containing three concentrations of Ca (6, 8 or 10 g/kg), two concentrations of non-phytate phosphorus (NPP) (3 or 4 g/kg) and two levels of exogenous microbial phytase (0 or 500 FTU/kg) from d 0 to 35. 2. Body weight (BW), feed intake (FI) and mortality records were collected. Two birds per replicate were killed at 24 d of age to obtain tibia samples. 3. Increasing Ca level significantly reduced the FI and body weight gain (BWG) between hatch and 10 and 24 d, especially with the phytase-supplemented diets. However, phytase supplementation of the diet containing 4 g NPP/kg improved the FI and BWG at d 10 and 24. At d 24, phytase supplementation improved feed conversion ratio (FCR) of birds that consumed diets containing high NPP. The overall FCR was better in birds offered the phytase-supplemented, medium-Ca diet. 4. There was a significant reduction in length, width and breaking strength of the tibia bone in birds fed on a diet with high Ca and low NPP. Phytase supplementation improved the tibia ash content and bone breaking strength of chicks fed on the diet containing 8 and 4 g/kg Ca and NPP, respectively. The Ca content of the tibia bone was low in birds fed on diets with 6 and 4 g/kg Ca and NPP, respectively, but this was counteracted by phytase supplementation. 5. Birds fed on diets with 4 g/kg NPP had the best carcass percentage and parts yield. Phytase supplementation to high-Ca diets significantly reduced the carcass yield of birds. 6. These results confirmed the detrimental effect of high dietary Ca on phytase activity and subsequent growth and bone development of birds, especially when NPP is in short supply.

  6. Efficacy of different commercial phytase enzymes and development of an available phosphorus release curve for Escherichia coli-derived phytases in nursery pigs.

    Science.gov (United States)

    Jones, C K; Tokach, M D; Dritz, S S; Ratliff, B W; Horn, N L; Goodband, R D; DeRouchey, J M; Sulabo, R C; Nelssen, J L

    2010-11-01

    In 2 experiments, a total of 184 pigs (PIC, initial BW of 10.3 and 9.7 kg for Exp. 1 and 2, respectively) were used to develop an available P (aP) release curve for commercially available Escherichia coli-derived phytases. In both experiments, pigs were fed a corn-soybean meal basal diet (0.06% aP) and 2 diets with added inorganic P (iP) from monocalcium phosphate (Exp. 1: 0.075 and 0.15% aP; Exp. 2: 0.07 and 0.14% aP) to develop a standard curve. In Exp. 1, 100, 175, 250, or 500 phytase units (FTU)/kg of OptiPhos 2000 or 200, 350, 500, or 1,000 FTU/kg of Phyzyme XP were added to the basal diet. In Exp. 2, 250, 500, 750, or 1,000 FTU/kg of OptiPhos 2000; 500, 1,000, or 1,500 FTU/kg of Phyzyme XP; or 1,850 or 3,700 FTU/kg of Ronozyme P were added to the basal diet. One FTU was defined as the amount of enzyme required to release 1 µmol of iP per minute from sodium phytate at 37°C. For all phytase products, the manufacturer-guaranteed phytase activities were used in diet formulation. All diets were analyzed for phytase activity using both the Phytex and AOAC methods. Pigs were blocked by sex and BW and allotted to individual pens with 8 pens per treatment. Pigs were killed on d 21, and fibulas were collected and analyzed for bone ash. In both experiments, increasing iP improved (linear, P < 0.01) G:F and percentage bone ash. Pigs fed increasing OptiPhos had improved (Exp. 1: linear, P < 0.001; Exp. 2: quadratic, P < 0.001) percentage bone ash, as did pigs fed increasing Phyzyme XP (linear, P < 0.001). In Exp. 2, increasing Ronozyme P improved (quadratic, P < 0.01) percentage bone ash. Using analyzed values from the AOAC method and percentage bone ash as the response variable, an aP release curve was developed for up to 1,000 FTU/kg of E. coli-derived phytases (OptiPhos 2000 and Phyzyme XP) in P-deficient diets. The prediction equation was Y = -0.000000125X(2) + 0.000236X + 0.016, where Y = aP release (%) and X = analyzed phytase (FTU/kg) in the diet.

  7. Novel phytase from Pteris vittata resistant to arsenate, high temperature, and soil deactivation.

    Science.gov (United States)

    Lessl, Jason T; Ma, Lena Q; Rathinasabapathi, Bala; Guy, Charles

    2013-03-01

    Arsenate interferes with enzymatic processes and inhibits inorganic phosphorus (Pi) uptake in many plants. This study examined the role of phytase and phosphatase in arsenate tolerance and phosphorus (P) acquisition in the arsenic hyperaccumulator Pteris vittata . Enzyme-mediated hydrolysis of phytate in P. vittata extracts was not inhibited by arsenate at 5 mM or by heating at 100 °C for 10 min. Root exudates of P. vittata exhibited the highest phytase activity (18 nmol Pi mg(-1) protein min(-1)) when available P was low, allowing its growth on media amended with phytate as the sole source of P. Phosphorus concentration in P. vittata gametophyte tissue grown on phytate was equivalent to plants grown with inorganic phosphate at 2208 mg kg(-1), and arsenic was increased from 1777 to 2630 mg kg(-1). After 2 h of mixing with three soils, P. vittata phytase retained more activity, decreasing from ∼ 26 to ∼ 25 nmol Pi mg(-1) protein min(-1), whereas those from Pteris ensiformis and wheat decreased from ∼ 18 to ∼ 1 nmol Pi mg(-1) protein min(-1). These results suggest P. vittata has a uniquely stable phytase enabling its P acquisition in P-limiting soil environments. Furthermore, the P. vittata phytase has potential use as a soil amendment, a transgenic tool, or as a feed additive supplement, reducing the need for nonrenewable, polluting P fertilizers.

  8. Degradation of phytate by the 6-phytase from Hafnia alvei: a combined structural and solution study.

    Science.gov (United States)

    Ariza, Antonio; Moroz, Olga V; Blagova, Elena V; Turkenburg, Johan P; Waterman, Jitka; Roberts, Shirley M; Vind, Jesper; Sjøholm, Carsten; Lassen, Søren F; De Maria, Leonardo; Glitsoe, Vibe; Skov, Lars K; Wilson, Keith S

    2013-01-01

    Phytases hydrolyse phytate (myo-inositol hexakisphosphate), the principal form of phosphate stored in plant seeds to produce phosphate and lower phosphorylated myo-inositols. They are used extensively in the feed industry, and have been characterised biochemically and structurally with a number of structures in the PDB. They are divided into four distinct families: histidine acid phosphatases (HAP), β-propeller phytases, cysteine phosphatases and purple acid phosphatases and also split into three enzyme classes, the 3-, 5- and 6-phytases, depending on the position of the first phosphate in the inositol ring to be removed. We report identification, cloning, purification and 3D structures of 6-phytases from two bacteria, Hafnia alvei and Yersinia kristensenii, together with their pH optima, thermal stability, and degradation profiles for phytate. An important result is the structure of the H. alvei enzyme in complex with the substrate analogue myo-inositol hexakissulphate. In contrast to the only previous structure of a ligand-bound 6-phytase, where the 3-phosphate was unexpectedly in the catalytic site, in the H. alvei complex the expected scissile 6-phosphate (sulphate in the inhibitor) is placed in the catalytic site.

  9. Potential of Phytase-Mediated Iron Release from Cereal-Based Foods: A Quantitative View

    Directory of Open Access Journals (Sweden)

    Anne S. Meyer

    2013-08-01

    Full Text Available The major part of iron present in plant foods such as cereals is largely unavailable for direct absorption in humans due to complexation with the negatively charged phosphate groups of phytate (myo-inositol (1,2,3,4,5,6-hexakisphosphate. Human biology has not evolved an efficient mechanism to naturally release iron from iron phytate complexes. This narrative review will evaluate the quantitative significance of phytase-catalysed iron release from cereal foods. In vivo studies have shown how addition of microbially derived phytases to cereal-based foods has produced increased iron absorption via enzyme-catalysed dephosphorylation of phytate, indicating the potential of this strategy for preventing and treating iron deficiency anaemia. Despite the immense promise of this strategy and the prevalence of iron deficiency worldwide, the number of human studies elucidating the significance of phytase-mediated improvements in iron absorption and ultimately in iron status in particularly vulnerable groups is still low. A more detailed understanding of (1 the uptake mechanism for iron released from partially dephosphorylated phytate chelates, (2 the affinity of microbially derived phytases towards insoluble iron phytate complexes, and (3 the extent of phytate dephosphorylation required for iron release from inositol phosphates is warranted. Phytase-mediated iron release can improve iron absorption from plant foods. There is a need for development of innovative strategies to obtain better effects.

  10. Growth performance, body lipid, brood amount, and rearing environment response to supplemental neutral phytase in zebrafish (Danio rerio) diet.

    Science.gov (United States)

    Liu, Liwei; Su, Jianmei; Liang, Xu-Fang; Luo, Yuliang

    2013-09-01

    The present study was to evaluate the effects of neutral phytase supplementation on growth performance, survival ratio (SR), body lipid, brood amount, and rearing environment in zebrafish. The control diet was not supplemented phytase, and three levels of phytase (500, 1000, or 1500 U kg(-1)) was added to the three other diets (named as PP500, PP1000, and PP1500). Triplicate groups (twelve 100-L tanks) of zebrafish (initial mean weight, 0.284±0.012 g) were fed twice daily (08:00 and 16:00 h) to satiation for 12 weeks. The results showed that supplemental phytase in the diet improved weight gain (60.49%, 86.63%, 99.06%, and 111.88% in control, PP500, PP1000, and PP1500) and the specific growth ratio of zebrafish (pzebrafish. The brood amounts (116, 123, and 124 eggs in PP500, PP1000, and PP1500) of fish fed with phytase-supplemented diets were little higher than the control (mean egg was 112). The ammonia-nitrogen concentration in water of fish fed with phytase-supplemented diet was significantly lower than the control. The nitrite concentration in water was also decreased in water of fish fed with phytase-supplemented diet. The SR was increased with the increasing of dietary phytase despite no significant difference was observed among each group. The present study first suggested that neutral phytase could be applied in the zebrafish diet. Furthermore, phytase addition increased the growth, body lipid, brood amount, and SR of zebrafish, and meanwhile decreased the ammonia-nitrogen and nitrite concentrations in rearing water.

  11. Influence of a microbial phytase and zinc oxide on young pig growth performance and serum minerals.

    Science.gov (United States)

    Walk, C L; Srinongkote, S; Wilcock, P

    2013-01-01

    Crossbred pigs (n=288; average age=21±3 d and BW=7.1±0.2 kg) were used in a 42-d trial to determine the influence of a microbial phytase and various doses of ZnO on growth performance and serum minerals. Pigs (6 castrated males or females/pen) were randomly allotted to treatments in a 2×3 factorial arrangement with 2 dietary levels of a microbial phytase (0 or 2,500 phytase units/kg) and 3 dietary levels of supplemental ZnO [0, 1750, or 3,500 mg/kg ZnO (72% Zn)] with 4 pens of castrated males and 4 pens of females per treatment. Diets were formulated to exceed all nutrient requirements, including Ca and P from d 0 to 21 (phase 1) and d 22 to 42 (phase 2). Growth performance, serum Zn, and serum P were not influenced (P>0.05) by a ZnO×phytase interaction during phase 1, phase 2, or overall (d 0 to 42). Phytase increased (P=0.01) ADFI and improved (P=0.02) ADG in phase 1 and improved (P=0.01) overall ADG, regardless of the level of ZnO supplemented. Zinc oxide supplementation linearly reduced (P=0.05) ADG, and ZnO at 3,500 mg/kg reduced (quadratic, P=0.04) G:F in pigs (phase 2). During phase 1, phytase increased serum Ca, but only in the absence of ZnO supplementation, which resulted in a ZnO×phytase interaction (P=0.02). Serum Zn was increased (linear, Psupplementation increased in the diet (phase 1). In phase 2, serum Ca was reduced (linear, P=0.04) and serum Zn was increased (linear, P=0.02) as ZnO supplementation increased in the diet. Phytase supplementation increased (P=0.009) serum Zn and increased (P=0.003) serum P (phase 1). There was no influence of phytase supplementation on serum minerals in phase 2. In summary, supplemental ZnO reduced growth performance in this experiment. Phytase supplementation improved ADG in Ca- and P-adequate diets regardless of the level of ZnO supplemented, which may be attributed to the reduction of phytate as an antinutrient. In addition, through phytate hydrolysis, phytase reduced phytate-Zn interactions and increased

  12. The Effect of Phytase Enzyme on the Performance of Broiler Flock (A-Review

    Directory of Open Access Journals (Sweden)

    Khan SA

    2013-12-01

    Full Text Available From the last few years, the inclusion of microbial phytase in poultry diets has increased significantly, mainly in response to heightened concerns over phosphorus pollution of the environment and as cheaper means to make phosphorus available to birds from phytate. Phytate is the major form of phosphorus, abundantly found in cereal grains, beans and oilseed meals used in poultry diet but the monogastric animals like poultry birds are unable to utilize this source of phosphorus due to lack of endogenous phytase enzyme. To meet the phosphorus requirements of poultry birds, inorganic phosphates are added to the poultry rations, which lead to the problem of environmental pollution as a large amount of phosphorus is excreted in the manure. Microbial phytase is used as an alternative of this, which has beneficial effects on the growth performance, feed efficiency, protein/amino acid digestibility, energy utilization, mineral retention, and bone growth of broilers due to the direct hydrolytic effects on phytate.

  13. Potential of phytase-mediated iron release from cereal-based foods: a quantitative view

    DEFF Research Database (Denmark)

    Nielsen, Anne Veller Friis; Tetens, Inge; Meyer, Anne S.

    2013-01-01

    The major part of iron present in plant foods such as cereals is largely unavailable for direct absorption in humans due to complexation with the negatively charged phosphate groups of phytate (myo-inositol (1,2,3,4,5,6)-hexakisphosphate). Human biology has not evolved an efficient mechanism...... to naturally release iron from iron phytate complexes. This narrative review will evaluate the quantitative significance of phytase-catalysed iron release from cereal foods. In vivo studies have shown how addition of microbially derived phytases to cereal-based foods has produced increased iron absorption via...... enzyme-catalysed dephosphorylation of phytate, indicating the potential of this strategy for preventing and treating iron deficiency anaemia. Despite the immense promise of this strategy and the prevalence of iron deficiency worldwide, the number of human studies elucidating the significance of phytase...

  14. The genomic complement of purple acid phosphatase phytases in the Triticeae

    DEFF Research Database (Denmark)

    Madsen, Claus Krogh; Dionisio, Giuseppe; Holme, Inger

    2011-01-01

    phytase activity in the mature grains, however varying significantly between the individual species. After multiple steps of chromatography, the phytase activity elutes as one peak, indicating that it consists of either one enzyme or more than one very similar enzymes. Recent work in our group** has...... demonstrated that these enzymes are purple acid phosphatase phytases (PAPhy’s) encoded by a few highly conserved mRNA’s expressed either during grain filling (PAPhy_a’s) or germination (PAPhy_b’s). In the present study, 15 genomic PAPhy sequences from wheat, barley, rye, einkorn and Aegilops taushii were......, PAPhy_b promoters contain elements typical of gibberellic acid induced germination related hydrolases. PAPhy_a promoters in contrast possess elements known from storage protein promoters. **Dionisio G, Madsen CK, Holm PB, Welinder KG, Jørgensen M, Stoger E, Arcalis E, Brinch-Pedersen H. Cloning...

  15. Phosphorus equivalence of a Consensus phytase produced by Hansenula polymorpha in diets for young turkeys.

    Science.gov (United States)

    Esteve-Garcia, Enric; Perez-Vendrell, Anna M; Broz, Jiri

    2005-02-01

    An experiment was conducted to determine the efficacy of a so-called consensus phytase preparation produced by Hansenula polymorpha on growth, tibia and toe ash and P retention of young turkeys. A total of 192 female turkeys (BUT 9 strain) were placed into 96 batteries at two per cage and assigned to one of eight diets: A negative control containing 2.5 g non-phytate P per kg feed (T-1); T-1 plus 125, 250, 500, 1000 or 10 000 phytase units (U) per kg feed, respectively, (T-2 to T-6); T-1 plus 0.5 or 1.0 g Pi/kg feed as dicalcium phosphate dihydrate (DCP), respectively, (T-7 and 8). The experiment lasted 32 d, and excreta were collected and weighed between 26 and 29 d of age. Feed was also weighed during this period in order to calculate P retention. Performance was calculated for the 0 to 32 d period. At the end of the experiment one bird per pen was killed for tibia and toe ash content determination, except for T-2, T-3, and T-7. Body weight, FCR, toe and tibia ash, and P retention responded to phytase or Pi supplementation. Using linear or quadratic models for comparing performance of the treatments containing supplemental Pi with phytase treatments, an equivalence between phytase and Pi was calculated. Body weight, toe ash, tibia ash and P retention showed a significant response to phytase supplementation. The values of equivalence for body weight, toe ash, tibia ash and P retention were 251, 597, 391 and 390 U to 1 g Pi/kg feed. At 10 000 U/kg feed there was a significant response in terms of weight gain and P retention, indicating that turkeys respond to levels greater than 1 000 U/kg feed.

  16. Effects of AsN1980 phytase on growth and phosphorus utilization of broilers

    Institute of Scientific and Technical Information of China (English)

    Ding Yi

    2003-01-01

    The experiment was conducted with 180 one-day-old AA broilers that were randomly divided into onecontrol group and one trial group to study the effects of AsN1980 phytase on broilers growth performance and P utilizationin low available phosphorus diets. Results showed that phytase supplement decreased F/W by 12.4%-13.4% andincreased phosphorus availability by 7.63%-8.27%, both of which were significantly superior to the control group(P <0.05).

  17. Redistribution of mineral elements in wheat grain when applying the complex enzyme preparations based on phytase

    Directory of Open Access Journals (Sweden)

    Elena Kuznetsova

    2016-01-01

    Full Text Available Biogenic minerals play an important role in the whole human nutrition, but they are included in the grain of the phytates that reduces their bioavailability. Whole wheat bread is generally considered a healthy food, but the presence of mineral elements in it is insignificant, because of weak phytate degradation. From all sources of exogenous phytase the most productive are microscopic fungi. To accelerate the process of transition hard mineral elements are mobilized to implement integrated cellulolytic enzyme preparation based on the actions of phytase (producer is Penicillium canescens. Phytase activity was assessed indirectly by the rate of release of phosphate from the substrate. It has been established that the release rate of the phosphoric acid substrate is dependent on the composition of the drug and the enzyme complex is determined by the presence of xylanase. The presented experimental data shows that a cellulase treatment of the grain in conjunction with the β-glucanase or xylanase leading to an increase in phytase activity could be 1.4 - 2.3 times as compared with the individual enzymes. As a result of concerted action of enzymes complex preparation varies topography grain, increase the pore sizes in seed and fruit shells that facilitate the penetration of the enzyme phytase in the aleurone layer to the site of phytin hydrolysis and leads to an increase in phytase activity. In terms of rational parameters of enzymatic hydrolysis, the distribution of mineral elements in the anatomical parts of the grain after processing complex enzyme preparation with the help of X-ray detector EMF miniCup system in a scanning electron microscope JEOL JSM 6390 were investigated. When processing enzyme preparation wheat trend in the distribution of mineral elements, characteristic of grain - the proportion of these elements in the aleurone layer decreases, and in the endosperm increases. Because dietary fiber and phytate found together in the

  18. The supplementation of low-P diets with microbial 6-phytase expressed in Aspergillus oryzae improves P digestibility in sows.

    Science.gov (United States)

    Torrallardona, D; Llauradó, L; Broz, J

    2012-12-01

    Two trials were conducted to evaluate a novel microbial 6-phytase expressed in Aspergillus oryzae (Ronozyme HiPhos; DSM Nutritional Products, Basel, Switzerland) in gestating and lactating sows. In the first trial, 24 sows (Duroc × Landrace; 223 kg BW) were offered, at 16 d of gestation, a low-P control diet (formulated to provide 4.0 g total P/kg; 1.5 g digestible P/kg) supplemented with 0, 500, or 1000 phytase activity (FYT)/kg of phytase. Two weeks later, fresh feces were sampled from all sows and the apparent total tract digestibility of P was measured using TiO(2) as indigestible marker. Phytase supplementation did not (P > 0.10) affect the total tract digestibility of P but reduced (P phytase. After 2 wk, fresh feces were sampled from all sows and the apparent total tract digestibility of P was measured using TiO(2) as indigestible marker. Phytase supplementation improved (P phytase tested increased the apparent total tract digestibility of P in sows and reduced P excretion in feces.

  19. The effect of phytase and fructooligosaccharide supplementation on growth performance, bone quality, and phosphorus utilization in broiler chickens.

    Science.gov (United States)

    Shang, Y; Rogiewicz, A; Patterson, R; Slominski, B A; Kim, W K

    2015-05-01

    An experiment was conducted to investigate the effects of phytase and 2 levels of fructooligosaccharide (FOS) supplementation on growth performance, bone mineralization, and P utilization of broiler chickens. A total of 210 day-old male broiler chickens (Ross) were randomly placed into 7 dietary treatments consisting of 6 replicates with 5 birds per pen. The experiment was designed as an augmented 2 × 3 factorial arrangement with 0 or 500 U/kg of phytase and 0, 0.5% or 1% of FOS added to a reduced Ca (0.8%) and available P (0.25%) negative control diet (NC). A positive control diet (PC) that contained 1% Ca and 0.45% available P was also included. During the entire experimental period, phytase supplementation significantly improved (P Phytase supplementation increased femur BMD (P Phytase alone and in combination with 0.5% FOS increased P utilization significantly when compared with other treatments (P phytase supplementation in low Ca and P diets improved growth performance, bone quality, and P utilization. However, supplementing NC diets with phytase and FOS did not result in bone mineralization values comparable with that of the PC diet. The application of dietary FOS alone had a negative effect on broiler bone quality.

  20. In vivo and in vitro digestibility of plant ingredients and diets by Bacillus phytases in tilapia, Oreochromis mossambicus

    Directory of Open Access Journals (Sweden)

    Rande B. Dechavez

    2012-12-01

    Full Text Available This study aimed to evaluate four Bacillus phytases for their efficacy in making plant-baseddiets bioavailable to tilapia (Oreochromis mossambicus using in vivo digestibility measurement and todetermine the in vitro level of dephosphorylation. The four Bacillus strains used were B. pumilus , B. megaterium , B. coagulans, and B. licheniformis. Phytase activities varied between bacterial sources aswell as between feed ingredients. For the cassava leaf meal, Pi released was highest in B. pumilus andwas not significantly different from those of B. megaterium and B. licheniformis. For the soybean meal, Pirelease was in this decreasing order: B. megaterium > B. pumilus > B. coagulans > B. licheniformisphytase. For the corn meal, addition of B. licheniformis phytase to the reaction mixture resulted insignificantly the highest Pi released followed by B. coagulans phytase which was not significantly differentfrom that of B. megaterium phytase which released the lowest Pi. Pi released by B. pumilus phytase fromcorn meal was not significantly different from the lowest Pi release of B. megaterium phytase. Theapparent digestibility coefficient (ADC values for the feed dry matter (DM ranged from 86.3 to 88.3%and were not significantly different from each other (p > 0.05.

  1. The interplay of dietary nutrient level and varying calcium to phosphorus ratios on efficacy of a bacterial phytase: 2. Ileal and total tract nutrient utilization.

    Science.gov (United States)

    Olukosi, O A; Fru-Nji, F

    2014-12-01

    A 14-d broiler experiment was conducted to assess the effects of 2 dietary variables on efficacy of a bacterial 6-phytase from Citobacter braakii on nutrient and phytate P (PP) utilization. Diets were formulated with or without nutrient matrix values (matrix) for phytase as negative control (NC) or positive control (PC), respectively, and with 2 Ca:total P (tP) levels (2:1 or 2.5:1). The diets were supplemented with 0, 1,000, or 2,000 phytase units (FYT)/kg of diet, thus producing a 2 × 2 × 3 factorial arrangement. Excreta were collected on d 19 to 21 and ileal digesta on d 21. There was no 3-way interaction on digestibility of any nutrient. There was matrix × phytase (P phytase interaction (P phytase increased (P phytase supplementation in diets with 2:1 Ca:tP, whereas there was no effect of phytase supplementation on PP disappearance or Ca retention in diets with 2.5:1 Ca:tP. Total P and Ca retention were reduced (P phytase supplementation on P utilization is reduced when diets contain adequate P as exemplified in the PC diets and that the negative impact of wide Ca:tP is more pronounced in diets with phytase matrix allowance as exemplified in the NC diets.

  2. The addition of a Buttiauxella sp. phytase to lactating sow diets deficient in phosphorus and calcium reduces weight loss and improves nutrient digestibility.

    Science.gov (United States)

    Wealleans, A L; Bold, R M; Dersjant-Li, Y; Awati, A

    2015-11-01

    Improving the efficiency of P use by pigs is especially important for lactating sows, whose metabolic requirements for P and Ca are high. The effect of a sp. phytase on lactating sow performance and nutrient digestibility was investigated using the combined data set for 6 studies. Treatments included a nutritionally adequate positive control diet (PC), a negative control diet (NC; with an average reduction of 0.16% available phosphorous and 0.15% Ca vs. PC), and NC supplemented with a sp. phytase at 250, 500, 1,000 or 2,000 phytase unit (FTU)/kg, respectively. Phosphorus and Ca deficiency in the NC resulted in significantly higher BW loss compared with the PC. All phytase treatments maintained BW loss at the same level as the PC. Increasing doses of phytase significantly ( phytase supplementation. Digestible P and Ca were significantly improved (linear, phytase supplementation. Significantly lower apparent total tract digestibility of energy, Ca, and P was found in the NC treatment vs. the PC treatment, whereas no significant differences were found between phytase treatment and the PC treatment. In conclusion, phytase supplementation at a level of 250 FTU/kg can replace 0.16% available phosphorous and 0.15% Ca; however, increasing the phytase dose can further reduce BW loss in sows fed P- and Ca- deficient diets.

  3. Comparison of 3 phytases on energy utilization of a nutritionally marginal wheat-soybean meal broiler diet.

    Science.gov (United States)

    Wu, D; Wu, S B; Choct, M; Swick, R A

    2015-11-01

    The net energy (NE) value may be a better measure than apparent metabolizable energy (ME) of the effect of supplemental phytase on energy utilization in broilers. The present study was conducted to assess the impact of 3 microbial phytases supplemented at an unconventionally high level (1,000 FTU/kg feed) on performance and NE of broilers using the indirect calorimetric method (IC). Four treatments included: 1) Control, formulated to be deficient in ME (12.35 MJ/kg in the starter diet; 12.56 MJ/kg in the grower diet), calcium (0.72% in the starter diet; 0.60% in the grower diet), and available phosphorus (0.25% in the starter diet; 0.20% in the grower diet); 2) control + intrinsically thermostable phytase A; 3) control + intrinsically thermostable phytase B; and 4) control + coated phytase C. A completely randomized design was employed. A total of 384 male broiler chicks were used, and each treatment had 6 replicates with 16 birds per replicate. The birds were reared until d 21 in floor pens with hardwood shavings. Thirty-two birds (8 birds per treatment) were randomly selected to determine heat production and NE (from 25-28 d) following a 3-d acclimatization in the respiratory chambers. Performance results at d 21 showed that supplementation with either of the 3 phytases improved body weight (P diet (P < 0.05). It may be concluded that the negative effects imposed by calcium and available phosphorus down-specification can be compensated by phytase supplementation in general, and intrinsically thermostable phytases improve the ME and NE value. However, phytase did not reduce heat production, heat increment, or increase NE:ME in birds.

  4. Isolation and Characterization of Potential Phytase-Producing Fungi from Environmental Samples of Antioquia (Colombia Aislamiento y Caracterización de Hongos Productores de Fitasa a partir de Muestras Ambientales de Antioquia (Colombia

    Directory of Open Access Journals (Sweden)

    Maritza Ocampo Betancur

    2012-06-01

    Full Text Available Abstract. Phytases are enzymes used as feed additive that enhance the phosphorus and mineral uptake in monogastric animals and reduce the level of phosphate excretion in their manure. Due to their easy cultivation and high production of extracellular enzymes, filamentous fungi are one of best sources of phytase for use in the feed industry. Phytase has been found principally in the genera Aspergillus, Penicillium, Mucor and Rhizopus. In this work, we report the isolation and characterization of environmental fungi producers of phytase with potential use as feed additives. Samples were collected from soils, fruits and cereals in Antioquia (Colombia. A total of 26 fungal strains were isolated and identified using ITS sequencing and morphological analysis. Strains belonged to the following genera: Penicillium, Aspergillus, Fusarium, Mortierella, Pestalotiopsis, Phoma, Paecilomyces and Rigidoporus. Fifty percent of isolates exhibited halos in phytase screening agar indicating that acidic phytases are common enzymes secreted by environmental fungi. Ten isolates were also able to grow in liquid phytase screening medium revealing their potential use for enzyme production in submerged fermentations. Molecular detection of the PhyA gene from Aspergillus was achieved. Partial sequence of the phyA gene from one A. niger isolate was obtained and analyzed.Resumen. Las fitasas son enzimas utilizadas como aditivo en productos de alimentación animal, con el fin de mejorar la asimilación de fósforo y minerales en animales monogástricos y disminuir la excreción de fósforo al ambiente. Los hongos filamentosos son una de las mejores fuentes de fitasas debido a su facilidad de cultivo y altos niveles de producción de enzimas extracelulares. Los principales productores de fitasas corresponden a miembros de los géneros Aspergillus, Penicillium, Mucor y Rhizopus. En este trabajo se reporta el aislamiento y caracterización de hongos ambientales productores de

  5. Effect of phytase supplementation on rumen fermentation characteristics and phosphorus balance in lactating dairy cows

    Directory of Open Access Journals (Sweden)

    Laura Winter

    2015-02-01

    Full Text Available This study aimed to evaluate the effects of exogenous phytase on rumen fermentation characteristics, the phosphorus (P-flow at the duodenum and the P-balance in lactating dairy cows. For this purpose ruminal and duodenally fistulated cows were assigned to one of three dietary treatments: high P (HP diet (n=7 provided a total of 45 g/d of P, archived by a supplementation of dicalcium phosphate to the diet; low P (LP diet (n=5 provided 34 g/d of P without supplementation; LP+phytase (LP+PHY diet (n=5 provided 34 g/d of P supplemented with an exogenous phytase. Dry matter intake and milk yield were recorded daily. In the first week of a sampling period Pbalance was determined. Samples of ruminal fluid were taken and duodenal chyme was collected in the second sampling week. Ruminal pH and the concentration of volatile fatty acids were not different between the treatments. The HP-group shows a higher P-flow at the duodenum than other groups. No differences in apparent total tract P-digestibility were found between the treatments. The P-balance in the HP-group (2.6 g/d was higher compared to the LP (-3.2 g/d and LP+PHY (-3.0 g/d group. Overall, phytase supplementation had no effect on P-digestibility in lactating dairy cows.

  6. Microbial phytase and liquid feeding increase phytate degradation in the gastrointestinal tract of growing pigs

    DEFF Research Database (Denmark)

    Blaabjerg, Karoline; Poulsen, Hanne Damgaard

    2010-01-01

    The quantitative degradation of inositol phosphates (InsP6 to InsP2) in the stomach and small intestine as influenced by microbial phytase and fermented liquid feeding was compared by combining the results from two experiments. Six pigs (49 kg) were fitted with gastric cannulas ( Exp. 1 ) and 3 p...

  7. OPTIMIZING THE PH PROFILE OF FUNGAL PHYTASE THROUGH PROTEIN ENGINEERING ALSO ENHANCES ACTIVITY

    Science.gov (United States)

    Phytic acid is the primary phosphorus storage compound in plants. As the amount of plant derived protein in animal feed has increased a problem developed. The concern is that monogastric animals lack a phytase in their digestive tract to hydrolyze this compound that is found in soybean and other p...

  8. PERFORMANCE OF LAYER HEN FED FERMENTED Jatropha Curcas L. MEAL SUPPLEMENTED WITH CELLULASE AND PHYTASE ENZYME

    Directory of Open Access Journals (Sweden)

    S. Sumiati

    2014-10-01

    Full Text Available The objective of the experiment was to study the effect of feeding fermented Jatropha curcas L.meal (JCM supplemented with cellulase and phytase on the performances of ISA-Brown laying henaged 25-30 weeks. The Jatropha curcas meal was fermented using Rizhopus oligosporus. In this study200 laying hens were used and distributed to 5 treatments and 4 replications in Completely RandomizedDesign. The diet treatments were: R0 = control diet (without JCM, R1; diet contained fermented JCM7.5%, R2; diet contained fermented JCM 7.5% + celullase 200 g/ton, R3; diet contained fermented JCM7.5% + phytase 200 g/ton and R4; diet contained fermented JCM 7.5% + cellulase 200 g/ton + phytase200 g/ton. The parameters observed were feed consumption, hen day egg production, egg massproduction, egg weight and feed conversion ratio. The results showed that feeding fermented JCM 7.5%,both enzyme supplemented as well as unsupplemented significantly decreased (P<0.05 the feedconsumption, hen day egg and egg mass production. However, the treatments did not influence the eggweight. Supplementation of cellulase (R2 or phytase (R3 improved the feed conversion ratio with thevalue as same as the R0 diet.

  9. Characterization of manure from conventional and phytase transgenic pigs by advanced solid-state NMR spectroscopy

    Science.gov (United States)

    Non-point phosphorus (P) pollution from animal manure is becoming a serious global problem. The current solution for the swine industry is including the enzyme phytase as a component of the cereal grain diet. A very real possibility in the future is the production of transgenic pigs that express phy...

  10. Purification and characterization of a thermostable extracellular phytase from Bacillus licheniformis PFBL-03.

    Science.gov (United States)

    Fasimoye, Feyisola O; Olajuyigbe, Folasade M; Sanni, Morakinyo D

    2014-01-01

    Extracellular phytase from Bacillus licheniformis PFBL-03 was purified in three steps by using ammonium sulfate precipitation, ion-exchange chromatography on a DEAE Sephadex A-50 column, and gel filtration chromatography on Sephadex G-100. The single protein band on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) suggested that the enzyme was homogeneous. The molecular mass determined from SDS-PAGE was 36 kD. The enzyme yield was 10% while the purification fold was 39. The purified phytase exhibited optimum activity at 55°C and was able to retain 55% of its original activity after 60 min of incubation at 80°C. The purified enzyme had optimum pH of 6.0 and was stable over a pH range of 4.0 to 7.5. The kinetic parameters K(m) and V(max) of the purified phytase for sodium phytate were 4.7 mM and 49.01 µmol/min. The activity of the enzyme was enhanced in the presence of Ca²⁺ but completely inhibited by Fe²⁺, Mn²⁺, and Cu²⁺. The exhibited characteristics of the purified phytase from Bacillus licheniformis PFBL-03 show that the enzyme has potential for applications in the animal feed industry.

  11. Characteristics and Applicability of Phytase of the Yeast Pichia anomala in Synthesizing Haloperoxidase.

    Science.gov (United States)

    Joshi, Swati; Satyanarayana, Tulasi

    2015-07-01

    The phytase of the yeast Pichia anomala is a histidine acid phosphatase based on signature sequences and catalytic amino acids identified by site-directed mutagenesis. Among modulators, N-bromosuccinimide and butanedione inhibit phytase, while Ca(2+) and Ni(2+) stimulate slightly. Vanadate exhibits competitive inhibition of phytase, making it bifunctional to act as haloperoxidase. Molecular docking supports vanadate to share its binding site with phytate. The T 1/2, activation energy (E a ), temperature quotient (Q 10), activation energy of thermal inactivation (Ed), and enthalpy (ΔH d (0) ) of the enzyme are 4.0 min (80 °C), 27.72 kJ mol(-1), 2.1, 410.62 kJ mol(-1), and ∼407.8 kJ mol(-1) (65-80 °C), respectively. The free energy of the process (ΔG d (o) ) increases from 49.56 to 71.58 kJ mol(-1) with rise in temperature, while entropy of inactivation (ΔS d (0) ) remains constant at ∼1.36 kJ mol(-1) K(-1). The supplementation of whole wheat dough with rPPHY resulted in 72.5 % reduction in phytic acid content of bread. These characteristics confirm that the phytase has adequate thermostability for its applicability as a food and feed additive.

  12. Diurnal variation in degradation of phytic acid by plant phytase in the pig stomach

    NARCIS (Netherlands)

    Kemme, P.A.; Jongbloed, A.W.; Mroz, Z.; Beynen, A.C.

    1998-01-01

    The effects of plant phytase on the gastric degradation of phytic acid and digestibilities of DM and P, and their diurnal variation were evaluated in pigs from 90 to 115 kg BW fitted with simple duodenal T-cannulas. Three diets were fed to three pigs in four collection periods according to a cross-o

  13. Increased understanding of the cereal phytase complement for better mineral bio-availability and resource management

    DEFF Research Database (Denmark)

    Brinch-Pedersen, Henrik; Madsen, Claus Krogh; Holme, Inger Bæksted

    2014-01-01

    for the bio-availability of micronutrients in human nutrition. The composition and levels of mature grain phytase activity (MGPA) in cereals is of central importance for efficient phytate hydrolysis. The MGPA varies considerably between species. Substantial activity is present in Triticeae tribe cereals like...

  14. The Effect of Phytase on the Oxygen Isotope Composition of Phosphate

    Science.gov (United States)

    von Sperber, C.; Tamburini, F.; Bernasconi, S. M.; Frossard, E.

    2013-12-01

    Plants and microorganisms under phosphorus (P) stress release extracellular phosphatases as a strategy to acquire inorganic phosphate (Pi) (1-2). These enzymes catalyze the hydrolysis of phosphoesters leading to a release of Pi. The enzymatic hydrolysis leads, via a nucleophilic attack, to the incorporation of one oxygen atom from the water into the newly formed Pi molecule. During the incorporation, an isotopic fractionation occurs, which might be used to identify the origin of Pi in the environment (3-6). While the effect of phosphomonoesterases and phosphodiesterases on the oxygen isotope composition of phosphate has been examined, there are, so far, no studies dealing with the effect of phytases (4-6). Phytases catalyze the hydrolysis of myo-inositol-hexakis-phosphate (IP6), which is an important component of organic P in many ecosystems (7). Enzymatic assays with phytase from wheat germ and Aspergillus niger were prepared under sterile and temperature controlled conditions in order to determine the effect of phytases on the oxygen isotope composition of phosphate, which has been liberated from IP6 via enzymatic hydrolysis. Assays with phytase from wheat germ lead to a turnover of the substrate close to 100%, while assays with phytase from Aspergillus niger lead to a turnover of the substrate close to 80%. In the case of the assays with phytase from wheat germ, our results indicate that one sixth of the total 24 oxygen which are associated to the phosphates in IP6 are exchanged with oxygen from water. From this we conclude that the incorporation of one oxygen atom from water occurs only at four phosphate molecules of IP6, while two phosphate molecules do not experience an incorporation of oxygen. This suggests that during the enzymatic hydrolysis, four P-O bonds and two C-O bonds are broken. Provided that, the isotopic fractionation can be calculated with an isotopic mass balance resulting in -8.4‰ (×3.6 SD). This is a value very similar to those reported

  15. Replacement P212H altered the pH-temperature profile of phytase from Aspergillus niger NII 08121.

    Science.gov (United States)

    Ushasree, Mrudula Vasudevan; Vidya, Jalaja; Pandey, Ashok

    2015-03-01

    Microbial phytase, a widely used animal feed enzyme, needs to be active and stable in the acidic milieu for better performance in the monogastric gut. Aspergillus niger phytases exhibit an activity dip in the pH range from 3.0 to 3.5. Replacement of amino acids, which changed the pKa of catalytic residues H82 and D362, resulted in alteration of the pH profile of a thermostable phytase from A. niger NII 08121. Substitution P212H in the protein loop at 14 Å distance to the active site amended the pH optimum from 2.5 to pH 3.2 nevertheless with a decrease in thermostability than the wild enzyme. This study described the utility of amino acid replacements based on pKa shifts of catalytic acid/base to modulate the pH profile of phytases.

  16. Nutritional Balance of broilers fed diets containing two calcium levels and supplemented with different phytase levels

    Directory of Open Access Journals (Sweden)

    DCZ Donato

    2013-12-01

    Full Text Available The objective of this study was to evaluate the effect of diets containing different calcium and phytase levels on the nutritional balance of broilers.A total of 108 male AG Ross 308 broilers were used in each of the replicates phases used in the study: starter (1-21 days, grower (29-35 days and finisher (36-42 days. A completely randomized experimental design in a 3 x 2 factorial arrangement, with three phytase levels (0, 600 or 1200 FTU/kg and two calcium levels (0.94 and 0.66%; 0.84 and 0.59%; e 0.78 and 0.54% in the starter, grower, and finisher phases, respectively, totaling six treatments with six replicates each.The experimental feeds also contained reduced available phosphorus levels and minimum crude protein level.The method of total excreta collection was applied to determine dry matter, nitrogen, gross energy, calcium, and phosphorus nutritional balance.Reducing dietary calcium levels to 0.66, 0.59, and 0.54%, and using 0.27, 0.22, and 0.20% available phosphorus, 18.50, 17.50, and 16.00% crude protein, and 600 FTU phytase/kg in the diets of the starter, grower, and finisher phases, respectively, allow higher nitrogen, phosphorus, and calcium retention in broilers.During the grower phase, there was positive linear effect of increasing phytase levels in high-calcium diets on AMEn utilization, and the optimal phytase levels for low-calcium diets was 598.71 FTU/kg.In the finisher phase, the best AMEn utilization was obtained with the high-calcium diets.

  17. Phytase supplementation improves blood zinc in rats fed with high phytate Iranian bread

    Directory of Open Access Journals (Sweden)

    Soodeh Shockravi

    2012-01-01

    Full Text Available Background: More than one third of energy intake in Iran is provided from bread. Therefore, improving bread mineral bioavailability through dephytinization can play an important role in decreasing the prevalence of many mineral deficiencies. In this study, effect of phytase supplementation on zinc, iron and calcium status in growing rats fed with a diet containing high phytate Iranian bread (Sangak was assessed. Methods: Thirty weanling Wistar male rats were assigned to phytase (Aspergillus niger or control group for 6 weeks. The diet was designed based on Iranian′s food pattern and 34.2% of the energy was supplied from Sangak bread. Food intake, body and organ weight and body height were measured. Zinc was measured in blood, liver and femur. Iron was assessed in blood and liver and calcium was titrated from femur bone. Statistical analyses were performed using SPSS software. Paired sample t-test, Wilcoxon signed-rank test and repeated measurement ANOVA were used for proper analysis of data. Results: Although weekly weight gain was not different between groups, final weight was in favor of control group. Food intakes, liver and femur bone weight did not differ between the two groups. However, the blood zinc was higher in the phytase group (26.2 ± 7.4 vs. 19.2 ± 5.2, P = 0.03. Thus positive effects of phytase supplementation on zinc, independent of growth was found. Other variables did not show any differences between groups. Conclusion: Addition of phytase to diet containing high phytate Iranian bread can improve blood zinc status in growing rats.

  18. Performance and Nutrient Utilization of Layers Fed Diet Supplemented with Microbial Phytase and Cellulase

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    A 31-week feeding trial was conducted to investigated the effects of dietary supplementation of microbial phytase and cellulase on performance,nutrients utilization and tibia quality of laying hens fed maize and soybean meal diets.192 18-week-old Hisex layers were used in the trial A 2×2×2 factorial design was used in the experiment with three factors of two levels each:0.38% and 0.16% of dietary non-phytate P(nP).0 and 300 U*kg-1 of phytase (Ph),and 0 and 0.10% of cellulase (Ce).The results showed that supplementation of 300 U*kg-1 phytase significantly improved utilization of dietary crude ash,CP,Ca,total P and copper (P<0.05),and improved tibia breaking strength (P<0.05).No effect of phytase on performance was observed.Addition of 0.10% cellulase decreased feed intake (P<0.05),increased utilization of CF (P<0.05) and Ca(P<0.01),and decreased total tibia ash weight (P<0.05).300 U*kg-1 phytase and 0.10% cellulase exhibited obvious positive interactions to enhance utilization of dietary phytic P and copper (P<0.05).0.16% nP did not reduce performance of the layers,but improved egg shell quality at 20 wks,increased utilization of dietary total P,phytic P and Copper (P<0.01),decreased utilization of dietary CP,increased tibia breaking strength and Ca,Mn contents of tibia(P<0.01)

  19. Characterization of a HAP-phytase from a thermophilic mould Sporotrichum thermophile.

    Science.gov (United States)

    Singh, Bijender; Satyanarayana, T

    2009-03-01

    The phytase of Sporotrichum thermophile was purified to homogeneity using acetone precipitation followed by ion-exchange and gel-filtration column chromatography. The purified phytase is a homopentamer with a molecular mass of approximately 456kDa and pI of 4.9. It is a glycoprotein with about 14% carbohydrate, and optimally active at pH 5.0 and 60 degrees C with a T(1/2) of 16h at 60 degrees C and 1.5h at 80 degrees C. The activation energy of the enzyme reaction is 48.6KJmol(-1) with a temperature quotient of 1.66, and it displayed broad substrate specificity. Mg(2+) exhibited a slight stimulatory effect on the enzyme activity, while it was markedly inhibited by 2,3-butanedione suggesting a possible role of arginine in its catalysis. The chaotropic agents such as guanidinium hydrochloride, urea and potassium iodide strongly inhibited phytase activity. Inorganic phosphate inhibited enzyme activity beyond 3mM. The maximum hydrolysis rate (V(max)) and apparent Michaelis-Menten constant (K(m)) for sodium phytate were 83nmolmg(-1)s(-1) and 0.156mM, respectively. The catalytic turnover number (K(cat)) and catalytic efficiency (K(cat)/K(m)) of phytase were 37.8s(-1) and 2.4x10(5)M(-1)s(-1), respectively. Based on the N-terminal and MALDI-LC-MS/MS identified amino acid sequences of the peptides, the enzyme did not show a significant homology with the known phytases.

  20. Recombinant, rice-produced yeast phytase shows the ability to hydrolyze phytate derived from seed-based feed, and extreme stability during ensilage treatment.

    Science.gov (United States)

    Hamada, Akira; Yamaguchi, Ken-Ichi; Harada, Michiko; Horiguchi, Ken-Ichi; Takahashi, Toshiyoshi; Honda, Hideo

    2006-06-01

    When fresh rice leaves producing yeast Schwanniomyces occidentalis phytase were grounded and mixed with the whole extract of seed-based feed for pigs, the release of orthophosphate increased significantly. More specifically, phytate, a major source of phosphorus in the seeds, was hydrolyzed by heterologous phytase. Moreover, when transgenic rice plants were ensiled for up to 12 weeks, no decrease in the phytase activity of the heterologous enzyme was observed. This result strongly suggests that transgenic rice plants producing yeast phytase can be stored as silage without any loss of enzyme activity until usage as a feed additive.

  1. Profile and bioavailability analysis of myo-inositol phosphates in rye bread supplemented with phytases: a study using an in vitro method and Caco-2 monolayers.

    Science.gov (United States)

    Duliński, R; Cielecka, E K; Pierzchalska, M; Byczyński, Ł; Żyła, K

    2016-06-01

    Commercial preparations of 6-phytase A alone and in combination with phytase B were used in rye breadmaking. Determination of bioavailability of myo-inositol phosphates from bread was performed by an in vitro digestion method followed by the measurement of an uptake by Caco-2 cells in culture. In bread supplemented with a combination of 6-phytase A and phytase B, a significant reduction in phytate content was observed from 3.62 μmol/g in the control to 0.7 μmol/g. Bioavailability of phytate estimated by an in vitro method simulating digestion in the human alimentary tract was 9% in the bread supplemented with phytase B, 7% (6-phytase A) and 50% in the control bread. In cell culture, the bioaccessibilities of inositol triphosphates from bread baked with the addition of 6-phytase A was higher by 36% as compared to the samples baked with phytase B and by 32% in breads baked with combination of both phytases.

  2. Mineral absorption and excretion as affected by microbial phytase, and their effect on energy metabolism in young piglets.

    Science.gov (United States)

    Kies, Arie K; Gerrits, Walter J J; Schrama, Johan W; Heetkamp, Marcel J W; van der Linden, Koos L; Zandstra, Tamme; Verstegen, Martin W A

    2005-05-01

    Positive effects of dietary phytase supplementation on pig performance are observed not only when phosphorus is limiting. Improved energy utilization might be one explanation. Using indirect calorimetry, phytase-induced changes in energy metabolism were evaluated in young piglets with adequate phosphorus intake. Eight replicates of 8 group-housed barrows each were assigned to either a control or a phytase-supplemented diet [1500 phytase units (FTU)/kg feed]. Piglets were fed a restricted amount of the control or phytase diet. The diets were made limiting in energy content by formulating them to a high digestible lysine:DE ratio. Fecal nutrient digestibility, portal blood variables, organ weights, and apparent absorption and urinary excretion of ash, Ca, P, Na, K, Mg, Cu, and Fe, were also measured. A model was developed to estimate energy required for absorption and excretion, which are partly active processes. Phytase tended to improve energy digestibility (P = 0.10), but not its metabolizability. Energy retention and heat production were not affected. At the end of the 3-wk period, pancreas weight (P mineral absorption (for most P minerals was estimated as 4.6 kJ/(kg(0.75) . d), which is 1% of the energy required for maintenance. The simultaneous existence of both increases and decreases in heat production processes resulted in the absence of a net effect on energy retention.

  3. Improved production of protease-resistant phytase by Aspergillus oryzae and its applicability in the hydrolysis of insoluble phytates.

    Science.gov (United States)

    Sapna; Singh, Bijender

    2013-08-01

    Among three hundred isolates of filamentous fungi, Aspergillus oryzae SBS50 secreted higher phytase activity at pH 5.0, 35 °C and 200 rpm after 96 h of fermentation. Starch and beef extract supported the highest phytase production than other carbon and nitrogen sources. A nine-fold improvement in phytase production was achieved due to optimization. Supplementation of the medium with inorganic phosphate repressed the enzyme synthesis. Among surfactants tested, Tween 80 increased fungal growth and phytase production, which further resulted in 5.4-fold enhancement in phytase production. The phytase activity was not much affected by proteases treatment. The enzyme resulted in the efficient hydrolysis of insoluble phytate complexes (metal- and protein-phytates) in a time dependent manner. Furthermore, the hydrolysis of insoluble phytates was also supported by scanning electron microscopy. The enzyme, being resistant to trypsin and pepsin, and able to hydrolyze insoluble phytates, can find an application in the animal food/feed industry for improving nutritional quality and also in combating environmental phosphorus pollution and plant growth promotion.

  4. Statistical optimization and mutagenesis for high level of phytase production by Rhizopus oligosporus MTCC 556 under solid state fermentation.

    Science.gov (United States)

    Suresh, S; Radha, K V

    2016-03-01

    The present study deals with production of phytase from Rhizopus oligosporus MTCC 556 by solid state fermentation (SSF) using different (ADT27, IR20, PAIYUR1, KG, and RASI) rice bran varieties, in which ADT27 rice bran yield maximum of 6.2 U gds⁻¹ phytase. Statistical optimization was employed by Central Composite Design (CCD); the results showed that 3.0 g dextrose, 2.5 g ammonium nitrate, substrate size of 80 mesh, 10 mg calcium chloride was 116 hr at optimal for phytase production by SSF, with maximum of 23.14 U gds'. Phytase production improved by 4 fold (31.3 U/gds) due to chemical mutagenesis (mutant Rhizopus oligosporus MTCC 1116) in optimized media composition. Partially purified phytase showed approximately 90 kDa of molecular mass and was optimally active at 5.5 pH and 50°C temperature. Substrate specificity exhibited in sodium phytic acid and phytase activity was stimulated by Zn²⁺ and Ca²⁺.

  5. Effect of Phytase Treatment on Phosphate Availability in the Potential Food Supplement Corn Distillers’ Grains with Solubles

    Directory of Open Access Journals (Sweden)

    Thomas P. West

    2014-01-01

    Full Text Available The most effective treatment of the potential food supplement corn distillers’ grains with solubles using a fungal phytase to degrade phytic acid and release phosphorus was explored. Compared to the untreated grains with solubles, the phytic acid level in the grains with solubles was reduced by phytase treatment but treatment with 4 units of enzyme/g was more effective than 2 units of enzyme/g after 2 h. At 30°C or 45°C, 4 units of phytase/g reduced the phytic acid content of the grains with solubles by at least 94% after 2 h. The available phosphate in the grains treated for 2 h with 4 units of enzyme/g was increased by at least 1.5-fold compared to the untreated grains. Although phytic acid levels in the grains with solubles was reduced at 28°C, 30°C, 37°C, 40°C, or 45°C using 4 units of phytase/g for 2 h, the greatest reduction (96% in phytic acid concentration occurred at 40°C. The available phosphate in the phytase-treated grains at 40°C was increased after 2 h by 1.9-fold compared to the control grains. Overall, phytase treatment of this potential food supplement improved it by increasing its phosphate content while reducing its phytic acid content.

  6. Efficiency of microbial phytase supplementation in diets formulated with different calcium:phosphorus ratios, supplied to broilers from 22 to 33 days old.

    Science.gov (United States)

    Naves, L de P; Rodrigues, P B; Teixeira, L do V; de Oliveira, E C; Saldanha, M M; Alvarenga, R R; Corrêa, A D; Lima, R R

    2015-02-01

    An experiment was conducted with broilers from 22 to 33 days of age to evaluate the efficiency of six microbial phytases supplemented in diets (1500 FTU/kg) that were formulated with three different calcium:available phosphorus (Ca:P(avail)) ratios (4.5:1.0, 6.0:1.0 and 7.5:1.0). A positive control diet without phytase was formulated with a Ca:P(avail) ratio of 7.5:3.4 to meet the nutritional requirements of the broilers. The P and ash contents of the tibia, magnesium in the plasma, performance, balance and retention of phytate phosphorus (P(phyt)), intake of total P and nitrogen (N), nitrogen-corrected apparent metabolizable energy and apparent digestibility of dry matter of the diets were not influenced (p > 0.05) by the type of phytase or the dietary Ca:P(avail) ratio. However, there was an interaction (p phytase type and the Ca:P(avail) ratio for the retention coefficients of total P, Ca and N. Phytase B resulted in the highest Ca deposition in the tibia (p Phytases D, E and F reduced the Ca concentrations in the tibia (p Phytase D increased the P level in the plasma and decreased the total P excretion (p Phytases E and F increased Ca excretion, while phytase A reduced it (p phytase type, increasing the dietary Ca:P(avail) ratio reduced (p phytase A, C, D or E/kg. This diet allows the maintenance of performance and adequate bone mineralization, and it improves the Ca, total P and P(phyt) utilization in addition to reducing the excretion of N and P into the environment.

  7. 新型中性植酸酶在大肠杆菌中的高效表达、纯化及酶学性质%Expression of a New Neutral Phytase in Escherichia coli and Its Purification and Enzymatic Characterization

    Institute of Scientific and Technical Information of China (English)

    路国伟; 许伟; 邵荣; 云志

    2012-01-01

    运用基因工程技术,将克隆到的淀粉液化芽孢杆菌Bacillus amyloliquefaciens DSM 1061新型中性植酸酶基因(GenBank登录号为HM747163)构建到表达载体pET22b(+)上,并在E.coli BL21(DE3)中进行高效表达。电泳结果表明该酶分子质量约为42kD,目的蛋白占大肠杆菌可溶性蛋白30%左右。利用Ni-NTA琼脂糖凝胶进行亲和纯化,酶学性质研究表明,其最适温度为60℃,最适pH值为7.0,最大酶比活力为15U/mg。60℃时以植酸钠为底物的Km值为0.30mmol/L。%A new neutral phytase gene(phyc) from Bacillus amyloliquefaciens DSM 1061(GenBank: HM747163) was cloned into expression vector pET22b(+).The phytase was expressed in Escherichia coli BL21(DE3).The molecule weight of the phytase protein was about 42 kD determined by SDS-PAGE.The expressed phytase was about 30% of the total soluble protein of E.coli.The recombinant protein was purified by 6×His-Tagged Protein Purification Kit.Optimal pH value and temperature of the phytase were 7.0 and 60 ℃,respectively.Under the optimum conditions,the activity was 15 U/mg.The Km value of the phytase for hydrolysis of sodium phytate under 60 ℃ was 0.30 mmol/L.

  8. The effect of combined preparation, probiotic and phytase on performance parameters and vitality of weaned piglets.

    Directory of Open Access Journals (Sweden)

    FESTIM SHYTAJ

    2014-06-01

    Full Text Available A combined preparation with probiotic Schaumalac F 80M BONVITAL 4b 1841 (2,5 x 1010 KBE Enterococcus faecium DSM 7134 and phytase was supplemented to a basal ration with 4%. The effects on growth performance on thirty five weaned piglets/each group were studied for thirty days experimental period. The supplementation of combined preparation improved slightly Daily Weight Gain (DWG, Feed Consumption (FC and Feed Conversion Ratio (FCR, kg feed/kg weight gain. At the end of experimental period, daily weight gain was increased 6% more and feed conversion ratio was decreased 3.4%, compared to the control group or without supplementation. The results indicate that the combined preparation may be less suppressive to the Escherichia coli. Overall a positive effect of the probiotic and phytase on growth performance was observed.

  9. Cloning and Characterization of Purple Acid Phosphatase Phytases from Wheat (Triticum aestivum L.), Barley (Hordeum vulgare L.), Maize (Zea maize L.) and Rice (Oryza sativa L.)

    DEFF Research Database (Denmark)

    Dionisio, Giuseppe; Madsen, Claus Krogh; Holm, Preben Bach;

    2011-01-01

    Barley (Hordeum vulgare L.) and wheat (Triticum aestivum L.) possess a significant phytase activity in the mature grains. Maize (Zea mays L.) and rice (Oryza sativa L.) possess little or virtually no pre-formed phytase activity in the mature grain and depend fully on de novo synthesis during...

  10. Dietary pharmacological or excess zinc and phytase effects on tissue mineral concentrations, metallothionein, and apparent mineral retention in the newly weaned pig.

    Science.gov (United States)

    Martínez, M M; Link, J E; Hill, G M

    2005-01-01

    Feeding pharmacological zinc (Zn) to weaned pigs improves growth, and dietary phytase improves P and Zn availability. Metallothionein (MT) increases in the duodenum, kidney, and liver of pigs fed 1000 mg Zn/kg with phytase or 2000 mg Zn/kg with or without phytase when fed for 14 d postweaning. The goal of this study was to determine the effects of feeding pharmacological Zn and phytase on tissue minerals, MT, mineral excretion, and apparent retention. Twenty-four newly weaned pigs (20 d; 7.2 kg) were individually fed twice daily, a basal diet supplemented with 0, 1000, or 4000 mg Zn/kg as Zn oxide, without or with phytase (500 phytase units [FTU]/kg) for 14 d, followed by a basal diet (100 mg Zn/kg) without phytase for 7 d. Pigs fed 4000 mg Zn/kg without phytase had higher (p=0.01) plasma, hepatic, renal Zn, renal Cu, and hepatic, renal, and jejunal MT than pigs fed the basal diet or 1000 mg Zn/kg. Duodenal MT was higher (p=0.0001) in pigs fed 1000 and 4000 mg Zn/kg than in pigs fed the basal diet. In pigs fed 1000 and 4000 mg Zn/kg, Zn loading occurred during the first 11 d of supplementation; by d 14, excess Zn was being excreted in the feces.

  11. Characterization of N-type glycosylation sites and glycan structures of Purple Acid Phosphatase Phytases from Wheat (Triticum aestivum L.)

    DEFF Research Database (Denmark)

    Dionisio, Giuseppe; Brinch-Pedersen, Henrik; Welinder, Karen Gjesing

    2011-01-01

    ., Skov L. Brinch-Pedersen H. (2011). The degradation of phytate by microbial and wheat phytases is dependent on the phytate matrix and the phytase origin. J. Sci. Food Agri. (in press). Dionisio G., Madsen C.K., Holm P.B., Welinder K.G., Jørgensen M., Stoger E., Arcalis E., Brinch-Pedersen H. (2011a...

  12. Effect of phytase supplementation to barley-canola meal and barley-soybean meal diets on phosphorus and calcium balance in growing pigs

    NARCIS (Netherlands)

    Sauer, W.C.; Cervantes, M.; He, J.M.M.; Schulze, H.

    2003-01-01

    Two metabolism experiments were carried out, to determine the effect of microbial phytase addition to barley-canola meal and barley-soybean meal diets on P and Ca balance in growing. pigs; In experiment 1, six barrows (29.6kg: initial LW) were fed a barley-canola meal diet, without or. with phytase

  13. Bioavailability and poultry fecal excretion of phosphorus from soybean-based diets supplemented with phytase

    Directory of Open Access Journals (Sweden)

    Bianca Almeida Brandão Martins

    2013-03-01

    Full Text Available Five hundred and four 1-day-old Cobb broiler chickens were randomly allotted to 18 treatments and fed a semi-purified basal diet (4 g/kg total phosphorus supplemented with 2 levels of supplemental P (0.5 or 1 g/kg during 21 days to evaluate performance and bone mineralization; to assess the relative bioavailability of P (RBP in wheat meal (WM and roasted (RS or extruded soybean meal (ES, with or without phytase (0 or 750 ftu/kg diet, using dicalcium phosphate (DC as standard; and to estimate phosphorus excretion. Weight gain (WG, feed intake (FI, feed to gain ratio (FG, and tibia ash weigh (AW were assessed. For each source, the RBP was calculated by the slope ratio method, when tibia ash weight was regressed on P ingested. Ingested and fecal P was determined to calculate P intake, and P excretion and excretion rate. Phytase addition increased overall FI, WG, and FG ratio by 3.3, 2.6 and 0.8%, respectively, but differences were not significant. Overall AW was increased by 10.8% after the enzyme inclusion in the diets. The relative bioavailability of phosphorus values of tested WM, RS, and ES were increased by 6.32, 64.32, 76.99%, respectively, in response to supplementation of phytase in diets containing WM, RS, and ES, respectively. Phytase reduced P excretion by 12.7% and the excretion rate by 15.0%, regardless of the phosphorus source or level.

  14. Effects of dietary supplementation of organic acids and phytase on performance and intestinal histomorphology of broilers

    Science.gov (United States)

    Mohammadagheri, Nasibeh; Najafi, Ramin; Najafi, Gholamreza

    2016-01-01

    The present experiment was conducted to evaluate the effects of organic acids and phytase enzyme supplementation on performance and intestinal histomorphology of broilers. The experiment was done in a factorial arrangement 2 × 2 × 2 based on completely randomized design with eight treatments, five replicates with 12 chicks in each until 42 days of age. Diets included natural vinegar (0 and 2%), citric acid (CA; 0.00 and 1.00%) and phytase enzyme (PHY; 0.00 and 500 FTU phytase per kg of feed). One bird from each treatment replicate was randomly selected and slaughtered to evaluate the small intestinal morphology on 42 days of age. Analysis of results showed that vinegar increased feed consumption and body weight gain in total experimental period (p ˂ 0.05), while CA significantly decreased feed consumption on 0-14 days of age (p ˂ 0.05). No effect was observed on performance in interaction of organic acids together and with PHY group (p > 0.05). In duodenum CA increased the villus height and width (p ˂ 0.05) and PHY enzyme increased villus width (p ˂ 0.05) and decreased crypt depth (p ˂ 0.05). On the other hand, CA along with PHY significantly decreased crypt depth (p ˂ 0.05). In jejunum PHY alone and in combination with vinegar increased the goblet cells numbers (p ˂ 0.05), whereas vinegar significantly increased the goblet cells numbers in ileum (p ˂ 0.05). The muscular thickness in duodenum, jejunum, and ileum was not affected among different treatment groups. The results showed that supplementation of organic acids and phytase together in this experiment, with no negative effects on each other, improved their effects on some parameters. PMID:27872714

  15. Effect of dietary NPP level and phytase supplementation on the laying performance over one year period

    Directory of Open Access Journals (Sweden)

    Annamaria Tischler

    2015-06-01

    Full Text Available Our trial was aimed to study the effect of different dietary non-phytin phosphorus (NPP levels with and without phytase enzyme supplementation on laying performance and eggshell quality of Tetra SL-LL in the last 25 weeks of the long-term (17 months egg production. A total of 69 Tetra SL-LL layers were allocated into 3 dietary treatments. Two diets with different levels of NPP (2.45 or 2.15 g/kg, HP and LP, respectively were formulated, and 0 or 300 FTU/kg phytase enzyme was added to low NPP feed (LP and LP+E, respectively. Dietary Ca was uniformly adjusted (38.2 g/kg to feed in each treatment. In the course of the trial, intensity of egg production (%, egg weight (g/egg, number of the broken eggs and feed intake (g/d/bird were recorded. Every 2 weeks 20 eggs per treatment were broken to determine the shell strength and thickness. Our results show that low NPP diet had detrimental effect on the intensity of egg production (P<0.05 and phytase added to the LP diet resulted the lowest number of broken eggs (P<0.05. In conclusion, NPP content of the layer diet can be reduced from 2.45 to 2.15 g/kg in the last 25 weeks of the elongated laying term (12-17 month of laying, if supplemented with 300 FTU/kg phytase enzyme without compromising the egg production, and in the same time it can improve eggshell quality and reduce the number of broken eggs.

  16. Substitution of fishmeal with soybean meal in humpback Grouper, Cromileptes altivelis juvenile diets supplemented with phytase

    Directory of Open Access Journals (Sweden)

    Rachman Syah

    2006-12-01

    Full Text Available Feeding experiments were conducted to evaluate the effects of replacing fishmeal with soybean meal in diet on growth of humpback grouper. Fifteen cages of 1 x 1 x 1.2 m3 each stocked with 16 humpback grouper juveniles (61.3 ± 0.4 g/pc were set up randomly in seawater. Fish were fed to satiation twice daily for 112 days. The control diet contained 61.9% fishmeal (63.34% crude protein. Four isonitrogenous (48% crude protein and isocaloric (4.7 kcal/g feed diets supplemented with commercial phytase “Rhonozyme-P” at 0.075% were formulated to contain different levels (8%, 16%, 24%, and 32% soybean meal (43.65% crude protein as a partial replacement for fishmeal. These diets contained total phosphorus levels between 3.6—4.5 (±0.4 % and 0.7—1.5 (±0.04 % available phosphorus. Replacement of fishmeal with soybean meal (8 to 32% replacement was not significantly different (P>0.05 to the control diet on daily growth rate (DGR, food conversion ratio (FCR, protein efficiency ratio (PER, and daily food consumption (DFC. However, the dietary levels of soybean meal significantly affected (P<0.05 whole body protein and phosphorus retention (Table 1. These data suggest that addition of phytase in diets could improve protein and phosphorus availability and reduce nitrogen and phosphorus loading in the environment. Phytase can therefore play an important role in formulating eco-friendly feed for humpback grouper. Based on P loading, supplementation of phytase enable up to 24% fishmeal replacement with soybean meal.

  17. Effect of particle size and microbial phytase on phytate degradation in incubated maize and soybean meal

    DEFF Research Database (Denmark)

    Ton Nu, Mai Anh; Blaabjerg, Karoline; Poulsen, Hanne Damgaard

    2014-01-01

    The objective of the study was to evaluate the effect of screen size (1, 2 and 3 mm) and microbial phytase (0 and 1000 FTU/kg as-fed) on phytate degradation in maize (100% maize), soybean meal (100% SBM) and maize–SBM (75% maize and 25% SBM) incubated in water for 0, 2, 4, 8 and 24 h at 38°C...

  18. Effect of intermittent feeding, structural components and phytase on performance and behaviour of broiler chickens.

    Science.gov (United States)

    Svihus, B; Lund, V B; Borjgen, B; Bedford, M R; Bakken, M

    2013-01-01

    1. Two experiments were conducted to study the effect of intermittent feeding on performance and the efficacy of an exogenous phytase, and to assess whether intermittent feeding changed the activity pattern of broiler chickens. 2. Broiler chickens were given, either ad libitum or intermittently, a phosphorus deficient pelleted diet containing either coarsely or finely ground oat hulls and either no enzyme or a phytase added from 10 d of age, in a 2 × 2 × 2 factorial arrangement. Ad libitum feeding consisted of continuous access to feed in a room with 18 h of light and 6 h of complete darkness, whereas birds on intermittent feeding had restricted access to feed from 7 d of age, with 4 1-h feeding bouts/d and one 2-h feeding bout/d from d 14. 3. Performance, characteristics of the anterior digestive tract and phosphorus retention were assessed in experiment 1, while in experiment 2, birds were observed during 4-h periods to quantify different behaviours. 4. Intermittent feeding and phytase improved performance, but intermittent feeding did not improve the efficacy of the enzyme added. Ad libitum fed broiler chickens ate and drank on average twice per hour, and spent close to three-quarters of their time resting. Apart from an increased standing and feed searching activity for intermittently fed birds compared to ad libitum fed birds during the last hour before feed was presented, no differences in activity was detected. 5. It was concluded that broiler chickens quickly adapt to intermittent feeding without reduction in final body weight and with improvements in feed efficiency, but without improving the efficacy of dietary phytase. Only small changes occur in the behaviour of intermittently fed birds compared to ad libitum fed birds.

  19. Effects of microbial phytase, produced by solid-state fermentation, on the performance and nutrient utilisation of broilers fed maize- and wheat-based diets.

    Science.gov (United States)

    Wu, Y B; Ravindran, V; Hendriks, W H

    2003-12-01

    1. The influence of a microbial phytase on the performance, toe ash contents and nutrient utilisation of male broilers fed diets based on maize and wheat was investigated. The experiment was conducted as 2 x 2 x 2 factorial arrangement of treatments. Within the factorial, two diet types (maize-soy or wheat-soy) containing two levels of non-phytate phosphorus (3.0 or 4.5 g/kg) were evaluated and each level of non-phytate phosphorus was supplemented with 0 or 500 PU phytase/kg diet. Each of the 8 dietary treatments were fed to 6 pens of 8 birds from d 1 to 21 post-hatching. 2. Main effects of diet type and phytase were observed for all parameters. Main effect of non-phytate phosphorus was significant only for feed/gain and toe ash contents. Phytase addition improved weight gains irrespective of diet type or non-phytate phosphorus level, but the magnitude of improvement in the phosphorus-deficient wheat-soy diet was greater, resulting in a diet type x non-phytate phosphorus interaction. Responses in toe ash contents were noted only in phosphorus-deficient diets, as indicated by a non-phytate phosphorus x phytase interaction. 3. Phytase addition improved apparent metabolisable energy values of wheat-based diets, but had little effect on the apparent metabolisable energy of maize-based diets as shown by a diet type x phytase interaction. The apparent metabolisable energy was not influenced by dietary non-phytate P. 4. Phytase improved ileal nitrogen digestibility in both diet types, but the responses to added phytase tended to be higher in wheat-based diets, as shown by a diet type x phytase interaction. 5. Increasing the dietary non-phytate phosphorus level reduced phosphorus digestibility and increased excreta phosphorus content. Addition of phytase improved phosphorus digestibility, but the increments were higher in low phosphorus diets resulting in a non-phytate phosphorus x phytase interaction. Phytase addition tended to lower the excreta phosphorus content, but

  20. Pulque, an alcoholic drink from rural Mexico, contains phytase. Its in vitro effects on corn tortilla.

    Science.gov (United States)

    Tovar, Luis Raul; Olivos, Manuel; Gutierrez, Ma Eugenia

    2008-12-01

    Pulque is made by fermenting the agave sap or aguamiel of Agave atrovirens with a whole array of microorganisms present in the environment including several lactic acid bacteria and yeasts such as Saccharomyces cerevisiae. Ascorbic acid was determined in pulque and aguamiel, respectively. Phytase activity in lees, liquid and freeze-dried pulque was assayed by measuring the appearance of phosphate from phytate by a colorimetric method likewise phosphate from phytate present in fresh corn tortilla was measured after in vitro incubation with pulque. Iron, zinc, calcium, magnesium and selenium contents were measured in pulque and corn tortilla as well as in nixtamalized corn flour (NCF), the latter is used to make instant tortilla, since corn provides most of the energy as well as most of the phytate in the Mexican rural diet. Pulque showed phytase activity but much less ascorbic acid and iron than previously reported; additionally, phytase in pulque hydrolyzed most of phytate's corn tortilla. Lees, which is mostly made of pulque's microbiota, significantly accumulated iron and zinc but no selenium. NCF was fortified with iron by the manufacturers but poorly blended. There were significant differences on selenium content between tortillas samples, apparently some soils in central Mexico are selenium deficient. Moderate pulque intake appears to increase the bioavailability of iron and zinc bound by phytate in corn.

  1. Purification and characterisation of an extracellular phytase from Aspergillus niger 11T53A9

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    Ralf Greiner

    2009-12-01

    Full Text Available An extracellular phytase from Aspergillus niger 11T53A9 was purified about 51-fold to apparent homogeneity with a recovery of 20.3% referred to the phytase activity in the crude extract. Purification was achieved by ammonium sulphate precipitation, ion chromataography and gel filtration. The purified enzyme behaved as a monomeric protein with a molecular mass of about 85 kDa and exhibited maximal phytate-degrading activity at pH 5.0. Optimum temperature for the degradation of phytate was 55°C. The kinetic parameters for the hydrolysis of sodium phytate were determined to be K M = 54 µmol l-1 and k cat = 190 sec-1 at pH 5.0 and 37°C. The purified enzyme was rather specific for phytate dephosphorylation. It was shown that the phytase preferably dephosphorylates myo-inositol hexakisphosphate in a stereospecific way by sequential removal of phosphate groups via D-Ins(1,2,4,5,6P5, D-Ins(1,2,5,6P4, D-Ins(1,2,6P3, D-Ins(1,2P2 to finally Ins(2P.

  2. Phytase production through response surface methodology and molecular characterization of Aspergillus fumigatus NF191.

    Science.gov (United States)

    Gangoliya, Shivraj Singh; Gupta, Raj Kishor; Singh, Nand Kumar

    2015-06-01

    Phytase play an important role in phytic acid catalysis that act as a food inhibitor in cereals. Here, we isolated high phytase producing isolates NF191 closely related to Aspergillus fumigatus sp. from piggery soil. DNA was isolated from the fungal culture and amplified the ITS region using ITS1 and ITS4 primer using PCR. The 400-900 bp amplicon was gel eluted and subjected to sequencing. The sequencing results were assembled and compared with NCBI data base which showed the 99% identity of Aspergillus fumigatus. Different carbon sources viz., fructose, galactose, lactose, dextrose, sucrose, maltose and different nitrogen sources (organic & inorganic) NH4Cl, NH4NO3, (NH4)2SO4, KNO3, NaNO3, urea, yeast extract, peptone, beef extract were tested for optimal production. The 0.3% dextrose, 0.5% NH4NO3 and 96 h incubation time showed the best production and enzyme activity at 45 degrees C incubation temperature. The selected parameters, dextrose, ammonium sulphate and incubation time, when employed with statistical optimization approach involving response surface optimization using Box Behnken Design, gave a 1.3 fold increase in phytase production compared to unoptimized condition.

  3. Isolation, purification and characterization of Bacillus subtilis Phytase from Holiwood Gresik

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    Leny Yuanita

    2012-01-01

    Full Text Available The aim of the research were isolation, purification and characterization of Bacillus subtilis phytase from Holiwood Gresik. The research was done in two stages; the first include enzyme isolation, precipitation with amonium sulphate, dialysis, gel filtration chromatography, SDS-PAGE analysis, while second determining optimum pH, optimum temperature, the effect of pH and temperature to enzim stability, the values of KM and Vmax Bacillus subtilis phytase from Holiwood Gresik. The first stage research design were One Shot Case Study and Post Test Only Control Group Design, while the second stage were Post Test Only Control Group Design and Factorial Design. The data being analyzed by one-way and two-way Anova. The results of research showed that Bacillus subtilis phytase has the molecular mass of 36.5 kDa, optimum pH at 6.5–7.0, optimum temperature at 41°C and it was found to be stable for 30 minute incubation at pH 7or 30° C with 2% or 3% lost of its activity respectively. KM value was 0.62 mM and VMax 0.393 mmol/ml/minute.

  4. Lowering of phytic acid content by enhancement of phytase and acid phosphatase activities during sunflower germination

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    Juliana da Silva Agostini

    2010-08-01

    Full Text Available The objective of this work was to investigate the germination of hybrid sunflowers BRS191 and C11 as a means of lowering phytic acid (PA content by enhancing the activity of endogenous phytase and acid phosphatase. The concentration of PA in hybrid sunflower achenes varied from 2.16 to 2.83g/100g of sample (p O objetivo deste trabalho foi investigar a germinação de girassóis híbridos BRS 191 e C11 com finalidade de reduzir o teor de AF e aumentar as atividades de phytases e fosfatases endógenas. A concentração do AF nos aquênios de girassóis híbridos variou de 2,16 a 2,83 g /100g de amostra (p< 0,005. As atividades de fitases e fosfatases de girassóis BRS191 e C11 foram elevadas no 4º e 5º dia de germinação, respectivamente, com liberação do fósforo necessário para o desenvolvimento da semente. Estes resultados indicam que o AF do girassol hibrido reduz e a atividade de phytase aumenta em períodos distintos da germinação, possibilitando assim a aplicação desta enzima no controle do teor de AF em cereais, melhorando o seu valor nutricional.

  5. Production, characteristics and applications of phytase from a rhizosphere isolated Enterobacter sp. ACSS.

    Science.gov (United States)

    Chanderman, Ashira; Puri, Adarsh Kumar; Permaul, Kugen; Singh, Suren

    2016-10-01

    Optimization of process parameters for phytase production by Enterobacter sp. ACSS led to a 4.6-fold improvement in submerged fermentation, which was enhanced further in fed-batch fermentation. The purified 62 kDa monomeric phytase was optimally active at pH 2.5 and 60 °C and retained activity over a wide range of temperature (40-80 °C) and pH (2.0-6.0) with a half-life of 11.3 min at 80 °C. The kinetic parameters K m, V max, K cat, and K cat/K m of the pure phytase were 0.21 mM, 131.58 nmol mg(-1) s(-1), 1.64 × 10(3) s(-1), and 7.81 × 10(6) M(-1) s(-1), respectively. The enzyme was fairly stable in the presence of pepsin under physiological conditions. It was stimulated by Ca(+2), Mg(+2) and Mn(+2), but inhibited by Zn(+2), Cu(+2), Fe(+2), Pb(+2), Ba(+2) and surfactants. The enzyme can be applied in dephytinizing animal feeds, and the baking industry.

  6. Production of phytase under solid-state fermentation using Rhizopus oryzae: novel strain improvement approach and studies on purification and characterization.

    Science.gov (United States)

    Rani, Richa; Ghosh, Sanjoy

    2011-11-01

    Present study introduces linseed oil cake as a novel substrate for phytase production by Rhizopus oryzae. Statistical approach was employed to optimize various medium components under solid state fermentation (SSF). An overall 8.41-fold increase in phytase production was achieved at the optimum concentrations (w/w, mannitol, 2.05%; ammonium sulfate, 2.84% and phosphate, 0.38%). Further enhancement by 59% was observed due to a novel strain improvement approach. Purified phytase (∼34 kDa) showed optimal temperature of 45 °C, dual pH optima at 1.5 and 5.5 and possesses high catalytic efficiency (2.38×10(6) M(-1) s(-1)). Characterization study demonstrates the phytase as highly thermostable and resistant to proteolysis, heavy metal ions, etc. Furthermore, an improved HPLC method was introduced to confirm the ability of phytase to degrade phytic acid completely and was found to be an efficient method.

  7. A Phytase Characterized by Relatively High pH Tolerance and Thermostability from the Shiitake Mushroom Lentinus edodes

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    Guo-Qing Zhang

    2013-01-01

    Full Text Available A monomeric phytase with a molecular mass of 14 kDa was acquired from fresh fruiting bodies of the shiitake mushroom Lentinus edodes. The isolation procedure involved chromatography on DEAE-cellulose, CM-cellulose, Q-Sepharose, Affi-gel blue gel, and a final fast protein liquid chromatography-gel filtration on Superdex 75. The purified phytase demonstrated the unique N-terminal amino acid sequence DPKRTDQVN, which exhibited no sequence similarity with those of other phytases previously reported. It expressed its maximal activity at pH 5.0 and 37°C. Phytase activity manifested less than 20% change in activity over the pH range of 3.0–9.0, considerable thermostability with more than 60% residual activity at 70°C, and about 40% residual activity at 95°C. It displayed a wide substrate specificity on a variety of phosphorylated compounds with the following ranking: ATP > fructose-6-phosphate > AMP > glucose-6-phosphate > ADP > sodium phytate > β-glycerophosphate. The phytase activity was moderately stimulated by Ca2+, but inhibited by Al3+, Mn2+, Zn2+, and Cu2+ at a tested concentration of 5 mM.

  8. Phytase production by Aspergillus oryzae in solid-state fermentation and its applicability in dephytinization of wheat bran [corrected].

    Science.gov (United States)

    Sapna; Singh, Bijender

    2014-08-01

    Aspergillus oryzae SBS50 secreted a high titre of phytase in solid-state fermentation (SSF) using wheat bran at 30 °C after 96 h at the initial substrate to moisture ratio of 1:2 and a water activity of 0.95. The production of phytase increased when wheat bran was supplemented with sucrose and beef extract. Further enhancement in enzyme production was recorded when the substrate was supplemented with the surfactant Triton X-100 (145 U/g of DMB). An overall 29-fold improvement in phytase production was achieved owing to optimization. Under optimized conditions, the mould secreted 9.3-fold higher phytase in SSF as compared to submerged fermentation (SmF). The mesophilic mould also secreted amylase, cellulase (CMCase), pectinase and xylanase along with phytase in SSF. Scanning electron microscopy revealed luxuriant growth of A. oryzae on wheat bran with abundant spores. The enzyme dephytinized wheat bran with concomitant liberation of inorganic phosphate.

  9. Mixed Substrate Fermentation for Enhanced Phytase Production by Thermophilic Mould Sporotrichum thermophile and Its Application in Beneficiation of Poultry Feed.

    Science.gov (United States)

    Kumari, Amit; Satyanarayana, T; Singh, Bijender

    2016-01-01

    The optimum values of the critical variables determined by the central composite design of response surface methodology (RSM) for maximum phytase production (1881.26 U g(-1) dry mouldy residue (DMR)) by Sporotrichum thermophile are 2.5 % Tween 80, 1.0 % yeast extract and 48 h of incubation period. Phytase production in the mixed substrate (sugarcane bagasse and wheat bran) fermentation enhanced 11.6-fold over the initial production as a consequence of optimization. Phytase titres are sustainable in flasks, trays and column bioreactor (1796 to 2095 U g(-1) DMR), thus validating the model and the process for large-scale phytase production. When the yeast extract was replaced with corn steep liquor (2 % w/v), a sustained enzyme titre (1890 U g(-1) DMR) was attained, making the process cost-effective. Among all the detergents, Tween 80 supported a higher phytase production than others. The enzyme efficiently liberated nutritional components from poultry feed (inorganic phosphate, soluble protein and reducing sugars) in a time-dependent manner.

  10. Concerted action of endogenous and heterologous phytase on phytic acid degradation in seed of transgenic wheat (Triticum aestivum L.).

    Science.gov (United States)

    Brinch-Pedersen, Henrik; Hatzack, Frank; Sørensen, Lisbeth D; Holm, Preben B

    2003-12-01

    Expression of heterologous phytases in crops offers a great potential for improving phosphate and mineral bioavailability in food and feed. In this context it is of relevance to describe the concerted action of endogenous and hetrologous phytases on the transgenic seed inositol phosphate profile. Here we report metal-dye detection HPLC analysis of inositol phosphate degradation in flour from transgenic wheat materials possessing wheat endogenous 6-phytase [EC 3.1.3.26] and Aspergillus 3-phytase [EC 3.1.3.8] activities under the control of the maize ubiquitin-1 promoter and the wheat high molecular weight glutenin subunit 1DX5 promoter respectively. During 50 min incubation there is an accumulation of InsP5 to InsP2 breakdown products in non-transgenic material. Aspergillus niger phytase specific breakdown products are transiently detected in transgenic material but after 50 min incubation virtually all InsP5, InsP4 and InsP3 isomers are hydrolysed.

  11. A phytase characterized by relatively high pH tolerance and thermostability from the shiitake mushroom Lentinus edodes.

    Science.gov (United States)

    Zhang, Guo-Qing; Wu, Ying-Ying; Ng, Tzi-Bun; Chen, Qing-Jun; Wang, He-Xiang

    2013-01-01

    A monomeric phytase with a molecular mass of 14 kDa was acquired from fresh fruiting bodies of the shiitake mushroom Lentinus edodes. The isolation procedure involved chromatography on DEAE-cellulose, CM-cellulose, Q-Sepharose, Affi-gel blue gel, and a final fast protein liquid chromatography-gel filtration on Superdex 75. The purified phytase demonstrated the unique N-terminal amino acid sequence DPKRTDQVN, which exhibited no sequence similarity with those of other phytases previously reported. It expressed its maximal activity at pH 5.0 and 37 °C. Phytase activity manifested less than 20% change in activity over the pH range of 3.0-9.0, considerable thermostability with more than 60% residual activity at 70 °C, and about 40% residual activity at 95°C. It displayed a wide substrate specificity on a variety of phosphorylated compounds with the following ranking: ATP > fructose-6-phosphate > AMP > glucose-6-phosphate > ADP > sodium phytate > β -glycerophosphate. The phytase activity was moderately stimulated by Ca(2+), but inhibited by Al(3+), Mn(2+), Zn(2+), and Cu(2+) at a tested concentration of 5 mM.

  12. Effect of a novel microbial phytase on production performance and tibia mineral concentration in broiler chickens given low-calcium diets.

    Science.gov (United States)

    Singh, A; Walk, C L; Ghosh, T K; Bedford, M R; Haldar, S

    2013-01-01

    1. In a 42-d feeding trial, 264 one-d-old, as hatched, Cobb 400 broiler chickens (6 pens per group, n = 11 per pen in a 2 × 2 factorial arrangement) were fed on two concentrations of dietary calcium (Ca) (9.0 and 7.5 g/kg in starter, 7.5 and 6 g/kg in grower phases) and supplemental phytase (0 and 500 U/kg diet). 2. During d 0-21, the high Ca + phytase diet improved body weight. During d 0-42, feed intake was increased by the low Ca diet and decreased by phytase supplementation. Feed conversion ratio during d 0-21 was improved by the high Ca + phytase diet. 3. At d 42, Ca in duodenal digesta was reduced by low dietary Ca and supplemental phytase. High dietary Ca reduced P in duodenal and jejunal digesta. Phytase reduced digesta P and increased serum P concentration. 4. Relative tibia length decreased with low dietary Ca and increased with phytase. The robusticity index of tibia was improved by the low Ca diet and phytase supplementation. Phytase supplementation increased tibia ash and concentrations of Ca, magnesium (Mg), manganese (Mn), copper (Cu), zinc (Zn) and iron (Fe) in tibia. The low Ca diet increased Mg, Mn and Fe and reduced Cu and Zn in tibia. 5. It was concluded that 7.5 g Ca/kg during weeks 0-3 and 6 g Ca/kg during weeks 3-6 sustained broiler performance and bone ash, while phytase supplementation facilitated tibia mineralisation, particularly during the grower phase.

  13. Extra-phosphoric effects of superdoses of a novel microbial phytase.

    Science.gov (United States)

    Walk, C L; Bedford, M R; Santos, T S; Paiva, D; Bradley, J R; Wladecki, H; Honaker, C; McElroy, A P

    2013-03-01

    An experiment was conducted to evaluate the influence of a novel microbial phytase on broiler performance from d 0 to 42 and tibia ash at d 21. Male Cobb 500 broilers (n = 2,016) were fed 1 of 7 experimental diets: positive control (PC) formulated to meet or exceed nutrient recommendations; PC plus dicalcium phosphate (PC+DCP) formulated to provide Ca and P at 0.10% above the PC; PC plus 500 U/kg of microbial phytase (PC+500); negative control (NC) with Ca and P reduced from the PC by 0.16 and 0.15%, respectively; and NC plus 500 (NC+500), 1,000 (NC+1,000), or 1,500 (NC+1,500) U/kg of microbial phytase. Diets were fed in 3 phases from d 0 to 21, d 22 to 42, and d 43 to 49 to 32 birds/pen and 9 replicate pens/diet. From d 0 to 21, broilers fed the NC diet had decreased (P ≤ 0.05) BW gain and tibia ash compared with broilers fed all other diets, except tibia ash in birds fed PC+500. Phytase supplementation at 500, 1,000, or 1,500 U/kg to the NC improved (P ≤ 0.05) BW gain and tibia ash comparable with the PC. Feed conversion ratio (FCR) was improved (P ≤ 0.05) in broilers fed NC+1,500 compared with broilers fed all other diets. From d 0 to 49, growth performance was not influenced (P > 0.05) by diet. However, FCR was improved (P ≤ 0.05) in broilers fed 1,500 U/kg of microbial phytase compared with broilers fed the PC, PC+DCP, and NC. There were no differences in performance or tibia ash between broilers fed the PC or PC+DCP, which would indicate the PC diet was sufficient in Ca and P. Therefore, the improvements in FCR in the NC+1500 may be associated with mitigation of the antinutrient effects of phytate rather than improved P utilization.

  14. Phytase activity of lactic acid bacteria and their impact on the solubility of minerals from wholemeal wheat bread.

    Science.gov (United States)

    Cizeikiene, Dalia; Juodeikiene, Grazina; Bartkiene, Elena; Damasius, Jonas; Paskevicius, Algimantas

    2015-01-01

    The objective of this study was to determinate phytase activity of bacteriocins producing lactic acid bacteria previously isolated from spontaneous rye sourdough. The results show that the highest extracellular phytase activity produces Pediococcus pentosaceus KTU05-8 and KTU05-9 strains with a volumetric phytase activity of 32 and 54 U/ml, respectively, under conditions similar to leavening of bread dough (pH 5.5 and 30 °C). In vitro studies in simulated gastrointestinal tract media pH provide that bioproducts prepared with P. pentosaceus strains used in wholemeal wheat bread preparation increase solubility of iron, zinc, manganese, calcium and phosphorus average 30%. Therefore, P. pentosaceus KTU05-9 and KTU05-8 strains could be recommended to use as a starter for sourdough preparation for increasing of mineral bioavailability from wholemeal wheat bread.

  15. Comparison of phosphate estimating methods in the presence of phytic acid for the determination of phytase activity.

    Science.gov (United States)

    Sanikommu, Suma; Pasupuleti, Mukesh; Vadalkonda, Lakshmipathi

    2014-01-01

    Phosphate released from phytic acid can be used as a measure of phytase activity. However, most of the phosphate estimation methods have not examined the interference or interaction of phytic acid in the assay. In this article, we report the kinetics and influence of unreduced phytic acid on phosphate estimation by three of the often-used methods for phytase estimation, the AOAC, Cooper-Gowing, and Fiske-Subbarow methods. Our results show that the AOAC method is most suitable to estimate the phytase activity in the presence of phytate in the medium. In the Fiske and Subbarow method, we noticed that the time factor plays a role in the interference of the phytic acid; especially the readings taken during the second hour of incubation are influenced by the presence of phytic acid. The method of Cooper and Gowing is labor-intensive and is prone to give error values at higher concentrations.

  16. Microbial Phytase as a Way to Improve Growth Performance of Weaned Piglets and to Reduce Phosphorus Excretion

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    REZANA PENGU

    2016-03-01

    Full Text Available The microbial phytase preparation (Aspergillus niger, NATUPHOS was supplemented to a basal ration 750 FTU/kg feed and the effects on growth performance of weaned piglets were studied. The supplementation of microbial phytase improved slightly Daily Weight Gain (DWG, Feed Conversion Ratio (FCR and increases the digestibility and bioavailability of phosphorus from phytate, reduces the amount of inorganic phosphorus needed to maximize growth and bone mineralization and markedly reduces fecal excretion of phosphorus. At the end of experimental period, Daily Weight Gain (DWG was increased 6.15% more and Feed Conversion Ratio (FCR was decreased 4.7%, compared to the control group. Overall a positive effect of the microbial phytase on performance parameters was observed. The P-excretion in the faeces was reduced by 25-30%.

  17. Effects of microbial phytase on apparent and standardized total tract digestibility of calcium in calcium supplements fed to growing pigs.

    Science.gov (United States)

    González-Vega, J C; Walk, C L; Stein, H H

    2015-05-01

    An experiment was conducted to test the hypothesis that differences in the apparent total tract digestibility (ATTD) and standardized total tract digestibility (STTD) of Ca exist among Ca supplements and that inclusion of microbial phytase increases the ATTD and STTD of Ca. One hundred and four growing barrows (average initial BW of 17.73 ± 2.53 kg) were allotted to a randomized complete block design with 13 dietary treatments and 8 pigs per treatment. A basal diet containing corn, cornstarch, potato protein isolate, soybean oil, calcium carbonate, monosodium phosphate, vitamins, and minerals was formulated. Five additional diets were formulated by adding monocalcium phosphate (MCP), dicalcium phosphate (DCP), calcium carbonate, Lithothamnium calcareum Ca, or a high-Ca sugar beet co-product to the basal diet at the expense of cornstarch. Six additional diets that were similar to the previous 6 diets with the exception that they also contained 500 units per kilogram of microbial phytase were also formulated. A Ca-free diet was used to determine basal endogenous losses of Ca. Feces were collected using the marker-to-marker approach. Results indicated that regardless of inclusion of microbial phytase, MCP had the greatest (P supplements. Regardless of inclusion of microbial phytase, the ATTD of P was greater ( Ppigs fed basal, MCP, or DCP diets than in pigs fed calcium carbonate, L. calcareum Ca, or the sugar beet co-product, but pigs fed calcium carbonate diets had greater ( Ppigs fed L. calcareumCa or the sugar beet co-product. Regardless of Ca source, inclusion of microbial phytase increased (P supplements used in this experiment, followed by DCP. Basal, MCP, and DCP diets had greater ATTD of P than the other diets, and inclusion of microbial phytase increased the ATTD and STTD of Ca and the ATTD of P in the diets.

  18. Purification and characterization of a novel cold-adapted phytase from Rhodotorula mucilaginosa strain JMUY14 isolated from Antarctic.

    Science.gov (United States)

    Yu, Peng; Wang, Xue-Ting; Liu, Jing-Wen

    2015-08-01

    A yeast producing a cold-adapted phytase was isolated from Antarctic deep-sea sediment and identified as a Rhodotorula mucilaginosa strain JMUY14 of basidiomycetous yeasts. It was cultured in fermentation optimized by a response surface methodology based on the Box-Behnken design. The maximum activity of phytase reached 205.447 U ml(-1), which was close to the predicted value of 201.948 U ml(-1) and approximately 3.4 times higher than its initial activity. The extracellular phytase was purified by 15.2-fold to homogeneity with a specific activity of 31,635 U mg(-1) by (NH4 )2 SO4 precipitation, and a combination of DEAE Sepharose Fast Flow, SP Sepharose Fast Flow, and Sephadex G-100. The molecular weight of the purified enzyme was estimated to be 63 kDa and its pI was 4.33. Its optimal temperature and pH were 50 °C and 5.0, respectively. Its activity was 85% at 37 °C, and showed good stability at pH 3.0 ∼ 7.0. When compared with mesophilic counterparts, the phytase not only exhibited a higher activity during 20 ∼ 30 °C but also had a low Km (247 µM) and high kcat (1394 s(-1)). The phytase activity was slightly stimulated in the presence of Mg(2+), Fe(2+), Fe(3+), K(+), Na(+), Ca(2+), EDTA, and EGTA and moderately inhibited by Cu(2+), Zn(2+), Mn(2+), Ag(+), PMSF, SDS, and phenylgloxal hydrate. It was resistant to both pepsin and trypsin. Since the phytase produced by the R. mucilaginosa JMUY14 showed a high specific activity, good pH stability, strong protease resistance, and high activity at low temperature, it has great potential for feed applications, especially in aquaculture.

  19. Effect of phytase supplementation on apparent phosphorus digestibility and phosphorus output in broiler chicks fed low-phosphorus diets

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    Xian-Ren Jiang

    2015-04-01

    Full Text Available This study was conducted to evaluate the effect of supplemental phytase in broiler chicks fed different low levels of total phosphorus (P on the apparent phosphorus digestibility (APD and phosphorus output (PO in the faeces and ileal digesta. After fed a standard broiler starter diet from day 0 to 14 post-hatch, a total of 144 male broiler chicks were allocated to 6 groups for a 7-d experiment with a 2 × 3 factorial design comparing phytase (supplemented without (CTR or with 400 FTU/kg phytase (PHY and total P levels (2.0, 2.5 and 3.0 g/kg. The faecal samples were collected from day 17 to 21 post-hatch. At 22 days of age, all the chicks were slaughtered and collected the ileal digesta. Phytase supplementation significantly (P < 0.01 increased APD and decreased PO in the faeces and ileal digesta in comparison with the CTR group. In addition, PO in the faeces expressed as g/kg DM diets and faeces (Diet × P level, P = 0.047 and < 0.01, respectively as well as PO in the ileal digesta expressed as g/kg DM digesta (Diet × P level, P = 0.04 were affected by diet and P level, which were due to the significant reduction (P < 0.01 by PHY supplementation to the diets with 3.0 g/kg total P. The results evidenced that supplemental phytase improved the APD and PO when chicks was fed 3.0 g/kg total P diet, while lower total P levels may limit exogenous phytase efficacy.

  20. Quantitative analysis of phytate globoids isolated from wheat bran and characterization of their sequential dephosphorylation by wheat phytase

    DEFF Research Database (Denmark)

    Bohn, Lisbeth; Josefsen, Lone; Meyer, Anne S.

    2007-01-01

    Wheat phytase was purified to investigate the action of the enzyme toward its pure substrate (phytic acid - myo-inositol hexakisphosphate) and its naturally occurring substrate (phytate globoids). Phytate globoids were purified to homogeneity from wheat bran, and their nutritionally relevant...... phytic acid was replaced with phytate globoids as substrate. Time course degradation of phytic acid or phytate globoids using purified wheat phytase was followed by HPIC identification of inositol phosphates appearing and disappearing as products. In both cases, enzymatic degradation initiated at both...

  1. Endosperm-specific co-expression of recombinant soybean ferritin and Aspergillus phytase in maize results in significant increases in the levels of bioavailable iron.

    Science.gov (United States)

    Drakakaki, Georgia; Marcel, Sylvain; Glahn, Raymond P; Lund, Elizabeth K; Pariagh, Sandra; Fischer, Rainer; Christou, Paul; Stoger, Eva

    2005-12-01

    We have generated transgenic maize plants expressing Aspergillus phytase either alone or in combination with the iron-binding protein ferritin. Our aim was to produce grains with increased amounts of bioavailable iron in the endosperm. Maize seeds expressing recombinant phytase showed enzymatic activities of up to 3 IU per gram of seed. In flour paste prepared from these seeds, up to 95% of the endogenous phytic acid was degraded, with a concomitant increase in the amount of available phosphate. In seeds expressing ferritin in addition to phytase, the total iron content was significantly increased. To evaluate the impact of the recombinant proteins on iron absorption in the human gut, we used an in vitro digestion/Caco-2 cell model. We found that phytase in the maize seeds was associated with increased cellular iron uptake, and that the rate of iron uptake correlated with the level of phytase expression regardless of the total iron content of the seeds. We also investigated iron bioavailability under more complex meal conditions by adding ascorbic acid, which promotes iron uptake, to all samples. This resulted in a further increase in iron absorption, but the effects of phytase and ascorbic acid were not additive. We conclude that the expression of recombinant ferritin and phytase could help to increase iron availability and enhance the absorption of iron, particularly in cereal-based diets that lack other nutritional components.

  2. Effect of phytic acid and microbial phytase on the flow and amino acid composition of endogenous protein at the terminal ileum of growing broiler chickens.

    Science.gov (United States)

    Cowieson, A J; Ravindran, V

    2007-10-01

    The effects of phytic acid and microbial phytase on the flow and composition of endogenous protein at the terminal ileum of broiler chickens were investigated using the peptide alimentation method. Phytic acid (fed as the sodium salt) was included in a synthetic diet at 8.5, 11.5 and 14.5 g/kg (or 2.4, 3.2 and 4.0 g/kg phytate-phosphorus) and each diet was fed without or with an Escherichia coli-derived microbial phytase at 500 phytase units/kg diet. A control containing no phytate was fed as a comparison to estimate basal endogenous flows. Ingestion of phytic acid increased (P < 0.05) the flow of endogenous amino acids and N by an average of 47 % at the lowest phytic acid concentration and 87 % at the highest. The addition of microbial phytase reduced (P < 0.05) the inimical effects of phytic acid on endogenous amino acid flow at all dietary phytic acid levels. The composition of endogenous protein was also influenced (P < 0.10-0.001) by increasing phytic acid concentrations and phytase addition. The effects of phytic acid and phytase on endogenous flow and composition of endogenous protein, however, varied depending on the amino acid. It is concluded that the effects of phytase on amino acid digestibility may be mediated, in part, through a route of reduced endogenous loss.

  3. The effect of microbial phytase and myo-inositol on performance and blood biochemistry of broiler chickens fed wheat/corn-based diets.

    Science.gov (United States)

    Cowieson, A J; Ptak, A; Mackowiak, P; Sassek, M; Pruszynska-Oszmalek, E; Zyla, K; Swiatkiewicz, S; Kaczmarek, S; Józefiak, D

    2013-08-01

    A total of 1,200 Ross broiler chickens were used in 2 separate feeding studies to explore the effect of myo-inositol (MYO) and phytase on performance and blood biochemistry of broilers fed diets formulated to be either adequate or insufficient in Ca and digestible P (dP). Supplementation of diets that were formulated to be insufficient in Ca and dP with MYO resulted in improved BW gain and feed conversion ratio in both experiments. However, these effects were most pronounced in the finisher phase, and moderate negative effects were observed during the starter period. Supplementation of the diet with microbial phytase improved BW gain and feed conversion ratio to a similar extent as was observed with MYO, and there was a degree of subadditivity between the 2 additives. Blood glucose concentrations were increased by both MYO and phytase, though possibly by different mechanisms, because insulin concentrations were not directly relatable to circulating glucose levels, especially when both MYO and phytase were applied simultaneously. The increase in blood glucose concentrations with MYO and phytase was most pronounced in the diet with a lower Ca and dP concentration. It can be concluded that dietary supplementation with MYO or phytase was effective in improving performance of commercial broiler chickens. However, further work is required to explore complex ontogenetic effects of MYO and possible involvement of both MYO and phytase in Na-dependent transport mechanisms.

  4. Effect of non-phytate phosphorus levels and phytase sources on the growth performance, serum biochemical and tibial parameters of broiler chickens

    Directory of Open Access Journals (Sweden)

    Xian R. Jiang

    2013-07-01

    Full Text Available A 3×3 fattorial arrangement with dietary non-phytate phosphorus (NPP levels and phytase sources (3- and 6-phytase was conducted to evaluate the effects of NPP levels, phytase sources and their possible interactions on growth performance, serum biochemical and tibia parameters of broiler chickens from hatch to 42 days of age. A total of 540 1-day-old Arbor Acres male broiler chicks were randomly allocated into nine dietary treatments, each containing 5 replicates pens with 12 birds per pen. Interaction was statistically significant in the performance till day 21 of trial, supplementation of low NPP diet decreased body weight (BW (P<0.001, depressed average daily gain (ADG (P<0.001 and deteriorated average daily feed intake (ADFI (P<0.001 over day 42. During the 8-to-21-day period, even if interaction between NPP levels and phytase sources was significant (P<0.01, BW, ADG and ADFI always increased due to dietary supplementation of phytase, with source not differing. Dietary high NPP enhanced serum calcium and P concentrations on day 21 and 42 (linear contrast, P<0.01, while decreased alkaline phosphatase (AKP activity on day 42 (linear contrast, P<0.001, and interaction was not significant. Both dietary sources of phytase decreased serum AKP activities on day 42 (P<0.001, and urea nitrogen content on day 21 (P<0.01 and 42 (P<0.001. Both phytase improved ash percentage on day 21 and P content in tibia at 21 and 42 days of age (P<0.001. The results confirmed that dietary supplementation of phytase may enhance P availability during the 8-to-21-day period. Nevertheless, no difference between the two phytase sources was observed.

  5. Influence of an Escherichia coli-derived phytase on nutrient utilization in broiler starters fed diets containing varying concentrations of phytic acid.

    Science.gov (United States)

    Ravindran, V; Morel, P C H; Partridge, G G; Hruby, M; Sands, J S

    2006-01-01

    The influence of an Escherichia coli-derived phytase, on nutrient utilization was investigated in broilers fed starter diets containing different concentrations of phytate. The study was conducted as a 3 x 4 factorial arrangement of treatments with 3 concentrations of phytic acid (10.4, 11.8, and 13.6 g/kg; equivalent to 2.8, 3.3, and 3.8 g of phytate P/kg) and phytase (0, 500, 750, and 1,000 FTU/kg). One unit of phytase (FTU) is defined as the quantity of enzyme that releases 1 micromol of inorganic phosphorus/min from 0.00015 mol/L of sodium phytate at pH 5.5 at 37 degrees C. The dietary phytic acid concentrations were manipulated by the inclusion of rice bran. Increasing dietary concentrations of phytic acid resulted in reductions (P phytate concentrations. Apparent ileal digestibility coefficients of protein and most amino acids were influenced by phytate (P phytate concentrations, but the trend in responses to increasing phytase additions was different at different phytate concentrations as shown by significant phytate x phytase interactions (P phytate concentration, the ileal digestibility coefficients increased with increasing phytase supplementation. At the medium and high phytate concentrations, the greatest responses were observed at 500 FTU/kg of phytase, with little improvement attributable to further additions. Ileal digestibility of P was lowered (P phytate concentrations and increased (P phytate x phytase interaction (P phytate concentrations. These data demonstrate the anti-nutritive effects of phytic acid and the potential of microbial phytase to improve energy utilization and the availability of P and amino acids in broilers fed starter diets.

  6. Effect of calcium level and phytase addition on ileal phytate degradation and amino acid digestibility of broilers fed corn-based diets.

    Science.gov (United States)

    Amerah, A M; Plumstead, P W; Barnard, L P; Kumar, A

    2014-04-01

    This study investigated the effect of dietary Ca to available P (AvP) ratio and phytase supplementation on bone ash, ileal phytate degradation, and nutrient digestibility in broilers fed corn-based diets. The experimental design was a 4 × 2 factorial arrangement of treatments evaluating 4 Ca:AvP ratios (1.43, 2.14, 2.86, and 3.57) and 2 levels of phytase (0 and 1,000 phytase units/kg of feed). The 4 Ca:AvP ratios were achieved by formulating all diets to a constant AvP level of 0.28% and varying Ca levels (0.4, 0.6, 0.8, and 1.0%). Each treatment was fed to 6 cages of 8 male Ross 308 broilers from 5 to 21 d. At 21 d, digesta from the terminal ileum was collected and analyzed for energy, phytate, P, Ca, and amino acids (AA) to determine digestibility. Digesta pH was measured in each segment (crop, gizzard, duodenum, and ileum) of the digestive tract. Data were analyzed by 2-way analysis of covariance. There was a significant interaction between dietary Ca:AvP ratio and phytase supplementation for weight gain (WG), feed intake (FI), and feed conversion ratio (FCR). In diets with no phytase, Ca:AvP ratio had a greater effect on WG, FI, and FCR compared with those fed diets without phytase. The orthogonal polynomial contrasts showed that the increase in dietary Ca:AvP ratio significantly decreased WG and FI in a quadratic manner, whereas FCR increased (P Phytase addition increased (P 0.05) between the main factors. Digestibility of AA was positively correlated (P phytase (1,000 phytase units/kg of feed) improved phytate, and P and AA digestibility at all Ca:AvP ratios evaluated in this study.

  7. Microbial phytase in finisher diets of White Pekin ducks: effects on growth performance, plasma phosphorus concentration, and leg bone characteristics.

    Science.gov (United States)

    Orban, J I; Adeola, O; Stroshine, R

    1999-03-01

    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

  8. 植酸酶研究进展及其在饲料工业中的应用%Research Advance and Application of Phytase in Feed Industry

    Institute of Scientific and Technical Information of China (English)

    马玺; 单安山

    2001-01-01

    The classes, sources and properties of phytase, test way of phytase activity and the research advance on highly productive strains as well as the effect of supplying phytase on nutritive value of feeds and production performance of animal were reviewed.%综述了植酸酶的种类、来源、酶学特性、酸活的测定方法、当前国内外对植酸酶高产菌株的研究进展,以及应用植酸酶制剂对饲料营养价值与畜禽生产性能的影响。

  9. Interactions between supplemented mineral phosphorus and phytase on phytate hydrolysis and inositol phosphates in the small intestine of broilers1,2.

    Science.gov (United States)

    Zeller, Ellen; Schollenberger, Margit; Witzig, Maren; Shastak, Yauheni; Kühn, Imke; Hoelzle, Ludwig E; Rodehutscord, Markus

    2015-05-01

    Phytate breakdown in the digestive tract of broilers is affected by supplements of mineral phosphorus (P) and phytase with unknown interactions between the 2 factors. It was the objective to study phytate hydrolysis and the presence of inositol phosphate isomers (InsPs) as affected by supplements of mineral P and phytase in the small intestine of broilers. Fifteen-day old broilers were assigned to 48 pens of 20 broilers each (n = 8 pens/treatment). Two low-P corn-soybean meal-based diets without (BD-; 4.4 g P/kg dry matter) or with monocalcium phosphate (MCP; BD+; 5.2 g P/kg dry matter) were supplied without or with added phytase at 500 or 12,500 FTU/kg. On d 24, digesta from the duodenum/jejunum and lower ileum was pooled per segment on a by-pen basis, freeze-dried, and analyzed for P, InsPs, and the marker TiO2. Another 180 broilers (n = 6 pens/treatment, 10 birds each) were fed the 3 BD+ diets from d 1 to 21 to assess the influence of supplemented phytase on tibia mineralization and strength. Significant interactions between MCP and phytase supplements on myo-inositol 1,2,3,4,5,6-hexakis (dihydrogen phosphate) (InsP6) hydrolysis (duodenum/jejunum: P ≤ 0.001; ileum: P = 0.004) and level of specific lower InsPs were detected. Supplementation with 12,500 FTU/kg phytase resulted in 92% InsP6 hydrolysis and strong degradation of InsP5. This treatment resulted in higher P net absorption, affirmed by higher BW gain, tibia strength, and mineralization compared to treatments without or with 500 FTU/kg phytase (P ≤ 0.05). MCP supplementation reduced the degradation of InsP6 and specific lower InsPs in birds fed diets without or with 500 FTU/kg of phytase (P ≤ 0.05), but did not reduce InsP6 hydrolysis or degradation of InsP5 at the high phytase dose. Effects of added MCP on phytase efficacy depend on the dose of supplemented phytase. Differences in the concentrations of lower InsPs indicated that the initial step of InsP6 hydrolysis is not the only

  10. Environmental Assessment of Ronozyme® P5000 CT Phytase as an Alternative to Inorganic Phosphate Supplementation to Pig Feed Used in Intensive Pig Production

    DEFF Research Database (Denmark)

    Nielsen, Per Henning; Wenzel, Henrik

    2007-01-01

    Ronozyme® P5000 CT is an industrially produced enzyme product (phytase) which is able to degrade naturally occurring phytate in animal feed and release the phytate's content of phosphorus for pig's growth. Ronozyme P5000 CT (hereafter called Ronozyme Phytase) can be used as an alternative...... of agricultural land is given very large relative weight. Hundreds of enzyme products are commercially available on the market today, each with a range of different applications. There are several indications that enzymes like Ronozyme Phytase can play an important role in a transition to a more sustainable...

  11. Effects of phytase-assisted processing method on physicochemical and functional properties of soy protein isolate.

    Science.gov (United States)

    Wang, Hongjian; Chen, Yeming; Hua, Yufei; Kong, Xiangzhen; Zhang, Caimeng

    2014-11-12

    Phytate is an important antinutritional factor in food products. In this study, a phytase-assisted processing method was used to produce low-phytate soybean protein isolate (SPI) samples, and their physicochemical and functional properties were examined. Hydrolysis condition at low temperature (room temperature) and pH 5.0 was better than that recommended by manufacturer (pH 5.0, 55 °C) at keeping the properties of SPI, so the former condition was selected to prepare SPI samples with phytate contents of 19.86-0.11 mg/g by prolonging hydrolysis time (0 (traditional method), 5, 10, 20, 40, and 60 min). Ash content (R(2) = 0.940), solubility (R(2) = 0.983), ζ-potential value (R(2) = 0.793), denaturation temperatures (β-conglycinin, R(2) = 0.941; glycinin, R(2) = 0.977), emulsifying activity index (R(2) = 0.983), foaming capacity (R(2) = 0.955), and trypsin inhibitor activity (R(2) = 0.821) of SPI were positively correlated with phytate content, whereas protein content (R(2) = 0.876), protein recovery (R(2) = 0.781), emulsifying stability index (R(2) = 0.953), foaming stability (R(2) = 0.919), gel hardness (R(2) = 0.893), and in vitro digestibility (R(2) = 0.969) were negatively correlated with phytate content. Simulated gastrointestinal digestion and subsequent dialysis showed that percentages of dialyzable Zn and Ca were increased with decreasing phytate content, whereas the amounts of dialyzable Zn and Ca revealed different behaviors: the former was increased and the latter was decreased. Circular dichroism spectra showed that secondary structure of SPI was changed by phytase. Compared with traditional processing method, the phytase-assisted processing method could produce SPI with lower phytate and higher protein contents, which had better in vitro digestibility and could be used to prepare gels with higher hardness by partially losing some other functional properties.

  12. Comparative Study on Different Expression Hosts for Alkaline Phytase Engineered in Escherichia coli.

    Science.gov (United States)

    Chen, Weiwei; Yu, Hongwei; Ye, Lidan

    2016-07-01

    The application of alkaline phytase as a feed additive is restricted by the poor specific activity. Escherichia coli is a frequently used host for directed evolution of proteins including alkaline phytase towards improved activity. However, it is not suitable for production of food-grade products due to potential pathogenicity. To combine the advantages of different expression systems, mutants of the alkaline phytase originated from Bacillus subtilis 168 (phy168) were first generated via directed evolution in E. coli and then transformed to food-grade hosts B. subtilis and Pichia pastoris for secretory expression. In order to investigate the suitability of different expression systems, the phy168 mutants expressed in different hosts were characterized and compared in terms of specific activity, pH profile, pH stability, temperature profile, and thermostability. The specific activity of B. subtilis-expressed D24G/K70R/K111E/N121S mutant at pH 7.0 and 60 °C was 30.4 U/mg, obviously higher than those in P. pastoris (22.7 U/mg) and E. coli (19.7 U/mg). Moreover, after 10 min incubation at 80 °C, the B. subtilis-expressed D24G/K70R/K111E/N121S retained about 70 % of the activity at pH 7.0 and 37 °C, whereas the values were only about 25 and 50 % when expressed in P. pastoris and E. coli, respectively. These results suggested B. subtilis as an appropriate host for expression of phy168 mutants and that the strategy of creating mutants in one host and expressing them in another might be a new solution to industrial production of proteins with desired properties.

  13. Iterative key-residues interrogation of a phytase with thermostability increasing substitutions identified in directed evolution.

    Science.gov (United States)

    Shivange, Amol V; Roccatano, Danilo; Schwaneberg, Ulrich

    2016-01-01

    Bacterial phytases have attracted industrial interest as animal feed supplement due to their high activity and sufficient thermostability (required for feed pelleting). We devised an approach named KeySIDE,  an iterative Key-residues interrogation of the wild type with Substitutions Identified in Directed Evolution for improving Yersinia mollaretii phytase (Ymphytase) thermostability by combining key beneficial substitutions and elucidating their individual roles. Directed evolution yielded in a discovery of nine positions in Ymphytase and combined iteratively to identify key positions. The "best" combination (M6: T77K, Q154H, G187S, and K289Q) resulted in significantly improved thermal resistance; the residual activity improved from 35 % (wild type) to 89 % (M6) at 58 °C and 20-min incubation. Melting temperature increased by 3 °C in M6 without a loss of specific activity. Molecular dynamics simulation studies revealed reduced flexibility in the loops located next to helices (B, F, and K) which possess substitutions (Helix-B: T77K, Helix-F: G187S, and Helix-K: K289E/Q). Reduced flexibility in the loops might be caused by strengthened hydrogen bonding network (e.g., G187S and K289E/K289Q) and a salt bridge (T77K). Our results demonstrate a promising approach to design phytases in food research, and we hope that the KeySIDE might become an attractive approach for understanding of structure-function relationships of enzymes.

  14. Efficacy of OptiphosTM phytase on mineral digestibility in diets for breeding sows: effect during pregnancy an lactation

    NARCIS (Netherlands)

    Jongbloed, A.W.; Diepen, van J.T.M.; Binnendijk, G.P.; Bikker, P.; Vereecken, M.; Bierman, K.

    2013-01-01

    Phosphorus in most diets for breeding sows is digested for 20 to 40%, thus leading to a relatively high amount of P in the manure. To enhance the P digestibility in diets for both lactating and gestating sows, two separate experiments were carried out to study the efficacy of OptiphosTM phytase deri

  15. Effect of graded doses and a high dose of microbial phytase on the digestibility of various minerals in weaner pigs

    NARCIS (Netherlands)

    Kies, A.K.; Kemme, P.A.; Sebek, L.B.J.; Diepen, van J.T.M.; Jongbloed, A.W.

    2006-01-01

    An experiment with 224 weaner pigs (initial BW of 7.8 kg) was conducted to determine the effect of dose of dietary phytase supplementation on apparent fecal digestibility of minerals (P, Ca, Mg, Na, K, and Cu) and on performance. Four blocks, each with 8 pens of 7 pigs, were formed. Eight dietary tr

  16. Treating thin stillage or condensed distillers solubles with phytase for production of low phytate co-products

    Science.gov (United States)

    Fuel ethanol production from grains is mainly based on dry grind processing, during which phytate is concentrated about three fold in distillers dried grains with solubles (DDGS), a major co-product. For reducing phyate in DDGS, two industrial phytase preparations (Natuphos and Ronozyme) were used ...

  17. Reduction of phytate content in unfermented whole grain wheat flour dough using permeabilized phytase active Candida versatilis mutants

    Directory of Open Access Journals (Sweden)

    Bindu Sadanandan*

    2012-08-01

    Full Text Available High phytic acid (PA content in whole wheat flour products reduces the bioavailability of nutrients, especially dietary minerals. Monogastric animals cannot breakdown PA, an organophosphorus compound as they do not produce the enzyme phytase. Hence phytate gets excreted from the system leading to phosphorus deficiency and accumulates in the environment. Accumulation of PA also causes serious soil and water pollution during animal husbandry. In non-fermented food products the benefit of activation of innate grain phytases or microbial phytases cannot be exploited. In the present study the use of freeze-thaw permeabilized phytase active mutant yeast cells to reduce PA content in unfermented foods has been successfully tested.  Candida versatilis mutants, UVY 505 and EMY 505 used in this process were able to bring about phytate reduction of 24.32 to 45%. The substantial but not complete removal of PA here also helps to derive the cardiovascular and other health benefits of PA acknowledged of late. The method developed is simple, rapid, chemical free, nontoxic, economical and ideal for food applications.

  18. Phenolics, phytic acid, and phytase in Canadian-grown low-tannin faba bean (Vicia faba L.) genotypes.

    Science.gov (United States)

    Oomah, B Dave; Luc, Geneviéve; Leprelle, Claire; Drover, John C G; Harrison, Judith E; Olson, Mark

    2011-04-27

    Thirteen low-tannin faba bean genotypes grown at two locations in north central Alberta in 2009 were evaluated to investigate the variation in seed characteristics, phenolic and phytate contents, and phytase and antioxidant activities and to elucidate the relationship of these components as a breeding strategy. Seed characteristics including color were predominantly genotype dependent. The faba bean genotypes with total phenolic content ranging from 5.5 to 41.8 mg of catechin equiv/g of sample was linearly related to tannin content and the best predictor of antioxidant activity. Phytic acid content and phytase activity varied significantly among genotypes and between locations, ranging from 5.9 to 15.1 g/kg and from 1606 to 2154 FTU/kg sample, respectively. Multivariate data analysis performed on 19 components analyzed in this study using principal component analysis (PCA) and cluster analysis demonstrate that differences in seed characteristics, phenolic components, phytic acid, and phytase are major factors in segregating faba bean genotypes. The relatively low phytic acid content and high phytase activity of these low-tannin faba bean genotypes are beneficial/essential traits for their use in human and animal nutrition.

  19. Nutrient balance of layers fed diets with different calcium levels and the inclusion of phytase and/or sodium butyrate

    Directory of Open Access Journals (Sweden)

    MM Vieira

    2011-06-01

    Full Text Available In this study, Hisex Brown layers in lay were evaluated between 40 and 44 weeks of age to evaluate the inclusion of bacterial phytase (Ph and sodium butyrate (SB to diets containing different calcium levels (CaL. Performance, average egg weight and eggshell percentage, in addition to nutrient metabolizability and Ca and P balance were evaluated for 28 days. Birds were distributed according to a completely randomized experimental design with a 3x2x2 factorial arrangement, with three calcium levels (2.8, 3.3, 3.8%; the addition or not of phytase (500PhU/kg and the addition or not of sodium butyrate (20mEq/kg, composing 12 treatments with eight replicates of one bird each. There was no additive effect of phytase or SB on the evaluated responses. Feed intake and feed conversion ratio were influenced by CaL, with the best performance obtained with 3.3% dietary Ca. Ca balance was positively affected by dietary Ca, and P balance by the addition of phytase. Ca dietary concentration, estimated to obtain Ca body balance, was 3.41%, corresponding to an apparent retention of 59.9% of Ca intake.

  20. The influence of dietary microbial phytase and calcium on the accumulation of cadmium in different organs of pigs

    Energy Technology Data Exchange (ETDEWEB)

    Zacharias, B.; Lantzsch, H.J.; Drochner, W. [Hohenheim Univ., Stuttgart (Germany). Inst. fuer Tierernaehrung

    2001-07-01

    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. (orig.)

  1. Tetrachlorofluorescein TInsP as a Substrate Analog Probe for Measuring Phytase Activity in Surface Water: Proof of Concept.

    Science.gov (United States)

    Berry, Duane F; Harich, Kim

    2013-01-01

    An innovative approach for measuring phytase activity (PA) in surface water is presented. A substrate analog of -inositol hexakis(dihydrogen) phosphate (InsP), commonly referred to as phytic acid, 1--5--(1-oxo-1-(2' ,4,7,7' -tetrachloro-3',6'-dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9'-xanthen]-6-yl)-5,8,11-trioxa-2-azatridecan-13-yl)-inositol 1,2,3,4,6-pentakis--(dihydrogen) phosphate, referred to as tetrachlorofluorescein (TET) tethered (T)InsP, has been developed that can be used to monitor the (phytase-catalyzed) phosphate ester bond-cleavage reaction. Test phytases, (wheat [4-] and [3-] phytase) sequentially remove phosphate groups from TET TInsP, producing dephosphorylated probe species that were readily separated by reversed-phase high-performance liquid chromatography (RP-HPLC). Because dephosphorylated probe species retain the TET group, highly sensitive quantification could be achieved using fluorescence detection (excitation/emission ' = 245/540 nm). Calibration curves for TET TInsP, which could be used as a standard for quantifying all probe species, were linear ( > 0.999) over the range of concentrations tested. Phytase-generated dephosphorylated probe species were characterized or identified using RP-HPLC with mass spectrometry. Results of mass spectrometry analysis show that the RP-HPLC system was capable of distinguishing between dephosphorylated probe species at the regioisomeric level. The TET TInsP molecular probe was used to successfully measure PA in pond water. We found that the PA associated with the particulate plus water-soluble fraction was greater than that observed for the water-soluble fraction alone. Moreover, it appeared that 4- and 3-phytase were active in pond water based on an analysis of the chromatographic profile (i.e., elution sequence) of dephosphorylated probe species produced. The advent of a fluorescent substrate analog of InsP affords environmental scientists with the means to unambiguously quantify an extremely small

  2. Isolation and characterization of potential phytase-producing fungi from environmental samples of antioquia (colombia) / aislamiento y caracterización de hongos productores de fitasa a partir de muestras ambientales de antioquia (colombia)

    OpenAIRE

    Maritza Ocampo Betancur; Luisa Fernanda Patiño Cervantes; Mauricio Marín Montoya; Mauricio Salazar Yepes; Pablo Andrés Gutiérrez Sánchez

    2012-01-01

    Abstract. Phytases are enzymes used as feed additive that enhance the phosphorus and mineral uptake in monogastric animals and reduce the level of phosphate excretion in their manure. Due to their easy cultivation and high production of extracellular enzymes, filamentous fungi are one of best sources of phytase for use in the feed industry. Phytase has been found principally in the genera Aspergillus, Penicillium, Mucor and Rhizopus. In this work, we report the isolation and characterization ...

  3. Detection of transgenic and endogenous plant DNA fragments and proteins in the digesta, blood, tissues, and eggs of laying hens fed with phytase transgenic corn.

    Directory of Open Access Journals (Sweden)

    Qiugang Ma

    Full Text Available The trials were conducted to assess the effects of long-term feeding with phytase transgenic corn (PTC to hens on laying performance and egg quality, and investigate the fate of transgenic DNA and protein in digesta, blood, tissues, and eggs. Fifty-week old laying hens (n = 144 were fed with a diet containing 62.4% PTC or non-transgenic isogenic control corn (CC for 16 weeks. We observed that feeding PTC to laying hens had no adverse effect on laying performance or egg quality (P>0.05 except on yolk color (P<0.05. Transgenic phyA2 gene and protein were rapidly degraded in the digestive tract and were not detected in blood, heart, liver, spleen, kidney, breast muscle, and eggs of laying hens fed with diet containing PTC. It was concluded that performance of hens fed diets containing PTC, as measured by egg production and egg quality, was similar to that of hens fed diets formulated with CC. There was no evidence of phyA2 gene or protein translocation to the blood, tissues, and eggs of laying hens.

  4. Detection of transgenic and endogenous plant DNA fragments and proteins in the digesta, blood, tissues, and eggs of laying hens fed with phytase transgenic corn.

    Science.gov (United States)

    Ma, Qiugang; Gao, Chunqi; Zhang, Jianyun; Zhao, Lihong; Hao, Wenbo; Ji, Cheng

    2013-01-01

    The trials were conducted to assess the effects of long-term feeding with phytase transgenic corn (PTC) to hens on laying performance and egg quality, and investigate the fate of transgenic DNA and protein in digesta, blood, tissues, and eggs. Fifty-week old laying hens (n = 144) were fed with a diet containing 62.4% PTC or non-transgenic isogenic control corn (CC) for 16 weeks. We observed that feeding PTC to laying hens had no adverse effect on laying performance or egg quality (P>0.05) except on yolk color (PTransgenic phyA2 gene and protein were rapidly degraded in the digestive tract and were not detected in blood, heart, liver, spleen, kidney, breast muscle, and eggs of laying hens fed with diet containing PTC. It was concluded that performance of hens fed diets containing PTC, as measured by egg production and egg quality, was similar to that of hens fed diets formulated with CC. There was no evidence of phyA2 gene or protein translocation to the blood, tissues, and eggs of laying hens.

  5. Phytase, phosphatase activity and p-nutrition of soybean as influenced by inoculation of bacillus.

    Science.gov (United States)

    Ramesh, A; Sharma, Sushil K; Joshi, O P; Khan, I R

    2011-01-01

    The efficiency of different Bacillus isolates on rhizosphere soil enzyme activities and P-nutrition of soybean was carried out under microcosm conditions. Significant increase in enzyme activities viz., fluorescein diacetate activity, phosphatase and phytase activity and consequent effects on P-nutrition were observed with the inoculation of Bacillus isolates over uninoculated control. Among the isolates, BD-3-1B, KHBD-6, BDKH-3, Bacillus amyloliquefacians, and Bacillus cereus were found to be promising. The phytic acid-P as a percentage of total P content in soybean seeds decreased with the inoculation of Bacillus isolates as compared to un-inoculated control. A decrease in phytic-P in soybean seeds not only results in better digestibility and increased feed efficiency. Pearson correlation studies revealed a significant positive association between acid, alkaline phosphatases, phytase activity on available P content in soil and P content in seeds with the inoculation of Bacillus isolates, indicating role of these enzymes in P mobilization and acquisition by soybean.

  6. The role played by phytase and metabolism in the accumulation of uranium in the poultry bones

    Energy Technology Data Exchange (ETDEWEB)

    Arruda-Neto, J.D.T.; Manso Guevara, M.V.; Vanin, V.R.; Deppman, A.; Likhachev, V.P.; Mesa, J.; Helene, O.A.M.; Martins, M.N.; Gouveia, A.N. [Sao Paulo Univ., SP (Brazil). Inst. de Fisica; Cestari, A.C.; Jorge, S.A.C. [Universidade de Santo Amaro (UNISA), SP (Brazil); Nogueira, G.P.; Fonseca, L.E.C. [UNESP, SP (Brazil). Faculdade de Medicina Veterinaria; Zamboni, C.B.; Saiki, M. [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil); Jorge, S.A.C. [Instituto Butantan, Sao Paulo, SP (Brazil). Lab. de Imunologia Viral; Rodriguez, O.; Guzman, F [Institute of Nuclear Sciences and Technology, Havana (Cuba); Garcia, F. [Universidade Estadual de Santa Cruz, BA (Brazil)

    2002-07-01

    Groups of seven days old Cobb broilers were fed with feed doped with uranyl nitrate at a fixed concentration of 20 ppm-U, and two concentrations of phytase (120 and 180 ppm). Two animals per group were sacrificed weekly up to their adultness. The uranium content in tibia was measured by neutron activation analysis. It was observed that the biokinetics of U does not change by administration of phytase, but the U concentration in the bones increased by up to a factor of 2, and in a non expected periodically time oscillating fashion. Quite surprising too, the concentration of uranium ({mu}g-U/g-bone) is decreasing all along the animal life spanning period of 14-42 days, meaning that the skeleton mass is growing faster than the corresponding accumulation of uranium is. This last finding is interpreted as a possible interplay between two metabolic peculiarities, associated both with U transfer to (uptake), and U removed from (clearance) the bones, respectively. (author)

  7. Comparative Proteomics of Leaves from Phytase-Transgenic Maize and Its Non-transgenic Isogenic Variety

    Science.gov (United States)

    Tan, Yanhua; Yi, Xiaoping; Wang, Limin; Peng, Cunzhi; Sun, Yong; Wang, Dan; Zhang, Jiaming; Guo, Anping; Wang, Xuchu

    2016-01-01

    To investigate unintended effects in genetically modified crops (GMCs), a comparative proteomic analysis between the leaves of the phytase-transgenic maize and the non-transgenic plants was performed using two-dimensional gel electrophoresis and mass spectrometry. A total of 57 differentially expressed proteins (DEPs) were successfully identified, which represents 44 unique proteins. Functional classification of the identified proteins showed that these DEPs were predominantly involved in carbohydrate transport and metabolism category, followed by post-translational modification. KEGG pathway analysis revealed that most of the DEPs participated in carbon fixation in photosynthesis. Among them, 15 proteins were found to show protein-protein interactions with each other, and these proteins were mainly participated in glycolysis and carbon fixation. Comparison of the changes in the protein and tanscript levels of the identified proteins showed that most proteins had a similar pattern of changes between proteins and transcripts. Our results suggested that although some significant differences were observed, the proteomic patterns were not substantially different between the leaves of the phytase-transgenic maize and the non-transgenic isogenic type. Moreover, none of the DEPs was identified as a new toxic protein or an allergenic protein. The differences between the leaf proteome might be attributed to both genetic modification and hybrid influence. PMID:27582747

  8. ISOLATION AND CHARACTERIZATION OF A LIPOLYTIC AND PHYTASE PRODUCING PROBIOTIC FOR POTENTIAL APPLICATION IN POULTRY FEED

    Directory of Open Access Journals (Sweden)

    K. RAZDAN

    2012-07-01

    Full Text Available In the current study a total of 35 bacterial isolates from 17 food and fecal samples were examined. Five among those were earmarked as putative probiotic candidates. All the selected isolates survived the low pH conditions of 2.0, and resisted the presence of bile salts (0.02-0.25% and NaCl (2-14 %, indicating their ability to survive in the gastrointestinal (GI tract conditions and hence making them suitable candidates for probiotic applications. The selected probiotic isolates showed considerable levels of hydrophobicity indicating their potential adhering properties with the gut epithelium. In addition the five selected probiotic candidates depicted substantial antagonistic action against potent pathogens like Bacillus subtilis, B. cereus, Escherichia coli, Pseudomonas aeruginosa, P. alcaligenes, Staphylococcus aureus and Streptococcus sp. The isolates CM-4 and KD-7 were most remarkable as they inhibited all the pathogens tested including S. aureus and Streptococcus sp. Extracellular enzymatic studies showed that all the five strains produced phytase whereas isolate CM-4 and KD-7 were the only lipase producers found. However no amylase protease activity was detected. Isolate CM-4 was found to be the best among all five as it showed all desirable probiotic features viz. bile salt, NaCl and pH tolerance, maximum hydrophobicity, antagonistic action against pathogens, phytase and lipase activity, therefore was identified by using 16S rDNA sequencing and MEGA BLAST.

  9. Effects of low-phytic acid corn, low-phytic acid soybean meal, and phytase on nutrient digestibility and excretion in growing pigs.

    Science.gov (United States)

    Hill, B E; Sutton, A L; Richert, B T

    2009-04-01

    Forty-eight grower pigs were used to evaluate the effects of feeding low phytic acid (LPA) corn, LPA soybean meal, normal corn (NC), normal soybean meal (NSBM), and phytase on nutrient digestibility and excretion. Barrows were blocked by BW (initial BW=45.3+/-1.6 kg) and randomly assigned to 1 of 8 dietary treatments in a 2 x 2 x 2 factorial arrangement (6 pigs/treatment). Pigs were fed twice daily (0700 and 1700 h) at 3 times the ME requirement for maintenance. Phytase was added to the diet at 510 phytase units/kg of feed (where 1 phytase unit is the quantity of enzyme that liberates 1 mumol of inorganic P/min from 0.005 mol/L of sodium phytate at pH 5.5 and 37 degrees C), at the expense of corn starch, and all diets were formulated to provide 0.39% total P, 0.50% Ca, and 1.0% lysine with no supplemental inorganic P. Pigs were adapted to metabolism crates and dietary treatments for 7 d, followed by a 3-d total collection of urine and feces. Total fecal DM excreted, percentage of DM of feces, and percentage of DM digested were not different (P>0.53) among treatments. Fecal P excretion was reduced for pigs fed LPA corn vs. NC (2.85 vs. 3.24+/-0.119 g/d; P=0.024), for pigs fed LPA soybean meal vs. NSBM (2.79 vs. 3.30+/-0.119 g/d; P=0.007), for pigs fed phytase vs. nonphytase diets (2.80 vs. 3.29+/-0.119 g/d; P=0.009), and for pigs fed LPA corn, LPA soybean meal, and phytase vs. NC and NSBM without phytase (2.16 vs. 3.70+/-0.237 g/d; Psoybean meal, and phytase diet vs. the NC and soybean meal diet (60.1 vs. 34.1+/-4.5%; Psoybean meal vs. NSBM (47.2 vs. 41.1+/-2.27%; P=0.075). Corn type and soybean meal type had no effect (P>0.11) on water-soluble P excretion. However, pigs fed diets containing phytase tended to excrete less total water-soluble P than those without phytase inclusion (1.99 vs. 2.27+/-0.099 g/d; Psoybean meal, and phytase was additive, significantly improving P digestibility and dramatically decreasing P excretion to reduce the potential impacts of P

  10. The effect of dietary phosphorus level and phytase supplementation on growth performance, bone-breaking strength, and litter phosphorus concentration in broilers.

    Science.gov (United States)

    Powell, S; Johnston, S; Gaston, L; Southern, L L

    2008-05-01

    This research was conducted to determine the effects of feeding different P levels with and without phytase supplementation on broiler growth performance, bone-breaking strength (BBS), and litter P concentration. An experiment with 4 trials was conducted with 7,840 Ross x Ross straight-run broilers. For each trial, 1,960 broilers were allotted on d 0 to treatments, with 7 replications each and with 70 broilers per replication. The broilers were fed a 4-period feeding program consisting of starter (0 to 14 d), grower (14 to 32 d), finisher (32 to 41 d), and withdrawal (41 to 50 d) periods. For each trial, the same pen was used continuously for each treatment-replication combination, and the litter was not removed between trials. Broilers were fed a control diet [0.43, 0.40, 0.36, or 0.32% nonphytate P (nPP)] in the starter, grower, finisher, and withdrawal periods, respectively, a low Ca and P (LCaP) diet with a 0.05% reduction in nPP in each period, and these 2 diets supplemented with phytase at 600 phytase units/kg (nPP and Ca were reduced by 0.094% in diets with phytase). Diet did not affect (P > 0.10) broiler performance in the starter or withdrawal periods. Generally, both phytase addition and the LCaP diet decreased some aspects of growth performance during the grower and finisher periods. There was no main effect of phytase on BBS, but BBS was decreased in the broilers fed the LCaP diet with phytase addition (nPP x phytase, P < 0.01) in the grower period, and BBS was decreased in the finisher (P < 0.02) and withdrawal (P < 0.01) periods for broilers fed the LCaP diet. Total P, soluble P (SP), and reactive soluble P (RSP) were decreased (P < 0.04) in the litter of broilers fed the LCaP diets. Total P, SP, and RSP were decreased (P < 0.01) in the litter of broilers fed phytase. These data indicate that phytase supplementation at 600 phytase units/kg reduces growth in some periods, has no effect on BBS, and reduces total P, SP, and RSP in the litter.

  11. Dietary calcium, phosphorus, and phytase effects on bird performance, intestinal morphology, mineral digestibility, and bone ash during a natural necrotic enteritis episode.

    Science.gov (United States)

    Paiva, Diego; Walk, Carrie; McElroy, Audrey

    2014-11-01

    The objective of this study was to evaluate the effects of dietary Ca, P, and phytase on performance, intestinal morphology, bone ash, and Ca and P digestibility during a necrotic enteritis (NE) outbreak. The 35-d trial was designed as a 2 × 2 × 2 factorial, which included 2 Ca levels (0.6 and 0.9%), 2 P levels (0.3 and 0.45%), and 2 levels of phytase [0 and 1,000 phytase units (FTU)/kg]. Birds were placed on litter from a previous flock that exhibited clinical signs of NE. Birds and feed were weighed on d 12, 19, and 35, and BW gain, feed intake, and feed conversion were calculated. Mortality was recorded daily, and gastrointestinal pH was measured. Tibias and ileal digesta were also collected. Birds began exhibiting clinical signs of NE on d 9, and NE-associated mortality persisted until d 26. Dietary Ca supplemented at 0.9% or inclusion of 1,000 FTU/kg of phytase significantly increased mortality compared with 0.6% Ca or 0 FTU/kg of phytase, respectively. From d 0 to 12, birds fed 0.9% Ca and 0.45% available P with phytase had greater BW gain compared with birds fed 0.6% Ca, 0.45% available P, and phytase. From d 0 to 19, birds fed diets with 0.9% Ca and 0.3% available P had decreased feed intake and improved feed conversion compared with birds fed 0.9% Ca and 0.45% available P. Calcium at 0.9% increased gizzard (d 19) and jejunum (d 12) pH. Phytase supplementation significantly increased Ca digestibility regardless of Ca and P levels of the diets. In addition, diets containing 0.6% Ca and 1,000 FTU/kg of phytase resulted in a significant increase in P digestibility. The results suggest that dietary Ca level may influence NE-associated mortality. In addition, bird performance was affected by interactions of Ca, P, and phytase during the exposure to Clostridium perfringens and the subsequent NE outbreak. Results showed improvements in bird performance when birds were fed 0.6% Ca and 0.3% P in diets supplemented with phytase, which was likely consequent to the

  12. Use of Zn concentration in the gastrointestinal tract as a measure of phytate susceptibility to the effect of phytase supplementation in broilers

    OpenAIRE

    Morgan, NK; Scholey, DV; Burton, EJ

    2016-01-01

    Zinc (Zn) is the most vulnerable cation to complexation with phytate. An experiment was conducted to evaluate the potential of measurements of Zn concentration in the gastrointestinal tract as a marker to assess the anti-nutritional impact of phytate and susceptibility of phytate to phytase in broilers. Ross 308 broilers (n = 180) were fed one of 5 experimental diets with differing phytase activity levels, analyzed at 605, 1150, 1804, 3954 and 5925 U/kg. Broiler performance and Zn concentrati...

  13. Effect of phytase supplementation to diets varying in chloride level on performance, litter moisture, foot pad score, and gait score of growing turkeys.

    Science.gov (United States)

    Farahat, M H; Abdel-Razik, W M; Hassanein, E I; Noll, S L

    2013-07-01

    Phytase was examined as a means to improve turkey performance in diets with high Cl by modifying dietary electrolyte balance. Nicholas turkey toms (10 poults/pen; 10 replicates per pen) were fed corn-soy-distillers dried grains with solubles-canola meal-based diets (DDGS/CM) with varying Cl (0.2, 0.3, 0.4, or 0.5%) and phytase (0 or 500 phytase units/kg of diet), making 8 treatments (4 × 2 factorial). A ninth treatment was a corn-soy based diet (CS) with 0.3% Cl and without phytase (positive control). The diets were formulated to contain similar AMEn and digestible amino acid levels for each of 5 feeding periods (2 to 5, 5 to 8, 8 to 11, 11 to 14, and 14 to 17 wk of age) and fed in mash form. Turkey BW, ADG, ADFI, and feed conversion ratio (FCR) were determined for each pen along with litter moisture, foot pad, and gait scores. An ANOVA was conducted to determine the effects of Cl, phytase, and their interaction. Turkey BW, ADG, and ADFI decreased in a linear or curvilinear manner with increasing diet Cl (P Phytase improved FCR during 8 to 11 wk and increased litter moisture at 11 wk of age (P Phytase modified the FCR response to Cl during 2 to 5 and 14 to 17 wk of age at the 2 lowest levels of dietary Cl (P phytase, and associated dietary electrolyte balance in turkey diets containing high levels of alternative byproducts.

  14. High Cell Density Process for Constitutive Production of a Recombinant Phytase in Thermotolerant Methylotrophic Yeast Ogataea thermomethanolica Using Table Sugar as Carbon Source.

    Science.gov (United States)

    Charoenrat, Theppanya; Antimanon, Sompot; Kocharin, Kanokarn; Tanapongpipat, Sutipa; Roongsawang, Niran

    2016-12-01

    The yeast Ogataea thermomethanolica has recently emerged as a potential host for heterologous protein expression at elevated temperature. To evaluate the feasibility of O. thermomethanolica as heterologous host in large-scale fermentation, constitutive production of fungal phytase was investigated in fed-batch fermentation. The effect of different temperatures, substrate feeding strategies, and carbon sources on phytase production was investigated. It was found that O. thermomethanolica can grow in the temperature up to 40 °C and optimal at 34 °C. However, the maximum phytase production was observed at 30 °C and slightly decreased at 34 °C. The DOT stat control was the most efficient feeding strategy to obtain high cell density and avoid by-product formation. The table sugar can be used as an alternative substrate for phytase production in O. thermomethanolica. The highest phytase activity (134 U/mL) was obtained from table sugar at 34 °C which was 20-fold higher than batch culture (5.7 U/mL). At a higher cultivation temperature of 38 °C, table sugar can be used as a low-cost substrate for the production of phytase which was expressed with an acceptable yield (85 U/mL). Lastly, the results from this study reveal the industrial favorable benefits of employing O. thermomethanolica as a host for heterologous protein production.

  15. Effect of super dosing of phytase on growth performance, ileal digestibility and bone characteristics in broilers fed corn-soya-based diets.

    Science.gov (United States)

    Manobhavan, M; Elangovan, A V; Sridhar, M; Shet, D; Ajith, S; Pal, D T; Gowda, N K S

    2016-02-01

    A feeding trial was designed to assess the effect of super dosing of phytase in corn-soya-based diets of broiler chicken. One hundred and sixty-eight day-old broilers were selected and randomly allocated to four dietary treatment groups, with 6 replicates having 7 chicks per treatment group. Two-phased diets were used. The starter and finisher diet was fed from 0 to 3 weeks and 4 to 5 weeks of age respectively. The dietary treatments were consisted of normal phosphorus (NP) group without any phytase enzyme (4.5 g/kg available/non-phytin phosphorus (P) during starter and 4.0 g/kg during finisher phase), three low-phosphorus (LP) groups (3.2 g/kg available/non-phytin P during starter and 2.8 g/kg during finisher phase) supplemented with phytase at 500, 2500, 5000 FTU/kg diet, respectively, to full fill their phosphorus requirements. The results showed that super doses of phytase (at 2500 FTU and 5000 FTU/kg) on low-phosphorus diet improved feed intake, body weight gain, ileal digestibility (serine, aspartic acid, calcium, phosphorus), blood P levels and bone minerals such as calcium (Ca), P, magnesium (Mg) and zinc (Zn) content. It could be concluded that super doses of phytase in low-phosphorus diet were beneficial than the normal standard dose (at 500 FTU/kg) of phytase in diet of broiler chicken.

  16. Development of a Rapid Immunochromatographic Lateral Flow Device Capable of Differentiating Phytase Expressed from Recombinant Aspergillus niger phyA2 and Genetically Modified Corn.

    Science.gov (United States)

    Zhou, Xiaojin; Hui, Elizabeth; Yu, Xiao-Lin; Lin, Zhen; Pu, Ling-Kui; Tu, Zhiguan; Zhang, Jun; Liu, Qi; Zheng, Jian; Zhang, Juan

    2015-05-06

    Phytase is a phosphohydrolase considered highly specific for the degradation of phytate to release bound phosphorus for animal consumption and aid in the reduction of environmental nutrient loading. New sources of phytase have been sought that are economically and efficiently productive including the construction of genetically modified (GM) phytase products designed to bypass the costs associated with feed processing. Four monoclonal antibodies (EH10a, FA7, AF9a, and CC1) raised against recombinant Aspergillus niger phyA2 were used to develop a highly specific and sensitive immunochromatographic lateral flow device for rapid detection of transgenic phytase, such as in GM corn. Antibodies sequentially paired and tested along lateral flow strips showed that the EH10a-FA7 antibody pair was able to detect the recombinant yeast-phytase at 5 ng/mL, whereas the AF9a-CC1 antibody pair to GM phytase corn was able to detect at 2 ng/mL. Concurrent to this development, evidence was revealed which suggests that antibody binding sites may be glycosylated.

  17. Effect of Wheat Middlings, Microbial Phytase, and Citric Acid on Phytate-Phosphorus, Calcium, and Protein Utilization of Broilers

    Institute of Scientific and Technical Information of China (English)

    GONG Yi-feng; LIAO He-rong; WANG Jin-fu; LI Hong-yan

    2006-01-01

    A corn-soybean meal diet (CSB) (or Diet 1) containing 23% crude protein (CP) was used as the positive control, and another corn-soybean meal diet containing 21% CP and 15% wheat middlings (WM) (or Diet 2) was used as the basal diet, which was treated with four different treatments. Digestibility experiment was employed to discuss the collective effect of citric acid, and intrinsic and microbial phytase. By comparing and analyzing effects of them in the low-nutrient broiler diets, the results showed five treatments had similar effects on Tibia ash (%) (mg) (P > 0.05). Under the supplementation of bacterial phytase or citric acid, the daily body weight gain (ADG), gain:feed (G:F) ratio, and calcium (Ca) utilization were similar to that of standard-nutrient CSB diet (Diet 1) (P > 0.05). And, fecal phosphorus (P) and CP utilization were lower than (P < 0.05) that of Diet 1. But P utilization was significantly higher than (P < 0.01) that of Diet 1.However, the ADG, G:F, and CP utilization produced by supplementation of intrinsic phytase were lower than those of Diet 1, but other aspects were similar to those produced by Diet 1 (P > 0.05). In Diet 5, citric acid, intrinsic and bacterial phytase were added to the diet, which produced a 1.4% decrease on fecal P, a 7.2% increase on Ca utilization, which was significantly higher than (P < 0.01) those of the other four Diets, a 3.9% increase on G:F, which was similar to that of Diet 1, and a 2.3% increase on CP utilization, which was higher than (P < 0.05) that of the other three diets. In summary, the results of this study indicated that citric acid, intrinsic and bacterial phytase might have some additive or synergistic effects, and low-nutrient CSB diets with 15% wheat middlings, 750 U kg-1 phytase, and 3% citric acid might substitute completely for standard CSB in broilers.

  18. The potential of sugar cane juice as the liquid supplement and phytase enzyme carrier for poultry by in vitro

    Directory of Open Access Journals (Sweden)

    Ermin Widjaja

    2011-12-01

    Full Text Available Most of the components of poultry feed (80% of grains and meal that contains phytic acid which has anti-nutritional factor because it can bind minerals and reduce its availability. Phytic acid can be hydrolyzed by the enzyme phytase. Phytase enzyme naturally found in sugar cane juice, but its use as poultry feed supplements have not been done. The study was conducted using sugar cane juice PS 851 from Jatiroto PTPN XI, Lumajang, East Java in order to get the information potential of sugar cane juice as a liquid supplement and phytase enzyme carrier for poultry viewed from the aspect of nutrient content of sugarcane juice and phytase activity in the release rate of phosphorus. Research conducted at the Faculty of Animal IPB for 10 months. The rate of hydrolysis of phytase on P was tested using rice bran as a substrate. Sugar cane juice is added to the 2.5% level, using 4-level incubation (1, 2, 3 and 4 hours, each level consisting of 37°C and 42°C; pH 2; pH 4.5 and pH 5 with three replications. Study using a Two Factors Experiments in Completely Randomized Design and it was continued by DMRT test. P release rate was measured by spectrophotometry. The results showed that the sugar cane juice has a phytase activity of 0.0766 U / ml, brix level of 22.15%, containing water 73.03%, protein 0.47%, crude fiber 6.43%, minerals Ca 0.03%, P 0,02%, Co 0.14 mg / l, Fe 1.8 mg/l, Mn 1.55 mg/l, Zn 1.37 mg/ l, Cu 0.19 mg/ l, Se 12.63 mcg/100 g, vitamins B3 5.26 mg/100 g, C 0.72 mg/100 g, E 0.08 mg/100 g, sucrose 32.42%, fructose 2.41%, galactose 2% and glucose 1.58%. Supplementation of 2.5% sugar cane juice can increase the P release rate of 112-235% at optimum conditions of pH 5, at 37°C with a long incubation period of 1-4 hours.

  19. Escherichia coli phytase improves growth performance of starter, grower, and finisher pigs fed phosphorus-deficient diets.

    Science.gov (United States)

    Jendza, J A; Dilger, R N; Adedokun, S A; Sands, J S; Adeola, O

    2005-08-01

    Corn-soybean meal-based diets, consisting of a high-P control (HPC) containing supplemental dicalcium phosphate (DCP), a basal diet containing no DCP, and the basal diet plus Escherichia coli phytase at 500 or 1,000 phytase units per kilogram (FTU/kg; as-fed basis) were fed to evaluate growth performance in starter, grower, and finisher pigs. Pigs were blocked by weight and gender, such that average weight across treatments was similar, with equal numbers of barrows and gilts receiving each treatment in each block. In Exp. 1, 48 pigs with an average initial BW of 11 kg, housed individually, with 12 pens per diet, were used to evaluate growth performance over 3 wk. Overall ADG and G:F were increased linearly (P < 0.05) by dietary phytase addition. Final BW and plasma P concentrations at 3 wk also increased linearly (P < 0.05). In Exp. 2, 128 pigs with an average initial BW of 23 kg, housed four pigs per pen, with eight pens per diet, were used to evaluate growth performance over 6 wk. A linear increase in response to phytase was noted for ADG and G:F in all three 2-wk periods, as well as overall (P < 0.05). Percentage of bone ash also showed a linear increase (P < 0.01). In Exp. 3, 160 pigs (53 kg), housed five pigs per pen, with eight pens per diet, were used to evaluate growth performance over 6 wk. A linear increase was detected for final BW, as well as ADG and G:F in the first and second 2-wk periods, and overall (P < 0.01). Twenty-four 15-kg individually housed pigs were used to evaluate total-tract nutrient digestibility in Exp. 4. Daily absorption of P linearly increased (P < 0.05) with phytase supplementation. Results of this research indicate that E. coli phytase is effective in liberating phytate P for uptake and utilization by starter, grower, and finisher pigs.

  20. Growth responses of native chicken Sentul G-3 on diet containing high rice-bran supplemented with phytase enzyme and ZnO

    Directory of Open Access Journals (Sweden)

    Cecep Hidayat

    2014-10-01

    Full Text Available This study was conducted to determine the effect of phytase enzymes and ZnO supplementation on the performance of native chicken Sentul-G3 fed high rice-bran diet. Two hundred and seventy day old chicks (DOC native chicken Sentul-G3 from three different hatcheries were used in this study. Factorial randomized block design (3 x 3 was applied in this study. The first factor was the enzyme phytase supplementation levels (0; 1000; 2000 U/kg, the second factor was the level of supplementation of ZnO (0; 1.5; 3.2 g/kg, so that there are nine treatment given, namely R1 = 50% commercial diet : 50% rice bran; R2 = R1 + 1.5 g ZnO/kg; R3 = R1 + 3.2 g ZnO/kg; R4 = R1 + phytase enzyme 1000 U/kg; R5 = R1 + (phytase enzyme 1000 U/kg + 1.5 g ZnO/kg; R6 = R1 + (phytase enzyme 1000 U/kg + 3.2 g ZnO/kg; R7 = R1 + phytase enzyme 2000 U/kg; R8 = R1 + (phytase enzyme 2000 U/kg + 1.5 g ZnO/kg; R9 = R1 + (phytase enzyme 2000 U/kg + 3.2 g ZnO/kg. Each experimental unit consisted of 6 head unsexed native chicken Sentul-G3. The experimental diet was fed for 10 weeks. The variables measured were body weight, body weight gain, feed intake, feed conversion ratio, mortality, mineral deposition of Ca, P, Zn in the tibia bone, alkaline phosfatase enzyme activity in serum. Results showed that there was significant interaction (P 0.05 between phytase enzyme and ZnO supplementation on feed intake, mortality, alkaline phosphatase enzyme activity in serum, and deposition of calcium and phosphorus in the tibia bone. It was concluded that supplementation of phytase enzyme and ZnO were not able to increase the growth of native chicken Sentul-G3 on fed diet containing high rice bran.

  1. Strategies to improve the nutritive value of rice bran in poultry diets. III. The addition of inorganic phosphorus and a phytase to duck diets.

    Science.gov (United States)

    Farrell, D J; Martin, E A

    1998-12-01

    1. In the first of 2 experiments ducklings grown from 2 to 19 d were given diets with 0, 200 or 400 g rice bran, with or without a phytase and with 1 or 3 g inorganic phosphorus (Pi) per kg for rice bran-based diets only. In the 2nd experiment rice bran concentrations were 0, 300 or 600 g rice bran per kg with or without a phytase and 1 g Pi/kg. Ducks were grown from 19 to 40 d of age. 2. In experiment 1, a response to phytase was observed for weight gain and food intake on most diets except those with 200 g rice bran (3 g Pi) and 4.00 g rice bran (1 g P)i/kg. Main effects showed that 400 g rice bran depressed growth rate and food conversion ratio (FCR); increasing Pi depressed food intake, while food phytase increased food intake and growth rate over 2 to 19 d. There were several interactions. Dry matter and P retention were reduced but N digestibility improved when rice bran was increased from 200 g to 400 g/kg at 2 to 10 d of age; apparent metabolisable energy (AME) and calcium retentions were improved, similar results being seen at 10 to 19 d of age. Calcium and P retentions increased with the addition of food phytase and, at 10 to 19 d of age, phytase increased dry matter digestibility. Increasing Pi improved calcium and P retention, but only at 2 to 10 d of age. 3. Tibia ash (g or g/kg) content of bone was lowest on the diet without rice bran and without phytase; Pi concentration had no effect but phytase increased tibia ash on diets with 0 and 200 g rice bran and 1 g Pi/kg. Retention of several minerals in tibia ash declined at the highest rice bran inclusion rate; Pi level and phytase both increased Mg retention. 4. In experiment 2, food intake and growth rate of ducks, but not FCR, declined as rice bran inclusion increased from 0 to 600 g/kg. Phytase improved growth rate but not food intake and FCR on all 3 diets. Dry matter digestibility declined with increasing rice bran inclusion, but AME increased; retention of P and Mg declined but those of Ca and Fe

  2. Isolation of a thermostable acid phytase from Aspergillus niger UFV-1 with strong proteolysis resistance

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    Paulo S. Monteiro

    2015-03-01

    Full Text Available An Aspergillus niger UFV-1 phytase was characterized and made available for industrial application. The enzyme was purified via ultrafiltration followed by acid precipitation, ion exchange and gel filtration chromatography. This protein exhibited a molecular mass of 161 kDa in gel filtration and 81 kDa in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE, indicating that it may be a dimer. It presented an optimum temperature of 60 °C and optimum pH of 2.0. The KM for sodium phytate hydrolysis was 30.9 mM, while the kcat and kcat/KM were 1.46 ×105 s−1 and 4.7 × 106s−1.M−1, respectively. The purified phytase exhibited broad specificity on a range of phosphorylated compounds, presenting activity on sodium phytate, p-NPP, 2- naphthylphosphate, 1- naphthylphosphate, ATP, phenyl-phosphate, glucose-6-phosphate, calcium phytate and other substrates. Enzymatic activity was slightly inhibited by Mg2+, Cd2+, K+ and Ca2+, and it was drastically inhibited by F−. The enzyme displayed high thermostability, retaining more than 90% activity at 60 °C during 120 h and displayed a t1/2 of 94.5 h and 6.2 h at 70 °C and 80 °C, respectively. The enzyme demonstrated strong resistance toward pepsin and trypsin, and it retained more than 90% residual activity for both enzymes after 1 h treatment. Additionally, the enzyme efficiently hydrolyzed phytate in livestock feed, liberating 15.3 μmol phosphate/mL after 2.5 h of treatment.

  3. Isolation of a thermostable acid phytase from Aspergillus niger UFV-1 with strong proteolysis resistance.

    Science.gov (United States)

    Monteiro, Paulo S; Guimarães, Valéria M; de Melo, Ricardo R; de Rezende, Sebastião T

    2015-03-01

    An Aspergillus niger UFV-1 phytase was characterized and made available for industrial application. The enzyme was purified via ultrafiltration followed by acid precipitation, ion exchange and gel filtration chromatography. This protein exhibited a molecular mass of 161 kDa in gel filtration and 81 kDa in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), indicating that it may be a dimer. It presented an optimum temperature of 60 °C and optimum pH of 2.0. The K M for sodium phytate hydrolysis was 30.9 mM, while the k cat and k cat / K M were 1.46 ×10 (5) s (-1) and 4.7 × 10 (6) s (-1) .M (-1) , respectively. The purified phytase exhibited broad specificity on a range of phosphorylated compounds, presenting activity on sodium phytate, p-NPP, 2- naphthylphosphate, 1- naphthylphosphate, ATP, phenyl-phosphate, glucose-6-phosphate, calcium phytate and other substrates. Enzymatic activity was slightly inhibited by Mg (2+) , Cd (2+) , K (+) and Ca (2+) , and it was drastically inhibited by F (-) . The enzyme displayed high thermostability, retaining more than 90% activity at 60 °C during 120 h and displayed a t 1/2 of 94.5 h and 6.2 h at 70 °C and 80 °C, respectively. The enzyme demonstrated strong resistance toward pepsin and trypsin, and it retained more than 90% residual activity for both enzymes after 1 h treatment. Additionally, the enzyme efficiently hydrolyzed phytate in livestock feed, liberating 15.3 μmol phosphate/mL after 2.5 h of treatment.

  4. Effects of phytase on growth and slaughter performance, digestibility and nitrogen and mineral balance in heavy pigs

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    Gianluca Galassi

    2012-10-01

    Full Text Available The study aimed at evaluating the effects of phytase (Ronozyme® NP to the diet of fattening pigs. In comparison with a control diet (C which included 0.5% calcium salt (CaHPO4, two diets without CaHPO4 were tested: 150Phy and 300Phy, which contained 150 and 300 mg phytase/kg, respectively. Sixty barrows of about 47 kg body weight (BW each were randomly divided into 3 groups of 20 animals each, every group split into 5 pens. Average daily gain (ADG, feed intake and feed conversion ratio (FCR were registered. The trial was divided into 2 phases: 0 to 68 days (47 to 100 kg BW and 69 to116 days (100 to 140 kg BW. During phase II, 6 pigs/diet were housed in individual metabolic cages for the digestibility-balance trial. Addition of phytase resulted in higher ADG (723, 783 and 772 g/d for C, 150Phy and 300Phy, respectively and improved FCR (2.84, 2.64 and 2.65 in phase I. No differences were registered in phase II, though a better FCR was overall observed. All slaughter parameters and the total tract apparent digestibility (TTAD of nutrients – except for calcium (Ca and sodium (Na –, were similar among treatments. Likewise, total nitrogen (N excretion did not differ across treatments. Phosphorous (P excreted for C, 150Phy and 300Phy was 47.1, 53.3 and 52.5% of the PI. Trace-minerals showed similar retentions among diets. Phytase effectively improved growth performance until 100 kg BW at both inclusion levels. Yet, it did not improve growth performance, digestibility, N and P balance in heavier animals.

  5. Phytase production by Aspergillus niger CFR 335 and Aspergillus ficuum SGA 01 through submerged and solid-state fermentation.

    Science.gov (United States)

    Shivanna, Gunashree B; Venkateswaran, Govindarajulu

    2014-01-01

    Fermentation is one of the industrially important processes for the development of microbial metabolites that has immense applications in various fields. This has prompted to employ fermentation as a major technique in the production of phytase from microbial source. In this study, a comparison was made between submerged (SmF) and solid-state fermentations (SSF) for the production of phytase from Aspergillus niger CFR 335 and Aspergillus ficuum SGA 01. It was found that both the fungi were capable of producing maximum phytase on 5th day of incubation in both submerged and solid-state fermentation media. Aspergillus niger CFR 335 and A. ficuum produced a maximum of 60.6 U/gds and 38 U/gds of the enzyme, respectively, in wheat bran solid substrate medium. Enhancement in the enzyme level (76 and 50.7 U/gds) was found when grown in a combined solid substrate medium comprising wheat bran, rice bran, and groundnut cake in the ratio of 2 : 1 : 1. A maximum of 9.6 and 8.2 U/mL of enzyme activity was observed in SmF by A. niger CFR 335 and A.ficuum, respectively, when grown in potato dextrose broth.

  6. Effect of microbial phytase on phytate P degradation and apparent digestibility of total P and Ca throughout the gastrointestinal tract of the growing pig.

    Science.gov (United States)

    Rutherfurd, S M; Chung, T K; Moughan, P J

    2014-01-01

    The effect of a dietary microbial phytase on mineral digestibility throughout the gastrointestinal tract (GIT) of the growing pig was studied. Thirty-two entire male pigs (~22 kg BW) were allocated equally to 4 corn-soybean meal diets. One diet was adequate in total P and Ca, the second diet was deficient in total P and Ca (low-P diet), and the third and fourth diets were the low-P diet with microbial phytase added at 1,107 U/kg or 2,215 U/kg, respectively. Titanium dioxide (3 g/kg) was included in the diets as an indigestible marker. The pigs were fed their respective diets for 42 d. Fecal samples were collected from d 38 to 41 and stomach chyme, terminal jejunal, and terminal ileal digesta samples were collected after euthanasia on d 42. Phytate P degradability and apparent total P and Ca digestibility were determined at the gastric, jejunal, ileal, and total tract levels. Phytate P degradation and apparent total P and Ca digestibility were not significantly different between the 2 microbial phytase inclusion levels. Across both microbial phytase-low-P diets, phytase supplementation increased (P phytase-supplemented, low-P (26% units greater) diets. The latter result was not reflected by the apparent total P digestibility estimate, which was not significantly different between the ileal and total tract levels for both the unsupplemented and phytase-supplemented, low-P diets. Consequently, there appeared to be considerable phytate degradation in the hindgut, presumably due to the action of hindgut microbes, but the phytate P released in the hindgut did not appear to be absorbed. Apparent Ca digestibility was not significantly different among jejunal, ileal, and total tract levels for any of the dietary treatments, but apparent Ca digestibility was greater (P phytase-supplemented diets compared with the unsupplemented diets. Overall, dietary microbial phytase supplementation led to greater phytate degradation to the end of the jejunum. Because the phytate P

  7. Effect of Phytase Superdoses and Citric Acid on Growth Performance, Plasma Phosphorus and Tibia Ash in Broilers Fed Canola Meal-Based Diets Severely Limited in Available Phosphorus

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    Taheri HR

    2015-06-01

    Full Text Available This experiment was conducted to investigate the effect of phytase superdoses alone or in combination with citric acid (CA in canola meal-based diets severely limited in available phosphorus (Pa on growth performance, plasma phosphorus (P, and tibia ash (TA in broilers from 22 to 42 d of age. Two hundreds and eighty 21-d-old male broilers were used in 28 pens of 10 birds per each. The experimental diets consisted of a positive control (PC diet and six negative control (NC diets which consisted of two levels of CA (0 and 20 g/Kg and three levels of phytase (0, 1000 and 4000 U/Kg in a 2 × 3 factorial arrangement. The PC diet contained 4.3 g/Kg Pa, but all NC diets contained 1.5 g/Kg Pa. Results indicated that the birds fed the PC diet had a significantly higher average daily gain (ADG, plasma P and TA, but a lower feed conversion ratio (FCR than those fed the NC diet. The ADG, FCR and plasma P values in birds fed NC diets supplemented with 4000 U/Kg phytase enzyme (with or without CA significantly reached those of birds fed the PC diet. But, addition of phytase enzyme at 1000 U/Kg only plus CA to the NC diet could significantly improve FCR and plasma P. A significant interaction was observed between phytase and CA for FCR and plasma P. Although TA values in NC + 1000 U/Kg phytase treatments (with or without CA were similar to the PC treatment, TA values of NC + 4000 U/Kg phytase treatments (with or without CA was greater than that of the PC treatment. Results of this study showed that, in severely limited Pa corn-canola meal-based diets, supplementing 4000 U/Kg phytase or also 1000 U/Kg phytase plus CA will be sufficient to obtain the comparable feed efficiency in broilers to those fed the adequate Pa diet.

  8. Influence of high dose of phytase and an emulsifier on performance, apparent metabolisable energy and nitrogen retention in broilers fed on diets containing soy oil or tallow.

    Science.gov (United States)

    Zaefarian, F; Romero, L F; Ravindran, V

    2015-01-01

    The effects of high dose of microbial phytase and an emulsifier on the performance, apparent metabolisable energy (AME) and nitrogen (N) retention in broilers fed on diets containing different fat sources were examined in a 5-week trial. Two fat sources (soy oil and tallow), two inclusion levels of E. coli phytase (500 or 1000 phytase units (FTU)/kg diet) and two inclusion levels of lysolecithin emulsifier (0 or 3.5 g/kg of diet) were evaluated in a 2 × 2 × 2 factorial arrangement of treatments. Throughout the 5-week trial, soy oil supplementation improved weight gain and feed per gain compared with tallow, but had no effect on feed intake. The high dose of phytase increased the weight gain and feed intake and lowered the feed per gain during d 1-21, but had no effect on performance parameters over the whole trial period. An effect of emulsifier was observed for feed intake during d 1-21 and over the whole trial period. Addition of emulsifier increased feed intake compared with diets without emulsifier. During weeks 1, 2, 3 and 5, birds fed on soy oil-based diets had higher nitrogen-corrected AME (AMEN) compared with those fed on tallow-based diets. During weeks 2, 3 and 5, the effect of phytase was significant for AMEN, with the high dose increasing the AMEN. During week 2, AMEN was increased with emulsifier addition. During weeks 1, 2, 3 and 5, birds fed on soy oil-based diets had higher fat retention compared with those fed on tallow-based diets. The high dose of phytase improved the retention of fat during week 5 and the addition of emulsifier resulted in higher fat retention during week 1. During weeks 2, 3 and 5, an interaction between fat source × phytase × emulsifier was observed for N retention. In soy oil-based diets, emulsifier plus 1000 FTU/kg phytase increased N retention compared with other groups, while in tallow-based diets, emulsifier addition increased N retention in diets with 500 FTU/kg, but not in 1000 FTU/kg diet. Overall, the

  9. Efficacy of New 6-Phytase from Buttiauxella spp. on Growth Performance and Nutrient Retention in Broiler Chickens Fed Corn Soybean Meal-based Diets.

    Science.gov (United States)

    Kiarie, E; Woyengo, T; Nyachoti, C M

    2015-10-01

    A total of 420 day-old male Ross chicks were weighed at d 1 of life and assigned to test diets to assess the efficacy of a new Buttiauxella spp. phytase expressed in Trichoderma reesei. Diets were: positive control (PC) adequate in nutrients and negative control (NC) diet (40% and 17% less available phosphorous (P) and calcium (Ca), respectively) supplemented with 6 levels of phytase 0, 250, 500, 750, 1,000, and 2,000 phytase units (FTU)/kg of diet. All diets had titanium dioxide as digestibility marker and each diet was allocated to ten cages (6 birds/cage). Diets were fed for 3 wk to measure growth performance, apparent retention (AR) on d 17 to 21 and bone ash and ileal digestibility (AID) on d 22. Growth performance and nutrient utilization was lower (pPhytase response in NC birds was linear (pphytase at ≥750 FTU resulted in AID of total AA commensurate to that of PC fed birds and at ≥1,000 FTU improved (pphytase and PC diet. In conclusion, the result from this study showed that in addition to increased P and Ca utilization, the new Buttiauxella phytase enhanced growth performance and utilization of other nutrients in broiler chickens in a dose-dependent manner.

  10. The supplementation of low-P diets with microbial 6-phytase expressed in Aspergillus oryzae increases P and Ca digestibility in growing pigs.

    Science.gov (United States)

    Torrallardona, D; Salvadó, R; Broz, J

    2012-12-01

    A trial was conducted to evaluate the dose response of a novel microbial 6-phytase expressed in Aspergillus oryzae (Ronozyme HiPhos; DSM Nutritional Products, Basel, Switzerland) in pigs. Forty-eight individually housed pigs (Landrace × Pietrain; 52 kg BW; 24 males and 24 females) were distributed among 6 experimental treatments consisting of a low-P diet (3.5 g P/kg; 1.1 g digestible P/kg), which was supplemented with 500, 1000, 2000, or 4000 units of phytase activity/kg, and a standard-P diet (4.5 g P/kg; 1.8 g digestible P/kg) that was supplemented with CaHPO(4). After 17 d, fresh feces were sampled from all pigs and the apparent total tract digestibility of DM, OM, ash, P, and Ca was measured using TiO(2) as indigestible marker. Blood samples were also obtained from each pig and serum was analyzed for P and Ca concentrations. The nonsupplemented low-P diet increased Ca and reduced P blood serum concentrations (P Phytase supplementation of the low-P diet reduced Ca (from 10.8 to 9.9 mg/dL; linear, P Phytase improved the total tract digestibility of P (from 29.0 to 62.3%; linear and quadratic, P phytase tested improves the apparent total tract digestibility of P in growing pigs and reduces P excretion in feces in a dose-dependent manner.

  11. Phytase production by solid-state fermentation of groundnut oil cake by Aspergillus niger: A bioprocess optimization study for animal feedstock applications.

    Science.gov (United States)

    Buddhiwant, Priyanka; Bhavsar, Kavita; Kumar, V Ravi; Khire, Jayant M

    2016-08-17

    This investigation deals with the use of agro-industrial waste, namely groundnut oil cake (GOC), for phytase production by the fungi Aspergillus niger NCIM 563. Plackett-Burman design (PBD) was used to evaluate the effect of 11 process variables and studies here showed that phytase production was significantly influenced by glucose, dextrin, distilled water, and MgSO4 · 7H2O. The use of response surface methodology (RSM) by Box-Behnken design (BBD) of experiments further enhanced the production by a remarkable 36.67-fold from the original finding of 15 IU/gds (grams of dry substrate) to 550 IU/gds. This is the highest solid-state fermentation (SSF) phytase production reported when compared to other microorganisms and in fact betters the best known by a factor of 2. Experiments carried out using dried fermented koji for phosphorus and mineral release and also thermal stability have shown the phytase to be as efficient as the liquid enzyme extract. Also, the enzyme, while exhibiting optimal activity under acidic conditions, was found to have significant activity in a broad range of pH values (1.5-6.5). The studies suggest the suitability of the koji supplemented with phytase produced in an SSF process by the "generally regarded as safe" (GRAS) microorganism A. niger as a cost-effective value-added livestock feed when compared to that obtained by submerged fermentation (SmF).

  12. Influence of dietary non-phytate phosphorous levels and phytase supplementation on the performance and bone characteristics of broilers

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    A Karimi

    2011-03-01

    Full Text Available Precise phosphorus nutrition is important for significant reductions in both P pollution and ration costs. The influence of different levels (% of dietary nPP fed from 0 to 20 d (0.45, 0.40, 0.35, 0.30, 0.25, compared with feeding 0.20 nPP with and without 500 F.T.U. of phytase per kg of diet and from 21 to 36 d of age (0.414, 0.364, 0.314, 0.264, 0.214, compared with 0.164 nPP with and without 500 F.T.U. of phytase per kg of diet were evaluate using a total of 588 day-old commercial broiler chicks. Each treatment was replicated four times in a completely randomized design. Body weight (BW, daily gain (DG, feed intake (DFI, feed conversion ratio, plasma P level and bone characteristics were determined, and from these data the P equivalency of the phytase was estimated. Feeding diets containing less than 0.40% of nPP to birds between 0 and 20d resulted in inferior BW, DFI, plasma P level and bone characteristics compared with the control. However, optimum FCR and mortality was supported at lower levels of nPP (0.25%. Between 21 and 36 d, 0.364% was enough to optimise BW, DFI, and femur ash (%; whilst only 0.314% or greater was needed to support optimum DG, toe and tibia ash and only 0.214 to 0.264% was required to optimise shank, femur and tibia lengths; FCR and survivability. A broken line analysis also showed that the nPP (% requirement ranged from 0.267 to 0.410 and 0.272 to 0.380% during 0 to 20 and 21 to 36 d, respectively. Phytase supplementation improved performance and bone criterion and its P equivalency, depending upon the response of interest and birds age, ranged from 0.00 to 0.110%. In conclusion, the results showed that the combination of a lower level of nPP and phytase may be used to increase dietary P utilization, without severe changes in performance and bone quality.

  13. The oxygen isotope composition of phosphate released from phytic acid by the activity of wheat and Aspergillus niger phytase

    Science.gov (United States)

    von Sperber, C.; Tamburini, F.; Brunner, B.; Bernasconi, S. M.; Frossard, E.

    2015-07-01

    Phosphorus (P) is an essential nutrient for living organisms. Under P-limiting conditions plants and microorganisms can exude extracellular phosphatases that release inorganic phosphate (Pi) from organic phosphorus compounds (Porg). Phytic acid (myo-inositol hexakisphosphate, IP6) is an important form of Porg in many soils. The enzymatic hydrolysis of IP6 by phytase yields available Pi and less phosphorylated inositol derivates as products. The hydrolysis of organic P compounds by phosphatases leaves an isotopic imprint on the oxygen isotope composition (δ18O) of released Pi, which might be used to trace P in the environment. This study aims at determining the effect of phytase on the oxygen isotope composition of released Pi. For this purpose, enzymatic assays with histidine acid phytases from wheat and Aspergillus niger were prepared using IP6, adenosine 5'-monophosphate (AMP) and glycerophosphate (GPO4) as substrates. For a comparison to the δ18O of Pi released by other extracellular enzymes, enzymatic assays with acid phosphatases from potato and wheat germ with IP6 as a substrate were prepared. During the hydrolysis of IP6 by phytase, four of the six Pi were released, and one oxygen atom from water was incorporated into each Pi. This incorporation of oxygen from water into Pi was subject to an apparent inverse isotopic fractionation (ϵ ~ 6 to 10 ‰), which was similar to that imparted by acid phosphatase from potato during the hydrolysis of IP6 (ϵ ~ 7 ‰), where less than three Pi were released. The incorporation of oxygen from water into Pi during the hydrolysis of AMP and GPO4 by phytase yielded a normal isotopic fractionation (ϵ ~ -12 ‰), similar to values reported for acid phosphatases from potato and wheat germ. We attribute this similarity in ϵ to the same amino acid sequence motif (RHGXRXP) at the active site of these enzymes, which leads to similar reaction mechanisms. We suggest that the striking

  14. The oxygen isotope composition of phosphate released from phytic acid by the activity of wheat and Aspergillus niger phytase

    Directory of Open Access Journals (Sweden)

    C. v. Sperber

    2015-03-01

    Full Text Available Phosphorus (P is an essential nutrient for living organisms. Under P-limiting conditions plants and microorganisms can exude extracellular phosphatases that release inorganic phosphate (Pi from organic phosphorus compounds (Porg. Phytic acid (IP6 is an important form of Porg in many soils. The enzymatic hydrolysis of IP6 by phytase yields plant available inorganic phosphate (Pi and less phosphorylated inositol derivates as products. The hydrolysis of organic P-compounds by phosphatases leaves an isotopic imprint on the oxygen isotope composition (δ18O of released Pi, which might be used to trace P in the environment. This study aims at determining the effect of phytase on the oxygen isotope composition of released Pi. For this purpose, enzymatic assays with histidine acid phytases from wheat and Aspergillus niger were prepared using IP6, adenosine 5'monophosphate (AMP and glycerophosphate (GPO4 as substrates. For a comparison to the δ18O of Pi released by other extracellular enzymes, enzymatic assays with acid phosphatases from potato and wheat germ with IP6 as substrate were prepared. During the hydrolysis of IP6 by phytase, four Pi are released, and one oxygen atom from water is incorporated into each Pi. This incorporation of oxygen from water into Pi is subject to an apparent inverse isotopic fractionation (ϵ ∼ 6 to 10‰, which is similar to that imparted by acid phosphatase from potato during the hydrolysis of IP6 (ϵ ∼ 7‰ where less than three Pi are released. The incorporation of oxygen from water into Pi during the hydrolysis of AMP and GPO4 by phytase yielded a normal isotopic fractionation (ϵ ∼ −12‰, again similar to values reported for acid phosphatases from potato and wheat germ. We attribute this similarity in ε to the same amino acid sequence motif (RHGXRXP at the active site of these enzymes, which leads to similar reaction mechanisms. We suggest that the striking substrate

  15. N-糖基化对植酸酶phy(QF)酶学性质的影响%The Effects of N-glycosylation on Enzymological Properties of Phytase Phy(QF)

    Institute of Scientific and Technical Information of China (English)

    王茜; 宋玛丽; 杜军; 付月君; 胡风云; 梁爱华

    2015-01-01

    为探讨N-糖基化对植酸酶phy( QF )酶学性质的影响,将植酸酶基因phy( QF )在毕赤酵母中进行了表达,利用脱糖基化酶Endo Hf去除重组植酸酶phy(QF)的糖链,对去除糖链前后的植酸酶phy(QF)进行了纯化和酶学性质研究与比较。结果表明,糖基化的植酸酶phy(QF)分子量约为53 kDa,酶活为3201 U/mg,Km =392μmol/L,Vmax =3267 U/mg;糖基化的植酸酶phy( QF )最适pH值为4.5,最适反应温度为55℃,与去糖基化的植酸酶phy( QF )的最适pH值无明显区别,比去糖基化的植酸酶phy(QF)最适温度提高了5℃,Tm值提高约2℃。经70℃孵育10 min后,去糖基化的植酸酶phy(QF)完全失活,而糖基化的植酸酶phy(QF)仍残留13%的活性,说明N-糖基化作为一种重要的翻译后修饰,对植酸酶phy( QF )的稳定性有促进作用。%In order to study the effects of N-glycosylation on enzymological properties of phytase phy (QF),the recombinant phytase gene phy(QF) was expressed in pichia pastoris GS115.Phytase phy(QF) was deglycosylated by endoglycosylase Endo H f .Enzymology properties of glycosylated phy (QF)and deglycosylated phy (QF)were an-alyzed.The results showed that the glycosylated phy (QF) had a specific activity of 3 201 U/mg with an MW of ap-proximately 53 kDa and the Km and Vmax of glycosylated phy(QF) were 392 μmol/L and 3 267 U/mg,respectively. The optimal pH value of glycosylated phy(QF) was 4.5 and was consistented with that of deglycosylated phy (QF). However,the optimal temperature of glycosylated phy (QF) was 55 ℃,5 ℃ higher than that of deglycosylated phy (QF) and 2 ℃increased in the melting temperature (Tm).The glycosylated phy(QF) exhibited 13%residual ac-tivity after treatment at 70 ℃for 10 min and deglycosylated phy (QF)was completely deactivated under the condi-tion.As one of posttranslational modifications ,N-glycosylation may play an important

  16. Expression of Exogenous Gene in Prokaryotes and Eukaryotes

    Institute of Scientific and Technical Information of China (English)

    雷正玉; 张晓

    2014-01-01

    [目的]研究外源基因在原核生物和真核生物中的表达。[方法]将带有目的基因的表达载体 pTYB2-WF转化大肠杆菌 ER2566,用 IPTG诱导植酸酶基因表达。用 SDS-PAGE验证植酸酶融合蛋白表达情况,并进一步对融合蛋白进行纯化。用ç赤酵母表达系统表达植酸酶基因 phyA;构建酵母整合载体pPIC9K-phyA,转化ç赤酵母 GS115,构建工程菌 GS115-pPIC9K-phyA。[结果]在甲醇的诱导下表达植酸酶,经酶活力的测定,其植酸酶活力达7.3μ/ml,构建了ç赤酵母工程菌GS115-pPIC9K-phyA。[结论]甲醇酵母表达机制在分子生物学以及工业应用领域发挥作用。%Objective This study aimed to investigate the expression of exogenous gene in prokaryotes and eukaryotes. [Method] The expression vector pTYB2-WF harboring target gene was transformed into E. coli ER2566. IPTG was employed to induce the expression of phytase gene. The expression of phytase fusion protein was detected by SDS-PAGE, and the fusion protein was further purified. Phytase gene phyA was expressed in Pichia pastoris expression system. Yeast recombinant vector pPIC9K-phyA was constructed and transformed into P. pastoris GS115 to construct engineering strain GS115-pPIC9K-phyA. [Result] Phytase protein was ex-pressed under methanol induction. Enzyme activity assay indicated that the activity of phytase was 7.3 U/ml. P. pastoris engineering strain GS115-pPIC9K-phyA was successful y constructed. [Conclusion] Methanol yeast expression mechanisms play a certain role in molecular biology and industrial applications.

  17. The Research Development of Production and Application of Phytase%植酸酶生产与应用的研究进展

    Institute of Scientific and Technical Information of China (English)

    李晓宇; 陈耀国; 柳志强

    2011-01-01

    The phytase can be used to degrade the phytate in the feedstuffs and release inorganic phosphate. It is important to increase the availability of phosphate, improve the absorption of the mineral nutrition in feedstuffs by monogastric animals and decrease the environmental phosphate pollution. The phytase can be also used as food additives, promote the absorption of minerals in human body. The phytate degradation product IP3 also has broad physiological functions in medical field. In order to embody the research significance and application value of phytase, this study reviewed phytase production from microorganism,transgenic plant and transgenic animals. The problems existed in the production of phytase were analyzed, and the prospect of phytase was also discussed.%植酸酶能降解植物性饲料中的植酸盐类,释放出无机磷酸,对于提高单胃动物饲料中磷的利用率,促进饲料中矿质元素的吸收利用,减轻高磷排泄物对环境的污染有重要作用.植酸酶也可以用作食品添加剂,促进人体对食品中矿质元素的吸收利用;植酸的降解产物IP3在医药领域也具有广泛的生理功能.为了体现植酸酶的研究意义和应用价值,本研究综述了国内外利用微生物、转基因植物和转基因动物生产植酸酶的概况,提出了植酸酶在生产过程中存在的一些问题,并对植酸酶的发展前景进行了展望.

  18. Insights into Broilers' Gut Microbiota Fed with Phosphorus, Calcium, and Phytase Supplemented Diets

    Science.gov (United States)

    Borda-Molina, Daniel; Vital, Marius; Sommerfeld, Vera; Rodehutscord, Markus; Camarinha-Silva, Amélia

    2016-01-01

    Phytase supplementation in broiler diets is a common practice to improve phosphorus (P) availability and to reduce P loss by excretion. An enhanced P availability, and its concomitant supplementation with calcium (Ca), can affect the structure of the microbial community in the digestive tract of broiler chickens. Here, we aim to distinguish the effects of mineral P, Ca, and phytase on the composition of microbial communities present in the content and the mucosa layer of the gastrointestinal tract (GIT) of broiler chickens. Significant differences were observed between digesta and mucosa samples for the GIT sections studied (p = 0.001). The analyses of 56 individual birds showed a high microbial composition variability within the replicates of the same diet. The average similarity within replicates of digesta and mucosa samples across all diets ranged from 29 to 82% in crop, 19–49% in ileum, and 17–39% in caeca. Broilers fed with a diet only supplemented with Ca had the lowest body weight gain and feed conversion values while diets supplemented with P showed the best performance results. An effect of each diet on crop mucosa samples was observed, however, similar results were not obtained from digesta samples. Microbial communities colonizing the ileum mucosa samples were affected by P supplementation. Caeca-derived samples showed the highest microbial diversity when compared to the other GIT sections and the most prominent phylotypes were related to genus Faecalibacterium and Pseudoflavonifractor, known for their influence on gut health and as butyrate producers. Lower microbial diversity in crop digesta was linked to lower growth performance of birds fed with a diet only supplemented with Ca. Each diet affected microbial communities within individual sections, however, no diet showed a comprehensive effect across all GIT sections, which can primarily be attributed to the great variability among replicates. The substantial community differences between digesta

  19. Recovery of phytase produced by solid-state fermentation on citrus peel

    Directory of Open Access Journals (Sweden)

    Daniel Ernesto Rodríguez-Fernández

    2010-12-01

    Full Text Available The extraction of phytase produced by solid-state fermentation of citrus peel was studied employing a multistage leaching process. It was observed that the extracts containing EDTA retained over 90% of phytase activity at room temperature after 24 h after the leaching. A fractional design 2² (with 4 replicates at the central point was carried out for testing the pH and agitation as process independent factors. Only the interaction between the pH and agitation showed a significant influence. These factors were optimized with a central composite design. Agitation at 300 rpm and pH at 5.0 were the best conditions to extract the enzyme from solid matrix. The modeling of the process indicated that diffusivity of the enzyme in the solvent was the controlling mechanism. The corresponding kinetic constant and saturation concentration in this process were 0.89 min-1 and 4.0 IU/mL, respectively. The multistage process indicated that after two steps, it was possible to recover 85% of total enzyme produced.A extração de fitases produzidas por fermentação em estado sólido de polpa cítrica foi estudada utilizando um processo de extração sólido-líquido em varias etapas. A adição de EDTA permite manter durante 24 horas a temperatura ambiente 90% da atividade inicial do caldo com a enzima extraída. Um planejamento fatorial 2², com 4 replicas no ponto central, foi desenvolvido para testar os valores de ph e agitação convenientes para a extração das enzimas. A interação entre ambos os fatores foi estadisticamente significativa. A atividade da enzima foi otimizada nos valores onde o pH (5.0 e a agitação (350 rpm resultaram ser as melhores condições para extrair a enzima da matriz sólida. O ajuste do modelo matemático obtido mostra que é possível considerar a difusividade como o mecanismo que controla o processo de transferência de massa. A constante cinética que descreve este processo e a concentração de saturação foram 0.039 min

  20. Assimilation of phytate-phosphorus by the extracellular phytase activity of tobacco (Nicotiana tabacum) is affected by the availability of soluble phytate

    OpenAIRE

    2006-01-01

    Phytate, the major organic phosphorus in soil, is not readily available to plants as a source of phosphorus (P). It is either complexed with cations or adsorbed to various soil components. The present study was carried out to investigate the extracellular phytase activities of tobacco (Nicotiana tabacum variety GeXin No.1) and its ability to assimilate external phytate-P. Whereas phytase activities in roots, shoots and growth media of P i-fed 14-day-old seedlings were only 1.3-4.9% of total a...

  1. Heat-treatment, phytase and fermented liquid feeding affect the presence of inositol phosphates in ileal digesta and phosphorus digestibility in pigs fed a wheat and barley diet

    DEFF Research Database (Denmark)

    Blaabjerg, Karoline; Jørgensen, Henry; Tauson, A H

    2010-01-01

    The aim was to evaluate the effect of heat-treatment, microbial phytase addition and feeding strategy (dry feeding v. fermented liquid feeding) on degradation of phytate (myo-inositol hexakisphosphate, InsP6) and formation and further degradation of lower inositol phosphates (myo-inositol pentaki......The aim was to evaluate the effect of heat-treatment, microbial phytase addition and feeding strategy (dry feeding v. fermented liquid feeding) on degradation of phytate (myo-inositol hexakisphosphate, InsP6) and formation and further degradation of lower inositol phosphates (myo...

  2. 植酸酶在畜禽养殖中的研究进展%Research Progress on Phytase in Animal Production

    Institute of Scientific and Technical Information of China (English)

    刘永超; 王建平

    2012-01-01

    When adding phytase to animal feed, the phosphorus in animal feed can be used effectively, and reducing the discharge of phosphorus. The article reviewed studies and development on phytase and its application in animal feed.%在饲料中添加植酸酶能够提高饲料利用率,增加动物对饲料中有效磷的利用率,减少磷的排出。文章综述了植酸酶的研究进展及在饲粮中的应用。

  3. Effect of high-dose phytase and citric acid, alone or in combination, on growth performance of broilers given diets severely limited in available phosphorus.

    Science.gov (United States)

    Taheri, H R; Jabbari, Z; Adibnia, S; Shahir, M H; Hosseini, S A

    2015-01-01

    1. Two trials were conducted to evaluate the effect of high-dose phytase alone or in combination with citric acid (CA) in the diet severely limited in available phosphorus (P) on performance, plasma P and plasma Ca of broilers from 22 to 42 d of age. 2. In Trial 1, 297 21-d-old female chicks were placed into 27 pens and allocated to 9 maize-soybean meal-based dietary treatments, which were a positive control [PC, 4.23 g/kg non-phytate P (NPP)] and 8 negative control (NC, 1.35 g/kg NPP) groups consisting of two concentrations of CA (0 and 20 g/kg) and 4 concentrations of phytase (0, 1000, 2000 and 4000 U/kg) in a 2 × 4 factorial arrangement. In Trial 2, 192 21-d-old male chicks were placed into 24 pens and allocated to 6 wheat-canola meal-based dietary treatments, which were a PC (4.2 g/kg NPP), a NC (1.68 g/kg NPP) and 4 NC groups consisting of two concentrations of CA (0 and 20 g/kg) and two concentrations of phytase (2000 and 4000 U/kg) in a 2 × 2 factorial arrangement. 3. In both trials, birds fed on the PC had significantly higher average daily gain (ADG), average daily feed intake (ADFI), plasma P and lower feed conversion ratio (FCR) and plasma Ca than those of birds fed on the NC. CA supplementation significantly increased ADG and ADFI. There was a significant interaction between CA and phytase on plasma P where CA improved the effect of phytase on plasma P. In Trial 1, phytase addition improved ADG, ADFI, FCR and plasma Ca linearly. 4. Briefly, this research showed the interaction effect between CA and phytase on plasma P when broilers were fed on diets based on maize-soybean meal or wheat-canola meal. The results showed that CA supplementation lowered the concentration of phytase that is needed in low NPP diets to increase plasma P.

  4. Influence of superdoses of a novel microbial phytase on growth performance, tibia ash, and gizzard phytate and inositol in young broilers.

    Science.gov (United States)

    Walk, C L; Santos, T T; Bedford, M R

    2014-05-01

    An experiment was conducted to evaluate the influence of a novel microbial phytase on performance, tibia ash, and the content of phytate, phytate esters, and inositol in the gizzard of young broilers. Male Cobb 500 broilers (n = 1,680) were fed 1 of 7 experimental diets: positive control (PC) formulated to meet or exceed nutrient recommendations; PC plus dicalcium phosphate (PC+DCP) formulated to provide Ca and P at 0.10% above the PC; PC plus 500 U/kg of microbial phytase (PC+500); negative control (NC) with Ca and P reduced from the PC by 0.16% and 0.15%, respectively; and the NC plus phytase at 500 (NC+500), 1,000 (NC+1,000), or 1,500 (NC+1,500) U/kg. Diets were fed in crumbled form to 20 birds/pen and 12 replicate pens/diet from d 0 to 21. On d 21, 4 birds/pen were euthanized for collection of right tibias and gizzard digesta for determination of tibia ash and gizzard phytate. In general, broilers fed the NC diet had reduced (P ≤ 0.05) feed intake and BW gain compared with broilers fed diets supplemented with phytase, but not different than the PC or PC+DCP. Phytase supplementation in the NC diet improved (P ≤ 0.05) BW gain comparable with or above that of the PC. Feed conversion ratio was improved in broilers fed the NC+1,000 or NC+1,500 compared with broilers fed all other diets. Tibia ash was reduced (P ≤ 0.05) in broilers fed the NC compared with broilers fed all other diets, and phytase supplementation improved tibia ash comparable with the PC. Phytase supplementation reduced (P ≤ 0.05) phytate (inositol hexa-phosphate) concentration in the gizzard. Inositol concentration in the gizzard was higher (P ≤ 0.05) in birds fed NC+1,000 or NC+1,500 compared with all other diets and this was correlated with growth performance (P ≤ 0.05) rather than tibia ash (P > 0.05). Improvements in feed conversion ratio associated with superdoses of phytase may be attributed to phytate destruction and the provision of inositol.

  5. Effects of an experimental phytase on performance, egg quality, tibia ash content and phosphorus bioavailability in laying hens fed on maize- or barley-based diets.

    Science.gov (United States)

    Francesch, M; Broz, J; Brufau, J

    2005-06-01

    A 24-week performance trial was conducted to evaluate the efficacy of an experimental phytase on performance, egg quality, tibia ash content and phosphorus excretion in laying hens fed on either a maize- or a barley-based diet. At the end of the trial, an ileal absorption assay was conducted in order to determine the influence of phytase supplementation on the apparent absorption of calcium and total phosphorus (P). Each experimental diet was formulated either as a positive control containing 3.2 g/kg non-phytate phosphorus (NPP), with the addition of dicalcium phosphate (DCP), or as a low P one, without DCP addition. Both low P diets (containing 1.3 or 1.1 g/kg NPP) were supplemented with microbial phytase at 0, 150, 300 and 450 U/kg. The birds were housed in cages, allocating two hens per cage as the experimental unit. Each of 10 dietary treatments was assigned to 16 replicates. Low dietary NPP (below 1.3 g/kg) was not able to support optimum performance of hens during the laying cycle (from 22 to 46 weeks of age), either in maize or barley diets. Rate of lay, daily egg mass output, feed consumption, tibia ash percentage and weight gain were reduced in hens fed low NPP diets. The adverse effects of a low P diet were more severe in hens on a maize diet than in those on a barley diet. Low dietary NPP reduced egg production, weight gain, feed consumption and tibia ash content and microbial phytase supplementation improved these parameters. Hens given low NPP diets supplemented with phytase performed as well as the hens on positive control diets containing 3.2 g/kg of NPP. A 49% reduction of excreta P content was achieved by feeding hens on low NPP diets supplemented with phytase, without compromising performance. Phytase addition to low NPP diets increased total phosphorus absorption at the ileal level, from 0.25 to 0.51 in the maize diet and from 0.34 to 0.58 in the barley diet. Phosphorus absorption increased linearly with increasing levels of dietary phytase

  6. Environmental assessment of Ronozyme (R) p5000 CT phytase as an alternative to inorganic phosphate supplementation to pig feed used in intensive pig production

    DEFF Research Database (Denmark)

    Nielsen, Per H.; Wenzel, Henrik

    2007-01-01

    Goal, Scope and Background. Ronozyme (R) P5000 CT is an industrially produced enzyme product (phytase) which is able to degrade naturally occurring phytate in animal feed and release the phytate's content of phosphorus for pig's growth. Ronozyme P5000 CT (hereafter called Ronozyme Phytase) can...... use for enzyme production is small and unimportant unless use of agricultural land is given very large relative weight. Recommendations and Perspectives. Hundreds of enzyme products are commercially available on the market today, each with a range of different applications. There are several...

  7. Carbohydrase and phytase supplementation in diets for semi-heavy laying hens

    Directory of Open Access Journals (Sweden)

    Adriano Geraldo

    2014-07-01

    Full Text Available This study was conducted in order to evaluate the association of phytase with an enzymatic complex comprised of carbohydrases (α-galactosidase, galactomannan, xylanase and β-glucanase in nutrition reduction diets for semi-heavy laying hens and its effect on egg performance and egg quality. Four hundred Isa Brown laying hens with 42 to 57 weeks of age were distributed in an entirely random experiment with five treatments and 8 repetitions, during five production periods of 21 days. Variables studied: egg production, feed intake, mean egg weight, feed conversion, Haugh unit, percentage of yolk, egg white and albumen, yolk color, eggshell thickness and specific gravity. There was a significant interaction (p 0.05 of treatment on production, egg weight or internal and external egg quality. Treatment effects on feed conversion showed better values for hens fed with the control diet. The levels of nutrient reduction used in the diets with or without enzyme supplementation did not provide good results with regard to feed conversion and feed intake. However, they did not affect the other parameters for egg production and internal and external egg quality.

  8. 植酸酶液体发酵条件研究%Optimum conditions for phytase liquid fermentation

    Institute of Scientific and Technical Information of China (English)

    汪世华; 杨燕凌; 胡开辉

    2006-01-01

    对绿色木霉LH374液体发酵生产植酸酶的条件作了研究,确立了所试因素的最佳组合:碳源为5%葡萄糖,氮源为3%胰蛋白胨和0.3%硝酸铵,金属离子为0.003%Fe2++0.003%Mn2+.每500 mL三角瓶装50 mL培养基,初始pH为6.5,接种量为5%.在最适条件下发酵72 h生成植酸酶的最高产量为2510 U·mL-1,平均产量为2490 U·mL-1.%Optimization of the fermentation medium for maximum phytase production was carried out using a new strain, Trichoderma viride LH374, and the test results were listed as follows: carbon source was 5% glucose, nitrogen source were 3% tryptone and 0.3% NH4NO3, and metallic ion concentration were 0.003% Fe2+ and 0.003% Mn2+. The 500 mL flasks were filled with 50 mL media with pH 6.5 to incubate 72 h, and inoculation amount was 5%. Under optimum condition, Trichoderma viride LH374 was able to yield the maximum phytase activity of 2510 U·mL-1 and the average of 2490 U·mL-1 phytase.

  9. Age and adaptation to Ca and P deficiencies: 2. Impacts on amino acid digestibility and phytase efficacy in broilers.

    Science.gov (United States)

    Li, W; Angel, R; Kim, S-W; Jiménez-Moreno, E; Proszkowiec-Weglarz, M; Plumstead, P W

    2015-12-01

    A total of 1,152 straight-run hatchling Heritage 56M×fast feathering Cobb 500F broiler birds were used to determine Ca, age, and adaptation effects on apparent ileal digestibility of crude protein (AID of CP), amino acids (AID of AA) and phytase efficacy. Twelve treatments with 8 replicates, each were fed from 7 to 9 d (6 birds per replicate), 7 to 21 d (6 birds per replicate) and 19 to 21 d (3 birds per replicate) d of age. Diets were prepared with 3 Ca (0.65, 0.80, and 0.95%) and 2 non-phytate P, (0.20 and 0.40%) concentrations. A 6-phytase was added at 500 or 1,000 FTU/kg to the 0.20% nPP diet at each Ca concentration. The age and adaptation effects were determined by comparing the responses between birds fed from 7 to 9 and 19 to 21 d of age, 19 to 21, and 7 to 21 d of age, respectively. An age effect was observed regardless of Ca, nPP, or phytase concentration, with older birds (19 to 21 d) having greater apparent ileal digestibility (AID) of amino acids (AA) and CP than younger birds (7 to 9 d; Pphytase concentrations. Constant lower AID of CP and AA was seen in adapted birds (7 to 21 d) compared to unadapted bird (19 to 21 d) when 0.20% nPP diets were fed at 0.95% Ca concentrations (PPhytase efficacy was significantly lower in younger (7 to 9 d) compared to older birds (19 to 21 d; PPhytase inclusion increased AID of CP and AA regardless of Ca (P<0.05). In conclusion, the AID of CP and AA can be affected by diet, age, and adaptation.

  10. Influence of phytase, EDTA, and polyphenols on zinc absorption in adults from porridges fortified with zinc sulfate or zinc oxide.

    Science.gov (United States)

    Brnić, Marica; Wegmüller, Rita; Zeder, Christophe; Senti, Gabriela; Hurrell, Richard F

    2014-09-01

    Fortification of cereal staples with zinc is recommended to combat zinc deficiency. To optimize zinc absorption, strategies are needed to overcome the inhibitory effect of phytic acid (PA) and perhaps polyphenols. Five zinc absorption studies were conducted in young adults consuming maize or sorghum porridges fortified with 2 mg zinc as zinc sulfate (ZnSO4) or zinc oxide (ZnO) and containing combinations of PA or polyphenols as potential inhibitors and EDTA and phytase as potential enhancers. Fractional absorption of zinc (FAZ) was measured by using the double isotopic tracer ratio method. Adding phytase to the maize porridge immediately before consumption or using phytase for dephytinization during meal preparation both increased FAZ by >80% (both P zinc molar ratio of 1:1 increased FAZ from maize porridge fortified with ZnSO4 by 30% (P = 0.01) but had no influence at higher EDTA ratios or on absorption from ZnO. FAZ was slightly higher from ZnSO4 than from ZnO (P = 0.02). Sorghum polyphenols had no effect on FAZ from dephytinized sorghum porridges but decreased FAZ by 20% from PA-rich sorghum porridges (P zinc absorption from zinc-fortified cereals, EDTA at a 1:1 molar ratio modestly enhanced zinc absorption from ZnSO4-fortified cereals but not ZnO-fortified cereals, and sorghum polyphenols inhibited zinc absorption in the presence, but not absence, of PA. This trial was registered at clinicaltrials.gov as NCT01210794.

  11. Optimization of heterologous expression of the phytase (PPHY) of Pichia anomala in P. pastoris and its applicability in fractionating allergenic glycinin from soy protein.

    Science.gov (United States)

    Joshi, Swati; Satyanarayana, T

    2014-06-01

    The phytase (PPHY) of Pichia anomala has the requisite properties of thermostability and acidstability, broad substrate spectrum, and protease insensitivity, which make it a suitable candidate as a feed and food additive. The 1,389-bp PPHY gene was amplified from P. anomala genomic DNA, cloned in pPICZαA, and expressed extracellularly in P. pastoris X33. Three copies of PPHY have been detected integrated into the chromosomal DNA of the recombinant P. pastoris. The size exclusion chromatography followed by electrophoresis of the pure rPPHY confirmed that this is a homohexameric glycoprotein of ~420 kDa with a 24.3 % portion as N-linked glycans. The temperature and pH optima of rPPHY are 60 °C and 4.0, similar to the endogenous enzyme. The kinetic characteristics K(m), V(max), K(cat), and K(cat)/K(m) of rPPHY are 0.2 ± 0.03 mM, 78.2 ± 1.43 nmol mg(-1) s(-1), 65,655 ± 10.92 s(-1), and 328.3 ± 3.12 μM(-1) s(-1), respectively. The optimization of medium components led to a 21.8-fold improvement in rPPHY production over the endogenous yeast. The rPPHY titer attained in shake flasks could also be sustained in the laboratory fermenter. The rPPHY accounts for 57.1 % of the total secreted protein into the medium. The enzyme has been found useful in fractionating allergenic protein glycinin from soya protein besides dephytinization.

  12. Comparative carcass and tissue nutrient composition of transgenic Yorkshire pigs expressing phytase in the saliva and conventional Yorkshire pigs.

    Science.gov (United States)

    Forsberg, C W; Meidinger, R G; Ajakaiye, A; Murray, D; Fan, M Z; Mandell, I B; Phillips, J P

    2014-10-01

    A transgenic line of Yorkshire (YK) pigs named the Cassie (CA) line was produced with a low copy number phytase transgene inserted in the genome. The transgenic line efficiently digests P, Ca, and other major minerals of plant dietary origin. The objectives of this study were to 1) compare carcass and tissue nutrient composition and meat quality traits for third generation hemizygous CA line market BW finisher pigs (n = 24) with age-matched conventional YK finisher pigs (n = 24) and 2) examine effects of outbreeding with high-index conventional YK boars on modifying carcass leanness from the third to sixth generations in CA line finisher boars (n = 73) and gilts (n = 103). Cassie boars (n = 12) and CA gilts (n = 12) were fed diets without supplemental P and comparable numbers of age-matched YK boars and gilts fed diets containing supplement P were raised throughout the finisher phase. The pigs were slaughtered and then fabricated into commercial pork primals before meat composition and quality evaluation. Proximate and major micronutrient composition was determined on tissues including fat, kidney, lean, liver, and skin. The main difference observed was greater (P = 0.033) crude fat content in CA boar carcasses and increased (P pigs. There were no substantive differences in tissue composition, except for CA boar kidneys. Numerous changes in the mineral, fatty acid, and indispensable AA composition for CA boar kidneys were not apparent in CA gilts. These changes may point to adaptive physiological changes in the boar kidney necessary for homeostatic regulation of mineral retention related to phytase action rather than to insertion of the transgene. However, from a meat composition perspective, transgenic expression of phytase in the CA line of YK pigs had little overall effect on meat composition. Outbreeding of high-index CA gilts with high-index commercial YK boars linearly reduced (P = 0.002) back fat thickness with a corresponding linear increase (P = 0.001) in

  13. Meta-analysis of phosphorus utilization by growing pigs: effect of dietary phosphorus, calcium and exogenous phytase.

    Science.gov (United States)

    Létourneau-Montminy, M P; Jondreville, C; Sauvant, D; Narcy, A

    2012-10-01

    Optimizing phosphorus (P) utilization in pigs requires improving our capacity to predict the amount of P absorbed and retained, with the main modulating factors taken into account, as well as precisely determining the P requirements of the animals. Given the large amount of published data on P utilization in pigs, a meta-analysis was performed to quantify the impact of the different dietary P forms, calcium (Ca) and exogenous phytases on the digestive and metabolic utilization criteria for dietary P in growing pigs. Accordingly, the amount of phytate P (PP) leading to digestible P (g/kg) was estimated to be 21%, compared with 73% for non-phytate P (NPP) from plant ingredients and 80% for NPP from mineral and animal ingredients (P < 0.001). The increase in total digestible dietary P following the addition of microbial phytase (PhytM) from Aspergillus niger (P < 0.001) was curvilinear and about two times higher than the increase following the addition of plant phytase, which leads to a linear response (P < 0.001). The response of digestible P to PhytM also depends on the amount of substrate, PP (PhytM(2) × PP, P < 0.001). The digestibility of dietary P decreased with dietary Ca concentration (P < 0.01) independently of phytase but increased with body weight (BW, P < 0.05). Although total digestible dietary P increased linearly with total NPP concentration (P < 0.001), retained P (g/kg), average daily gain (ADG, g/day) and average daily feed intake (ADFI, g/day) increased curvilinearly (P < 0.001). Interestingly, whereas dietary Ca negatively affected P digestibility, the effect of dietary Ca on retained P, ADG and ADFI depended on total dietary NPP (NPP × Ca, P < 0.01, P < 0.05 and P < 0.01, respectively). Increasing dietary Ca reduced retained P, ADG and ADFI at low NPP levels, but at higher NPP concentrations it had no effect on ADG and ADFI despite a positive effect on retained P. Although the curvilinear effect of PhytM on digestible P increased with PP (P < 0

  14. Consequences of phosphorus interactions with calcium, phytase, and cholecalciferol on zootechnical performance and mineral retention in broiler chickens.

    Science.gov (United States)

    Delezie, E; Maertens, L; Huyghebaert, G

    2012-10-01

    The objective was to determine the effect of calcium (Ca), total phosphorus (Ptot), cholecalciferol, and phytase level in the diet on the performance, tibia ash percentage, and Ca and P retention in broilers until slaughter age. Broilers were randomly assigned to 12 treatments, each with 6 replicates, comprising 3 diets differing in Ca and P level: 1) normal Ca and Ptot level (NN); 2) normal Ca and low Ptot level (NL), 3) low Ca and Ptot level (LL). Broilers were also given 2 levels of cholecalciferol and 2 levels of phytase. The normal levels of Ca and Ptot for the starter, grower, and finisher phases were 0.90, 0.82, 0.74% and 0.67, 0.62, 0.57%, respectively. The low Ca and Ptot levels for the 3 phases were 0.67, 0.60, 0.52% and 0.57, 0.51, 0.46%, respectively. Broilers of the NL treatment obtained the lowest BW, whereas BW of the NN and LL groups were comparable. Cholecalciferol significantly affected the BW, with differences up to 2.6 and 1.2% for the starter and grower phases, respectively. The highest cholecalciferol effect was found in combination with the NN treatment. The percentage of retained Ca increased from 33% to 41% and 48% when the imbalanced diet was replaced by the NN and LL balanced diets, respectively. P release from phytate was 64 and 67% for the NL and LL diets, respectively. Phytase and cholecalciferol had significantly favorable effects on retention values but these effects were dependent on Ca and Ptot levels and their ratio. In conclusion, both diets with the balanced Ca/Ptot ratio resulted in the best performance, highest tibia ash percentage and P release from phytate. A reduction of the Aviagen (2009) recommended P requirements by 25 to 30% and Ca by 15 to 20% over the various phases did not negatively affect performance, bone development, and improved Ca and Ptot retention. The effects of supplementing cholecalciferol and phytase were additive but not significant and no synergism between both was present.

  15. Phytase production by Rhizopus microsporus var. microsporus biofilm: characterization of enzymatic activity after spray drying in presence of carbohydrates and nonconventional adjuvants.

    Science.gov (United States)

    Sato, Vanessa Sayuri; Jorge, João Atílio; Oliveira, Wanderley Pereira; Souza, Claudia Regina Fernandez; Guimarães, Luis Henrique Souza

    2014-02-28

    Microbial phytases are enzymes with biotechnological interest for the feed industry. In this article, the effect of spray-drying conditions on the stability and activity of extracellular phytase produced by R. microsporus var. microsporus biofilm is described. The phytase was spray-dried in the presence of starch, corn meal (>150 μm), soy bean meal (SB), corn meal (drying adjuvants. The residual enzyme activity after drying ranged from 10.7% to 60.4%, with SB and CM standing out as stabilizing agents. Water concentration and residual enzyme activity were determined in obtained powders as a function of the drying condition. When exposed to different pH values, the SB and CM products were stable, with residual activity above 50% in the pH range from 4.5 to 8.5 for 60 min. The use of CM as drying adjuvant promoted the best retention of enzymatic activity compared with SB. Spray drying of the R. microsporus var. microsporus phytase using different drying adjuvants showed interesting results, being quite feasible with regards their biotechnological applications, especially for poultry diets.

  16. Investigation of the interaction between separate calcium feeding and phytase supplementation on growth performance, calcium intake, nutrient digestibility and energy utilisation in broiler starters

    NARCIS (Netherlands)

    Abdollahi, M.R.; Duangnumsawang, Y.; Kwakkel, R.P.; Steenfeldt, S.; Bootwalla, S.M.; Ravindran, V.

    2016-01-01

    The interaction between separate calcium (Ca) feeding and phytase supplementation on performance, coefficient of apparent ileal digestibility (CAID) of nitrogen (N), starch, fat and phosphorus (P), total tract retention (TTR) of Ca and P, and apparent metabolisable energy (AME) in broiler starter

  17. Effect of Phytase on Apparent Total Tract Digestibility of Phosphorus in Corn-Soybean Meal Diets Fed to 100 kg Pigs

    Science.gov (United States)

    Five experiments were conducted to investigate the ability of different sources of phytase supplemented to the diet at graded levels to improve apparent P digestibility in finishing pigs. A corn-soybean meal basal diet containing 0.50% Ca and 0.32% P (0.06% available P) was used in all experiments a...

  18. A single mutation in the hepta-peptide active site of Aspergillus niger PhyA phytase leads to myriad of biochemical changes

    Science.gov (United States)

    The active site motif of proteins belonging to ‘Histidine Acid Phosphatase’ (HAP) contains a hepta-peptide region, RHGXRXP. A close comparison among fungal and yeast HAPs has revealed the fourth residue of the hepta-peptide to be E instead of A, which is the case with A. niger phyA phytase. However,...

  19. Gene

    Data.gov (United States)

    U.S. Department of Health & Human Services — Gene integrates information from a wide range of species. A record may include nomenclature, Reference Sequences (RefSeqs), maps, pathways, variations, phenotypes,...

  20. In vitro versus in situ evaluation of the effect of phytase supplementation on calcium and phosphorus solubility in soya bean and rapeseed meal broiler diets.

    Science.gov (United States)

    Morgan, N K; Walk, C L; Bedford, M R; Burton, E J

    2014-01-01

    1. In vitro assays provide a rapid and economical tool to evaluate dietary effects, but have limitations. In this study, the effect of phytase supplementation on solubility, and presumed availability, of calcium (Ca) and phosphorus (P) in soya bean meal (SBM) and rapeseed meal (RSM) based diets were evaluated both in situ and by a two-step in vitro digestion assay that simulated the gastric and small intestine (SI) phases of digestion. 2. Comparison of the in vitro findings to in situ findings was used to evaluate the in vitro assay. Ross 308 broilers (n = 192) were fed on one of 6 SBM or RSM diets supplemented with 0, 500 or 5000 FTU/kg phytase from 0 to 28 d post hatch. The 6 diets and raw SBM and RSM were exposed to a two-step in vitro assay. Ca and P solubility and pH in the gizzard and jejunal digesta and in the gastric and SI phase of in vitro digestion were measured. 3. Both in vitro and in situ analyses detected that Ca solubility was lowest when diets were supplemented with 500 FTU/kg phytase, compared to the control diets and diets supplemented with 5000 FTU/kg phytase. Phosphorus solubility increased with increasing phytase level. Both methods also identified that mineral solubility plateaus in the gastric phase. 4. Overall relationship of the two methods was strong for both determination of gastric phase Ca and P solubility (r = 0.96 and 0.92, respectively) and also SI phase Ca and P solubility (r = 0.71 and 0.82, respectively). However, mineral solubility and pH were higher when measured in vitro than in situ, and the in situ assay identified an interaction among the effects of phase, protein source and phytase inclusion level on Ca solubility that the in vitro assay did not detect. 5. This two-step in vitro assay successfully predicted phytase efficacy, but to determine detailed response effects in the animal, in situ data is still required.

  1. Performance, carcass characteristics, meat quality and plasma constituents of meat type drakes fed diets containing different levels of lysine with or without a microbial phytase.

    Science.gov (United States)

    Attia, Y A

    2003-02-01

    This experiment was conducted to study growth performance, carcass characteristics, meat quality and plasma constituents of Campbell drakes fed diets containing different levels of lysine with or without a microbial phytase. Basal vegetable duck all-mash diets were fed during the growing (1-35 d of age), and finishing period (36-56 d of age) and were formulated to contain 0.90% and 0.73% lysine (negative control), respectively. These diets were supplemented or not with L-lysine HCl, which resulted in a dietary lysine level of 0.90, 0.95, 1.01 and 1.06% and 0.73, 0.80, 0.87 and 0.94%, during the growing and finishing period, respectively. Furthermore, the diets were fed with or without 600 FTU phytase (Natuphos) except for those containing 1.06 and 0.94% lysine during the growing and finishing period, respectively (positive control). A lysine level of 1.01/0.87% in the growing/finishing diet significantly increased BWG and improved FCR of drakes by 2.1 and 1.8%, respectively. Phytase significantly increased BWG by 2.1% and 3.5% after feeding the basal diet and 1.01/0.87% lysine, respectively. Also, FCR was significantly improved by 2.2 and 1.8% of groups fed 0.95/0.80, and 1.01/0.87% lysine, respectively. Phytase as an independent variable increased BWG by 1.8, and improved FCR by 1.0%. Lysine and/or phytase did not affect carcass yield, and meat quality treats as well as plasma constituents of drakes. However, lysine level at 0.95/0.80% and 1.01/0.87% significantly decreased abdominal fat deposition compared to either the negative or the positive control. In conclusion, a lysine level of 1.01/0.87% in the growing/finishing diets for drakes is adequate. After phytase supplementation of the basal diet the BWG at a lysine level of 0.90/0.73% were similar to the positive control (1.06/0.94% lysine). However, the best FCR was obtained after feeding diets containing 1.01/0.87% lysine supplemented with phytase.

  2. Effect of reduced dietary calcium concentration and phytase supplementation on calcium and phosphorus utilization in weanling pigs with modified mineral status.

    Science.gov (United States)

    Létourneau-Montminy, M P; Narcy, A; Magnin, M; Sauvant, D; Bernier, J F; Pomar, C; Jondreville, C

    2010-05-01

    The present study was conducted to assess the effect of 2 dietary Ca concentrations on P and Ca digestive and metabolic utilization in weanling pigs fed diets providing practical concentrations of P, with or without phytase. The responses of pigs fed diets adequate or moderately deficient in Ca and P postweaning were compared. A total of 60 pigs weaned at 28 d of age were used. Two groups of 30 pigs with differing mineral status resulted from a 10-d depletion period, during which the animals received depletion diets (DD) that consisted of corn-soybean meal with either 1.42% Ca and 0.80% P (DD+) or 0.67% Ca and 0.43% P (DD-), designed to achieve the same Ca:digestible P ratio. At the end of the depletion period, a plasma sample was taken from each pig and 12 pigs (6 from each group) were slaughtered for bone assessment to establish the baseline mineral status. The animals fed the DD- diet had signs of P deficiency with reduced plasma P (13%; P pigs from each group were fed 1 of 4 repletion diets: 1) 0.56% P, 1.06% Ca; 2) 0.56% P, 0.67% Ca; 3) diet 1 + 1,000 phytase units (FTU) of Natuphos phytase/kg; and 4) diet 2 + 1,000 FTU of Natuphos phytase/kg. Total feces and urine were collected from d 5 to 11, and a blood sample was taken from each pig at d 11 and 25. The initial moderate P deficiency (DD-) stimulated Ca absorption (5%; P pigs achieved full recovery of femur DM and ash weight when they received phytase, whereas ash concentration tended to remain reduced by 3% (P = 0.08). Phosphorus digestibility was improved in the diets supplemented with phytase (73.0 vs. 56.0%; P dietary Ca decreased P digestibility (65.6 vs. 63.4%; P dietary Ca reduced equally P digestibility with and without phytase and did not influence the efficiency of phytase in releasing P in the digestive tract. In pigs fed diets with phytase, however, the reduction of Ca (Ca:P from 1.9 to 1.3) increased urinary P losses 5-fold. Those extra losses were due to a lack of Ca for skeleton ash

  3. 植酸酶单克隆抗体的制备及鉴定%Preparation and Characteristic of Monoclonal Antibody against Phytase

    Institute of Scientific and Technical Information of China (English)

    胡骁飞; 魏凤仙; 李青梅; 职爱民; 王方雨; 孙亚宁; 柴书军; 付晓鹤

    2012-01-01

    The hybridoma lines that secrete phytase monoclonal antibody (mAb) were established through cell fusion. Phytase mAb was prepared by inducing ascites in mice. The immunological characteristics of the mAb were I-dentified. The result indicated that two hybridomas were selected. The subtype of mAb generated by 5C3 hybridoma was IgGl ,the titer was 1: 1. 02 × 106 ,IC50 was 100. 07 ng/mL and Kaff was 6. 70 x 109 L/mol. Phytase mAb IC50 to normal, coated and concentrated phytase was 130.43,126. 16,125.33 ng/mL, and cross-reactivity was 76. 72% , 79.32% and 79. 84% .respectively. Phytase mAb IC50 to inactivated (boiled for 5 min) wheat bran phytase,normal,coated and concentrated phytase was 4 205. 45 ,4 015. 86 ,3 593. 19 ,3 610. 40 ng/mL,and the cross-reactivity was 2. 38% ,2. 49% ,2. 78% and 2. 77% , respectively. In conclusion, hybridoma lines that secrete phytase mAb were established, phytase mAb with high affinity and specificity and sensitive was prepared. The phytase mAb may lay a foundation for the subsequent ELISA kit and strip development.%通过细胞融合建立分泌抗植酸酶单克隆抗体(mAb)的杂交瘤细胞株.采用体内诱生腹水法制备单克隆抗体,并对其免疫学特性进行鉴定.结果表明,筛选出2株杂交瘤细胞,其中,5C3细胞株产生单抗亚型为IgG1型,效价为1:(1.02x106),IC50=100.07 ng/mL,亲和常数为6.70×109L/mol,与市售常规植酸酶、包被植酸酶及浓缩植酸酶的IC50分别达到130.43,126.16,125.33 ng/mL,交叉反应率分别为76.72%,79.32%和79.84%.与高温失活(100℃水煮5 min)的麸皮植酸酶、普通植酸酶、包被植酸酶及浓缩植酸酶IC50分别为4 205.45,4 015.86,3 593.19,3 610.40ng/mL,交叉反应率分别为2.38%,2.49%,2.78%,2.77%.说明成功筛选了杂交瘤细胞株,该细胞株能分泌抗植酸酶mAb,该植酸酶mAb对植酸酶亲和力高、特异性强、灵敏度好,为植酸酶ELISA检测试剂盒及试纸条研制奠定了基础.

  4. Microbial phytase addition resulted in a greater increase in phosphorus digestibility in dry-fed compared with liquid-fed non-heat-treated wheat-barley-maize diets for pigs

    DEFF Research Database (Denmark)

    Blaabjerg, Karoline; Thomassen, Anne-Marie; Poulsen, Hanne Damgaard

    2015-01-01

    and fed one of four diets for 12 days; 5 days for adaptation and 7 days for total, but separate collection of feces and urine. The basal diet was composed of wheat, barley, maize, soybean meal and no mineral phosphate. Dietary treatments were: basal dry-fed diet (BDD), BDD with microbial phytase (BDD......The objective was to evaluate the effect of microbial phytase (1250 FTU/kg diet with 88% dry matter (DM)) on apparent total tract digestibility (ATTD) of phosphorus (P) in pigs fed a dry or soaked diet. Twenty-four pigs (65±3 kg) from six litters were used. Pigs were housed in metabolism crates......+phy), BDD soaked for 24 h at 20°C before feeding (BDS) and BDS with microbial phytase (BDS+phy). Supplementation of microbial phytase increased ATTD of DM and crude protein (N×6.25) by 2 and 3 percentage units (P

  5. The efficacy of a new 6-phytase obtained from Buttiauxella spp. expressed in Trichoderma reesei on digestibility of amino acids, energy, and nutrients in pigs fed a diet based on corn, soybean meal, wheat middlings, and corn distillers' dried grains with solubles.

    Science.gov (United States)

    Adedokun, S A; Owusu-Asiedu, A; Ragland, D; Plumstead, P; Adeola, O

    2015-01-01

    Sixteen cannulated pigs were used to evaluate the effect of a new 6-phytase derived from Buttiauxella spp. and expressed in Trichoderma reesei on apparent ileal digestibility (AID) of AA and apparent total tract digestibility (ATTD) of DM, N, Ca, P, Na, Mg, K, Cl, and energy. Pigs were fed 4 diets for 2 periods in a crossover design. Within each period, there were 4 blocks of 4 pigs per block with each diet represented within each block. The average initial BW in periods 1 and 2 were 22 and 30 kg, respectively. Each period lasted 9 d with fecal collection on d 5 and 6 and a 12-h ileal digesta collection on d 7, 8, and 9. Pigs received a daily feed allowance of approximately 4.5% of their BW. The experimental diets were based on corn, soybean meal, wheat middlings, and corn distillers dried grain with solubles. Phytase was added at 0; 500; 1,000; or 2,000 phytase units/kg of diet to a basal diet that contained 205, 15, 5.4, and 10 g of CP, Lys, total P (1.6 g of nonphytate P), and Ca/kg diet, respectively. The addition of phytase improved (P phytase supplementation linearly and quadratically increased (P Phytase supplementation of the basal diet improved (P Phytase supplementation increased (P phytase supplementation of the basal diet increased (P phytase supplementation to the basal diet showed a tendency (P phytase supplementation. Increasing the level of phytase supplementation resulted in linear increases (P phytase expressed in Trichoderma reesei enhanced ileal digestibility of N and several AA in growing pigs in a dose-dependent manner.

  6. Dephytinisation with intrinsic wheat phytase and iron fortification significantly increase iron absorption from fonio (Digitaria exilis) meals in West African women.

    Science.gov (United States)

    Koréissi-Dembélé, Yara; Fanou-Fogny, Nadia; Moretti, Diego; Schuth, Stephan; Dossa, Romain A M; Egli, Ines; Zimmermann, Michael B; Brouwer, Inge D

    2013-01-01

    Low iron and high phytic acid content make fonio based meals a poor source of bioavailable iron. Phytic acid degradation in fonio porridge using whole grain cereals as phytase source and effect on iron bioavailability when added to iron fortified fonio meals were investigated. Grains, nuts and seeds collected in Mali markets were screened for phytic acid and phytase activity. We performed an iron absorption study in Beninese women (n = 16), using non-dephytinised fonio porridge (FFP) and dephytinised fonio porridge (FWFP; 75% fonio-25% wheat), each fortified with (57)Fe or (58)Fe labeled FeSO4. Iron absorption was quantified by measuring the erythrocyte incorporation of stable iron isotopes. Phytic acid varied from 0.39 (bambara nut) to 4.26 g/100 g DM (pumpkin seed), with oilseeds values higher than grains and nuts. Phytase activity ranged from 0.17±1.61 (fonio) to 2.9±1.3 phytase unit (PU) per g (whole wheat). Phytic acid was almost completely degraded in FWFP after 60 min of incubation (pH≈5.0, 50°C). Phytate∶iron molar ratios decreased from 23.7∶1 in FFP to 2.7∶1 in FWFP. Iron fortification further reduced phytate∶iron molar ratio to 1.9∶1 in FFP and 0.3∶1 in FWFP, respectively. Geometric mean (95% CI) iron absorption significantly increased from 2.6% (0.8-7.8) in FFP to 8.3% (3.8-17.9) in FWFP (Pporridge with intrinsic wheat phytase increased fractional iron absorption 3.2 times, suggesting it could be a possible strategy to decrease PA in cereal-based porridges.

  7. Dephytinisation with intrinsic wheat phytase and iron fortification significantly increase iron absorption from fonio (Digitaria exilis meals in West African women.

    Directory of Open Access Journals (Sweden)

    Yara Koréissi-Dembélé

    Full Text Available Low iron and high phytic acid content make fonio based meals a poor source of bioavailable iron. Phytic acid degradation in fonio porridge using whole grain cereals as phytase source and effect on iron bioavailability when added to iron fortified fonio meals were investigated. Grains, nuts and seeds collected in Mali markets were screened for phytic acid and phytase activity. We performed an iron absorption study in Beninese women (n = 16, using non-dephytinised fonio porridge (FFP and dephytinised fonio porridge (FWFP; 75% fonio-25% wheat, each fortified with (57Fe or (58Fe labeled FeSO4. Iron absorption was quantified by measuring the erythrocyte incorporation of stable iron isotopes. Phytic acid varied from 0.39 (bambara nut to 4.26 g/100 g DM (pumpkin seed, with oilseeds values higher than grains and nuts. Phytase activity ranged from 0.17±1.61 (fonio to 2.9±1.3 phytase unit (PU per g (whole wheat. Phytic acid was almost completely degraded in FWFP after 60 min of incubation (pH≈5.0, 50°C. Phytate∶iron molar ratios decreased from 23.7∶1 in FFP to 2.7∶1 in FWFP. Iron fortification further reduced phytate∶iron molar ratio to 1.9∶1 in FFP and 0.3∶1 in FWFP, respectively. Geometric mean (95% CI iron absorption significantly increased from 2.6% (0.8-7.8 in FFP to 8.3% (3.8-17.9 in FWFP (P<0.0001. Dephytinisation of fonio porridge with intrinsic wheat phytase increased fractional iron absorption 3.2 times, suggesting it could be a possible strategy to decrease PA in cereal-based porridges.

  8. Characterization of a Thermotolerant Phytase Produced by Rhizopus microsporus var. microsporus Biofilm on an Inert Support Using Sugarcane Bagasse as Carbon Source.

    Science.gov (United States)

    Sato, Vanessa Sayuri; Jorge, João Atílio; Guimarães, Luis Henrique Souza

    2016-06-01

    The Rhizopus microsporus var. microsporus biofilm was able to produce increased levels of an extracellular thermotolerant phytase using polyethylene and viscose as an inert support in both modified NBRIP medium and modified Khanna medium containing sugarcane bagasse as the carbon source. The enzyme production was strictly regulated by the phosphorus content with optimal production at 0.5 mM of sodium phytate and KH2PO4. The extracellular phytase, RMPhy1, was purified 4.18-fold with 4.78 % recovery using DEAE-cellulose and CM-cellulose. A single protein band with a molecular mass of 35.4 kDa was obtained when the samples were subjected to 10 % SDS-PAGE. The optimum temperature for activity was 55 °C and the optimum pH was 4.5. R. microsporus var. microsporus phytase exhibited high stability at 30 and 40 °C with a half-life of 115 min at 60 °C. The enzyme activity increased in the presence of Ca (2+) and was inhibited by Zn(2+), arsenate, and sodium phosphate. Phytase demonstrated high substrate specificity for sodium phytate with K m = 0.72 mM and V max = 94.55 U/mg of protein and for p-NPP with K m = 0.04 mM and V max = 106.38 U/mg of protein. The enzyme also hydrolyzed ATP, AMPc, glucose 6-phosphate, glucose 1-phosphate, and UDPG. This is the first report on phytase characterization delivered with biofilm technology. The properties of the enzyme account for its high potential for use in biotechnology and the possibility of application in different industrial sectors as feed in the future.

  9. Phytase-Producing Potential and Other Functional Attributes of Lactic Acid Bacteria Isolates for Prospective Probiotic Applications.

    Science.gov (United States)

    Andrabi, Syed Tabia; Bhat, Bilqeesa; Gupta, Mahak; Bajaj, Bijender Kumar

    2016-09-01

    Wide variations among multifaceted-health benefitting attributes of probiotics fueled investigations on targeting efficacious probiotics. In the current study, lactic acid bacteria (LAB) isolated from poultry gut, feces of rat, chicken, human infants, and fermented foods were characterized for desired probiotic functional properties including the phytase-producing ability which is one of the wanted characteristics for probiotics for potential applications for upgrading animal nutrition, enhancing feed conversion, and minimizing anti-nutritional properties. Among 62 LAB isolates Weissella kimchii R-3 an isolate from poultry gut exhibited substantial phytase-producing ability (1.77 U/ml) in addition to other functional probiotic characteristics viz. hydrophobicity, autoaggregation, coaggregation with bacterial pathogens, and antimicrobial activity against pathogens. Survival of W. kimchii R-3 cells (in free and calcium alginate encapsulated state) was examined sequentially in simulated gastric and intestinal juices. Encapsulated cells exhibited better survival under simulated gut conditions indicating that encapsulation conferred considerable protection against adverse gut conditions. Furthermore, simulated gastric and intestinal juices with pepsin and pancreatin showed higher survival of cells than the juices without pepsin and pancreatin. W. kimchii R-3 due to its significant functional probiotic attributes may have prospective for commercial applications in human/animal nutrition.

  10. Nutrient utilization and manure P excretion in growing pigs fed corn-barley-soybean based diets supplemented with microbial phytase.

    Science.gov (United States)

    Emiola, Adewale; Akinremi, Oluwole; Slominski, Bogdan; Nyachoti, C Martin

    2009-02-01

    The effect of high levels of microbial phytase supplementation in diets for growing pigs was studied in a 2-week performance and nutrient digestibility trial involving 28 growing pigs weighing 16.4 +/- 1.06 (mean +/- SD) kg. Seven corn-barley-soybean meal-based diets consisting of a positive control (PC) formulated to meet or exceed NRC nutrient requirements; a negative control (NC) with non-phytate P reduced by 0.1% unit from NRC requirement and fed without or with 500 or 1000 U/kg; a doubled negative control (DNC) with no added inorganic P and fed without or with 2000 or 4000 U/kg. Chromic oxide was added as an indigestible marker and all diets were fed as mash. Pigs fed the PC diet had a higher P digestibility compared with those fed the NC (P DNC (P DNC diet resulted in linear (P DNC diets. Apparent total tract digestibility of N, OM and DM were higher (P DNC diet, but not the NC diet (P DNC diet resulted in a linear increase (P DNC diets linearly decreased fecal P (P DNC diets (P < 0.05). The results of this study show that complete removal of inorganic P from growing pig diets coupled with phytase supplementation improves digestibility and retention of P and N, thus reducing manure P excretion without any negative effect on pig performance.

  11. Purification and characterization of phytase with a wide pH adaptation from common edible mushroom Volvariella volvacea (Straw mushroom).

    Science.gov (United States)

    Xu, Lijing; Zhang, Guoqing; Wang, Hexiang; Ng, Tzi Bun

    2012-02-01

    A novel phytase with a molecular mass of 14 kDa was isolated from fresh fruiting bodies of the common edible mushroom Volvariella volvacea (Straw mushroom). The isolation procedure involved successive chromatography on DEAE-cellulose, CM-cellulose, Affi-gel blue gel, Q-Sepharose and Superdex-75. The enzyme was a monomeric protein and was unadsorbed on DEAE-cellulose, CM-cellulose and Affi-gel blue gel, but was adsorbed on Q-Sepharose. The enzyme was purified 51.6-fold from the crude extract with 25.9% yield. Its N-terminal amino acid sequence GEDNEHDTQA exhibited low homology to the other reported phytases. The optimal pH and temperature of the purified enzyme was 5 and 45 degrees C, respectively. The enzyme was quite stable over the pH range of 3.0 to 9.0 with less than 30% change in its activity, suggesting that it can be used in a very wide pH range. The enzyme exhibited broad substrate selectivity towards various phosphorylated compounds, but lacked antifungal activity against tested plant pathogens.

  12. Engineering the residual side chains of HAP phytases to improve their pepsin resistance and catalytic efficiency

    Science.gov (United States)

    Niu, Canfang; Yang, Peilong; Luo, Huiying; Huang, Huoqing; Wang, Yaru; Yao, Bin

    2017-01-01

    Strong resistance to proteolytic attack is important for feed enzymes. Here, we selected three predicted pepsin cleavage sites, L99, L162, and E230 (numbering from the initiator M of premature proteins), in pepsin-sensitive HAP phytases YkAPPA from Yersinia kristensenii and YeAPPA from Y. enterocolitica, which corresponded to L99, V162, and D230 in pepsin-resistant YrAPPA from Y. rohdei. We constructed mutants with different side chain structures at these sites using site-directed mutagenesis and produced all enzymes in Escherichia coli for catalytic and biochemical characterization. The substitutions E230G/A/P/R/S/T/D, L162G/A/V, L99A, L99A/L162G, and L99A/L162G/E230G improved the pepsin resistance. Moreover, E230G/A and L162G/V conferred enhanced pepsin resistance on YkAPPA and YeAPPA, increased their catalytic efficiency 1.3–2.4-fold, improved their stability at 60 °C and pH 1.0–2.0 and alleviated inhibition by metal ions. In addition, E230G increased the ability of YkAPPA and YeAPPA to hydrolyze phytate from corn meal at a high pepsin concentration and low pH, which indicated that optimization of the pepsin cleavage site side chains may enhance the pepsin resistance, improve the stability at acidic pH, and increase the catalytic activity. This study proposes an efficient approach to improve enzyme performance in monogastric animals fed feed with a high phytate content. PMID:28186144

  13. A simplified model for A. Niger FS3 growth during phytase formation in solid State fermentation

    Directory of Open Access Journals (Sweden)

    Michele Rigon Spier

    2009-11-01

    Full Text Available A simplified model to describe fungal growth during citric pulp fermentation for phytase production was described for the first time. Experimental data for biomass growth were adjusted to classical mathematical growth models (Monod and Logistic. The Monod model predictions showed good agreement with the experimental results for biomass concentration during 96 hours of fermentation. Parameters such as yield of biomass from oxygen (Y X/O, maintenance coefficient (m and specific growth rate (µ were compared showing a good correlation between the data and the model. An alternative method for biomass determination in this process was developed since a great correlation was found between biomass growth and enzyme formation.Um modelo simplificado para descrever o crescimento fúngico durante a fermentação em polpa citric para a produção da fitase foi descrita pela primeira vez. Dados experimentais para a formação de biomassa foram ajustados a modelos clássicos de crescimento microbiano (Monod e Logístico. O modelo Monod previsto mostrou boa correlação aos resultados experimentais para a concentração de biomassa até 96 horas de fermentação. Parâmetros como rendimento de biomassa a partir de oxigênio (Y X/O, coeficiente de manutenção (m e taxa específica de crescimento (µ foram comparados mostrando uma boa correlação entre os dados e o modelo. Um método alternativo para a determinação de biomassa neste processo foi desenvolvido a partir de uma excelente correlação encontrado entre o crescimento microbiano e a formação da enzima.

  14. Effect of citric acid and microbial phytase on small intestinal morphology in broiler chicken

    Directory of Open Access Journals (Sweden)

    Rouhollah Nourmohammadi

    2013-01-01

    Full Text Available An experiment was carried out to investigate the effects of citric acid (CA (0, 3 and 6% and microbial phytase (MP (0, 500 and 1000 IU/kg on morphology of different segments of small intestine (duodenum, jejunum and ileum in broiler chickens fed on corn and soybean meal based diets. The effect of 9 experimental treatments (3×3 factorial design were assessed using 270 7-d-old Ross 308 male broiler chicks in a randomized complete block design in three replicates of 10 birds each. The mean villi length (VL, crypt depth (CD and goblet cell number (GCN in duodenum, jejunum and ileum and villi width (VW and VL:CD ratio in duodenum was significantly greater for the birds fed on acidified diets compared to the control birds at day 42 of age (P<0.01. Inclusion of 3% CA in diet significantly decreased the epithelial thickness (ET in duodenum, jejunum and ileum (P<0.01. The birds received diets with 1000 IU/kg of MP showed significant increase in CD (P<0.01 and GCN in jejunum (P<0.05, and significant decrease in VL:CD ratio and ET in the duodenum (P<0.01, jejunum (P<0.05 and ileum (P<0.01 segments. No variable of interest were affected by CA × MP interaction. It was concluded that CA and MP independently exhibit positive impact on morphometery of small intestine, toward facilitating the nutrient absorption and reducing the metabolic demands of the intestinal tract in broiler chickens.

  15. The effect of addition high rape cake and phytase on nutritive value of diets for broiler chickens

    Directory of Open Access Journals (Sweden)

    Banaszkiewicz Teresa

    2013-01-01

    Full Text Available The aim of this study was to evaluate the effect of high rape cake content and phytase added to phosphorus and calcium deficient diets on the nutritive value for broiler chickens. Two experiments were performed: a growth experiment on four groups of one-day-old broilers Ross 308, 30 birds per group (6 replications x 5 birds and a digestibility experiment on 60 chickens divided into four groups of 20 birds (4 replications of 5 birds. The digestibility of the starter diets were evaluated on chickens at 7 days of age and of grower diets at 28 days of age. The diets used in the digestibility tests were the same for the growth trial. In the growth experiment four diets were prepared for the due periods: starters (1-21 day and growers (22-49 day of chicken life. The control diet (SBM did not contain rape cake, while experimental diets contained 15% (starters and 20% (growers rape cake of Lirajet cultivar. The experimental diet denoted RC HP had P and Ca contents equal the control diet (phosphorus about 7.5 g and calcium about 10 g.kg-1, while the diet denoted as RC LP contained less P and Ca (5.8 g and 6.8 g.kg-1 respectively than the control and RC HP diets. The diet denoted RC LP+ Phy was supplemented with an enzyme preparation containing phytase at a quantity of 875 FYT.kg-1. Application of 15% of rape cake into starter and 20% into grower diets (RC HP allowed for similar body weights and feed conversion ratio as the control group, whereas reduction of phosphorus and calcium content in the starter diet (RC LP significantly decreased body weight at day 21. The addition of phytase to the starter diet with low level of phosphorus and calcium showed the tendency to improve body weight in this period. Application of rape cake into starter and grower diets had poor effects on fat digestibility in all groups, whereas supplementation of grower diets with a low level of phytase phosphorus and calcium improved the digestibility of total phosphorus in

  16. Effect of high-dose phytase supplementation in broilers from 22 to 42 days post-hatch given diets severely limited in available phosphorus.

    Science.gov (United States)

    Taheri, H R; Heidari, A; Shahir, M H

    2015-01-01

    1. Two trials were conducted from 22 to 42 d post-hatch to evaluate the effectiveness of high concentrations of supplemental phytase in maize-soya bean meal-based diets severely limited in available phosphorus (P). Growth performance, plasma P and tibia ash (TA) were measured. 2. Each trial used 220 21-d-old male broilers in 20 pens with 11 birds per pen. Dietary treatments included a positive control [PC, 4.3 g/kg nonphytate P (NPP)], negative control [NC, 2.3 g/kg NPP (Trial 1) or 1.4 g/kg NPP (Trial 2)] and NC plus 1000, 2000 or 4000 phytase U/kg of the diet. 3. Birds fed on the PC diet had higher average daily gain (ADG), gain to feed ratio (G:F), plasma P (Trials 1 and 2) and TA (Trial 2) than those fed on the NC. 4. In Trial 1, ADG and G:F values of the NC plus 1000, 2000 or 4000 phytase U/kg reached those of the PC. Plasma P values of the NC plus 2000 or 4000 phytase U/kg reached that of the PC. Although TA values of the NC, NC + 1000 or NC + 2000 reached that of the PC, TA of the NC + 4000 was more than that of the PC. 5. In Trial 2, ADG and G:F values of the NC plus 4000 phytase U/kg reached those of the PC; nevertheless, plasma P values of the NC diets did not come up to that of the PC. While TA values of the NC, NC + 1000 or NC + 2000 did not reach that of the PC, TA of the NC + 4000 was greater than that of the PC. 6. Results of this study showed that, in the diets with 2.3 and 1.4 g/kg NPP, respectively, 1000 and 4000 phytase U/kg can be sufficient to obtain a comparable performance in broilers to those given diets adequate in available P.

  17. Interactive effects of phytase and xylanase supplementation with extractable salt-soluble protein content of corn in diets with adequate calcium and nonphytate phosphorus fed to broilers.

    Science.gov (United States)

    Gehring, C K; Bedford, M R; Dozier, W A

    2013-07-01

    The objective was to determine the effects of extractable salt-soluble protein content of corn (PS) and exogenous enzyme supplementation on N, starch, and energy digestibility in broilers fed diets adequate in Ca and nonphytate P. Broilers were randomly distributed into floor pens (6 replicate pens per treatment) with 28 birds per pen at 1 d of age. Treatments consisting of 4 sources of corn varying in PS (A, 58.1; B, 54.2; C, 53.7; and D, 30.6 mg of BSA equivalent values) with or without phytase (0 and 1,000 phytase units/kg) and xylanase (0 and 16,000 units of xylanase activity/kg) were randomly assigned to each pen. Different sources of corn were provided from 1 to 9 and 24 to 29 d of age. However, enzyme treatments were provided throughout the experiment. From 1 to 9 d of age, no interactions were observed. Apparent ileal N digestibility (AIND) and apparent ileal digestible energy (IDE) of diets with the lowest PS (based on corn D) were lower (P ≤ 0.05) than those of diets with a higher PS. Phytase increased (P ≤ 0.01) AIND and IDE by 5 and 16%, respectively, and xylanase exerted the opposite effect (P ≤ 0.03). From 24 to 29 d of age, phytase and xylanase in combination resulted in reduced (P ≤ 0.05) AIND of diets with a low PS (based on corn D) compared with the basal diet in broilers. Broilers fed diets with the highest or lowest PS (based on corn A or D) had lower (3-way interaction; P ≤ 0.05) IDE when phytase and xylanase were supplemented in combination compared with either enzyme alone. In conclusion, responses to exogenous enzyme supplementation are not constant and are influenced by the source of ingredients as well as the age of broilers. The magnitudes of the responses to phytase on nutrient and energy digestibility were greater at 9 compared with 29 d of age.

  18. Microbial phytase addition resulted in a greater increase in phosphorus digestibility in dry-fed compared with liquid-fed non-heat-treated wheat-barley-maize diets for pigs.

    Science.gov (United States)

    Blaabjerg, K; Thomassen, A-M; Poulsen, H D

    2015-02-01

    The objective was to evaluate the effect of microbial phytase (1250 FTU/kg diet with 88% dry matter (DM)) on apparent total tract digestibility (ATTD) of phosphorus (P) in pigs fed a dry or soaked diet. Twenty-four pigs (65±3 kg) from six litters were used. Pigs were housed in metabolism crates and fed one of four diets for 12 days; 5 days for adaptation and 7 days for total, but separate collection of feces and urine. The basal diet was composed of wheat, barley, maize, soybean meal and no mineral phosphate. Dietary treatments were: basal dry-fed diet (BDD), BDD with microbial phytase (BDD+phy), BDD soaked for 24 h at 20°C before feeding (BDS) and BDS with microbial phytase (BDS+phy). Supplementation of microbial phytase increased ATTD of DM and crude protein (N×6.25) by 2 and 3 percentage units (Pphytase and soaking (P=0.02). This was due to a greater increase in ATTD of P by soaking of the diet containing solely plant phytase compared with the diet supplemented with microbial phytase: 35%, 65%, 44% and 68% for BDD, BDD+phy, BSD and BSD+phy, respectively. As such, supplementation of microbial phytase increased ATTD of P in the dry-fed diet, but not in the soaked diet. The higher ATTD of P for BDS compared with BDD resulted from the degradation of 54% of the phytate in BDS by wheat and barley phytases during soaking. On the other hand, soaking of BDS+phy did not increase ATTD of P significantly compared with BDD+phy despite that 76% of the phytate in BDS+phy was degraded before feeding. In conclusion, soaking of BDS containing solely plant phytase provided a great potential for increasing ATTD of P. However, this potential was not present when microbial phytase (1250 FTU/kg diet) was supplemented, most likely because soaking of BDS+phy for 24 h at 20°C did not result in a complete degradation of phytate before feeding.

  19. Over-expression of the bacterial phytase US417 in Arabidopsis reduces the concentration of phytic acid and reveals its involvement in the regulation of sulfate and phosphate homeostasis and signaling.

    Science.gov (United States)

    Belgaroui, Nibras; Zaidi, Ikram; Farhat, Ameny; Chouayekh, Hichem; Bouain, Nadia; Chay, Sandrine; Curie, Catherine; Mari, Stéphane; Masmoudi, Khaled; Davidian, Jean-Claude; Berthomieu, Pierre; Rouached, Hatem; Hanin, Moez

    2014-11-01

    Phytic acid (PA) is the main phosphorus storage form in plant seeds. It is recognized as an anti-nutrient for humans and non-ruminant animals, as well as one of the major sources of phosphorus that contributes to eutrophication. Therefore, engineering plants with low PA content without affecting plant growth capacity has become a major focus in plant breeding. Nevertheless, lack of knowledge on the role of PA seed reserves in regulating plant growth and in maintaining ion homeostasis hinders such an agronomical application. In this context, we report here that the over-expression of the bacterial phytase PHY-US417 in Arabidopsis leads to a significant decrease in seed PA, without any effect on the seed germination potential. Interestingly, this over-expression also induced a higher remobilization of free iron during germination. Moreover, the PHY-over-expressor lines show an increase in inorganic phosphate and sulfate contents, and a higher biomass production after phosphate starvation. Finally, phosphate sensing was altered because of the changes in the expression of genes induced by phosphate starvation or involved in phosphate or sulfate transport. Together, these results show that the over-expression of PHY-US417 reduces PA concentration, and provide the first evidence for the involvement of PA in the regulation of sulfate and phosphate homeostasis and signaling.

  20. Improving organic phosphate utilization in transgenic white clover by overexpression of Aspergillus niger PhyA gene

    Institute of Scientific and Technical Information of China (English)

    HAN Shengfang; GU Juntao; XIAO Kai

    2007-01-01

    Using the cotyledon of white clover as explants,the transgenic white clover lines ectopic expression of the PhyA gene were established based on Agrobacterium tumefaciens-mediated transformation method.It was found that the tested transgenic lines were all Polymerase Chain Reaction (PCR) positive.The transgenic lines 1 to 4 were used for further Southern blot and Northern blot analysis.The lines 1 and 3 with higher level of PhyA expression were used to assay the phytase activities in root and its intercellular space.When the phytate was the sole phosphorus source,the phytase activities in root in lines 1 and 3 were 31.43% and 44.76% higher than those in control (CK),respectively.Meanwhile,the phytase activities in the root intercellular space in lines 1 and 3 were 3.3-fold and 5.12-fold higher than those in CK,respectively.The phosphorus concentration of plants,the accumulative P amount per plant,plant fresh weight,and plant dry weight were all much higher in lines 1 and 3 than in CK.Thus,it is clearly shown that ectopic expression of Aspergillus niger PhyA gene could significantly increase the ability for white clover to utilize organic phosphate under inorganic phosphate (Pi)-deficient condition.

  1. Effect of amount and source of manganese and/or phytase supplementation on productive and reproductive performance and some physiological traits of dual purpose cross-bred hens in the tropics.

    Science.gov (United States)

    Attia, Y A; Qota, E M; Bovera, F; Tag El-Din, A E; Mansour, S A

    2010-04-01

    1. The effect of different amounts of added manganese (Mn) (0, 10, 20 and 40 mg/kg) in inorganic and organic form and phytase (0 and 300 U) was investigated on productive, reproductive and haematological traits on 480 hens and 60 cocks (Inchas breeds) divided into 12 groups: 10 and 20 Mn x 0 and 300 phytase x 2 Mn sources (8 groups), two negative controls (0 Mn x 2 levels of phytase) and two positive controls (40 Mn x 2 Mn sources). 2. Phytase supplementation increased laying rate by 1.1% and egg weight by 0.4 g. 3. Manganese supplementation at 10 mg/kg over dietary sources improved hatchability, at 20 mg/kg decreased death embryos and abnormality as those of hens supplemented with 40 mg/kg Mn. Inorganic Mn at 10 mg/kg significantly increased egg mass compared to the organic form. Inorganic Mn was more efficient in decreasing abnormal chicks than organic Mn. Phytase supplementation significantly increased hatchability of fertile eggs and decreased the number of abnormal chicks of groups fed on diets unsupplemented with Mn and those supplemented with 10 mg/kg Mn. 4. Mn supplementation at 10 mg/kg over dietary sources significantly improved sperm mass motility and decreased abnormal sperm. Phytase significantly decreased lymphocyte cells and plasma AST. 5. Mn supplementation of the control diet (containing only 16 mg/kg from raw materials) with 20 mg/kg of Mn from either organic or inorganic source is adequate to support egg production traits, egg quality, reproductive traits and economic efficiency of dual purpose cross-bred hens; however, phytase supplementation may reduce the required Mn supplementation to 10 mg/kg.

  2. Impacts of dietary calcium, phytate, and nonphytate phosphorus concentrations in the presence or absence of phytase on inositol hexakisphosphate (IP6) degradation in different segments of broilers digestive tract.

    Science.gov (United States)

    Li, W; Angel, R; Kim, S-W; Brady, K; Yu, S; Plumstead, P W

    2016-03-01

    A total of 1,440 straight-run Heritage 56M × fast-feathering Cobb 500F broiler birds were fed from 11 to 13 d of age to determine the impacts of calcium (Ca), phytate phosphorus (PP), nonphytate P (NPP) and phytase concentrations on the myo-inositol hexakisphosphate (IP6) flow through the different parts of gastrointestinal tract (GIT). The experiment was a 2×2×2×3 randomized block design with 2 Ca (0.7 and 1.0%), 2 PP (0.23 and 0.34%), 2 nPP (0.28 and 0.45%) and 3 phytase (0-, 500-, and 1,000-phytase unit (FTU)/kg) concentrations. The experiment was replicated twice (block) with 3 replicates per treatment (TRT) of 10 birds per block. Concentration of IP6 in crop, proventriculus (PROV) plus (+) gizzard (GIZ) and distal ileum digesta as well as the ileal IP6 disappearance was determined at 13 d of age. In crop, higher IP6 concentration was seen with increased Ca (P phytase, higher dietary PP led to greater IP6 concentration (P phytase were also seen in Prov+Giz and ileum (P phytase improved IP6 degradation but the degree of impact was dependent on nPP and PP (P phytase inclusion significantly reduced IP6 concentration and IP6 disappearance in distal ileum regardless of GIT segments or diet composition, but impacts of dietary Ca, nPP, and PP differed depending on GIT segment examined.

  3. A multi-factors rational design strategy for enhancing the thermostability of Escherichia coli AppA phytase.

    Science.gov (United States)

    Fei, Baojin; Xu, Hui; Cao, Yu; Ma, Shuhan; Guo, Hongxiu; Song, Tao; Qiao, Dairong; Cao, Yi

    2013-05-01

    Despite recent advances in our understanding of the importance of protein surface properties for protein thermostability,there are seldom studies on multi-factors rational design strategy, so a more scientific, simple and effective rational strategy is urgent for protein engineering. Here, we first attempted to use a three-factors rational design strategy combining three common structural features, protein flexibility, protein surface, and salt bridges. Escherichia coli AppA phytase was used as a model enzyme to improve its thermostability. Moreover, the structure and enzyme features of the thermostable mutants designed by our strategy were analyzed roundly. For the single mutants, two (Q206E and Y311K), in five exhibited thermostable property with a higher success rate of prediction (40 %). For the multiple mutants, the themostable sites were combined with another site, I427L, we obtained by directed evolution, Q206E/I427L, Y311K/I427L, and Q206E/Y311K/I427L, all exhibited thermostable property. The Y311K/I427L doubled thermostability (61.7 %, and was compared to 30.97 % after being heated at 80 °C for 10 min) and catalytic efficiency (4.46 was compared to 2.37) improved more than the wild-type AppA phytase almost without hampering catalytic activity. These multi-factors of rational design strategy can be applied practically as a thermostabilization strategy instead of the conventional single-factor approach.

  4. Display of phytase on the cell surface of Saccharomyces cerevisiae to degrade phytate phosphorus and improve bioethanol production.

    Science.gov (United States)

    Chen, Xianzhong; Xiao, Yan; Shen, Wei; Govender, Algasan; Zhang, Liang; Fan, You; Wang, Zhengxiang

    2016-03-01

    Currently, development of biofuels as an alternative fuel has gained much attention due to resource and environmental challenges. Bioethanol is one of most important and dominant biofuels, and production using corn or cassava as raw materials has become a prominent technology. However, phytate contained in the raw material not only decreases the efficiency of ethanol production, but also leads to an increase in the discharge of phosphorus, thus impacting on the environment. In this study, to decrease phytate and its phosphorus content in an ethanol fermentation process, Saccharomyces cerevisiae was engineered through a surface-displaying system utilizing the C-terminal half of the yeast α-agglutinin protein. The recombinant yeast strain, PHY, was constructed by successfully displaying phytase on the surface of cells, and enzyme activity reached 6.4 U/g wet biomass weight. Ethanol productions using various strains were compared, and the results demonstrated that the specific growth rate and average fermentation rate of the PHY strain were higher 20 and 18 %, respectively, compared to the control strain S. cerevisiae CICIMY0086, in a 5-L bioreactor process by simultaneous saccharification and fermentation. More importantly, the phytate phosphorus concentration decreased by 89.8 % and free phosphorus concentration increased by 142.9 % in dry vinasse compared to the control in a 5-L bioreactor. In summary, we constructed a recombinant S. cerevisiae strain displaying phytase on the cell surface, which could improve ethanol production performance and effectively reduce the discharge of phosphorus. The strain reported here represents a useful novel engineering platform for developing an environment-friendly system for bioethanol production from a corn substrate.

  5. Research on the progress of phytase in livestock-poultry husbandry%植酸酶在畜禽养殖上的研究进展

    Institute of Scientific and Technical Information of China (English)

    欧阳增理; 杨红兰

    2012-01-01

    Phytase is a kind of feed additive for livestock and poultry, which can degrade phytate into inosito and inorganic phosphorus effectively. It can remote the antiaction factor of phytate in diets. This article reviewed the sources, influence factor of enzyme action and application of phytase in livestock and poultry industry.%植酸酶作为一种畜禽饲料添加剂,能有效地将植酸磷降解为肌醇和无机磷,消除植酸的抗营养作用。本文综述了植酸酶的.来源、影响酶作用的因素及其在畜禽生产中的应用。

  6. Investigation of the interaction between separate calcium feeding and phytase supplementation on growth performance, calcium intake, nutrient digestibility and energy utilisation in broiler starters

    DEFF Research Database (Denmark)

    Abdollahi, M.R.; Duangnumsawang, Y.; Kwakkel, R.P.

    2016-01-01

    The interaction between separate calcium (Ca) feeding and phytase supplementation on performance, coefficient of apparent ileal digestibility (CAID) of nitrogen (N), starch, fat and phosphorus (P), total tract retention (TTR) of Ca and P, and apparent metabolisable energy (AME) in broiler starters...... assigned to 10 dietary treatments. Birds were also provided with a source of Ca in a separate feed trough. Increasing dietary Ca concentration decreased (P supplemented......, but the total Ca intake in phytase-supplemented diets were similar (P > 0.05). Birds fed 1.3 g Ca/kg diet showed the lowest (P 0.05) to the diets with 4.3 g Ca/kg. No significant (P > 0.05) effect of dietary Ca concentration on the CAID of starch, fat and GE was found...

  7. Application of Phytase in Corn Grain-to- ethanol Fermentation%植酸酶在玉米酒精发酵中的应用研究

    Institute of Scientific and Technical Information of China (English)

    李娜; 吕伟民; 赵姗姗; 夏海华

    2013-01-01

      探讨植酸酶在酒精发酵中的应用。实验室小试结果表明:在糖化时加入植酸酶,添加量为每克原料11个单位,作用 pH 为4.5,发酵温度为58~60℃,淀粉出酒率可提高1.5%,发酵周期可缩短4h。%The application of phytase in ethanol fermentation was discussed. The experimental results on laboratory scale showed that the ethanol yield was increased by 1.5% and the fermentation period was shorted for 4 hours, when 11u phytase per gram raw material was added under the pH 4.5 and temperature at 58-60℃ during saccharification.

  8. 植酸酶颗粒剂生产工艺的研究%Study on Production Technology of Phytase Granules

    Institute of Scientific and Technical Information of China (English)

    张渊舒; 罗琴; 攸德伟

    2011-01-01

    Through optimizing the external conditions influential to role of phytase, selecting low- cost coating materials and simple coating technology, the author developed the coated and temperature - tolerant type phytase granule Ruifeng P 5000 ( Reyphon P 5000 -CT).The main technical innovations of Reyphon P 5000 -CT are: using the coating technology to indirectly improve the heat resistance of phytase; optimizing the formula of the assisted materials.This product has obviously better temperature resistance, stability of granule - preparing, storage stability and granule uniformity than the similar domestic products.%笔者采用优化影响植酸酶发挥作用的外部条件的方法,通过选择成本低廉的包衣材料和简单的包衣工艺,研制开发了包衣耐温型植酸酶颗粒剂瑞丰磷5000(Reyphon P 5000-CT),其主要技术创新点为:采用包衣技术间接提高植酸酶的耐热性、优化辅料配方.该产品在耐温性、制粒稳定性、储藏稳定性以及颗粒均匀度等方面都远远优于国内同类产品.

  9. Glycosylation of fungal phytase affects its resistance to trypsin%糖基化影响真菌植酸酶的胰蛋白酶耐受性

    Institute of Scientific and Technical Information of China (English)

    朱庆锋; 王志林; 崔百元; 张琪; 陈庄; 晏石娟; 贝锦龙

    2016-01-01

    通过重叠聚合酶链式反应,将黑曲霉植酸酶(Aspergillus niger NRRL 3135 phytase)第238~337位与第382~444位多肽片段的基因编码序列替换为烟曲霉植酸酶(Aspergillus fumigatus ATCC 13073 phytase)中的对应序列,构建片段置换植酸酶(Fragments-replaced phytase)基因.将插入该基因的pGAPZα A重组质粒转化入毕赤酵母.通过离子交换与凝胶过滤纯化,获得重组酵母分泌的活性片段置换植酸酶.与黑曲霉植酸酶相比,片段置换显著提高了片段置换植酸酶的胰蛋白酶耐受性.而通过脱糖基化酶PNGase彻底去除N-糖基化,可恢复片段置换植酸酶对胰蛋白酶的敏感性.上述结果表明烟曲霉植酸酶对应片段上的N-糖基化修饰可显著提高毕赤酵母表达的黑曲霉植酸酶的胰蛋白酶耐受性.

  10. 耐高温植酸酶的热稳定性评估方法分析%Assessments of the thermostability of thermostable phytase

    Institute of Scientific and Technical Information of China (English)

    王海燕; 汤海鸥; 王晓睿; 杨禄良; 刘影

    2013-01-01

    耐高温植酸酶因热稳定性好、经饲料制粒后保存率高而应用日益广泛,但是目前尚无评估耐高温植酸酶热稳定性的统一的、公认的标准方法。文章结合目前评估植酸酶热稳定性的方法及本实验室的实际检测结果,分别介绍了在实验室和制粒条件下的评估情况,以便为植酸酶的热稳定性评估提供参考依据。%Thermostable phytase has become increasingly prevalent with its high heat-resistance and higher survival rate after feed pelleting, but currently there was no unified and accepted standard for assessing the thermostability of thermostable phytase. In this article we introduced the assessment under the lab and granulation condition, combined with the assessment for the thermostability of phytase and our actual detection results, we wish to arouse further communica-tion and exploration of this kind of assessments.

  11. Research Advance in Classification and Enzymological Characteristics of Microbial Phytase in the Rumen%瘤胃微生物植酸酶的分类与酶学性质研究进展

    Institute of Scientific and Technical Information of China (English)

    沈莎莎; 杨红建; 张晓明

    2011-01-01

    Ruminants have the ability to utilize phytate phosphate by rumen microorganisms. Until now, domestic research on ruminal microbe phytase is insufficient. In this paper, phytase origin and activity in the rumen was briefly reviewed, and five kinds of rumen bacteria were capable of producing phytase. Among these bacteria, phytase activity was particularly prevalent in Sele-nomonas ruminantium (199. 1-703. 6 U ? mL-1). Based on their optimal pH and catalytic mechanisms, ruminal phytases are classified into acid phytase and cysteine phosphatases phytase, respectively. According to the stereospecificity of phytase, rumen microbes may exist 3-phytase and 5-phytase. Furthermore, kinetics of enzymatic reaction characteristics of phytases were summarized and discussed comprehensively. In a brief, phytase of S. Ruminantium has a optimal temperature of 50-55 'C , an optimal pH range of 4. 0-5. 5, and exhibits the highest specificity constant for wryo-inositol hexakisphosphate and ATP. The optimal phytase activity of Megasphaera elsdenii is displayed at pH5. 0 and 60 °C , and it also displays strict specificity for mjyo-inositol hexakisphosphate.%反刍动物能很大程度上依靠瘤胃微生物来消化利用饲料中的植酸盐,但目前国内有关瘤胃微生物植酸酶的研究报道较少.本文对瘤胃微生物植酸酶的主要来源和活性进行了综述分析,发现主要有5种瘤胃细菌能分泌植酸酶,其中,反刍兽新月形单胞菌(Selenomonas ruminantium)分泌的植酸酶活性最高(199.1~703.6 U·mL-1);根据酶促反应的最适pH和催化机理,可将瘤胃微生物植酸酶分别归类为酸性植酸酶和半胱氨酸磷酸酶植酸酶;根据植酸酶水解植酸时的立体专一性,发现瘤胃微生物植酸酶中可能同时存在3-植酸酶和5-植酸酶2类植酸酶.最后本文论述了瘤胃微生物植酸酶酶促反应动力学特性,发现反刍兽新月形单胞菌植酸酶的最适pH为4.0~5.5、最适温度为50~55

  12. Available phosphorus levels in diets supplemented with phytase for male broilers aged 22 to 42 days kept in a high-temperature environment

    Directory of Open Access Journals (Sweden)

    Tarciso Tizziani

    2016-02-01

    Full Text Available ABSTRACT This study was conducted to evaluate the effect of reduction of the available phosphorus (avP in diets supplemented with 500 FTU/kg phytase on performance, carcass characteristics, and bone mineralization of broilers aged 22 to 42 days kept in a high-temperature environment. A total of 336 Cobb broilers with an average initial weight of 0.883±0.005 kg were distributed in a completely randomized design with six treatments - a positive control (0.354 and 0.309% avP without addition of bacterial phytase for the phases of 22 to 33 and 34 to 42 days, respectively, and another five diets with inclusion of phytase (500 FTU and reduction of the level of avP (0.354, 0.294, 0.233, 0.173, and 0.112%; and 0.309, 0.258, 0.207, 0.156, and 0.106% for the phases of 22 to 33 and 34 to 42 days, respectively - eight replicates, and seven birds per cage. The experimental diets were formulated to meet all nutritional requirements, except for avP and calcium. Birds were kept in climatic chambers at a temperature of 32.2±0.4 °C and air humidity of 65.3±5.9%. Phytase acted by making the phytate P available in diets with reduction in the levels of avP, keeping feed intake, weight gain, feed:gain, and carcass characteristics unchanged. Treatments affected ash and calcium deposition and the Ca:P ratio in the bone; the group fed the diets with 0.112 and 0.106%, from 22 to 33 and 34 to 42 days of age, respectively, obtained the lowest values, although the phosphorus deposition in the bone was not affected. Diets supplemented with 500 FTU of phytase, with available phosphorus reduced to 0.173 and 0.156%, and a fixed Ca:avP ratio of 2.1:1, meet the requirements of broilers aged 22 to 33 and 34 to 42 days, respectively, reared in a high-temperature environment.

  13. Influence of dietary calcium level, calcium source, and phytase on bird performance and mineral digestibility during a natural necrotic enteritis episode.

    Science.gov (United States)

    Paiva, D M; Walk, C L; McElroy, A P

    2013-12-01

    The objective of this study was to determine the influence of Ca source [highly soluble calcified seaweed (HSC) or limestone], phytase supplementation, and dietary levels of Ca on bird performance and mineral digestibility (Ca and P) during a necrotic enteritis (NE) episode. Cobb 500 male broilers were weighed and randomized into 8 treatment groups (9 pens/treatment; 30 birds/pen) at day of hatch. The 21-d trial was designed as a 2 × 2 × 2 factorial, which included 2 dietary levels of Ca (0.6 and 0.9%), 2 Ca sources (limestone or HSC), and 2 levels of an Escherichia coli phytase (0 or 1,000 FTU/kg). One unit of phytase (FTU) is defined as the quantity of enzyme that releases 1 μmol of inorganic phosphorus/min from 0.00015 mol/L of sodium phytate at pH 5.5 at 37° C. Birds were placed on used litter from a previous flock that exhibited clinical signs of NE. Birds and feed were weighed on d 7, 14, and 21, and BW gain, feed intake, and feed conversion were calculated for each of these periods and cumulatively. Mortality was recorded daily and pH of the gizzard and duodenum were measured on d 7, 14, and 21. Ileal digesta (8 birds/pen) was collected on d 7, 14, and 21. Significance is reported at P phytase were heavier than the other treatments regardless of Ca source. Broilers fed diets formulated with HSC had significantly higher feed conversion then broilers fed diets formulated with limestone. The gizzard of broilers fed 0.9% Ca in the diet was significantly less acidic than the gizzard of broilers fed 0.6% Ca in the diet. Broilers fed 0.6% Ca in diets supplemented with phytase showed significant improvements in P and Ca digestibility. In conclusion, higher dietary Ca (0.9% vs. 0.6%) had a negative effect on mortality associated with NE and on bird performance.

  14. Qualitative and Quantitative PCR Detection of the Phytase (phyA2)Transgenic Corn%转植酸酶基因(phyA2)玉米定性、定量PCR检测

    Institute of Scientific and Technical Information of China (English)

    于彩虹; 田少亭; 路兴波; 李凡; 杨淑珂; 孙红炜

    2012-01-01

    The detection of phytase transgenic corn is important to supervise its release and provides technical support. According to phytase gene (phyA2) sequences in transgenic phyA2 corn (Zea mays), the primers for qualitative and quantitative were designed. The qualitative PCR results showed that the primers were only effective to detect phyA 2 transgenic corn, which indicated that the primers were highly specific. The SYBR Green I fluorescence quantitative PCR of endogenous reference zSSIIb zea mays starch synthase isoform zSTSII-2 and phyA2 genes were studied in this paper. Transgenic contens were calculated according to the standard Ci-copies linear graphs of the two genes. The results showed that the standard curves of zSSIIb and phyA2 genes had higher R2 value as 0.998 and 0.995, respectively, and the melting curves had single peak which mean the primers were specific. The mixed corn samples were detected to be 1.1%, 0.54% and 0.17%, respectively, which were close to the true values. The qualitative and quantitative PCR methods were developed in this study which can be effective at detecting phyA 2 transgenic corn.%转植酸酶基因玉米检测是对转植酸酶基因玉米释放进行有效监管的重要前提和技术支撑.本研究根据转植酸酶基因玉米(Zea mays)外源植酸酶基因phyA2序列设计基因特异性引物,进行了定性、定量PCR检测.定性结果显示,该引物可有效检测植酸酶玉米中phyA2基因,而在其他不含植酸酶基因的材料中则检测不到相应基因,表明该引物具有很高的特异性.以玉米内标准基因玉米淀粉合酶异构体zSTSII-2基因(zSSIIb)和植酸酶基因phyA2为对象,进行SYBR Green Ⅰ染料法荧光定量PCR反应,绘制2种基因扩增循环数-拷贝数标准曲线图,根据混合样品中(zSSIIb)和植酸酶基因phyA2的拷贝数计算样品中的转基因含量,并做熔解曲线分析.结果表明,zSSIIb和phyA2标准曲线相关系数分别为0.998和0.995,相关性

  15. Preliminary Investigation of Myo-Inositol Phosphates Produced by ASUIA279 Phytase on MCF-7 Cancer Cells

    Directory of Open Access Journals (Sweden)

    N. Mohd. Yusoff

    2011-12-01

    Full Text Available Phytate or myo-inositol hexakisphosphates (IP6 is widely distributed in plants like rice brans. The production of myo-inositol phosphate intermediates has received much attention due to the remarkable potential health benefits offered by the compounds. In this study, the cytotoxicity of the partially purified myo-inositol phosphate fractions and commercial IP1 and IP6 were investigated against MCF-7 breast cancer cell lines. The study showed that the commercial standard IP1 and IP6 showed good inhibition towards the MCF-7 cell line. The MCF-7 cells growth was inhibited in minimum concentration of myo-inositol phosphates (<1000 µg/ml. However, no inhibition observed on the MCF-7 cell line by the myo-inositol phosphates fractions partially purified from rice bran at concentration <1000 ?g/ml. The inhibition of MCF-7 was only observed at concentration more than 30 mg/ml with more than 40% cells were inhibited. This indicates that the partially purified rice bran myo-inositol phosphates degraded by ASUIA279 phytase on MCF-7 breast cancer cells exhibit positive results towards the inhibition of cancer cells growth at relatively high concentration..KEYWORDS: myo-inositol phosphates, phytase, MCF-7,  cancerABSTRAK: Fitat atau myo-inositol hexakisphosphate (IP6 dikenali umum teragih di dalam tumbuhan seperti dedak padi. Penghasilan perantaraan fosfat myo-inositol mendapat perhatian memandangkan ia berpotensi tinggi dalam kesihatan. Dalam kajian ini, kesitotoksikan sebahagian daripada fosfat myo-inositol separa tulen, IP1 komersil dan IP6 komersil dikaji terhadap produk yang berupa sel kekal (cell lines kanser payu dara MCF-7. Tumbesaran sel MCF-7 direncatkan dalam pekatan minima fosfat myo-inositol (<1000 μg/ml. Tetapi, tidak ada perencatan dilihat terhadap sel kekal MCF-7 oleh sebahagian fosfat myo-inositol separa tulen daripada dedak padi pada kepekatan <1000 mg/ml. Perencatan MCF-7 hanya dilihat pada kepekatan lebih daripada 30 mg/ml dengan lebih

  16. Effects of Different Germination Conditions on Phytase Activity on Faba Bean (Vicia faba L.)%不同发芽条件对蚕豆植酸酶活性的影响

    Institute of Scientific and Technical Information of China (English)

    罗羽洧; 解卫华; 马娟; 郁志芸

    2012-01-01

    以蚕豆为试材,研究发芽过程中植酸酶在蚕豆不同部位的变化和性质.发芽蚕豆不同部位植酸酶活性呈现先高后低的变化趋势,子叶植酸酶在发芽第6天达最高值.超声波处理对发芽蚕豆胚根、子叶和胚芽植酸酶活性有一定促进作用,其中以15min超声波处理对胚根和胚芽植酸酶活性影响最大,5min处理、10min处理和15min处理均对子叶植酸酶活性有显著影响.在发芽过程中不同光照对蚕豆进行处理,不论胚根、子叶还是胚芽,黑暗处理植酸酶活性高于光照处理.核酸抑制剂放线菌素(Act-D)和蛋白质抑制剂环已酰胺(CHM)对发芽蚕豆胚根、子叶和胚芽中的植酸酶活性有一定影响,其中以CHM的抑制效果最为明显.%Phytase activity and property in different faba bean fractions during germination were investigated with faba bean as the testing piece. The phytase activity appeared a change trend of increasing first then decreased with the maximum value at the sixth day during germination in cotyledon. Ultransonic treatments had facilitations on phytase activity in cotyledon, radicle and plumule among which 15 min treatment has the biggest effect on phytase activity in radicle and plumule. 5 min treatment, 10 min treatment and 15 min treatment had obvious effects on phytase activity in cotyledon. Whether cotyledon,radicle and plumule,faba bean during darkness treatment had higher phytase activity than that of different illuminations during germination. Act - D and CHM had certain effects on cotyledon, radicle and plumule of phytase activity in faba bean, among which, the CHM had the largest inhibition effect on phytase activity.

  17. The gene engineering of feed phytase%饲用植酸酶的基因工程

    Institute of Scientific and Technical Information of China (English)

    刘梅英; 彭健

    2006-01-01

    植酸酶即肌醇六磷酸水解酶,是催化植酸及植酸盐水解成肌醇与磷酸(或磷酸盐)的一类酶的总称(Ullah AH.,1996)。植酸酶广泛存在于微生物、植物和动物中,但猪与家禽等单胃动物体内缺乏该酶。目前商品化的植酸酶均来源于微生物,主要有黑曲霉(Aspergillus niger)(EC3.1.3.8)、无花果曲霉(Aspergillus ficuum)(EC 3.1.3.5)和米曲霉(Aspergillus oryzae)(EC3.1.3.26)。另报道还有聚多曲霉菌(Aspergillus sydowl)、枯草芽孢杆菌(Bacillus subtills)及隔孢伏革菌(Peniphora lycii)等,但以黑曲霉和无花果曲霉的效率较高(张若寒,2002)。

  18. In vitro probiotic evaluation of phytase producing Lactobacillus species isolated from Uttapam batter and their application in soy milk fermentation.

    Science.gov (United States)

    Saraniya, Appukuttan; Jeevaratnam, Kadirvelu

    2015-09-01

    Probiotic lactic acid bacteria are health promoters and have been traditionally consumed without the knowledge that they have beneficial properties. These bacteria mainly involve in secreting antimicrobials, enhance immune-modulatory effects, and preserve the intestinal epithelial barrier by competitively inhibiting the pathogenic organisms. The aim of this study was to investigate the in vitro probiotic properties of Lactobacillus pentosus, Lactobacillus plantarum, Lactobacillus plantarum ssp. argentoratensis, and Lactobacillus plantarum ssp. plantarum isolated from fermented Uttapam batter. The isolates produced bacteriocins that were effective against several pathogens. All the isolates exhibited tolerance to bile, gastric, and intestinal conditions. Beneficial properties like cholesterol assimilation and production of enzymes such as β-galactosidase, phytase and bile hydrolase varied among the isolates. Four isolates from each sub-species effectively adhered to Caco-2 cells and prevented pathogen adhesion. Using these strains, the soy milk was fermented, which exhibited higher antioxidant activity, 2,2-diphenylpicrylhydrazyl (DPPH) scavenging activity and decreased phytate content when compared to unfermented soy milk. Thus, these probiotic isolates can be successfully used for formulation of functional foods that thereby help to improvise human health.

  19. Phytase enzyme in diets containing defatted rice bran for growing swine Enzima fitase dietas com farelo de arroz desengordurado para suínos em crescimento

    Directory of Open Access Journals (Sweden)

    José Aparecido Moreira

    2003-12-01

    Full Text Available Organic phosphorus is poorly utilized by monogastric animals because they lack phytase, the enzyme that cleaves the ortho-phosphate groups from the phytate molecule. Diets fed to pigs are supplemented with inorganic P, and this can increase environmental pollution and diet costs. Sixty mixed sex, half-breed pigs, were used to evaluate the effect of increasing dietary levels of phytase (253, 759, 1265 and 1748 PU kg-1 feed on animal performance as compared to a control without phytase but supplemented with dicalcium phosphate. Enzyme levels did not affect daily feed intake, food conversion, average daily weight gain, plasma P and Ca, calcium and phosphorus in bone ash, and the calcium/phosphorus ratio in the plasma and bones. A quadratic relationship between phytase levels and the percentages of P and Ca in bone ash was observed, reaching a maximum at the 880 and 879 PU levels, respectively. Animals fed diets containing phytase presented low plasma P values when compared to the control, but no effects were observed for the regression analysis. Using 759 PU phytase in rations containing corn, soybean bran and defatted rice bran for growing pigs can eliminate the use of traditional sources of P.Os animais monogástricos não aproveitam eficientemente o P orgânico das dietas, pois não sintetizam a enzima fitase, sendo necessária a suplementação das rações com P inorgânico, podendo elevar o custo das dietas e a poluição ambiental. Foram utilizados 60 leitões mestiços (machos castrados e fêmeas para avaliar a eficácia dos níveis dietéticos crescentes da enzima fitase (253, 759, 1265 e 1748 UF kg-1 de ração sobre os parâmetros de desempenho e comparar com o tratamento testemunha que diferia dos demais por não conter fitase e por conter fosfato bicálcico. Os níveis da enzima fitase não afetaram o consumo diário de ração, conversão alimentar, ganho diário de peso, P e Ca no plasma, cinzas no osso e relação cálcio e f

  20. The effects of sow parity on digestibility of proximate components and minerals during lactation as influenced by diet and microbial phytase supplementation.

    Science.gov (United States)

    Kemme, P A; Radcliffe, J S; Jongbloed, A W; Mroz, Z

    1997-08-01

    Ninety-six (Finnish Landrace x Dutch Landrace) reproductive sows were used at parities 1, 3, 5, and 7 + 8 from d 107 of gestation to d 21 of lactation to investigate the effects of diet and parity on apparent total tract digestibility (ATTD). Animals were randomly assigned to one of four dietary treatments. Dietary treatments were 1) a P-deficient (1.1 g digestible P [dP]/kg) Dutch semipractical diet; 2) Diet 1 supplemented with 400 FTU Aspergillus niger phytase per kilogram of diet (1.7 g dP/kg); 3) a corn-soybean meal-based diet (1.3 g dP/kg); and 4) Diet 3 supplemented with extra monocalcium phosphate (MCP; 2.4 g dP/kg of diet). Animals were fed twice daily at 2.8 times maintenance (418 kJ ME/ BW75) from d 8 to the end of lactation. Feces and urine were collected during d 11 to 13 and d 18 to 20 of lactation. The ATTD of DM, OM, ash, CP, Ca, Mg, and total P (P supplemented with phytase. Addition of MCP enhanced total P ATTD by an average of 6.7 percentage units. Addition of microbial phytase improved Ca, Mg, and total P ATTD, but the effects were dependent on the stage of lactation. Lower ATTD of OM and CP were seen for first parity animals compared with higher parity sows. The ATTD of Mg increased with increasing parity. Parity had little effect on the ATTD of minerals during lactation, and dietary effects were prominent and followed a similar trend to those seen in growing-finishing pigs.

  1. Screening of strain producing neutral phytase and its enzymatic characteristics%中性植酸酶菌株的筛选及性质

    Institute of Scientific and Technical Information of China (English)

    朱启忠; 王文君; 黄梅娜; 马敏; 徐海峰

    2012-01-01

    About 20 neutral phytase-producing strains out of natural soil were screened. A new strain phy7 was isolated. It was identified as Pseudomonas aeruginosa by the phosphorus molybdenum blue method and 16S rDNA sequence analysis. The characteristics of the phytase were investigated. The new strain produced phytase activity and the optimal conditions were pH 7. 0 and 40 ℃. For substrate dodeca-sodium phytate, Km was 0. 26 mmol/L and vmax was 0. 050 6 nmol/min at 40 ℃. The enzymatic activity was inhibited by Zn2+ , Al3+ , Cu 2+ , and Mn 2+ , but activated by Fe.%从天然土壤中筛选出产胞外中性植酸酶的细菌20余株,通过钼蓝法对菌种进行复筛,确定phy7为研究菌.通过16S rDNA测序分析方法鉴定该菌株属于绿脓假单胞菌属.结果表明:该菌株分泌中性植酸酶,其最适反应pH为7.0、最适反应温度为40℃;在37℃时以植酸钠为底物的Km为0.26 mmol/L,vmax为0.050 6 nmol/min.金属离子Zn2+、Al3+、Cu2+和Mn2+等对该酶有抑制作用,而Fe2+等则有一定的激活作用.

  2. Effect of citric acid and microbial phytase enzyme on ileal digestibility of some nutrients in broiler chicks fed corn-soybean meal diets

    Directory of Open Access Journals (Sweden)

    Moslem Bashtani

    2012-01-01

    Full Text Available A total of 270 Ross 308 male broilers were used in a randomized complete block design for a period of five weeks. The experiment included nine treatments, three replicates with 10 birds in each replicate of 3×3 factorial design. Two main factors of citric acid (CA (0, 3 and 6% and a microbial phytase (MP (0, 500 and 1000 IU/kg were used. The results indicated that diets containing 3% CA caused significant increase in P content in tibia ash, ileal digestibility of crude protein, apparent metabolizable energy, total phosphorus and body weight (P

  3. Glycosylations and truncations of functional cereal phytases expressed and secreted by Pichia pastoris documented by mass spectrometry

    DEFF Research Database (Denmark)

    Dionisio, Giuseppe; Jørgensen, Malene; Welinder, Karen Gjesing;

    2012-01-01

    been expressed in Pichia pastoris from constructs encoding the alpha-mating factor at the N-termini and a His6 tag before the stop codon in all constructs. A protein chemical study of a C-terminally truncated recombinant wheat phytase, r-TaPAPhy_b2, was carried out to clarifying the posttranslational...... processing of proteins secreted from P. pastoris. Extensive mass spectrometric sequencing of tryptic, chymotryptic and AspN derived peptides of both the native and endoH deglycosylated forms showed: (i) All mating factor derived sequence had been removed and further unspecific proteolysis left highly...

  4. Assessing changes in the composition of broiler litters from commercial poultry units in Northern Ireland following the adoption of phytase in diets.

    Science.gov (United States)

    Foy, R H; Ball, M E E; George, J

    2014-11-01

    Microbial phytases increase the bioavailability of phytate P in poultry diets, and a survey was undertaken to determine if their use had lowered the P composition of broiler litter in Northern Ireland compared with standard values of litter composition listed in the current United Kingdom fertilizer recommendations. Litter samples were collected from a total of 20 units across Northern Ireland in 2010 and analyzed for DM, N, phosphate (P2O5), potash (K2O), magnesium oxide (MgO), water-soluble P (WSP), ammonium N (NH4N), and uric acid N. Dry matter of litter was positively correlated (P light of the changing composition of broiler litter.

  5. Digestibility of nutrients in growing-finishing pigs is affected by Aspergillus niger phytase, phytate and lactic acid levels. 2. Apparent total tract digestibility of phosphorus, calcium and magnesium and ileal degradation of phytic acid

    NARCIS (Netherlands)

    Kemme, P.A.; Jongbloed, A.W.; Mroz, Z.; Kogut, J.; Beynen, A.C.

    1999-01-01

    In growing-finishing pigs, the effects of supplemental microbial phytase, lactic acid and Na phytate in a maize-soybean meal based diet on the apparent total tract digestibility (ATTD) of ash, total P, Ca and Mg and the ileal degradation (AID) of phytic acid were studied. The experimental design was

  6. Effects of Supplemental Dietary Phytase and 25-Hydroxycholecalciferol on the Performance Characteristics of Commercial Layers Inoculated before or at the Onset of Lay with the F-Strain of Mycoplasma gallisepticum

    Science.gov (United States)

    The effects of dietary supplementation with phytase and 25-hydroxycholecalciferol on the performance characteristics of commercial layers that were inoculated prelay (12 wk of age) or at the onset of lay (22 wk of age) with F-strain Mycoplasma gallisepticum were assessed. Experimental layer diets, w...

  7. Influence of supplemental dietary poultry fat, phytase, and 25-hydroxycholecalciferol on the egg characteristics of commercial layers inoculated before or at the onset of lay with F-strain Mycoplasma gallisepticum

    Science.gov (United States)

    The effects of 2 supplemental levels of dietary poultry fat (PF) and the combination of PF, phytase (PHY) and 25-hydroxycholecalciferol (D3) on the egg characteristics of commercial layers inoculated with F-strain Mycoplasma gallisepticum (FMG) were investigated in 2 trials. Sham and FMG inoculatio...

  8. Effects of Supplemental Dietary Phytase and 25-Hydroxycholecalciferol on the Digestive and Reproductive Organ Characteristics of Commercial Layers Inoculated Before or at the Onset of Lay with the F-Strain of Mycoplasma galli

    Science.gov (United States)

    In 3 trials, the effects of dietary supple mentation with phytase (PHY) and 25-hydroxycholecalciferol (25-D3) on the digestive and reproductive organ characteristics of commercial layers that were inoculated pre-lay (12 wk of age) or at the onset of lay (22 wk of age) with F-strain Mycoplasma gallis...

  9. Influence of Supplemental Dietary Poultry Fat, Phytase, and 25-Hydroxycholecalciferol on the Performance of Commercial Layers Inoculated Before or at the Onset of Lay with F-Strain Mycoplasma gallisepticum

    Science.gov (United States)

    The effects of 2 levels of supplemental dietary poultry fat (PF) and the combination of PF, phytase (PHY) and 25-hydroxycholecalciferol (D3) on the performance of commercial layers inoculated with F-strain Mycoplasma gallisepticum (FMG) were investigated in 2 trials. Sham and FMG inoculations were ...

  10. Dietary poultry fat, phytase, and 25-hydroxycholecalciferol influence the digestive and reproductive organ characteristic of commercial...at the onset of lay with F-strain Mycoplasma gallisepticum 1 , 2

    Science.gov (United States)

    ABSTRACT Effects of 2 supplemental concentrations of dietary poultry fat (PF) and the combination of PF, phytase (PHY) and 25-hydroxycholecalciferol [25(OH)D] on the gross digestive and reproductive organ characteristics of commercial layers inoculated with F-strain Mycoplasma gallisepticum (FMG) w...

  11. Influence of supplemental dietary poultry fat, phytase, and 25-hydroxycholecalciferol on the blood characteristics of commercial layers inoculated before or at the onset of lay with F-strain Mycoplasma gallisepticum

    Science.gov (United States)

    The effects of 2 supplemental levels of dietary poultry fat (PF) and the combination of PF, phytase (PHY) and 25-hydroxycholecalciferol [25(OH)D] on the blood characteristics of commercial layers inoculated with F-strain Mycoplasma gallisepticum (FMG) were investigated in 2 trials. Sham and FMG ino...

  12. Effect of transgenic phytase corn on growth performance and Ca-P metabolism in broiler%转植酸酶基因玉米对肉仔鸡生长性能及钙磷代谢的影响

    Institute of Scientific and Technical Information of China (English)

    张军民; 邓丽青; 陈茹梅; 马永喜

    2011-01-01

    The effect of transgenic phytase corn on growth performance and Ca-P metabolism were investigated in broilers. 480 AA broilers (1-day-old) were divided into four treatments of three replicates each. The control group was fed CaHPO4 without phytase;the test groups were fed half of CaHPO4 ,test Ⅰ , Ⅱ and Ⅲ were fed 500 U/kg of exogenous phytase,500 U/kg and 750 U/kg phytase which was from transgenic phytase corn,respectively. The results showed that broilers' health status in each group were normal during the trial,there was no significant difference (P>0.05) in the average daily gain and FCR (feed conversion rate) ;each group of calcium,phosphorus and nitrogen in serum and tibia was no significant difference (P>0.05) ;the retention of phosphorus in faeces in test group at days 21 and 42 after treatment were lower than that of the control group by 49.43% ,55.17% ,54.02% and 31.50%, 32.28 %, 14.96 % respectively, which was significantly different (P<0.01) ;there was no significant difference (P>0.05) between transfer phytase corn group and the microbial phytase. Transgenic phytase corn as a source of phytase can replace the exogenous phytase in broiler diets,not only does not affect the production performance, but also can reduce the use level of CaHPO4 and N-P emissions.%选择480只AA商品代肉仔鸡,随机分为4组,每组3个重复,对照组添加磷酸氢钙,试验各组磷酸氢钙减半,分别添加500 U/kg的外源植酸酶和500、750 U/kg转植酸酶基因玉米来源的植酸酶.试验结果表明:肉仔鸡试验期间健康状况正常,各组平均日增重、饲料转化率均无显著差异(P>0.05);各组间血清和胫骨中钙、磷、氮水平无显著差异(P>0.05);试验组21和42日龄时粪中磷的存留量分别比对照组降低49.43%、55.17%、54.02%和31.50%、32.28%、14.96%,差异极显著(P<0.01);转植酸酶基因玉米组与微生物植酸酶组相比差异不显著(P>0.05).转

  13. Effect of germination conditions on the phytate content and phytase activity of barley%不同发芽条件对大麦中植酸含量及植酸酶活力的影响

    Institute of Scientific and Technical Information of China (English)

    张善飞; 董亮; 李欣; 张利; 赵长新

    2012-01-01

    The key factors of germinating temperature, pH value of leaching water and the varieties and content of metal ions in leaching water in the process of barley sprout affected the content of phytic acid and the activities of phytase notably. Result showed that,at the germination temperature of 16X2 , phytase was activated earlier than higher or lower temperature, phytase activity reached to 4. 032 8 U/g dry base ,which was more conducive to the decomposition of phytic acid. When leaching water was under neutral conditions, phytase activity increased rapidly. Adding Ca2+ , Mg2+ , Fe3+ and other metal ions during leaching peried had a greater inhibitory effect on phytase activity.%大麦发芽过程中,发芽温度、浸麦水pH值及浸麦水中金属离子的种类和含量是影响大麦中植酸含量及植酸酶活力的重要因素.实验发现,当发芽温度为16℃时,植酸酶较高温和低温发芽更早的被激活,其活力达到4.032 8 U/g(绝干),更加有利于植酸的分解;在浸麦水为中性条件下浸麦,植酸酶活力上升较快,植酸含量迅速下降;在浸麦期间添加Ca2+、Mg2和Fe3+等金属离子对植酸酶的活力有较大抑制作用.

  14. Effect of phytate, microbial phytase, fiber, and soybean oil on calculated values for apparent and standardized total tract digestibility of calcium and apparent total tract digestibility of phosphorus in fish meal fed to growing pigs.

    Science.gov (United States)

    González-Vega, J C; Walk, C L; Stein, H H

    2015-10-01

    Two experiments were conducted to determine the effects of phytate, phytase, fiber, and soybean oil on apparent total tract digestibility (ATTD) and standardized total tract digestibility (STTD) of Ca and on ATTD of P in fish meal fed to growing pigs. In Exp. 1, 40 growing pigs (initial average BW: 19.16 ± 2.04 kg) were randomly allotted to 1 of 5 diets with 8 pigs per treatment and placed in metabolism crates. Four diets were used in a 2 ´ 2 factorial design with 2 levels of phytate (0 or 0.7%) and 2 levels of microbial phytase (0 or 500 phytase units/kg). The diet containing no phytate was based on sucrose, cornstarch, fish meal, casein, and soybean oil, and the diet containing 0.7% phytate was based on corn, corn germ, fish meal, casein, and soybean oil. A Ca-free diet was used to determine basal endogenous losses of Ca. Feces were collected from d 6 to 13 after a 5-d adaptation period. Results indicated that the ATTD and STTD of Ca in fish meal and the ATTD of P increased ( phytase was used and were greater ( phytase and fiber increased the ATTD and STTD of Ca and the ATTD of P in fish meal, but inclusion of soybean oil did not affect digestibility of Ca or P. The observation that values for the ATTD and STTD of Ca and ATTD of P are greater in corn-based diets than in cornstarch-based diets indicates that values for the digestibility of Ca and P obtained in cornstarch-based diets may not always be representative for the digestibility in practical corn-based diets.

  15. Fitase na alimentação da tilápia do Nilo (Oreochromis niloticus. Desempenho e digestibilidade Phytase as feeding for Nile tilapia (Oreochromis niloticus. Performance and digestibility

    Directory of Open Access Journals (Sweden)

    Wilson Massamitu Furuya

    2001-06-01

    Full Text Available Este experimento foi realizado para avaliar os efeitos da adição de diferentes níveis de fitase (0, 500, 1500 e 3000 unidades de fitase ativa [UFA]/kg de ração em dietas para a tilápia do Nilo (8,88 ± 0,02 g. Todas as rações foram fornecidas até saciedade durante 45 dias. O nível de fitase baseou-se nos parâmetros de desempenho e digestibilidade e foi estimado pelos modelos quadrático e/ou LRP ("broken line". De acordo com os resultados obtidos com os níveis de fitase, o modelo LRP apresentou o melhor ajustamento de dados. A suplementação de fitase aumentou o desempenho, a retenção de minerais nos ossos, a digestibilidade da proteína e a disponibilidade de cálcio e fósforo. Os melhores resultados de desempenho, retenção de minerais nos ossos e digestibilidade foram obtidos com 700 UFA/kg de ração.This experiment was carried out to evaluate the effects of addition of different levels of phytase (0, 500, 1500 and 3000 units of active phytase [UFA]/kg diet in the diets for Nile tilapia (8.88 ± 0.02 g. All diets were fed to satiation daily for 45 days. The phytase level determination was based on the performance and digestibility parameters and was estimated by the quadratic and/or the broken line models. The broken line model showed a better adjustment for phytase levels according to the observed results. Phytase supplementation increased performence, bone minerals, protein digestibility and calcium and phosphorus availability. The best results of performance, bone mineral retentions and digestibility were obtained with 700 UFA/kg of diet.

  16. Effect of phytase application during high gravity (HG) maize mashes preparation on the availability of starch and yield of the ethanol fermentation process.

    Science.gov (United States)

    Mikulski, D; Kłosowski, G; Rolbiecka, A

    2014-10-01

    Phytic acid present in raw materials used in distilling industry can form complexes with starch and divalent cations and thus limit their biological availability. The influence of the enzymatic hydrolysis of phytate complexes on starch availability during the alcoholic fermentation process using high gravity (HG) maize mashes was analyzed. Indicators of the alcoholic fermentation as well as the fermentation activity of Saccharomyces cerevisiae D-2 strain were statistically evaluated. Phytate hydrolysis improved the course of the alcoholic fermentation of HG maize mashes. The final ethanol concentration in the media supplemented with phytase applied either before or after the starch hydrolysis increased by 1.0 and 0.6 % v/v, respectively, as compared to the control experiments. This increase was correlated with an elevated fermentation yield that was higher by 5.5 and 2.0 L EtOH/100 kg of starch, respectively. Phytate hydrolysis resulted also in a statistically significant increase in the initial concentration of fermenting sugars by 14.9 mg/mL of mash, on average, which was a consequence of a better availability of starch for enzymatic hydrolysis. The application of phytase increased the attenuation of HG media fermentation thus improving the economical aspect of the ethanol fermentation process.

  17. Effects of phytase on enhancement of phosphorus utilization from plant materials in bybrid catfish [Clarias macrocephalus (Gunther x Clarias gariepinus (Burchell

    Directory of Open Access Journals (Sweden)

    Phromkunthong,W.

    2005-01-01

    Full Text Available Hybrid catfish of initial weight 5.57-5.72g were studied in 235-1 glass tanks filled with 1801 water and closed-recirculation system with 0.8 l/m flow rate was attached. Complete randomized design was employed in which there were three replications (20 fingerlings each in each of nine treatments. The experimental period was 10 weeks. Formula 1 feed was the control in which fishmeal and soybean meal were used as protein source and was nutritionally suitable for catfish. In formulae 2-6 Feeds, only plant materials were used as feed components and sprayed phytase was aplied at 0,250,500,750 and 1,000 unit/kg feed. For formula 7-9 feeds, phosphorus from dicalcium phosphate (DCP at 0.1,0.2 and 0.3% was supplemented. The results showed that the supplementation of 500 unit/kg feed as well as of 0.2% phosphorus were the most effective in improving the weight gain, specific growth rate and feed efficiency (FCR, PER and ANPU. The supplementation of phytase as well as of DCP elevated the bone as well as in the whole body while the blood parameters were not markedly affected.

  18. Bioprocess for efficient production of recombinant Pichia anomala phytase and its applicability in dephytinizing chick feed and whole wheat flat Indian breads.

    Science.gov (United States)

    Joshi, Swati; Satyanarayana, T

    2015-10-01

    The phytase of the yeast Pichia anomala (PPHY) is a suitable biocatalyst as a food and feed additive because of its adequate thermostability, acid stability, protease insensitivity and broad substrate spectrum. The cell-bound nature and low phytase titres are the main bottlenecks for its utility in food and feed industries. In this investigation, we have overcome the problems by constitutive secretory expression of PPHY under glyceraldehyde phosphate dehydrogenase (GAP) promoter. A ~44-fold increase in rPPHY titre has been achieved after optimization of cultural variables by one-variable-at-a-time approach and two factorial statistical design. The use of GAP promoter makes the cultivation of the recombinant P. pastoris straight forward and eliminates the requirement of methanol for induction and hazards associated with its storage. Among metal-phytate complexes, Ca(2+) phytate is hydrolyzed more efficiently by rPPHY than Co(2+), Mn(2+), Mg(2+), Fe(3+) and Zn(2+) phytates. The enzyme is effective in dephytinizing whole wheat unleavened flat Indian breads (naan and tandoori) and different broiler feeds, thus mitigating anti-nutritional effects of phytates.

  19. Single-step purification and immobilization of MBP-phytase fusion on starch agar beads: application in dephytination of soy milk.

    Science.gov (United States)

    Ushasree, Mrudula Vasudevan; Gunasekaran, Paramasamy; Pandey, Ashok

    2012-07-01

    Periplasmic phytase, appA from E. coli has been noticed as a superior feed and food additive owing to its high specific activity, acidic pH optimum and resistance to gastric proteases. E. coli phytase was expressed as a fusion protein with maltose-binding protein, affinity-purified to homogeneity and, subsequently, immobilized in one step using a cost-effective matrix prepared from starch agar bead. Immobilized enzyme revealed an activity optimum at pH 6, while that of free enzyme was observed at pH 4. Both the immobilized and free enzyme showed a temperature optimum at 60 °C. Cleavage of 87 kDa fusion protein using factor Xa released 45 kDa appA. Hydrolysis of soy milk using immobilized enzyme led to 10% increase in release of inorganic phosphate at 50 °C relative to free fusion protein. This study suggests the usability of MBP as an immobilizing linker to other food enzymes for economical use in industry.

  20. Plausible exploitation of Jatropha de-oiled seed cake for lipase and phytase production and simultaneous detoxification by Candida parapsilosis isolated from poultry garbage.

    Science.gov (United States)

    Kannoju, Balakrishna; Ganapathiwar, Swaruparani; Nunavath, Hanumalal; Sunkar, Bindu; Bhukya, Bhima

    2017-02-01

    Jatropha de-oiled seed cake was explored to utilize as a basic nutrient source for Candida parapsilosis, isolated from poultry garbage and selected based on the production of lipase and phytase enzymes under submerged fermentation. At optimized parameters under solid-state fermentation, lipase and phytase activities were recorded as 1056.66±2.92 and 833±2.5U/g of substrate (U/g), respectively. Besides enzyme production, complete elimination of phorbol esters and significant phytate reduction from 6.51±0.01 to 0.43±0.01g/100g of seed cake were noted after 3days incubation. Curcin and trypsin inhibition activity were reduced significantly from 26.33±0.43 to 0.56±0.02mg/100g and 229.33±2.02 to 11.66±0.28U/g, respectively after 5days incubation. Saponins were reduced from 5.56±0.19 to 1.95±0.01g/100g of seed cake after 7days incubation.

  1. Cloning and expression analysis of recombination activating genes (RAG1/2) in red snapper (Lutjanus sanguineus).

    Science.gov (United States)

    Zhang, X L; Lu, Y S; Jian, J C; Wu, Z H

    2012-04-01

    Recombination activating genes (RAG1 and RAG2), involved in the V(D)J recombination of immunoglobulin and T-cell receptor genes play a crucial role in the adaptive immune response in vertebrates. The expression of these genes was required for the proper development and maturity of lymphocytes so that they can be used as useful markers to evaluate the development of lymphoid organ. In this paper, the cDNA of RAG1 and RAG2 in red snapper, Lutjanus sanguineus were cloned by homological cloning and rapid amplification of cDNA ends (RACE) methods. Results showed the full length of RAG1 cDNA was 3944 bp, containing a 5' untranslated region (UTR) of 200 bp, a 3'-UTR of 561 bp and an open reading frame of 3183 bp encoding 1060 amino acids. Three important structural motifs, a RING/U-box domain, a RING/FYVE/PHD-type domain and a RAG Nonamer-binding domain were detected in the deduced amino acid sequence of RAG1 by InterProScan analysis. The full length of RAG2 cDNA was 2200 bp, consisting of a 141 bp 5'-UTR, a 457 bp 3'-UTR and an open reading frame of 1602 bp encoding 533 amino acids. Two important structural motifs, a Galactose oxidase/kelch, beta-propeller domain and a kelch-type beta-propeller domain were detected in the deduced amino acid sequence of RAG2 by InterProScan analysis. BLAST analysis revealed that the RAG1 and RAG2 in red snapper shared a high homology with other known RAG1 and RAG2 genes, while the greatest degree of identity was observed with Hippoglossus hippoglossus RAG1 at 82% and Takifugu rubripes RAG2 at 87%, respectively. The differential expressions of RAG1 and RAG2 in various tissues of red snapper were analyzed by fluorescent quantitative real-time PCR. The overall expression pattern of the two genes was quite similar. In healthy red snappers, the RAGs transcripts were mainly detected in thymus, following head kidney, spleen, intestine, liver and brain. After vaccinated with inactivated Vibrio alginolyticus 48 h later, the RAGs m

  2. Isolation of Inositol Hexaphosphate (IHP)-Degrading Bacteria from Arbuscular Mycorrhizal Fungal Hyphal Compartments Using a Modified Baiting Method Involving Alginate Beads Containing IHP

    Science.gov (United States)

    Hara, Shintaro; Saito, Masanori

    2016-01-01

    Phytate (inositol hexaphosphate; IHP)-degrading microbes have been suggested to contribute to arbuscular mycorrhizal fungi (AMF)-mediated P transfer from IHP to plants; however, no IHP degrader involved in AMF-mediated P transfer has been isolated to date. We herein report the isolation of IHP-degrading bacteria using a modified baiting method. We applied alginate beads as carriers of IHP powder, and used them as recoverable IHP in the AM fungal compartment of plant cultivation experiments. P transfer from IHP in alginate beads via AMF was confirmed, and extracted DNA from alginate beads was analyzed by denaturing gradient gel electrophoresis targeting the 16S rRNA gene and a clone library method for the beta-propeller phytase (BPP) gene. The diversities of the 16S rRNA and BPP genes of microbes growing on IHP beads were simple and those of Sphingomonas spp. and Caulobacter spp. dominated. A total of 187 IHP-utilizing bacteria were isolated and identified, and they were consistent with the results of DNA analysis. Furthermore, some isolated Sphingomonas spp. and Caulobacter sp. showed IHP-degrading activity. Therefore, we successfully isolated dominant IHP-degrading bacteria from IHP in an AMF hyphal compartment. These strains may contribute to P transfer from IHP via AMF. PMID:27383681

  3. 天然蒙脱石/植酸酶复合载体制备及其植酸磷水解活性研究%Preparation of Phytase/Natural Montmorillonite Composite and Its Phytate Hydrolysis Activity

    Institute of Scientific and Technical Information of China (English)

    张闻中; 徐芳; 徐建雄

    2013-01-01

      通过吸附过程制备了天然蒙脱石/植酸酶复合载体并系统探讨其催化水解植酸磷活性.结果表明,天然蒙脱石固载植酸酶符合Elovich 动力学模型,固载等温曲线满足 Langmuir 方程,植酸酶最大固载容量为1.48 mg/mg;天然蒙脱石/植酸酶复合载体作为微型固定化酶反应器催化水解植酸磷,同自由植酸酶相比较,温度耐受性、抗蛋白酶水解能力均有效改善;适宜的 pH 值适用范围为4.0~4.5;重复利用5次后酶活保留40%以上.天然蒙脱石/植酸酶复合载体作为饲料添加剂可有效减少植酸酶制剂投加量,提高植酸磷有效利用率,溯源性降低畜禽养殖业“磷”污染排放.%Phytase/natural montmorillonite composite was prepared by adsorption process and the composite’s enzymatic activity for phytate hydrolysis was studied. The results showed that: the kinetic for adsorption of phytase onto natural montmorillonite could be best described by Elovich model, and the adsorptive isotherms fit for Langmuir model, with the maximum monolayer adsorption capacity of 1.48 mg/mg. The phytase/natural montmorillonite composites served as micro enzyme immobilized reactor for phytate hydrolysis. In comparison to free phytase, the relevant properties of the composites were effectively enhanced in view of the temperature tolerance and anti-proteolytic capability. The suitable pH media range for the composites in real practice was from 4.0 to 4.5. The enzymatic activity retained more than 40% after five testing cycles. Overall, the development of phytase/natural montmorillonite composite as feed additives could improve the availability of phosphorus from phytate and reduce the dosage of phytase, which contribute to the reduction of phosphorus emissions from livestock and poultry waste.

  4. 耐高温植酸酶毕赤酵母工程菌发酵中试条件优化%Optimization of Pilot-Scale Conditions of Thermostable Phytase Fermentation by Engineered Pichia pastoris Strain

    Institute of Scientific and Technical Information of China (English)

    黄魁英; 夏枫耿; 黄乐天; 明飞平; 杜少平

    2011-01-01

    The phytase fermentation conditions of the genetic-engineered Pichia pastoris strain PEY-2 were studies in this paper. The optimal phytase fermentation conditions were: induction time 72 h, enrichment time before induction 48 h, inoculum size 10%, seed age 24 h, pH for cell growth and inducement at 5.5 and 6.0 respectively. In scale-up culture in 50 L fermenter, phytase fermentation of high density is achieved and the activity of phytase highly reached 5.0×103 IU/mL by constant dissolve oxygen fed-batch way of glucose. Thermal stability tests showed that after 20-minute incubation at 90℃, the remaining activity of the phytase was 69% at pH 2.5 and 75% at pH 5.5 respectively,. The phytase was thermostable and could meet requirements of feed processing.%本文对植酸酶毕赤酵母基因工程菌PEY-2的发酵条件进行研究,结果表明:其摇瓶最佳产酶条件为诱导时间72h,诱导前适宜增殖时间48 h,接种量10%,种龄24h,诱导初始pH 6.0,生长阶段初始pH 5.5.在此基础上进行了50 L罐的发酵中试,50 L罐诱导产酶量达5.0×103 IU/mL,实现了高密度发酵.热稳定性试验表明:植酸酶液pH 2.5,90℃热处理20 min残留酶活69%,pH 5.5,90℃热处理20 min残留酶活75%,热稳定性良好,可以满足饲料高温制粒的要求.

  5. Effects of phytase on the performance and broiler meat qualityEfeitos da fitase no desempenho e na qualidade da carne de frangos de corte

    Directory of Open Access Journals (Sweden)

    Alexandre Oba

    Full Text Available The effects of phytase on the performance, carcass yield and on the meat quality (lipid oxidation of drumstick and breast muscle were evaluated on broilers submitted to diets containing corn, soybean and defatted corn germ meal, an ingredient with high level of phytic acid. Three hundred male broilers, with 4 d of age, were distributed in a completely randomized design with three treatments (represented by three levels of phytase in the diet: 0; 750 e 1500 FTU/kg of ration, with ten replicates and ten broilers per experimental unit. The feeding program was divided in three periods, according the age, starting (4 to 21 d, growing (22 to 35 d and finishing phases (36 to 42 d. There were no phytase´s effects on growth performance and carcass yield. There was an increasing the TBARS values to the refrigerated drumstick and the breast muscles, corresponding a negative effect on the lipid oxidation of meat.Foram avaliados os efeitos da enzima fitase sobre o desempenho, o rendimento de carcaça e a qualidade da carne (oxidação lipídica dos músculos da coxa a e peito de frangos de corte submetidos a dietas formuladas à base de milho, farelo de soja e farelo de gérmen de milho desengordurado (FGMD, um ingrediente com alta concentração de ácido fítico. Foram utilizados 300 frangos de corte machos, com quatro dias de idade, distribuídos em um delineamento experimental inteiramente casualizado, com três tratamentos (correspondentes a três níveis dietéticos de fitase: 0; 750 e 1500 FTU/kg de ração e dez repetições com dez aves cada. Foi utilizado um programa alimentar em três fases, inicial (4 a 21 dias de idade, crescimento (22 a 35 dias e terminação (36 a 42 dias. Os níveis de fitase não afetaram o desempenho e o rendimento das carcaças e suas partes, mas aumentaram os valores de TBARS do músculo da coxa e do peito refrigerados, indicando piora da oxidação lipídica da carne.

  6. Getting living phytase from the culture medium of rumen contents%从瘤胃内容物培养物得到活性植酸酶

    Institute of Scientific and Technical Information of China (English)

    郭冬生; 彭小兰

    2003-01-01

    @@ 植酸酶(phytate)是一种有机分子,大量存在于植物来源的饲料组分中.植物性饲料组分中磷大多以植酸形式存在,植酸也可能与微量元素、维生素、氨基酸和其它有机成分互相螯合(chelate)以螯合物形式存在.大量研究报道,植酸酶(phytase enzymes)能使以上螯合的养分分解释放出来,使它们能被鸡所消化利用.

  7. 单胃动物应用植酸酶研究进展%Advances on the application of phytase in monogastic animals

    Institute of Scientific and Technical Information of China (English)

    冯志华; 赵国先

    2003-01-01

    @@ 植酸酶(六磷酸肌醇磷酸脂水解酶,Phytase,myo-inositol-hexaphosphate phosphohydrolase)是一种能降解植物性饲料中植酸及其盐类的磷酸酯酶.畜禽饲料以植物性饲料为主,其常规植物性饲料中约有60 %~80 %的磷是以植酸磷的形式存在于稳定的植酸盐复合物中.由于猪、鸡等单胃动物体内都缺乏内源植酸酶系统,不能充分利用植酸磷中贮藏的大量磷和肌醇.

  8. Effect of several germination conditions on total P, phytate P, phytase, and acid phosphatase activities and inositol phosphate esters in rye and barley.

    Science.gov (United States)

    Centeno, C; Viveros, A; Brenes, A; Canales, R; Lozano, A; de la Cuadra, C

    2001-07-01

    Two assays were conducted to study the evolution of rye and barley phosphatases (phytase and acid phosphatase) and the degradation of its substrates (inositol phosphate esters) during seed germination. In this manner we could obtain a low-phytate, endogenous phosphatase rich ingredient to be used in animal nutrition. In the first assay, the seeds were soaked for 1 and 14 h and germinated for 3 and 5 days with and without the addition of gibberellic acid (GA3). In the second assay, the seeds were soaked for 1 h and germinated for 1, 3, and 5 days with GA3. Phytase (up to 5739 and 3151 U x kg(-1)) and acid phosphatase (up to 18288 and 3151 U x g(-1)) activities, and IP6 (6.09 and 6.01 mg x g(-1)), IP5 (0.48 and 0.48 mg x g(-1)), and IP4 (0.13 and 0.06 mg x g(-1)) were detected in ungerminated rye and barley, respectively. The germination process caused a significant increase of Phy and AcPh activities in rye (up to 112 and 213%) and barley (up to 212 and 634%) and a reduction in the phytate phosphorus content (up to 84 and 58%, respectively). Phytate phosphorus content was affected only by soaking time in the case of rye. Finally, during the course of germination, IP6 and IP5 were rapidly degraded in rye (88 and 79%) and barley (67 and 52%), and IP4 was only a short-living intermediate, which was increased during hydrolysis and degraded to IP3. In conclusion, a marked increase of Phy and AcPh activities in rye and barley with a concomitant decrease in phytate phosphorus content and an increase in the content of lower inositol phosphates were observed during the rye and barley germination.

  9. Research on preparing instant phytase by using freeze-drying method%冷冻干燥法制备速溶植酸酶的研究

    Institute of Scientific and Technical Information of China (English)

    马俊孝; 刘伟; 史衍鲁

    2011-01-01

    采用毕赤酵母发酵制备酶液,膜超滤浓缩、硫酸铵盐析法沉淀酶蛋白,结合冷冻干燥法制得速溶型植酸酶.结果表明,对酶活力达19 500 U/ml的发酵液进行浓缩后,以70%饱和度的硫酸铵进行盐析,分离后的酶蛋白经过冷冻干燥得到活力为500 000 U/g的植酸酶,速溶性好,溶解后清亮透明.酶的冻干粉50 ℃保存90 d内酶活力仍能维持96%以上的活力.为高酶活速溶植酸酶的工业化生产提供了一条可行的思路.%The freeze-drying technology for producing instant phytase was studied. The crude preparation was obtained by Pichia pastoris high cell density fermentation, microporous membrane ultrafiltration,ammonium sulfate precipitation and freeze-drying. Fermentation liquid, activity achieved 19 500 U/ml,was managed by ultrafiltration membrane, concentrated with ammonium sulphate in 70% saturation. After freeze-drying, the final product was obtained with up to 500 000 U/g. The preparation could soluble in water quickly with no turbidity, no precipitation and good transparency. The powder remained over 96%of its activity under 50 ℃ within 90 days. Our studies provide a feasible method for preparing instant phytase with higher activity.

  10. Effect of supplemented fungal phytase on performance and phosphorus availability by phosphorus-depleted juvenile rainbow trout (Oncorhynchus mykiss), and on the magnitude and composition of phosphorus waste output

    DEFF Research Database (Denmark)

    Dalsgaard, Anne Johanne Tang; Schøn Ekmann, K.; Pedersen, Per Bovbjerg

    2009-01-01

    of 11 °C. Two factors comprised of two dietary fungal phytase levels (0 or 1400 U kg− 1 feed− 1), and three dietary total phosphorus levels (0.89, 0.97 or 1.12%). All fish were acclimated to the lowest total phosphorus diet for 16 days, which included 0.29% phytate-phosphorus and no supplemental fungal...... supplementation, reaching an upper level of 74% at an available dietary phosphorus concentration of 0.71%. Adding fungal phytase to the diets improved the availability of phytate-phosphorus from an average of 6 to 64%. The fish retained 53–79% of the ingested phosphorus, while 24–44% was recovered in the faeces...

  11. 正交实验法优化转基因玉米中植酸酶的提取工艺%Optimization of the Extract Technology on the Phytase from Transgenic Corn by Orthogonal Experiment

    Institute of Scientific and Technical Information of China (English)

    刘娜; 柳鸿敏; 于壮; 张欣芳; 盖春宇; 邢珍娟; 陈奕冰; 李静; 张明

    2011-01-01

    Orthogonal test method was used to study the ultrasonic extraction conditions of the phytase from transgenic corn. It was found that the content of the phytase extracted was the highest under the conditions of ultrasonic extraction for 15min, using pH 5.50 buffer liquid at a liquid-solid ratio of 4 tol, which guaranteed for the evaluation of the transgenic corn.%采用正交实验法研究pH值、料液比和提取时间等因素对酶提取效率的影响,探索超声提取转基因玉米中植酸酶最佳提取条件.结果显示,最佳提取条件为提取液pH值5.5,料液比为1:4,提取时为15 min.

  12. Function-Based Metagenomic Library Screening and Heterologous Expression Strategy for Genes Encoding Phosphatase Activity.

    Science.gov (United States)

    Villamizar, Genis A Castillo; Nacke, Heiko; Daniel, Rolf

    2017-01-01

    The release of phosphate from inorganic and organic phosphorus compounds can be mediated enzymatically. Phosphate-releasing enzymes, comprising acid and alkaline phosphatases, are recognized as useful biocatalysts in applications such as plant and animal nutrition, bioremediation and diagnostic analysis. Metagenomic approaches provide access to novel phosphatase-encoding genes. Here, we describe a function-based screening approach for rapid identification of genes conferring phosphatase activity from small-insert and large-insert metagenomic libraries derived from various environments. This approach bears the potential for discovery of entirely novel phosphatase families or subfamilies and members of known enzyme classes hydrolyzing phosphomonoester bonds such as phytases. In addition, we provide a strategy for efficient heterologous phosphatase gene expression.

  13. Isolation and Identification of a High Phytase-Producing Bacillus Strain from Soil%一株土壤源高产植酸酶芽孢杆菌的分离与鉴定

    Institute of Scientific and Technical Information of China (English)

    孔志园; 丁轲; 余祖华; 王天奇; 闫文朝; 刘一尘

    2012-01-01

    对河南省不同地点采集的45份土样,利用分离培养基对产植酸酶芽孢杆菌进行分离和纯化,采用钒钼酸铵法测定其酶活,结合菌落和显微形态、生理生化试验和16SrDNA序列鉴定其种属.鉴定结果表明:从分离的80余株菌中筛选获得一株有较大植酸酶酶活的菌株B.Ld2-C,植酸酶酶活为552U/mL.经鉴定为纺锤形赖氨酸芽孢杆菌(B.sphaericus),能形成芽孢,而且该菌株能够耐受较强的酸、胆盐和高温,为研究其作为饲料添加剂菌种奠定了良好基础.%45 soil samples were collected from different areas in Henan province. More than 80 phytase-producing Bacillus strains were isolated and purified by isolation medium. The phytase activity of Bacillus was detected with vanadium-ammonium molybdate colorimetry, and the high phytase producing Bacillus was screened and identified by the colony, morphology, microscopic morphology in combination with the physiological and biochemical feature and 16S rDNA sequence. The results showed that the strain with the highest phytase activity of 552 U/mL,named B. Ld2-C,is obtained and identified as Bacillus sphaericus(B. spaericus) ,which can form spore and have the ability of anti-acid,ati-bile salt and high temperature. It has laid a good foundation as a probiotics feed additives.

  14. Production Conditions of Phytase from Candida tropicafis ypy06 using Response Surface Methodology%响应面法优化热带假丝酵母ypy06植酸酶发酵条件的研究

    Institute of Scientific and Technical Information of China (English)

    李晓宇; 柳志强; 田富; 陈耀国

    2011-01-01

    从海南洋浦近海分离到1株产植酸酶的酵母菌株,经鉴定为热带假丝酵母(Candida tropicalis),利用Plackett-Burman设计对其发酵条件进行了筛选。结果表明,豌豆粉浓度、硫酸铵浓度和初始pH对酶产量具有显著的影响;利用响应面法对3个因子进行了优化,获得了最佳发酵条件:豌豆粉1.0%,葡萄糖2.0%,硫酸铵1.7%,氯化钠1.0%,初始pH 5.2,培养72 h,植酸酶产量达到727.5 U/mL。%A phytase producing yeast Candida tropicalis ypy06 was isolated from the coastal water of Yangpu,Hainan.Plackett-Burman design and Response Surface Methodology(RSM) were used to optimize the production conditions of phytase.The results showed that the concentrations of pea,ammonium sulfate and initial pH were the most significant factors to influence phytase production.The optimized composition of medium was 1.0% pea,2.0% glucose,1.7% ammonium sulfate,1.0% NaCl,initial pH 5.2.After optimization,the phytase activity was 727.5 U/mL.

  15. Optimization of Phytase Production by Fermentation of Wheat Bran Using Response Surface Methodology%响应面法优化麦麸发酵产植酸酶条件的研究

    Institute of Scientific and Technical Information of China (English)

    刘玲玲; 陈钧

    2009-01-01

    Response surface methodology was applied for the optimization of phytase production by Bacillus sub-tilis ( natto) JSU -2 in solid - substrate fermentation using wheat bran as sole substrate. In the first step of optimization, with 2~(4-1) fractional factorial design, wheat bran to water ratio and cultivation time were found to be the important factors affecting phytase production. The two major variables were subsequently optimized using the central composite experimental design. Results; The obtained optimal conditions for phytase production are particle size 3 mm, wheat bran to water ratio 1:9.6, inoculum size 3 mL and cultivation time 25 h. Under these optimal conditions, the phytase production from B. subtilis (natto) JSU -2 reaches 595 IU/g wheat bran.%以纳豆芽孢杆菌JSU-2为供试菌株,麦麸皮为唯一固体发酵基质,运用响应面法对其产植酸酶的条件进行优化.首先用2~(4-1)部分因子试验设计对影响植酸酶活性的主要变量进行了评价,发现主要影响因子为麸皮与水的比例和培养时间.然后用中心组合试验设计确定主要变量的最佳水平.最佳产酶发酵条件为:粒径为3 mm、麸皮与水的比例为1:9.6、接种量为3 mL和培养时间为25 h.在最佳产酶条件下进行发酵,得到的植酸酶活性为595 U/g麦麸.

  16. Genotypic variation in the ability of landraces and commercial cereal varieties to avoid manganese deficiency in soils with limited manganese availability: is there a role for root-exuded phytases?

    Science.gov (United States)

    George, Timothy S; French, Andrew S; Brown, Lawrie K; Karley, Alison J; White, Philip J; Ramsay, Luke; Daniell, Tim J

    2014-07-01

    The marginal agricultural-systems of the Machair in the Western Isles of Scotland often have limited micronutrient availability because of alkaline soils. Traditional landraces of oats, barley and rye are thought to be better adapted to cope with the limited manganese (Mn) availability of these soils. When commercial cultivars are grown on the Machair, limited Mn-availability reduces crop yield and quality. We hypothesised that traditional cereal landraces selected on the Machair acquire Mn more effectively and that this could be linked to exudation of phytase from roots which would release Mn complexed with inositol phosphates. Growth and Mn-acquisition of five landraces and three commercial cultivars of barley and oats were determined in Machair soil. In addition, root phytase activities were assayed under Mn-starvation and sufficiency in hydroponics. In Machair soil, landraces had greater capacity for acquiring Mn and a greater ability to achieve maximum yield compared to the commercial cultivars. Under Mn-starvation, root phytase exudation was upregulated in all plants, suggesting that this trait might allow cereals to acquire more Mn when Mn-availability is limited. In the landraces, exuded phytase activity related positively to relative Mn-accumulation, whereas in the commercial cultivars this relationship was negative, suggesting that this trait may be secondary to an efficiency trait that has been lost from commercial germplasm by breeding. This research shows that cereal landraces possess traits that could be useful for improving the Mn-acquisition of commercial varieties. Exploiting the genetic diversity of landraces could improve the sustainability of agriculture on marginal calcareous lands globally.

  17. Effects of a 6-phytase on the apparent ileal digestibility of minerals and amino acids in ileorectal anastomosed pigs fed on a corn-soybean meal-barley diet.

    Science.gov (United States)

    Guggenbuhl, P; Waché, Y; Simoes Nunes, C; Fru, F

    2012-12-01

    Phosphorus of plant-based feedstuffs for monogastric animals is mainly in the form of phytic P, which has a very low bioavailability. The nondigested phytic P may contribute to P pollution. Furthermore, phytic acid may reduce digestibility of other minerals and protein. This study evaluated effects of the microbial 6-phytase RONOZYME HiPhos on apparent ileal digestibility of P, phytic acid, Ca, CP, energy, and AA in six 60-d-old ileorectal anastomosed pigs. In a duplicated 3 × 3 Latin square design, pigs had free access to alternatively a corn (Zea mays)-soybean (Glycine max) meal-barley (Hordeum vulgare)-based diet or this diet supplemented with RONOZYME HiPhos at either 500 units/kg (RH500) or 1000 units/kg (RH1000). Pigs fed diets supplemented with RH500 or RH1000 increased (P phytase increased apparent ileal digestibility of these indispensable minerals and phytate. The phytase increased digestibility of CP and indispensable AA indicating a better availability of plant-based proteins.

  18. 一株产植酸酶酵母菌株的分类鉴定与诱变育种%Identification and Mutation of the Phytase-producing Yeast

    Institute of Scientific and Technical Information of China (English)

    马忠友; 程飞; 邱亮; 包佳源; 樊美珍; 黄勃

    2011-01-01

    根据酵母形态学特征、生理生化特性和ITS区序列的测定结果把产植酸酶酵母RCEF4907菌株分类鉴定为东方伊萨酵母Issatchenkia orientalis.利用亚硝基胍、紫外线、亚硝酸和氯化锂4种诱变剂对酵母RCEF4907菌株进行复合诱变处理,将经过处理的菌悬液用植酸钙琼脂平板进行初筛,然后取发酵上清液在37℃,pH2.5的条件下,利用钒钼酸铵比色法测定植酸酶活力进行复筛.经过多次诱变处理后,最终从大量的突变菌株中筛选出高产植酸酶的G3297突变株,其植酸酶活性达到409 U/mL,是出发菌株RCEF4907的5.76倍.%The phytase-producing yeast RCEF4907 were identified as lssatchenkia orientalis based on its sequence, the morphological and physiological characteristics. A large number of mutants were qualitatively screened by calcium phytate agar medium after I. orientalis RCEF4907 isolate was mutated with Nitrosoguanidine, Ultraviolet, Nitrous acid and Lithium chloride. The ratio of clear zones to colony diameter of the mutants was used to estimate extracellular phytase production. The high-yield phytase mutant was isolated by quantitative analysis of phytase activity of fermentation broth with Vanadium-Ammonium Molybdate colorimetric method at 37℃ and pH2.5. The phytase activity of the mutant G3297 obtained from a great amount of mutants was 409U/mL, which was 5.76 times than the I. orientalis RCEF4907.

  19. Activity of fungal phytases stored in two ways in response to the period of storage at room temperature Atividade de fitases fúngicas armazenadas de duas maneiras em resposta ao período de armazenamento à temperatura ambiente

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    Luciana de Paula Naves

    2012-01-01

    Full Text Available Two distinct experiments were conducted simultaneously with phytases of Aspergillus oryzae and A. niger for determining enzyme activity in response to storage period (up to 180 days at room temperature - RT. In the first experiment, enzymes were stored as they were acquired (pure form and the activity was measured periodically during 180 days of storage at RT. In the second experiment, the phytases were incorporated to a supplement containing vitamins, minerals, and amino acids; and then this supplement was stored at RT up to 180 days, so that every 30 days of storage was collected one aliquot from each replicate for determining enzymatic activity. In conclusion, the phytase activity is affected by storage duration. To ensure 80% of the initial activity, the phytases of A. oryzae and A. niger can be stored in the pure forms for up to 53 and 135 days at RT, respectively. However, if the phytases of A. oryzae and A. niger are incorporated to a supplement containing vitamins, minerals, and amino acids then the storage period at RT should not exceed 67 and 77 days, respectively.Dois experimentos distintos foram realizados simultaneamente com fitases de Aspergillus oryzae e A. niger para a determinação da atividade enzimática em resposta ao período de armazenamento (por até 180 dias em temperatura ambiente - TA. No primeiro experimento, as enzimas foram armazenadas como adquiridas (forma pura e a atividade foi determinada periodicamente durante 180 dias de armazenamento em TA. No segundo experimento, as fitases foram incorporadas a um suplemento contendo vitaminas, minerais e aminoácidos. Então este suplemento foi armazenado em TA por até 180 dias, de modo que, a cada 30 dias de armazenamento, uma alíquota de cada repetição foi coletada para a determinação da atividade enzimática. Conclui-se que a atividade da fitase é afetada pela duração do armazenamento. Para assegurar 80% da atividade inicial, as fitases de A. oryzae e A. niger

  20. Effect of microbial phytase on spontaneous peritonitis of patients with hepatic cirrhosis%益生菌预防肝硬化自发性腹膜炎的疗效

    Institute of Scientific and Technical Information of China (English)

    周红宇; 周国华; 陈叶青; 颜君

    2011-01-01

    Objective To discuss the preventive measures of spontaneous peritonitis in patients with hepatic cirrhosis. Methods 43 patients with spontaneous peritonitis of hepatic cirrhosis were randomly divided into three groups:microbial phytase group,antibiotic group and control group. All patients were treated with anti-infectious treatment,liver treatment,diureses for 10 - 14 d. Then microbial phytase group was given Bifid-triple viable capsule(0. 63 g ,3 times a day)for 6 months, antibiotic group was given norfloxacin capsule (200 mg, once a day )for 6 months, control group was not given any treatment. The curative effect and small intestinal bacterial overgrowth were observed. Resuits ①Curative effect: statistical differences were found between patients of microbial phytase group and control group( P < 0. 05 ) ,antibiotic group and control groug( P < 0. 05 ), there was no significant difference between antibiotic group and microbial phytase group( P >0. 05). ②Small intestinal bacterial overgrowth:the rates were descent in microbial phytase group and antibiotic group,statistical differences were found between microbial phytase or antibiotic group and control group ( P < 0. 05 ). Conclusion Microbial phytase and antibiotic are effective for preventing spontaneous peritonitis in patients with hepatic cirrhosis.%目的 探讨预防肝硬化腹水并自发性膜腹炎的方法与效果.方法 选择肝硬化腹水并自发性腹膜炎患者共43例,随机分为益生菌组15例,抗生素组15例,对照组13例.三组患者住院后均给予抗感染、护肝、利尿、对症支持等常规治疗10~14 d.此后,益生菌组给予双歧三联活菌胶囊0.63 g/d,6个月;抗生素组给予诺氟沙星胶囊200 mg,口服,每日1次;对照组未做特殊处理.三组均于用药内6个月进行定期复查,评价其有效率、自发性腹膜炎再发率及小肠细菌过生长情况.结果 (1)疗效:6个月内益生菌组总有效率为86.7%,再生率为13.3%,

  1. Effect of γ-amino Butyric Acid and Phytic Acid Content in Germinated Brown Rice by Adding Exogenous Phytase%外源植酸酶对发芽糙米γ一氨基丁酸和植酸含量的影响

    Institute of Scientific and Technical Information of China (English)

    刘玮; 曹龙奎

    2015-01-01

    Effects of adding exogenous phytase on germinated brown rice γ-amino butyric acid and phytic acid content were studied.γ-amino butyric acid content in germinated brown rice could be increase, and levels of phytase could be reduce by adding exogenous phytase, thus nutritional value of brown rice germination could be improve.%本文研究了添加外源植酸酶对发芽糙米γ一氨基丁酸和植酸含量的影响,通过添加外源植酸酶,可以增加发芽糙米γ一氨基丁酸含量,降低植酸酶含量,从而提高发芽糙米的营养价值。

  2. FITASE EM DIETAS PARA SUÍNOS EM CRESCIMENTO: (I IMPACTO AMBIENTAL PHYTASE IN DIETS FOR GROWING PIGS: (I ENVIRONMENTAL IMPACT

    Directory of Open Access Journals (Sweden)

    Maria do Carmo Mohaupt Marques Ludke

    2002-02-01

    Full Text Available O objetivo deste trabalho foi determinar o nível adequado de fitase (0, 300, 600 e 900UF/kg da dieta a ser adicionado em dietas de suínos que continha dois níveis de proteína bruta (PB, um na exigência do animal (18%; NRC, 1988 e outro em nível menor (16%, para a redução do nitrogênio, fósforo e cálcio excretados, com o intuito de evitar poluição ambiental. Foram utilizados 24 leitões com peso médio inicial de 18,4kg alojados em gaiolas metabólicas para ensaio de metabolismo através do método de coleta total usando o marcador óxido férrico. Dietas isoenergéticas com 3295kcal de EM/kg foram fornecidas para os animais de acordo com o peso metabólico. Os parâmetros avaliados foram: quantidade de nitrogênio consumido e excretado total; quantidade de fósforo consumido e excretado nas fezes e urina; quantidade de cálcio consumido e excretado nas fezes e urina. Foi observado um aumento na biodisponibilidade dos nutrientes avaliados ao ser adicionado fitase nos níveis entre 421 - 466UF/kg da dieta.The objective of this experiment was determine the appropriate level of phytase (0, 300, 600 and 900PU/kg of the diet to be added in swine diets containing two levels of crude protein (CP, one in the demand of the animal (18% - NRC, 1988 and another in lower level (16%, for evaluation of the most effective reduction in excretion of nitrogen, phosphorus and calcium, to avoid larger environmental problems. Twenty four pigs with an initial avearage weight of 18.4kg were housed in metabolic cages for determination of the nitrogen, phosphorus and calcium metabolism through the method of total colection of faeces using ferric oxide as a marker. The diets were supplied for the animals related to their metabolic weight, and were isoenergetic with 3295kcal of ME/kg. The parameters evaluated were the amount of consumed and total excreted nitrogen; amount of phosphorus and calcium consumed and excreted in the feces and in the urine. An increase

  3. Suplementação de fitase microbiana na dieta de alevinos de jundiá: efeito sobre o desempenho produtivo e as características de carcaça Dietary microbial phytase supplementation of silver catfish fingerlings: effect of performance and carcass characteristics

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    Cleber Bastos Rocha

    2007-12-01

    Full Text Available Este estudo foi realizado para avaliar os efeitos da adição de diferentes níveis de fitase na dieta sobre o desempenho produtivo e as características de carcaça de alevinos de jundiá (Rhamdia quelen. Foram utilizados 208 alevinos com peso vivo inicial médio de 2,92±0,59g, distribuídos em um delineamento inteiramente casualizado com quatro tratamentos (0, 500, 1.000 e 1.500FTU kg-1 e quatro repetições. Na composição das dietas, foram utilizados somente ingredientes de origem vegetal e o alimento foi fornecido duas vezes ao dia. A cada quinzena, foram avaliados os parâmetros produtivos e, ao término do período experimental (45 dias, foram ainda realizadas avaliações nas carcaças. O ganho de peso médio foi significativamente crescente (PThis study evaluated the effects of adding increasing levels of microbial phytase to diets on performance and carcass traits of silver catfish fingerlings (Rhamdia quelen. Two-hundred and eight silver catfish fingerlings (average initial weight: 2.92±0.59g were randomly allotted to 4 treatments (0, 500, 1000 and 1500 phytase units kg-1 of diet with 4 replications, in a completely randomized design. No animal protein was added to the diets. The experiment was conducted for 45 days and fish were fed twice daily. Effects of phytase on performance were measured each 15 days and carcass traits were evaluated at the end of trial. Increasing levels of phytase linearly improved weight gain, from 1.24±0.27g (no phytase to 1.73±0.23g (1500PU kg-1 of diet (P<0.05; and rate of specific growth, from 0.78±0.12% (no phytase to 1.07±0.11% (1500PU kg-1 of diet (P<0.05. However, ether extract of carcass was linearly reduced, from 13.54±0.02% (no phytase to 10.59±0.10% (1500PU kg-1 of diet (P<0.05. Increasing levels of dietary microbial phytase, up to 1500PU kg-1 of diet, improve performance and carcass traits of silver catfish fingerlings.

  4. IRON AND HUMORAL IMMUNE RESPONSE IN PIGS FED PHYTASE-ADDED RATIONS WITH LOWER PHOSPHORUS LEVELS FERRO E IMUNIDADE HUMORAL EM SUÍNOS ALIMENTADOS COM FITASE E NÍVEIS REDUZIDOS DE FÓSFORO

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    Luiz Augusto Batista Brito

    2007-12-01

    Full Text Available

    Endogenous enzymes such as phytase have been widely used in swine production to increase phosphorus, amino acid and energy availability from feeds. This study aimed to evaluate the immune system through quantification of blood components associated to iron metabolism and determine humoral immune response elements in pigs fed phytase-added diets without micro minerals and vitamins and partial or total deletion of inorganic phosphorus. Forty-eight crossbred females with initial weight of 60 kg were randomly sorted into six groups of eight animals each, as follows: G1 -standard (complete ration (control group; G2 - standard ration except that micromineral and vitamin supplement was deleted; G3 - group 2 ration with phytase, G4 - group 2 ration less 1/3 of inorganic P with phytase, G5 -  group 2 ration less 2/3 of inorganic P with phytase and G6 - group 2 ration without inorganic P with phytase. Statistical difference (p>0,05 was not recorded neither in white cells and platelet counts nor hemoglobin, serum iron levels, considering all the animals in all treatments. Nevertheless, pigs up to 100 kg that consumed diet without micro minerals and vitamins, total deletion of inorganic P and phytase addition presented increased ferritin levels (p>0,05 when compared to animals fed similar diet with inorganic phosphorus and phytase. The enzyme guaranteed maintenance of iron stocks even in the absence of supplementation. Such difference was not recorded with 120-kg animals fed similar rations. Average total protein, IgG and IgM levels were not influenced by phytase, mineral and vitamin supplementation or inorganic phosphorus levels. The results demonstrate that decrease of inorganic phos-phorus, withdrawal of vitamin and mineral supplements and phytase addition in diets of finishing pigs do not lead to significant changes in hematological, biochemical and humoral immune response parameters.

  5. Genes and Gene Therapy

    Science.gov (United States)

    ... correctly, a child can have a genetic disorder. Gene therapy is an experimental technique that uses genes to ... or prevent disease. The most common form of gene therapy involves inserting a normal gene to replace an ...

  6. Isolation and characterization of a β-propeller gene containing phosphobacterium Bacillus subtilis strain KPS-11 for growth promotion of potato (Solanum tuberosum L.

    Directory of Open Access Journals (Sweden)

    Kashif eHanif

    2015-06-01

    Full Text Available Phosphate-solubilizing and phytate-mineralizing bacteria collectively termed as phosphobacteria provide a sustainable approach for managing P-deficiency in agricultural soils by supplying inexpensive phosphate to plants. A phosphobacterium Bacillus subtilis strain KPS-11 (Genbank accession no. KP006655 was isolated from potato (Solanum tuberosum L. rhizosphere and characterized for potato plant growth promoting potential. The strain utilized both Ca-phosphate and Na-phytate in vitro and produced 6.48 µg mL-1 indole-3-acetic acid in tryptophan supplemented medium. P-solubilization after 240 h was 66.4 µg mL-1 alongwith the production of 19.3 µg mL-1 gluconic acid and 5.3 µg mL-1 malic acid. The extracellular phytase activity was higher (4.3 x 10-10 kat mg-1 protein than the cell-associated phytase activity (1.6 x 10-10 kat mg-1protein. B. subtilis strain KPS-11 utilized 40 carbon sources and showed resistance against 20 chemicals in GENIII micro-plate system demonstrating its metabolic potential. Phytase-encoding gene β-propeller (BPP showed 92% amino acid similarity to BPP from B. subtilis (accession no.WP_014114128.1 and 83% structural similarity to BPP from B. subtilis (accession no 3AMR_A. Potato inoculation with B. subtilis strain KPS-11 increased the root/shoot length and root/shoot weight of potato as compared to non-inoculated control plants. Moreover, rifampicin-resistant derivative of KPS-11 were able to survive in the rhizosphere and on the roots of potato up to sixty days showing its colonization potential. The study indicates that B. subtilis strain KPS-11 can be a potential candidate for development of potato inoculum in P-deficient soils.

  7. Influence of Phytase Transgenic Corn on the Intestinal Microflora and the Fate of Transgenic DNA and Protein in Digesta and Tissues of Broilers.

    Science.gov (United States)

    Lu, Lin; Guo, Jiang; Li, Sufen; Li, Ang; Zhang, Liyang; Liu, Zhenhua; Luo, Xugang

    2015-01-01

    An experiment was conducted to investigate the effect of phytase transgenic corn (PTC) on intestinal microflora, and the fate of transgenic DNA and protein in the digesta and tissues of broilers. A total of 160 1-day-old Arbor Acres commercial male broilers were randomly assigned to 20 cages (8 chicks per cage) with 10 cages (replicates) for each treatment. Birds were fed with a diet containing either PTC (54.0% during 1-21 days and 61.0% during 22-42 days) or non-transgenic isogenic control corn (CC) for a duration of 42 days. There were no significant differences (P>0.05) between birds fed with the PTC diets and those fed with the CC diets in the quantities of aerobic bacteria, anaerobic bacteria, colibacillus and lactobacilli, or microbial diversities in the contents of ileum and cecum. Transgenic phyA2 DNA was not detected, but phyA2 protein was detected in the digesta of duodenum and jejunum of broilers fed with the PTC diets. Both transgenic phyA2 DNA and protein fragments were not found in the digesta of the ileum and rectum, heart, liver, kidney, and breast or thigh muscles of broilers fed with the PTC diets. It was concluded that PTC had no adverse effect on the quantity and diversity of gut microorganisms; Transgenic phyA2 DNA or protein was rapidly degraded in the intestinal tract and was not transferred to the tissues of broilers.

  8. Influence of Phytase Transgenic Corn on the Intestinal Microflora and the Fate of Transgenic DNA and Protein in Digesta and Tissues of Broilers.

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    Lin Lu

    Full Text Available An experiment was conducted to investigate the effect of phytase transgenic corn (PTC on intestinal microflora, and the fate of transgenic DNA and protein in the digesta and tissues of broilers. A total of 160 1-day-old Arbor Acres commercial male broilers were randomly assigned to 20 cages (8 chicks per cage with 10 cages (replicates for each treatment. Birds were fed with a diet containing either PTC (54.0% during 1-21 days and 61.0% during 22-42 days or non-transgenic isogenic control corn (CC for a duration of 42 days. There were no significant differences (P>0.05 between birds fed with the PTC diets and those fed with the CC diets in the quantities of aerobic bacteria, anaerobic bacteria, colibacillus and lactobacilli, or microbial diversities in the contents of ileum and cecum. Transgenic phyA2 DNA was not detected, but phyA2 protein was detected in the digesta of duodenum and jejunum of broilers fed with the PTC diets. Both transgenic phyA2 DNA and protein fragments were not found in the digesta of the ileum and rectum, heart, liver, kidney, and breast or thigh muscles of broilers fed with the PTC diets. It was concluded that PTC had no adverse effect on the quantity and diversity of gut microorganisms; Transgenic phyA2 DNA or protein was rapidly degraded in the intestinal tract and was not transferred to the tissues of broilers.

  9. Structural and biochemical analysis of a unique phosphatase from Bdellovibrio bacteriovorus reveals its structural and functional relationship with the protein tyrosine phosphatase class of phytase.

    Directory of Open Access Journals (Sweden)

    Robert J Gruninger

    Full Text Available Bdellovibrio bacteriovorus is an unusual δ-proteobacterium that invades and preys on other Gram-negative bacteria and is of potential interest as a whole cell therapeutic against pathogens of man, animals and crops. PTPs (protein tyrosine phosphatases are an important class of enzyme involved in desphosphorylating a variety of substrates, often with implications in cell signaling. The B. bacteriovorus open reading frame Bd1204 is predicted to encode a PTP of unknown function. Bd1204 is both structurally and mechanistically related to the PTP-like phytase (PTPLP class of enzymes and possesses a number of unique properties not observed in any other PTPLPs characterized to date. Bd1204 does not display catalytic activity against some common protein tyrosine phosphatase substrates but is highly specific for hydrolysis of phosphomonoester bonds of inositol hexakisphosphate. The structure reveals that Bd1204 has the smallest and least electropositive active site of all characterized PTPLPs to date yet possesses a unique substrate specificity characterized by a strict preference for inositol hexakisphosphate. These two active site features are believed to be the most significant contributors to the specificity of phytate degrading enzymes. We speculate that Bd1204 may be involved in phosphate acquisition outside of prey.

  10. Nutrient reduction in rations with phytase for growing pigs Redução de nutrientes em rações com fitase para suínos em crescimento

    Directory of Open Access Journals (Sweden)

    Valéria Vânia Rodrigues

    2011-02-01

    Full Text Available Two experiments were conducted to evaluate the performance, the energy and nutrient balance and economic viability of production of growing pigs fed diets with reduced levels and phytase. In experiment 1, 60 barrows and gilts (initial weight 25 ± 1.72 kg were housed in a randomized block design with three diets, ten replicates and two animals per experimental unit (one male and one female. One of the diets was formulated with a reduced level with phytase and evaluated in comparison to a positive-control diet that was formulated according to the nutritional recommendations for animals of this breed, and a negative-control diet without phytase. The different diets did not affect feed intake, but the diet with phytase promoted better final weight, daily weight gain and feed conversion, although it did not differ from the positive control diet. In experiment 2, 27 barrows (initial weight 49 ± 3.67 kg were housed in random blocks with three diets and nine replicates with experimental plot represented by one animal. The diets did not affect the balance of energy and nutrients but had a significant effect on the efficiency of utilization of calcium and phosphorus, which was the lowest in animals that received the positive-control diet. Economic viability improved when nutrients were reduced and phytase was added in the ration. The addition of phytase (500 ftu/kg in rations formulated with nutritional levels reduced for growing swines improves rearing productivity and it is economical viable.Dois experimentos foram conduzidos para avaliar o desempenho, o balanço de energia e nutrientes e a viabilidade econômica da produção de suínos em crescimento alimentados com rações com níveis reduzidos e fitase. No experimento 1, 60 suínos machos castrados e fêmeas (peso inicial de 25 ± 1,72 kg foram alojados em delineamento de blocos casualizados, com três dietas, dez repetições e dois animais por unidade experimental (macho e fêmea. Uma das

  11. Radioisotope and mathematical modeling in the assessment of supplementation of Phytase in diets for growing pigs; Radioisotopo e modelagem matematica na avaliacao da suplementacao de fitase em dietas para suinos em crescimento

    Energy Technology Data Exchange (ETDEWEB)

    Mota, L.C.; Moreira, J.A.; Oliveira, R.L.R., E-mail: japmoreira@bol.com.br [Universidade Federal do Rio Grande do Norte (UFRN), Natal, RN (Brazil); Vitti, D.M.S.S. [Centro de Energia Nuclear na Agricultura (CENA/USP), Piracicaba, SP (Brazil); Patino, R.M. [Universidade de Sucre, Bogota (Colombia)

    2014-02-15

    The purpose of this study was to evaluate the effects of including increasing levels of phytase enzyme in pig diets for growing pigs, using the mathematical model. Data from 20 crossbred male piglets, castrated and weighing 26.80 kg on average was used. The animals were housed in individual metabolic cages to collect feces and urine in a 17 day period. A randomized block experimental design containing five treatments and four repetitions was used. The experimental diet provided to piglets contained corn and soybean and was supplemented with five increasing levels of phytase enzyme (0, 250, 500, 750 and 1000 UF/kg), corresponding to 0.00 %, 0.01 %, 0.02 %, 0.03 % and 0.04 % respectively. The variables evaluated were: intake, excretion, output flow of P in the digestive tract, bloodstream, bones and soft tissues. The phytase enzyme did not affect the P intake (P>0.05 (F{sub 10}), the P excreted in urine (F{sub 02}) and the output flow of P in the bones (F{sub 32} e F{sub 23}) and soft tissue (F{sub 42} e F{sub 24}), however, there was a reduction in P excreted in feces (F{sub 01}) of 8.92 %, 26.76 %, 22.53 % and 28.64 % to the levels 0, 250, 500, 750 e 1000UF/kg, respectively and showed a positive linear effect (P<0.08) for the endogenous P (F{sub 12}). Corn and soybean meal based diets can be used with 50% of P by dicalcium phosphate, adding 250UF/kg diet for growing pigs, and may cause a reduction of 27 % of P excretion in feces. (author)

  12. A simple and accurate two-step long DNA sequences synthesis strategy to improve heterologous gene expression in pichia.

    Directory of Open Access Journals (Sweden)

    Jiang-Ke Yang

    Full Text Available In vitro gene chemical synthesis is a powerful tool to improve the expression of gene in heterologous system. In this study, a two-step gene synthesis strategy that combines an assembly PCR and an overlap extension PCR (AOE was developed. In this strategy, the chemically synthesized oligonucleotides were assembled into several 200-500 bp fragments with 20-25 bp overlap at each end by assembly PCR, and then an overlap extension PCR was conducted to assemble all these fragments into a full length DNA sequence. Using this method, we de novo designed and optimized the codon of Rhizopus oryzae lipase gene ROL (810 bp and Aspergillus niger phytase gene phyA (1404 bp. Compared with the original ROL gene and phyA gene, the codon-optimized genes expressed at a significantly higher level in yeasts after methanol induction. We believe this AOE method to be of special interest as it is simple, accurate and has no limitation with respect to the size of the gene to be synthesized. Combined with de novo design, this method allows the rapid synthesis of a gene optimized for expression in the system of choice and production of sufficient biological material for molecular characterization and biotechnological application.

  13. A simple and accurate two-step long DNA sequences synthesis strategy to improve heterologous gene expression in pichia.

    Science.gov (United States)

    Yang, Jiang-Ke; Chen, Fang-Yuan; Yan, Xiang-Xiang; Miao, Li-Hong; Dai, Jiang-Hong

    2012-01-01

    In vitro gene chemical synthesis is a powerful tool to improve the expression of gene in heterologous system. In this study, a two-step gene synthesis strategy that combines an assembly PCR and an overlap extension PCR (AOE) was developed. In this strategy, the chemically synthesized oligonucleotides were assembled into several 200-500 bp fragments with 20-25 bp overlap at each end by assembly PCR, and then an overlap extension PCR was conducted to assemble all these fragments into a full length DNA sequence. Using this method, we de novo designed and optimized the codon of Rhizopus oryzae lipase gene ROL (810 bp) and Aspergillus niger phytase gene phyA (1404 bp). Compared with the original ROL gene and phyA gene, the codon-optimized genes expressed at a significantly higher level in yeasts after methanol induction. We believe this AOE method to be of special interest as it is simple, accurate and has no limitation with respect to the size of the gene to be synthesized. Combined with de novo design, this method allows the rapid synthesis of a gene optimized for expression in the system of choice and production of sufficient biological material for molecular characterization and biotechnological application.

  14. 植酸酶对水貂氮、磷等营养物质利用及生长发育的影响%Effects of Microbial Phytase Levels on Availability of Nitrogen and Phosphorus Utilized and Performance in Growing Minks

    Institute of Scientific and Technical Information of China (English)

    王凯英; 徐超; 鲍坤; 刘晗璐; 李光玉; 纪银玲

    2013-01-01

    研究鲜料中添加植酸酶水平对生长发育期水貂氮、磷利用及生长发育性能的影响。配制Ⅰ~Ⅳ组日粮,Ⅰ组为对照组(基础日粮),Ⅱ~Ⅳ组在基础日粮中分别添加450IU/kg、900IU/kg、4500IU/kg的植酸酶,进行各组日粮干物质、蛋白质、脂肪、钙、磷消化代谢试验和生长发育期饲养试验。结果表明,水貂鲜料添加植酸酶可显著提高干物质、脂肪、粗蛋白、P消化率及N代谢率( P<0.01),N、P环境排放量降低极显著( P<0.01);可显著提高其体增重和平均日增重( P<0.05),其中添加900IU/kg的Ⅳ组生长发育性能最好。%Effects of microbial phytase levels on the availabitility of nutriments and growth performance in Growing Mink were studied .Sixty black male minks were randomly divided into 4 groups ,15 minks per each .The control was fed conventional fresh diet ,the testing groups were applied phytase ,respectively ,phytase 450IU/kg (Ⅱ) ,900IU/kg (Ⅲ) and 4 500IU/kg (Ⅳ) .The results showed that there were significant dif-ference among groups ,the values of DMD、CPD、EED、PD were significant higher than the control ( P0.05) .There was clearly trend that phytase could significantly reduce the nitrogen and phosphorus excretion of growing minks ( P<0.01) ,and increase dietary nitrogen metablic rates in minks ,the 900IU/kg phytase was the best in all groups .There were a significantly increase in the in-dex of body weight gain and average daily gain the groups of phytase than control group ( P<0.05) .The results showed that it could significant-ly improve the growth performance and availabitility of DM、CP、EE、Ca、P ,applied 900IU/kg of phytase during growing period in mink diets .

  15. Phytase-producing capacity of yeasts isolated from traditional African fermented food products and PHYPk gene expression of Pichia kudriavzevii strains

    DEFF Research Database (Denmark)

    Greppi, Anna; Krych, Lukasz; Costantini, Antonella

    2015-01-01

    Phytate is known as a strong chelate of minerals causing their reduced uptake by the human intestine. Ninety-three yeast isolates from traditional African fermented food products, belonging to nine species (Pichia kudriavzevii, Saccharomyces cerevisiae, Clavispora lusitaniae, Kluyveromyces...

  16. Desempenho de frangos de corte alimentados com rações contendo diferentes níveis de cálcio suplementadas com fitase Performance of broilers fed diets with different calcium levels and supplemented with phytase

    Directory of Open Access Journals (Sweden)

    Daniella Carolina Zanardo Donato

    2011-10-01

    Full Text Available Um experimento foi conduzido para avaliar o efeito de rações contendo diferentes níveis de cálcio e suplementadas com fitase sobre o desempenho de frangos de corte nas fases inicial (1 a 21 dias, de crescimento (22 a 35 dias e final (36 a 42 dias. Foram utilizados 2.160 pintos de corte machos da linhagem AG Ross 308, de 1 a 42 dias de idade, distribuídos em 6 tratamentos, cada um com 6 repetições. Foi utilizado um delineamento inteiramente casualizado em esquema fatorial 3 × 2, com 3 níveis de fitase (0, 600 e 1.200 ftu/kg e 2 de cálcio (0,94 e 0,66%; 0,84 e 0,59%; e 0,78 e 0,54% para as fases inicial, de crescimento e final, respectivamente. As rações experimentais ainda tinham níveis reduzidos de fósforo disponível e níveis mínimos de proteína bruta. Ao final de cada fase, avaliaram-se o ganho de peso, o consumo de ração, a conversão alimentar e a mortalidade. Houve interação entre os níveis de fitase e de cálcio estudados para ganho de peso, consumo de ração e mortalidade nas três fases avaliadas. Os melhores resultados foram obtidos com a suplementação de fitase no nível de 1.200 ftu/kg de ração em associação aos menores níveis de cálcio. Nenhum resultado significativo foi observado para conversão alimentar. Os níveis nutricionais de cálcio podem ser reduzidos em 30% em dietas com baixos níveis de fósforo disponível e níveis mínimos de proteína bruta, desde que as dietas sejam suplementadas com 1.200 ftu/kg de fitase.An experiment was carried out to evaluate the effect of diets with different calcium levels and supplemented with phytase on the performance of broiler chickens in initial (1 to 21 days of age, growing (22 to 35 days of age and final (36 to 42 days of age phases. Two thousand one hundred and sixty male broilers from AG Ross 308 line, from 1 to 42 days old were distributed in 6 treatments, with 6 replicates each. A completely randomized design in a 3 × 2 factorial scheme was used

  17. Effects of Phytase on Tibia Indicators, Calcium and Phosphorus Utilization in Broilers%植酸酶对肉鸡胫骨指标和钙磷利用率的影响

    Institute of Scientific and Technical Information of China (English)

    张旭; 蒋桂韬; 王向荣; 李昊帮; 戴求仲

    2012-01-01

    :通过对胫骨指标的测定和代谢试验,研究了微生物植酸酶在两个不同磷酸氢钙添加水平下对肉仔鸡胫骨指标和钙、磷利用率的影响试验结果表明,与正常磷酸氢钙添加水平(1-10%)的对照组相比,降低磷酸氢钙用量并添加植酸酶,可以提高24日龄肉仔鸡的胫骨长度,提高24和42日龄肉仔鸡的胫骨重量/体积;0.44%磷酸氢钙水平添加植酸酶组的24日龄肉鸡的脱脂干骨重和粗灰分含量较对照组显著降低(P〈0.05);降低磷酸氢钙水平并添加植酸酶不会对肉鸡胫骨的钙、磷含量造成影响(P〉0.05);降低磷酸氢钙水平并添加植酸酶能够显著提高磷的消化利用率(P〈0.05),极显著提高24日龄肉仔鸡的钙表观消化率(P〈O.01),提高42日龄肉仔鸡的钙表观消化率,但差异不显著(P〉O.05),%To study the impacts of microbial phytase added in two different CaHP04 addition levels of the broiler tibia indicators and calcium and phosphorus utilization through determination of tibia indicators and broiler metabolic test. The results showed that, compared with the control group of normal CaHP04 addition level(1.10%), reduce CaHP04 dosage and phytase supplementation increased the length of the 24-day-old broiler tibia, improved the tib- ia weight / volume ratio of 24-day-old and 42-day-old broilers; nonfat dry bone weight and crude ash content of 24-day-old broilers at the CaHP04 level of 0.44% phytase group were lower than the control group(P〈0.05); re- duce CaHPO~ levels and adding phytase had no impact on calcium and phosphorus content of broiler tibia(P〉0.05); reduce the CaHP04 level and add phytase c ouht significantly improved the digestibility of apparent phosphorus(P〈 0.05), and a very significant increase in the 24 days broilers'calcium apparent digestibility(P〈0.01), and increased in the 42 days broilers calcium apparent digestibility(P〉0.05).

  18. Application of the Taguchi method in poultry science: estimation of the in vitro optimum intrinsic phytase activity of rye, wheat and barley.

    Science.gov (United States)

    Sedghi, M; Golian, A; Esmaeilipour, O; Van Krimpen, M M

    2014-01-01

    1. In poultry investigations, the main interest is often to study the effects of many factors simultaneously. Two or three level factorial designs are the most commonly used for this type of investigation. However, it is often too costly to perform when number of factors increase. So a fractional factorial design, which is a subset or a fraction of a full factorial design, is an alternative. The Taguchi method has been proposed for simplifying and standardising fractional factorial designs. 2. An experiment was conducted to evaluate the applicability of the Taguchi method to optimise in vitro intrinsic phytase activity (IPA) of rye, wheat and barley under different culture conditions. 3. In order to have a solid base for judging the suitability of the Taguchi method, the results of the Taguchi method were compared with those of an experiment that was conducted as a 3(4) full factorial arrangement with three feed ingredients (rye, wheat and barley), three temperatures (20°C, 38°C and 55°C), three pH values (3.0, 5.5 and 8.0) and three incubation times (30, 60 and 120 min), with two replicates per treatment. 4. After data collection, a Taguchi L 9 (3(4)) orthogonal array was used to estimate the effects of different factors on the IPA, based on a subset of only 9 instead of 81 treatments. The data were analysed with both Taguchi and full factorial methods and the main effects and the optimal combinations of these 4 factors were obtained for each method. 5. The results indicated that according to both the full factorial experimental design and the Taguchi method, the optimal culture conditions were obtained with the following combination: rye, pH = 3, temperature = 20 °C and time of incubation = 30 min. The comparison between the Taguchi and full factorial results showed that the Taguchi method is a sufficient and resource saving alternative to the full factorial design in poultry science.

  19. Effects of Fermentation Product Containing Phytase on Productive Performance, Egg Quality, and Phosphorous Apparent Metabolism of Laying Hens Fed Different Levels of Phosphorus

    Institute of Scientific and Technical Information of China (English)

    WANG Zhi-hong; DONG Xiao-fang; TONG Jian-ming; XU Shang-zhong

    2014-01-01

    This study investigated the effects of fermentation product containing phytase (FPP) that was fermented using waste vinegar residue (WVR) as substrate from Aspergillus ifcuum NTG-23 on productive performance, egg quality, and phosphorus apparent metabolism of laying hens. First, 375 22-wk-old Jinghong hens were allocated into 5 treatments (5 replicates of 15 hens each) in an 8-wk experiment for evaluating the parameters of productive performance, egg quality, serum, and tibia. Experimental diets contained 4%FPP and 96%corn-soybean diet. The levels of dicalcium phosphate (DCP) were 1.34, 1.01, 0.67, 0.34 and 0%. Next, thirty 31-wk-old Jinghong hens were fed 5 types of diets for evaluating phosphorous apparent metabolism rate. Egg productive rate, egg weight, feed conversion ratio, Haugh unit, egg albumen height, serum calcium, tibia ash, tibia ash calcium and tibia breaking strength were not different signiifcantly among 5 treatments. The signiifcant difference of average daily feed intake was not appeared when the DCP content of corn-soybean-FPP diet was reduced to 0.67%;the eggshell hardness, eggshell thickness and serum phosphorus were not reduced signiifcantly until the DCP content of corn-soybean-FPP diet was reduced to 0.34%. The yolk color was improved when the laying hens fed deifcient DCP corn-soybean-FPP diet. A 22.14%reduction in excreta phosphorus was observed when the laying hens fed low phosphorus (0.67%DCP) corn-soybean-FPP diet. A 30%elevation of phosphorus apparent metabolism rate was obtained when the DCP content of corn-soybean-FPP diet was decreased from 1.34 to 1.01%. The reducing cost of layer diet was totalized about 120 CNY 1 000 kg-1 diet when the content of DCP was 0.67%in corn-soybean-FPP diet. These results indicated that FPP could be applied in laying hen as a potential, cost-effective and rational application of WVR.

  20. Aproveitamento de nutrientes e de energia da ração de codornas japonesas em postura com o uso de fitase Nutrient and energy utilization from the feedings of laying Japanese quails by using phytase

    Directory of Open Access Journals (Sweden)

    Heder José D'Avila Lima

    2010-07-01

    Full Text Available O experimento foi realizado para avaliar o efeito da adição de fitase no aproveitamento dos ingredientes da ração para codornas. Foram utilizadas 200 codornas japonesas fêmeas com 251 dias de idade, peso médio de 187,2 ± 6,0 g e taxa de produção de ovos de 84,8%, distribuídas em delineamento inteiramente casualizado com quatro dietas experimentais e cinco repetições de dez aves por unidade experimental. As dietas experimentais consistiram de uma ração basal à base de milho e farelo de soja, formulada para atender às exigências nutricionais das codornas e às recomendações da matriz da fitase, suplementada com 0, 200, 400 ou 600 unidades de fitase (uf/kg. Utilizando-se o método de coleta total de excretas, foram determinados os valores de matéria seca ingerida, matéria seca excretada, energia metabolizável aparente (EMA, energia metabolizável aparente corrigida pela retenção de nitrogênio (EMAn e os coeficientes de metabolizabilidade da energia corrigida ou não pelo balanço de nitrogênio. Foi calculada também a quantidade de fósforo, cálcio e nitrogênio retido por ave/dia. De maneira geral, houve melhora no aproveitamento da energia das rações com a suplementação de fitase. Os níveis de 195 e 186 uf/kg resultaram em maior aproveitamento da EMA e EMAn. Não houve diferença na retenção de fósforo e cálcio. O nível de 600 (uf/kg promove menor excreção de nitrogênio, porém o de 368 uF/kg é suficiente para máxima retenção de nitrogênio pelas codornas.This experiment was conducted to evaluate the effect of phytase addition on the use of feeding ingredients for laying quails. It was used 200 female Japanese quails at 251 days of age at an average weight of 187.2 ± 6.0 g and egg production rate of 84.8%. The quails were distributed in a complete random design with four experimental diets and five repetitions of ten birds per experimental unity. The experimental diets were consisted of corn and

  1. Fitase líquida em dieta extrusada para juvenis de tilápia do Nilo (Oreochromis niloticus = Liquid phytase in extruded diet for juvenile Nile tilapia (Oreochromis niloticus

    Directory of Open Access Journals (Sweden)

    Tarcila Souza de Castro Silva

    2007-10-01

    Full Text Available Este trabalho foi realizado para avaliar a utilização de fitase líquida sobre o desempenho de juvenis de tilápia do Nilo. Foram utilizados 176 peixes com peso inicial de 45,84 ± 3,31 g, alimentados com dietas sem e com 250; 500 e 1.000 unidades de fitase (UF kg-1 da dieta, durante 62 dias. Foi utilizada dieta-controle com 3012 kcal de energia kg-1, 29% de proteína bruta e 0,23% de fósforo disponível. Os peixes foram distribuídos em delineamento de blocos ao acaso, com quatro tratamentos e quatro repetições. Não foi observado efeito dos níveis de inclusão de fitase sobre o consumo, índice hepatossomático,gordura visceral, sobrevivência, retenção de cálcio nos ossos e teores de umidade, proteína bruta e cinzas na carcaça. Foi observado efeito linear crescente dos níveis de fitase sobre a retenção de fósforo e sobre os coeficientes de digestibilidade aparente da proteína bruta e do fósforo das dietas. Foi observado efeito quadrático dos níveis de suplementação de fitase sobre o ganho de peso, conversão alimentar, taxa de eficiência protéica, rendimento decarcaça e gordura na carcaça, em que foram estimados os valores de 350; 500; 500; 516 e 647 UF kg-1 de dieta para melhor valor destas variáveis, respectivamente. Conclui-se que a utilização 500 UF kg-1 é adequada em dietas para juvenis de tilápia do Nilo.This study was undertaken to evaluate the effects of liquid phytase on the performance of juvenile Nile tilapia. It was used 176 fish (45.84 ± 3.31 g. The fish were fed diets without phytase and with 250; 500 and 1,000 phytase units (PU kg-1 of diet, during62 days. A control diet was used containing 3,012 kcal digestible energy kg-1, 29% crude protein and 0.23% available phosphorus. The fish were assigned to a randomized block design, with four treatments and four replications. No effect was observed on the levels ofdietary phytase on feed intake, hepatosomatic index, visceral fat, survival, as well as on

  2. Effect of phytase supplementation on performance, bone densitometry and carcass yield in broilers chicks = Efeito da adição da enzima fitase sobre o desempenho, densitometria óssea e rendimento de carcaça de frangos de corte

    Directory of Open Access Journals (Sweden)

    Otto Mack Junqueira

    2011-07-01

    Full Text Available An experiment was conducted to evaluate the performance, bonedensitometry and carcass yield of broilers chicks, using different levels of phytase enzyme. Nine hundred and sixty male one-day-old broiler chicks were used. The birds were distributed in a completely randomized experimental design, involving five treatments andsix replications of 32 chicks each. The treatments consisted of a control diet for each phase, and four other diets were formulated adding growing levels of the phytase enzyme (250, 500, 750 and 1,000 FTU of phytase kg-1 feed. When adding the phytase enzyme, thenutritional matrix was valued to guarantee the same nutritional levels as the control diet. In general, the addition of phytase enzyme determined a linear decrease on the performance of the birds. However, the performance obtained with the level of 250 FTU phytase kg-1 feed were no different from the control treatment. The best bone density results were observed in the control treatment with no phytase, and the highest leg and thigh yield were obtainedat the level of 514 FTU phytase kg-1.Um experimento foi realizado com o objetivo de avaliar a inclusão de diferentes níveis da enzima fitase, sobre o desempenho, a densitometria óssea e o rendimento de carcaça e partes de frangos de corte nas diferentes fases de criação. Foram utilizados 960 pintainhos de corte com um dia de idade, da marca comercial Ross, distribuídos em um delineamento inteiramente casualizado, com quatro tratamentos mais uma testemunha (quatro níveis da enzima fitase -250, 500, 750 e 1.000 FTU de fitase kg-1 de ração + testemunha e seis repetições de 32 aves cada. Ao adicionar a enzima fitase, a matriz nutricional da mesma foi valorizada para garantir os mesmos níveis nutricionais da dieta testemunha. Foram avaliados os índices dedesempenho, de densitometria óssea e o rendimento de carcaça e de partes nas diferentes fases de criação. De maneira geral o aumento na inclusão da enzima

  3. Effect of K148 and K149 on the Binding between Aspergillus Niger NRRL 3135 Phytase and Phytate%K148与K149对黑曲霉NRRL3135植酸酶A与植酸结合机制的影响

    Institute of Scientific and Technical Information of China (English)

    康伟; 马宝全

    2013-01-01

    In order to clarify that K148 and K149 play a role in binding between Aspergillus niger NRRL 3135 phytase and phytate,a single mutant (K148E) and a double mutant (K148E and K149E) were constructed.Both of the mutant phytases showed a decrease of catalytic activity in different extent.With the help of InsightⅡ,the 3-D structures of the two mutant phytases were modeled and optimized.Some obvious changes of the electrostatic potential surfaces were observed in the two mutant phytases.The results illustrated that changing two sites in the α-helix (141~160) reduces phytate concentration around catalytic pocket and plays a retarded role in transferring phytate to binding sites of catalytic active center.%为了研究黑曲霉NRRL 3135植酸酶A K148与K149对底物与酶的结合反应的影响,构建了一个单位点突变体(K148E)和一个双位点突变体(K148E和K149E),由于带电的氨基酸残基发生变化,造成这2个突变体的酶活性都有不同程度的减少,利用InsightⅡ软件模拟了2种突变体酶的3-D结构,发现酶分子突变位置的表面电势有明显的改变.这2个α-螺旋(141~160)位点的改变可能降低了活性中心附近的底物浓度,减缓了底物与活性中心的反应速度.

  4. 植酸酶对0~3周樱桃谷鸭生长性能、钙、磷表观存留率影响%Effects of Phytase on Growth Performance and Cacium Phosphorus Apparent Retention Ratio of Duckling in 0~3 Weeks

    Institute of Scientific and Technical Information of China (English)

    贾振全; 顾惠明; 金岭梅; 赵枝新

    2001-01-01

    为了在负对照组(钙、磷水平各减少0.1%)日粮基础上添加植酸酶对肉鸭生长、钙、磷表观存留率的影响,用64只1日龄樱桃谷鸭进行试验,按体重随机分为4组,代谢期6天。试验组鸭分别添加植酸酶300和500PU/kg。结果表明:添加植酸酶显著提高试鸭的日采食量、日增重、钙、磷的表观存留率,植酸酶可以部分替代磷酸氢钙。%An experiment was conducted to investigate the effects of addition phytase in negative control group (cacium, phosphorus level reducing by 0.1% each) on growth, cacium, phosphorus apparent retention ration of ducklings which diet based on soybean meal and sorghum. 64 ducklings of l day old were randomly allotted by body weight to one of four groups The balance lasted for 6 days. The two test group diet contain 300and 500 phytase unit PU each. The results showed mean feed conversion rate, average daily feed intake, daily gain and cacium, phosphorus apparent retention ratio were affected by addition phytase. The phytase can replace part CaHPO4.2H2O in ducking's diet.

  5. Characterization and expression analysis of a gene cluster for nitrate assimilation from the yeast Arxula adeninivorans.

    Science.gov (United States)

    Böer, Erik; Schröter, Anja; Bode, Rüdiger; Piontek, Michael; Kunze, Gotthard

    2009-02-01

    In Arxula adeninivorans nitrate assimilation is mediated by the combined actions of a nitrate transporter, a nitrate reductase and a nitrite reductase. Single-copy genes for these activities (AYNT1, AYNR1, AYNI1, respectively) form a 9103 bp gene cluster localized on chromosome 2. The 3210 bp AYNI1 ORF codes for a protein of 1070 amino acids, which exhibits a high degree of identity to nitrite reductases from the yeasts Pichia anomala (58%), Hansenula polymorpha (58%) and Dekkera bruxellensis (54%). The second ORF (AYNR1, 2535 bp) encodes a nitrate reductase of 845 residues that shows significant (51%) identity to nitrate reductases of P. anomala and H. polymorpha. The third ORF in the cluster (AYNT1, 1518 bp) specifies a nitrate transporter with 506 amino acids, which is 46% identical to that of H. polymorpha. The three genes are independently expressed upon induction with NaNO(3). We quantitatively analysed the promoter activities by qRT-PCR and after fusing individual promoter fragments to the phytase (phyK) gene from Klebsiella sp. ASR1. The AYNI1 promoter was found to exhibit the highest activity, followed by the AYNT1 and AYNR1 elements. Direct measurements of nitrate and nitrite reductase activities performed after induction with NaNO(3) are compatible with these results. Both enzymes show optimal activity at around 42 degrees C and near-neutral pH, and require FAD as a co-factor and NADPH as electron donor.

  6. Effects of replacing monocalcium phosphate with phytase on growth and phosphorus utilization in black sea bream, Acanthopagrus schlegelii%中性植酸酶替代磷酸二氢钙对黑鲷生长和磷利用的影响

    Institute of Scientific and Technical Information of China (English)

    徐树德; 王树启; 游翠红; 张敏智; 张婷; 杜红方; 史宝军

    2014-01-01

    Because of a decrease in production and increase in the cost of fishmeal, there is increased interest in the use of plant-based feed in aquaculture. However, one of the major problems associated with the use of plant protein in fish feed is the presence of anti-nutritional factors, such as phytate, which is the primary storage form of phosphorus. Phy-tate is essentially unavailable to monogastric or agastric aquatic animals as they lack intestinal phytase that promote phytate hydrolysis during digestion. To address this issue, diets for monogastric or stomachless animals are often sup-plemented with monocalcium phosphate (MCP) to meet their phosphorus requirements. However, MCP is derived from fossils so it is a non-renewable resource. Furthermore, the cost of rock phosphorus has increased dramatically over the past several years. Alternatively, exogenous phytase can be added to hydrolyze phytate, and thus improve the use of phytate phosphorus. Thus, the addition of phytase may lower the need for MCP supplementation, thereby lower feed costs. This strategy may also lower the concentration of phosphorus excreted, resulting in lower eutrophication of the surrounding waters. We evaluated whether neutral phytase could partially replace MCP, and thus reduce the dietary in-clusion of MCP. The fish (initial body weight:11.52 ± 0.12 g) were fed one of four diets for 8 weeks:a control diet (designated P1.5) was prepared with 1.5%MCP but without phytase and three other diets (designated as PP1.5, PP1.0, and PP0.5) were supplemented with 1.5%, 1.0%, or 0.5%MCP, respectively, and 200 mg/kg (diet) phytase. Fish fed PP1.0 and PP0.5 had no significant change in weight gain rate(WGR), specific growth rate (SGR), protein efficiency rate (PER), or feed conversion ratio (FCR) compared with the control (P>0.05), whereas fish fed PP0 had significantly lower growth performance in the above parameters (P>0.05). Phytase addition did not affect body composition. The apparent digestibility

  7. Production, characterization and gene cloning of the extracellular enzymes from the marine-derived yeasts and their potential applications.

    Science.gov (United States)

    Chi, Zhenming; Chi, Zhe; Zhang, Tong; Liu, Guanglei; Li, Jing; Wang, Xianghong

    2009-01-01

    In this review article, the extracellular enzymes production, their properties and cloning of the genes encoding the enzymes from marine yeasts are overviewed. Several yeast strains which could produce different kinds of extracellular enzymes were selected from the culture collection of marine yeasts available in this laboratory. The strains selected belong to different genera such as Yarrowia, Aureobasidium, Pichia, Metschnikowia and Cryptococcus. The extracellular enzymes include cellulase, alkaline protease, aspartic protease, amylase, inulinase, lipase and phytase, as well as killer toxin. The conditions and media for the enzyme production by the marine yeasts have been optimized and the enzymes have been purified and characterized. Some genes encoding the extracellular enzymes from the marine yeast strains have been cloned, sequenced and expressed. It was found that some properties of the enzymes from the marine yeasts are unique compared to those of the homologous enzymes from terrestrial yeasts and the genes encoding the enzymes in marine yeasts are different from those in terrestrial yeasts. Therefore, it is of very importance to further study the enzymes and their genes from the marine yeasts. This is the first review on the extracellular enzymes and their genes from the marine yeasts.

  8. Effect of phytase on growth,body composition and phosphorus content in tissues of juvenile grass carp(Ctenopharyngodon idellus) fed plant protein based diet%饲料中添加植酸酶对草鱼生长、体组成及各组织磷含量的影响

    Institute of Scientific and Technical Information of China (English)

    马恒甲; 叶金云; 郭建林; 王友慧; 沈斌乾; 陈建明; 潘茜

    2011-01-01

    An 8-week laboratory trial was designed to study the effect of phytase on growth,body composition and phosphorus content in tissues of juvenile grass carp(C.idellus) fed plant protein based diet.Seven trial diets were formulated to feed fish(with mean initial body weight of 39.68±3.05 g) that were divided into 5 experimental groups and 2 control groups.Five triplicate experimental groups were supplemented with five different levels of phytase(500 U/kg,750 U/kg,1 000 U/kg,1 250 U/kg,2 500 U/kg).The positive control group was supplemented with 1.5% monobasic calcium phosphate;and the negative control group was not supplemented with phytase and monobasic calcium phosphate.Through 60 d experiment,the effects of phytase were examind on growth,body composition and phosphorus content in tissues of juvenile grass carp(C.idellus)fed plant protein based diet.The results showed that,the feed conversion rate(FCR) was significantly lower,whereas protein efficiency ratio(PER) and specific growth rate(SGR) were significantly higher than those of other groups(P0.05),when only monobasic calcium phosphate was supplemented.Compared with negative control group,after added phytase in diet,when the supplemented level of phytase was higher than 1 000 U/kg,the FCR of experimental group 3(1 000 U/kg) was significantly lower than that of negative control group(P0.05),and PER was significantly higher than that of negative control group(P0.05);The phytase has no effcets on the HSI,VSI,and CF of fish;the content of crude fat decreased in body,when the phytase content was higher than 1000U/kg,when the supplemented level of phytase content was 1 000 U/kg,the crude fat in muscle was significantly improved;The phosphorus content was enhanced to some degree in body,muscle and spine after added phytase,and when the content was 1 250 U/kg,the phosphorus content were significantly improved in muscle and spine.The result indicated that,when the supplemented

  9. Fluxo biológico do fósforo no metabolismo de suínos alimentados com dietas contendo fitase Biological P flow on metabolism of pigs fed diets containg phytase

    Directory of Open Access Journals (Sweden)

    José Aparecido Moreira

    2004-12-01

    Full Text Available A pesquisa foi conduzida para avaliar o fluxo biológico do P em suínos, na fase de crescimento, alimentados com dietas à base de milho, farelo de soja, farelo de arroz desengordurado e óleo de soja, isentas de P inorgânico e suplementadas com níveis crescentes de fitase (253, 759, 1.265 e 1.748 UF/kg de dieta, e destacar o melhor nível de utilização da enzima, usando modelos matemáticos. O modelo utilizado foi determinístico e compartimental, em que o trato gastrintestinal (C1, o plasma C2, os ossos (C3 e os tecidos moles (ossos, coração, figado, rins e músculos, estudados em conjunto (C4, representaram os compartimentos. Foram utilizados dados de metabolismo e cinética do P nos tecidos, obtidos pela técnica de diluição isotópica. Os parâmetros estimados foram: absorção, retenção, P endógeno que retorna ao trato gastrintestinal, P dietético absorvido, incorporação, reabsorção, balanço de P, P proveniente do osso, dos tecidos moles e do total absorvido que retorna ao trato gastrintestinal. o modelo biomatemático utilizado mostrou-se eficiente em explicar o fluxo do fósforo no organismo de suínos em crescimento. A fitase interferiu no fluxo biológico do P do compartimento C1 para o C3 e no refluxo dos compartimentos C3 e C4 para o C1. O nível 759 UF/kg de ração disponibilizou mais eficientemente o fósforo orgânico para o metabolismo dos suínos.The study was conducted to evaluate the biological flow of P in pigs fed diets based on corn, soybean meal, defatted rice bran (DRB and soybean oil, with increasing phytase levels (253, 759, 1265 and 1748 PU/kg of diet, using mathematics models. The model was deterministic and compartimental, in which the gastrintestinal tract (GIT (C1, the plasma C2, the bones (C3 and the soft tissues (liver, heart, kidney and muscle (C4 represented the compartments. Metabolism data and kinetics of P in tissues were used in the model, obtained by the isotopic dilution technique. The

  10. Utilização de ácidos orgânicos e fitase em dietas para leitões na creche Utilization of organic acids and phytase in diets for weaned piglets

    Directory of Open Access Journals (Sweden)

    E.V.H. Rocha

    2008-06-01

    Full Text Available Verificou-se o efeito da inclusão de um complexo de ácidos orgânicos (AO e diferentes níveis de fitase em dietas de leitões na creche. Foram utilizados 40 leitões desmamados (7,8±0,8kg, distribuídos em delineamento de blocos inteiramente ao acaso, com cinco tratamentos e quatro repetições, durante um período experimental de 28 dias. Os tratamentos foram: T1- ração basal com 500UF/kg (controle; T2- ração basal com 500UF/kg + 0,2% do complexo de AO; T3- ração basal com 450UF/kg + 0,2% do complexo de AO; T4- ração basal com 400UF/kg + 0,2% do complexo de AO; e T5- ração basal com 350UF/kg + 0,2% do complexo de AO. A adição de AO à ração que continha 500UF propiciou maior (P0,05 no ganho de peso. Para o consumo de ração médio diário, observou-se uma regressão quadrática (PThe effect of inclusion of organic acids complex (OAC and different levels of phytase in piglet diets were evaluated. Forty weaned piglets (7.8±0.8kg were allotted in a completely randomized block design with five treatments and four repetitions, during an experimental period of 28 days. The treatments were: T1-basal diet with 500 phytase unit/kg (control; T2-basal diet with 500PU/kg + 0.2% of OAC; T3-basal diet with 450PU/kg + 0.2% of OAC; T4-basal diet with 400PU/kg + 0.2% of OAC; and T5-basal diet with 350PU/kg + 0.2% complex of OA. The addition of OAC at the 500PU diet increased (P0.05 in the weight gain. For the daily average feed intake, it was observed a quadratic effect (P<0.05, and the 411PU level caused lesser feed intake. The level of 500PU + OAC resulted in higher percentage of hard feces compared to the control group during the two last weeks of the experimental period. It was concluded that the effect of the OAC resulted positive when phytase was supplemented to the diets of piglets from eight to 20kg.

  11. Immunoglobulin genes

    Energy Technology Data Exchange (ETDEWEB)

    Honjo, T. (Kyoto Univ. (Japan)); Alt, F.W. (Columbia Univ., Dobbs Ferry, NY (USA). Hudson Labs.); Rabbitts, T.H. (Medical Research Council, Cambridge (UK))

    1989-01-01

    This book reports on the structure, function, and expression of the genes encoding antibodies in normal and neoplastic cells. Topics covered are: B Cells; Organization and rearrangement of immunoglobin genes; Immunoglobin genes in disease; Immunoglobin gene expression; and Immunoglobin-related genes.

  12. Digestibility of the cottonseed meal with or without addition of protease and phytase enzymes in swine diet - doi: 10.4025/actascianimsci.v34i3.12360

    Directory of Open Access Journals (Sweden)

    Maria do Carmo Mouhaupt Marques Ludke

    2012-05-01

    Full Text Available This study evaluated the digestibility of cottonseed meal with or without addition of enzymes (phytase and protease for growing pigs. It was used 18 barrows, housed in metabolism cages, distributed in a completely randomized design, standardizing body weight (bw with average of 25.8 ± 3.6 kg, with three treatments and six repetitions. The treatments consisted of a reference diet based on corn and soybean meal, the second treatment with replacement of 30% of the reference diet by cottonseed meal without enzymes, and the third with 30% of the reference diet replaced by cottonseed meal with added enzymes. Was determined the digestible protein, digestible energy, digestibility of dry matter, energy and protein. It was also registered the balance of nitrogen and phosphorus. The use of cottonseed meal with the addition of enzymes in diets for growing pigs has no effect on the digestibility of dry matter, gross energy and crude protein, but improved the absorption of phosphorus, consequently reducing its excretion in the feces. There was no improvement in nitrogen balance in the diets containing cottonseed meal with enzymes.

  13. Iron bioavailability from a lipid-based complementary food fortificant mixed with millet porridge can be optimized by adding phytase and ascorbic acid but not by using a mixture of ferrous sulfate and sodium iron EDTA.

    Science.gov (United States)

    Cercamondi, Colin I; Egli, Ines M; Mitchikpe, Evariste; Tossou, Felicien; Hessou, Joamel; Zeder, Christophe; Hounhouigan, Joseph D; Hurrell, Richard F

    2013-08-01

    Home fortification with lipid-based nutrient supplements (LNSs) is a promising approach to improve bioavailable iron and energy intake of young children in developing countries. To optimize iron bioavailability from an LNS named complementary food fortificant (CFF), 3 stable isotope studies were conducted in 52 young Beninese children. Test meals consisted of millet porridge mixed with CFF and ascorbic acid (AA). Study 1 compared iron absorption from FeSO4-fortifed meals with meals fortified with a mixture of FeSO4 and NaFeEDTA. Study 2 compared iron absorption from FeSO4-fortifed meals without or with extra AA. Study 3 compared iron absorption from FeSO4-fortified meals with meals containing phytase added prior to consumption, once without or once with extra AA. Iron absorption was measured as erythrocyte incorporation of stable isotopes. In study 1, iron absorption from FeSO4 (8.4%) was higher than that from the mixture of NaFeEDTA and FeSO4 (5.9%; P porridge.

  14. 放射型根瘤杆菌植酸酶对麸皮中植酸的脱磷作用%Dephosphorization of Phytic Acid in Wheat Bran by Phytase from Agrobacterium radiobacter

    Institute of Scientific and Technical Information of China (English)

    李文; 王陶; 杨克

    2011-01-01

    Crude phytase solution separated from Agrobacterium radiobacter fermentation broth was used to hydrolyze wheat bran for phytic acid dephosphorization.Using one-factor-at-a-time combined with orthogonal array design method,such process conditions as enzyme amount,substrate concentration,pH,temperature and hydrolysis time were optimized to be 750 U/g,40 mg/mL,7,45 ℃,and 5 h,respectively.Under the optimized conditions,the hydrolysis rate of phytic acid was up to 81.50%.%采用放射型根瘤杆菌植酸酶粗酶液水解麸皮中的植酸,研究其脱磷的最适条件。依据单因素试验和正交试验获得脱磷的最优条件为加酶量750U/g、底物质量浓度40mg/mL、温度45℃、pH7、时间5h。最优条件下,麸皮中植酸的水解率在5h达最大,为81.50%。

  15. Study on enzymology characteristics of phytase secreted by efifcient phytate-degrading rhizobacteria Rahnella aquatilis JZ-GX1%高效植酸盐降解细菌水拉恩氏菌JZ-GX1的植酸酶特性研究

    Institute of Scientific and Technical Information of China (English)

    李桂娥; 吴小芹

    2014-01-01

    The efficient phytate-degrading rhizobacteria Rahnella aquatilis JZ-GX1 were used as the research object to study the locating expression of phytase secreted by JZ-GX1. The results show as follews:there were significantly differences among phytase specific activities of different parts of the bacteria culture incubated for 48 hours. the phytase specific activities of extracellular supernatant, intercellular substance and inside cell were 2.5037 U/mg, 0.0015 U/mg, 0.0002 U/mg respectively, which indicated that the phyratse secreted by JZ-GX1 was a kind of extracellular enzyme. Learned from the studies on enzymology characteristics of phytase of JZ-GX1, it showed that the phytase secreted by JZ-GX1 had excellent resistance to acid, but not alkali resistance, its optimal pH value was about 5.5, optimum reaction temperature was about 37℃, when temperature was less than 50 ℃, the phytase had good thermal stability. The results of this study could provide a useful reference for using R. aquatilis JZ-GX1 as bio-fertilizer in agricultural production in the future.%以高效植酸盐降解细菌水拉恩氏菌(Rahnella aquatilis)JZ-GX1为研究对象,对其分泌的植酸酶在细胞内的定域进行研究,结果表明:在该菌48 h的培养物中,菌株细胞不同部位的植酸酶比活力存在极显著差异,分别为:胞外上清液2.5037 U/mg,胞间质0.0015 U/mg,胞内0.0002 U/mg,说明JZ-GX1菌株所产植酸酶为胞外酶;通过研究pH值和温度对该菌分泌的胞外植酸酶活性的影响表明,JZ-GX1菌株所产植酸酶为酸性植酸酶,最适pH值为5.5,具有一定的耐酸性,但不耐碱;最适温度为37℃,当温度小于50℃时,该酶具有良好的热稳定性。本研究为水拉恩氏菌JZ-GX1将来作为微生物菌肥应用于农林业生产,提供了有价值的参考依据。

  16. Effects of low-available phosphorus diet supplemented with phytase on laying performance,digestive enzymes activities and nutrient utilization in laying hens%低磷日粮中添加植酸酶对蛋鸡产蛋性能、内源消化酶及养分消化率的影响

    Institute of Scientific and Technical Information of China (English)

    邱梅平; 温超; 张旭晖; 王恬

    2011-01-01

    To evaluate the effects of low-available phosphorus (AP) diet supplemented with phytase on the laying performance, the activity of digestive enzyme and nutrient utilization in laying hens, 18-week-old laying hens (n = 840)were randomly allocated into 7 groups.Group Ⅰ , as the positive control, was fed with the basal diet.Group Ⅱ, as the negative control, was fed with low-AP diet.Groups Ⅲ, Ⅳ and Ⅴ were fed with low-AP diet adding coated dosage-form phytase at three levels (300 U/kg, 1 000 U/kg and 10 000 U/kg, respectively).Groups Ⅵ and Ⅶ were supplemented with 300 U/kg phytase of powdery dosage-form and granular dosage-form, respectively.The results indicated that compared with the negative control, the addition of phytase to the low-AP diet did not significantly affect egg weight, broken eggshell rate, the indexes of egg quality,the activity of amylase, the activity of disaccharidase and the apparent digestibility of dry matter, ash and energy (P>0.05), but significantly enhanced the egg preduction, laying rate, the activity of trypsin in group Ⅳ and the apparent digestibility of Ca, P and protein ( P<0.05 ).It was concluded that low-AP diet supplemented with phytase could increase laying performance, the activity of digestive enzyme and nutrient utilization in laying hens.The 300 U/kg coated phytase was enough to meet the nutrient requirement of laying hens for growth and production.%为探明在低磷的玉米一豆粕型日粮中添加植酸酶对蛋鸡产蛋性能、内源消化酶及养分消化率的影响,该研究选用18周龄体重、产蛋周龄相近的伊莎褐蛋鸡840羽,随机分成7组:对照1( Ⅰ组,基础日粮,未添加植酸酶)、对照2(Ⅱ组,有效磷水平降低0.12个百分点)及在对照2基础上分别添加300 U/kg、1 000 U/kg和10 000U/kg的包衣状植酸酶、300 U/kg的粉状植酸酶及300 U/kg的微丸状植酸酶(试验Ⅲ、Ⅳ、Ⅴ、Ⅵ和Ⅶ组).结果表明:与对照2相比,除Ⅴ组外,各试验

  17. Modelos matemáticos para o estudo do fluxo biológico do fósforo em suínos alimentados com dietas suplementadas com níveis crescentes de fitase Mathematical models for the study of the biological flow of phosphorus in pigs fed with diets containing increasing levels of phytase

    Directory of Open Access Journals (Sweden)

    J.A. Moreira

    2009-04-01

    Full Text Available A pesquisa foi realizada para avaliar o fluxo biológico do fósforo entre os compartimentos, fisiológicos ou anatômicos, de suínos mantidos em dietas que continham níveis crescentes de fitase, usando o P-32 como traçador. O delineamento experimental foi inteiramente ao acaso, sendo os tratamentos constituídos por níveis de enzima fitase, 253, 759, 1265 e 1748 UF/kg, nas dietas. As variáveis avaliadas foram: ingestão, excreção, atividades específicas e fluxo de P nos compartimentos (trato digestivo, corrente sanguínea, tecidos moles e ossos. A enzima fitase não interferiu nas excreções fecais e urinárias, nas atividades específicas, na incorporação e na reabsorção de P nos ossos e nos tecidos moles e no fluxo bidirecional do trato digestivo e na corrente sanguínea, mas afetou linearmente a absorção e a retenção nos ossos. As respostas mais evidentes dos efeitos da adição da fitase são observadas nos níveis mais baixos.The biological flow of P was evaluated among the physiologic or the anatomical compartments of pigs fed diets with increasing phytase levels, using P-32 as tracer. The experimental design was completely randomized. Treatments consisted of phytase levels in diets (253, 759, 1265, and 1748UF/kg. The evaluated variables were: feed intake, excretion, specific activities and flow of P in the compartments (gut, blood, bone, and soft tissus. Phytase level did not interfere with fecal and urinary excretion, specific activities, incorporation and resorption of bone, and soft tissue P and in the bidirectional flow of gut and blood, but linearly affected P absorption and bone P retention. The best response was observed with the lowest phytase levels.

  18. Effects of phytase on growth, digestive enzyme activity and digestibility in juvenile redfin puffer Takifugu rubripes%植酸酶对红鳍东方鲀幼鱼生长、消化酶及消化率的影响

    Institute of Scientific and Technical Information of China (English)

    孟祥科; 孙阳; 屈菲; 姜志强; 毛明光; 李艳秋; 吴洪

    2013-01-01

    A 60-day feeding trail was conducted to study the influence of phytase on growth performance,digestive enzyme activity and apparent digestibility in juvenile fedfin puffer Takifugu rubripes with body weight of(4.58 ±0.05) g.The fish were fed diets containing fish meal protein (control group D1),and 70% of fish meal protein and 30% of soybean meal protein (control group D2).In the experimental group,the fish were fed the 30% fish meal diets supplemented with phytase at a rate of 500 (S1),1 000 (S2),1 500 (S3),and 2 000 (S4) U/kg with triplicate.The results showed that there were no significant effects on the survival rate of the juveniles in different groups(P>0.05).The best growth performance was observed in the juveniles in the control group D1,significantly better than that in the control group D2 (P < 0.05).There were significantly higher final body weight (FBW) and specific growth rate (SGR) and lower food conversion rate (FCR) in the fish fed the diets containing phytase at a rate over 1 000 U/kg (P<0.05).The fish fed the diets containing phytase had higher intestinal activities of protease,lipase and amylase,and showed more dry matter digestibility without significant difference (P>0.05).The maximum dietary protein digestibility was observed in the juveniles fed the diets supplemented with phytase at a dose of 1 000 U/kg diet,and significantly higher activity of lipase was found in the juveniles fed the diets supplemented with phytase at a dose of 1 000 and 1500 U/kg diet (P<0.05).Increase in dietary phytase level from 500 U/kg to 2 000 U/kg led to significantly elevated digestibility of phosphorus by the fish (P<0.05).In conclusion,it is suggested that the optimal level of dietary phytase be from 1 000 U/kg diet to 1 500 U/kg diet since the phytase will promote the growth and improve digestibility of the puffer.%以初始体质量为(4.58±0.05)g的红鳍东方鲀Takifgu rubripes幼鱼为对象,进行为期60 d的饲养试验,研究植酸

  19. Effects of phytase on vertebral shapes and non-specific immune related enzymes activities in channel%植酸酶对斑点叉尾鮰脊椎形态和非特异性免疫相关酶活力的影响

    Institute of Scientific and Technical Information of China (English)

    华雪铭; 王世忠; 陈瑶琴; 钟国防; 周洪琪

    2014-01-01

    ( myo-inositol-hexaphosphate phosphohydrolase) was the main storage form of phosphorus ( P) in plant proteins and not available for fish due to the lack of intestinal phytase for efficient phytate hydrolysis during digestion . Therefore , most of the phytate-P was excreted into the water which contributed to eutrophication effects . Phytase could be added to plant-based diets to hydrolyze non-digestible phytate into available-P during passing through the fish gut and improve the P availability , which reduced P excretion and its loss into the environment in fish diets . Furthermore , phytase could also improve protein digestibility by hydrolyze the form of phytate-protein complexes that are resistant to proteolytic digestion . In the present , most researches on phytase were directed toward determining its effect on bioavailability of nutrients ( P and protein) and the grow th performance of fish . Based on the fact that nutrients ( P and protein) were essential to maintain bone health and non-specific immune responses in fish , it was necessary to explore the effects of phytase on the bone health and non-specific immune responses in fish . This experiment was conducted to evaluate the effects of different levels of liquid phytase on vertebral shapes and non-specific immune responses in channel catfish ( Ictalurus punctatus ) . Six isonitrogenous ( 37.50% crude protein) and isocaloric ( 17.00 kJ/g gross energy) diets were formulated , a negative control ( NC) diet :a basal diet was supplemented with 1% dicalcium phosphate ( CaHPO4 ) , a positive control ( PC ) diet : the NC diet was supplemented with another 1% dicalcium phosphate (CaHPO4) ,in addition ,four phytase supplementation groups:the NC diets with graded levels ( 250 , 500 , 1 000 , 2 000 U/kg) of phytase were assigned and fed channel catfish (5.78 ± 0.07) g/individual with 5 replicates for 60 days after a foster period . Results showed that the mass gain rate and feed conversion ratio were significantly

  20. High Doses of Microbial Phytase Affect Nutrient Utilization in Broiler Chicks Fed Corn-soybean Meal Diets%玉米-豆粕型日粮中添加高剂量植酸酶对肉鸡养分利用率的影响

    Institute of Scientific and Technical Information of China (English)

    孙宏选; 高秀华; 杨禄良

    2011-01-01

    本试验研究了玉米-豆粕型日粮中添加高剂量不同水平的植酸酶对0~3和4~6周肉鸡养分利用率的影响.选用1日龄AA肉鸡公雏576只,随机分为6组,每组8个重复,每个重复12只.试验日粮分0~3和4~6周两个阶段,6个处理组:正对照组(前期为总磷0.69%,0.45%NPP;后期为总磷0.59%,0.35%NPP);负对照组(前期总磷0.60%,0.35%NPP;后期为总磷0.49%,0.25%NPP);4个试验组在负对照日粮中分别添加500、2000、8000和32000 FTU/kg 4种植酸酶水平.结果显示:①日粮添加高剂量不同水平的植酸酶显著影响生长前期(17~21日龄)肉鸡的钙、磷的消化利用率(P<0.05),随着植酸酶添加水平增加,钙的消化利用率显著增加(P<0.05).②日粮添加高剂量不同水平的植酸酶显著影响38~42日龄肉鸡磷的消化利用率(P<0.05).在负对照组日粮中随着植酸酶水平添加的提高,钙的消化利用率呈现先增加后降低的趋势,磷的养分利用率反而显著减少.③日粮添加高剂量不同水平植酸酶显著增加生长前期(17~21日龄)肉鸡的粗蛋白质的消化利用率和表观代谢能(P<0.05),在生长后期(38~42日龄)对肉鸡的粗蛋白质的消化利用率和表观代谢能提高达到显著水平(P<0.05).因此,高剂量不同水平的植酸酶显著提高肉鸡AME和粗蛋白质,钙、磷的消化利用率,添加高水平植酸酶可以适当调低日粮能量和蛋白质水平,日粮中总磷(tP)水平应低于NRC标准,减少无机磷和钙的添加量,以提高钙、磷在体内的存留量.%An experiment was conducted to determine the effect of high levels of microbial phytase supplementation in cornsoy-based diets on nutrient availability of broiler chickens. A totalof 576 one-day old male Arbor Acres broilers were assigned to 6 dietary treatments,consisting of 8 pens of 12 birds each,and were fed two periods experimental diets from 1 to 21 d and 22 to 42 d. The diets include a positive control

  1. MICROMINERAL-VITAMIN SUPPLEMENT, INGARNIC PHOSPHORUS REDUCTION AND PHYTASE ADDITION IN FINISHING RATIONS FOR SWINE RETIRADA DO SUPLEMENTO MICROMINERAL-VITAMÍNICO, REDUÇÃO DE FÓSFORO INORGÂNICO E ADIÇÃO DE FITASE EM RAÇÕES DE SUÍNOS NA FASE DE TERMINAÇÃO

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    Suelson Pereira dos Santos

    2008-10-01

    Full Text Available The experiment aimed to evaluate the effect of mineral-vitamin supplement withdrawal associated to inorganic phosphorus and phytase addition on swine performance and carcass characteristics in finishing phase.

    48 hybrid swine females with initial average weight of 66,34 kg and final average weight of 106,14 kg were housed in 24 stalls for 38 days and allotted to a completely randomized experimental design with six treatments (T1 – basal feed; T2 – basal feed without mineral vitamin supplement; T3 – basal feed without mineral vitamin supplement with phytase; T4 – basal feed without mineral vitamin supplement less1/3 of inorganic phosphorus with phytase; T5 - basal feed without mineral vitamin supplement less 2/3 of inorganic phosphorus with phytase and T6 – basal feed without mineral vitamin supplement and inorganic phosphorus with phytase with four replications of two animals each. Daily weight gain, total average weight gain and feed:gain ratio were determined and carcass characteristics were evaluated: loin eye area, backfat depth, carcass length, carcass weight and carcass yield. Although reduction in inorganic phosphorus levels or phytase addition had no influence in carcass characteristics (P>0,05, there was significant effect in performance (P<0,05, resulting in less average feed intake, weight gain and feed:gain ratio. Likewise, reduction in inorganic phosphorus levels or phytase addition under the conditions of this trial did not improve economic efficiency when compared to basal feed.

    KEY WORDS: Finishing phase pig, phosphorus, phytase.

    Para avaliar o efeito da retirada do suplemento micromineral-vitamínico associada à redução dos níveis de fósforo inorgânico e adição de fitase em rações, sobre o desempenho e as características de carcaça de suínos na fase de terminação, realizou-se um experimento na Escola de Veterin

  2. Optimization of phytase hydrolysis of soybean protein isolated by response surface methodology%响应面法优化植酸酶水解大豆分离蛋白的工艺

    Institute of Scientific and Technical Information of China (English)

    张仁楠; 姚磊; 富校轶

    2011-01-01

    以大豆分离蛋白为实验材料,在酶解温度、酶添加量、pH、水解时间等单因素实验的基础上,根据Box-Behnken中心组合实验设计原理,采用四因素三水平响应曲面分析法,依据回归分析确定各工艺条件的主要影响因素,以大豆分离蛋白中植酸的去除率为响应值作响应面。植酸去除的最佳工艺条件为:反应温度44.87℃,酶添加量为8.24%,pH为5.54,反应时间为3.03h。在最佳工艺条件下,植酸的去除率为95.15%,验证实验结果为94.98%。%With soybean protein isolated as experiment material,on the base of single factor experiments(hydrolysis time,hydrolysis temperature and so on),then through the experimental design of response surface methodology with four factors and three levels based on the principle of Box-Behnken design,the conditions of hydrolysis were studied.The optimal hydrolysis conditions:hydrolysis temperature was 44.87℃,exterior phytase addition about 8.24%,pH was 5.54,hydrolysis time was 3.03h.Experiments that checked the optimal hydrolysis time conditions showed that phytate of soybean protein isolated was degraded about 95.15%,proof test result was 94.98%.

  3. Quick Purification of Phytase from Aspergillus oryzae A-1 and Enzymatic Characterization%米曲霉A-1植酸酶快速纯化及酶学性质

    Institute of Scientific and Technical Information of China (English)

    何海燕; 张健; 覃拥灵

    2013-01-01

    In order to know phy A-l in deep,a phytase (Phy Al) was purified from the enzyme extract of A. oryzae A-l through QuixStandTM benchtop hollowfiber ultrafiltration system and Mono Q anion-exchange chromatography. The optimal working temperature for Phy Al was 45 ℃ and enzyme was stable under 60 ℃. The optimal pH for Phy Al was 5. 5, and enzyme was stable at pH 4. 0~10. 0, Mg2+ and Mn2+ had significant stimulative effects. The Km of Phy Al determined from Lineweaver-Burk plots was 42 μmol/L with sodium phytate as the substrate.%为进一步了解米曲霉A-1植酸酶,利用超滤及Mono Q阴离子纯化柱纯化,分离出米曲霉A-1的植酸酶Phy A1,分子量约为45 kDa,Phy A1在pH5.5时酶活最高,pH4.0~9.0较稳定,最适温度为45℃,在60℃以下内切酶稳定性较好.Mg2+、Mn2+对Phy A1植酸酶活性有较好的激活作用.以植酸钠为底物,应用Lineweaver-Burk双倒数作图法得到米氏常数Km为42 μmol/L.

  4. Fitase na alimentação da tilápia do Nilo (Oreochromis niloticus, durante o período de reversão de sexo - DOI: 10.4025/actascianimsci.v26i3.1778 Phytase as feeding for Nile tilapia (Oreochromis niloticus, during sex reversion period - DOI: 10.4025/actascianimsci.v26i3.1778

    Directory of Open Access Journals (Sweden)

    Lilian Carolina Rosa Silva

    2004-04-01

    Full Text Available Este trabalho foi realizado para avaliar a influência de níveis de fitase sobre o desempenho da tilápia do Nilo (Oreochromis niloticus no período de reversão de sexo. Foram utilizadas 1000 larvas de tilápia do Nilo com peso inicial 0,02 ± 0,002g, alimentadas com rações contendo 0; 500; 1000; 2000 e 4000 unidades de fitase (UF/kg, durante 31 dias. A ração referência continha 3143 kcal de energia digestível/kg, 30% de proteína bruta e 0,85 e 0,35% de fósforo (P total e disponível, respectivamente. Não foram observados efeitos (P>0,05 dos valores de inclusão de fitase sobre o ganho de peso e taxa de sobrevivência. O aumento nos níveis de fitase nas rações elevou linearmente (PThis work was carried out to examine the influence of dietary levels of phytase on Nile tilapia (Oreochromis niloticus performance during the sex reversion period. Were used 1,000 fries of Nile tilápia with initial weight 0.02 ± 0.002g, fed with diets containing 0; 500; 1000; 2000 and 4000 phytase units (PU/kg of diet 31 days. The reference diet provided 3,143 kcal of digestible energy/kg, 30% of crude protein and 0,85 and 0,35% of total and disponible phosphorus (P, respectively. No effects (P>0.05 were observed of phytase inclusion on weight gain and survival rate. Dietary phytase levels increase linearly (P<0.05 the calcium retention in the carcass. There was observed quadratic effect (P<0.05 of dietary phytase supplementation on P retention in the carcass, estimated the values of 1,990 PU/kg for the highest value for this variable. The results of this work indicate that a dietary level of 1,990 of PU provide maximal P retention in the carcass of Nile tilapia, during sex reversal period.

  5. Utilização da fitase em dietas com ou sem farelo de arroz desengordurado para suínos em crescimento/terminação Use of phytase in diets with or without defatted rice bran for growing/finishing pigs

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    Maria do Carmo Mohaupt Marques Ludke

    2002-09-01

    Full Text Available Este trabalho foi conduzido com o objetivo de comparar a eficiência de diferentes níveis de fitase em relação à suplementação de fosfato inorgânico (fosfato bicálcico, quando adicionados em dietas sem e com farelo de arroz desengordurado (FAD. Foram utilizados 128 suínos com peso inicial médio de 21,2 ± 2,4 kg, alojados em baias coletivas, para avaliar o desempenho nas fases de crescimento e terminação. Os suínos foram abatidos com peso médio de 95,4 ± 5,3 kg e coletado o osso metacarpo para determinação do cálcio e do fósforo. Foram usados oito tratamentos, sendo quatro dietas sem FAD (milho e farelo de soja e quatro dietas com 30% de inclusão de FAD, nas quais foram adicionados níveis de 0, 750 ou 1000 UF (Unidades de Fitase/kg. Foi observado que adicionar fitase, tanto em dietas sem FAD quanto em dietas com FAD, melhora a disponibilidade do fósforo orgânico. Com os níveis de 750 e 1000 UF/kg adicionados em ambas as dietas, sem suplementação de fosfato bicálcico, não houve prejuízo sobre o desempenho dos animais. Entretanto, as características ósseas são prejudicadas se as dietas com fitase não forem suplementadas com a fonte de fósforo inorgânico.The objective of this work was to compare the efficiency of different levels of phytase with the use of inorganic phosphate (dicalcium phosphate when added to diets with or without defatted rice bran (DRB. One hundred and twenty eight pigs with average initial weight of 21.6 ± 2.4 kg were housed in collective boxes for the pig performance evaluation in the growing and finishing phase. The pigs were slaughtered with an average weight of 95.42 ± 5.3 kg and the metacarpus bones were collected to determine the calcium and phosphorus content. Eight treatments were used with four diets being formulated without DRB (corn-soybean meal and the other four diets with DRB. The phytase levels added in the diets were 0, 750 or 1000 PU/kg of the diet. It was observed that

  6. Associação de carboidrase e fitase em dietas valorizadas para poedeiras semipesadas - doi: 10.4025/actascianimsci.v34i3.12735 Association of carbohydrases and phytase in enriched diets for semi heavy layers - doi: 10.4025/actascianimsci.v34i3.12735

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    Jerônimo Ávito Gonçalves de Brito

    2012-05-01

    Full Text Available O objetivo do estudo foi avaliar o efeito da associação de carboidrases e fitase em dietas de poedeiras. Foram utilizadas 300 poedeiras Isa Brown com 26 semanas, submetidas aos tratamentos: Controle Positivo: sem enzimas e valorização dos nutrientes; T2: valorização em 1,5 e 6% a EMA (kcal kg-1, do milho e o farelo de soja, respectivamente, e redução nos níveis nutricionais conforme a matriz nutricional para a fitase; T3: valorização da EMA do milho e o farelo de soja (idem T2 e redução nos níveis de cálcio e fósforo conforme matriz nutricional da fitase; T4: T2 suplementado com 100 g t-1 de carboidrases e 30 g t-1 de fitase; T5: T3 suplementado com 100 g t-1 de carboidrases e 30 g t-1 de fitase. Foi utilizado delineamento inteiramente casualizado, seis repetições e parcela subdividida no tempo (6 períodos de 21 dias. Não houve efeito significativo dos tratamentos sobre a produção, consumo, peso dos ovos e qualidade dos ovos. A valorização dos nutrientes preconizada com a utilização das enzimas foi eficiente tecnicamente para manutenção do desempenho e qualidade dos ovos de poedeiras semipesadas nas condições avaliadas, necessitando de outras pesquisas com maior redução nos níveis energéticos para avaliação do efeito das enzimas sobre a disponibilidade de energia.The aim of this study was to evaluate the effect of the combination of carbohydrases and phytase in diets of laying hens. A total of 300 Isa Brown hens at 26 weeks were submitted to the following treatments: Positive Control: without enzyme and nutrient recovery, T2: enrichment by 1.5 and 6% AME (kcal kg-1 of corn and soybean meal, respectively, and reduction in nutrient levels as the nutrient matrix for the phytase; T3: EMA enrichment of corn and soybean meal (T2 identification and reduced levels of calcium and phosphorus as the phytase nutrient matrix; T4: T2 supplemented with 100 g ton.-1 of carbohydrases and 30 g ton.-1 of phytase; T5: T3

  7. Effect of different inclusion levels of defatted corn germ meal in diets supplemented with phytase for laying hensEfeito de diferentes níveis de farelo de gérmen de milho desengordurado em dietas suplementadas com fitase para poedeiras comerciais

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    Sandra Regina Brunelli

    2012-10-01

    Full Text Available The experiment was designed to evaluate the performance, egg quality and minerals on tibia of laying hens from 28 to 44 weeks of age subjected to diets containing varying levels of DCGM (0, 12 and 30% and phytase (0, 500 and 1000 FTU/kg of diet. The experiment was carried in four cycles of 28 days and the variables studied were: the performance (feed intake, egg production, egg weight, egg mass and feed conversion and egg quality (egg specific gravity, yolk color, shell thickness and Haugh unity value. In the finish of experiment, five birds per treatment were abated for determination of the calcium and phosphorus in tibia. There was no interaction between DCGM and phytase on evaluated parameters. Inclusion of DCGM resulted in significantly higher feed intake and worst yolk pigmentation, than the 0% treatment. Increasing the phytase dose to 1000 FTU/kg improves yolk pigmentation. For the others variables no effect were observed for levels of DCGM or phytase. Based on the results from the present study, DCGM could be included at a rate of up to 30% of a commercial layer diets without any significant detrimental effects on the egg production, but with yolk pigment addition. The phytase improve the yolk color.O experimento foi realizado com objetivo de avaliar o desempenho, a qualidade dos ovos e a concentração de minerais na tíbia de poedeiras leves alimentadas com rações contendo farelo de gérmen de milho desengordurado (FGMD e fitase. Foram utilizadas 360 poedeiras de 28 a 44 semanas de idade. As rações experimentais continham diferentes inclusões de FGMD (0%, 12% e 30% e de fitase (0; 500 e 1000 FTU/kg, distribuídos em um delineamento experimental inteiramente casualizado em fatorial 3X 3, totalizando nove rações, com cinco repetições e oito aves por repetição. O experimento teve a duração de quatro ciclos de 28 dias cada, em que foram avaliados consumo de ração, produção, massa e peso médio dos ovos, conversão alimentar

  8. Gene therapy

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    2005147 CNHK200-hA-a gene-viral therapeutic system and its antitumor effect on lung cancer. WANG Wei-guo(王伟国),et al. Viral & Gene Ther Center, Eastern Hepatobilli Surg Instit 2nd Milit Univ, Shanghai 200438. Chin J Oncol,2005:27(2):69-72. Objective: To develop a novel vector system, which combines the advantages of the gene therapy,

  9. Trichoderma genes

    Science.gov (United States)

    Foreman, Pamela [Los Altos, CA; Goedegebuur, Frits [Vlaardingen, NL; Van Solingen, Pieter [Naaldwijk, NL; Ward, Michael [San Francisco, CA

    2012-06-19

    Described herein are novel gene sequences isolated from Trichoderma reesei. Two genes encoding proteins comprising a cellulose binding domain, one encoding an arabionfuranosidase and one encoding an acetylxylanesterase are described. The sequences, CIP1 and CIP2, contain a cellulose binding domain. These proteins are especially useful in the textile and detergent industry and in pulp and paper industry.

  10. Effects of Phytase at Different Levels of Energy and Non-phytate-phosphorus on Performance and Tibial Mineralization of Broilers%不同能量和非植酸磷水平下植酸酶对肉仔鸡生产性能和胫骨矿化的影响

    Institute of Scientific and Technical Information of China (English)

    李宗付; 余冰; 陈代文; 张克英

    2007-01-01

    试验选用640只1日龄爱拔益加(AA)肉仔鸡公雏,采用2×2×2因子设计,通过在不同能量水平(11.90、11.70 MJ/kg)和非植酸磷(NPP)水平(0~3周0.35%,0.25%;3~6周0.30%,0.20%)的肉鸡饲粮中添加植酸酶0、500 FTU/kg,考察了植酸酶对肉鸡生产性能和胫骨矿化的影响.试验共8个处理,每个处理8个重复,每个重复10只鸡,试验期42 d.结果表明:(1)饲粮添加植酸酶降低了肉仔鸡前期的平均日采食量(ADFI)和全期的料重比(F/G)(P<0.05),分别下降2.52%、1.81%;降低了前期胫骨灰分(P<0.05)、后期胫骨Ca(P<0.05)和P含量(P<0.01);但对日增重无显著影响(P>0.05).(2)植酸酶与饲粮能量和NPP水平对生产性能没有显著的交互作用(P>0.05);植酸酶和能量对前期胫骨Ca、P和后期胫骨Ca含量存在极显著的交互作用(P<0.01),表现为低能加酶组前后期胫骨Ca含量极显著高于低能组,但前期胫骨P含量极显著低于低能组.结论:以生产性能为标识,本试验条件下低磷或低能或低磷低能日粮添加植酸酶可以获得和对照组相似的生产性能.%A trial of 2×2×2 factorial design was conducted to evaluate the effects of phytase at the level of 0,500 FTU/kg added in the broiler diets with different energy levels (11.90, 11.70 MJ/kg) and different NNP levels (0.35%, 0.25%) on growth performance and tibial mineralization. A total of 640 arbor acres (AA) broilers at 1 d age were allotted into 8 treatments with 8 replicates of 10 birds each. Experiment was divided into 2 periods which lasted 7 weeks. The results indicated that: (1) In diets with phytase supplementation, ADFI of first period was decreased by 2.52% (P<0.05) and F/G of whole period was decreased by 1.81% (P<0.05) ; the contents of tibial ash (P<0.05) in the first period, tibial Ca (P<0.05) and P (P<0.01) in the last period were all decreased. But phytase did not exhibit significant influence on ADG (P>0.05). (2) Phytase and the

  11. METABOLIC EVALUATIONS IN PIGS WITH MICRO MINERAL AND VITAMINS WITHDRAWAL, INORGANIC PHOSPHORUS REDUCTION AND PHYTASE SUPPLEMENTATION AVALIAÇÃO METABÓLICA DA REDUÇÃO DE FÓSFORO INORGÂNICO E SUPRESSÃO DE MICROMINERAIS E VITAMÍNAS EM DIETAS COM FITASE PARA SUÍNOS

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    Moema Pacheco Chediak Matos

    2008-04-01

    Full Text Available The experiment aimed to evaluate the effects of diets without micro minerals and vitamins, reduced inorganic phosphorus levels and phytase addition in bone metabolism and renal and hepatic functions in finishing pigs. Forty eight 105-day old females were randomly allotted in six groups: standard diets (T1, composed of corn, soybean meal and wheat meal; standard ration without micro mineral and vitamin supplement (T2; T2 ration with phytase (T3 T2 ration reducing 1/3 of inorganic P with phytase (T4, T2 ration reduced 2/3 of inorganic P with phytase (T5 and T2 ration with complete reduction of inorganic P with phytase (T6. The experiment was carried out in two phases: phase 1: 24 animals slaughtered at 150 days of age (106.65 kg; and phase 2: 24 animals slaughtered at 175 days of age (123.87 kg. The serum parameters evaluated were: total and ionic calcium, phosphorus, calcium/phosphorus ratio, total protein, albumin, AST, GGT, alkaline phosphatase, urea and creatinine. All biochemical parameters were analyzed using commercial kits in an automatic analyzer. The levels obtained for all analyzed parameters ranged within normal established values for this species and age. Inorganic phosphorus decrease in phytase-added diets did not interfere in bone metabolism. Vitamin and mineral supplements withdrawal and progressive decrease of inorganic phosphorus and phytase addition in diets of finishing pigs did not change hepatic or renal function. Micro minerals and vitamin restriction for 60 days during finishing phase in pigs did not lead to clinical changes.KEY WORDS: Bone metabolism, hepatic function, renal function. 植酸酶对斑点叉尾鮰生长性能、营养物质表观消化率及氮、磷排泄的影响%Influence of Phytase on Growth Performance, Apparent Digestibility of Nutrients, Excretion of Nitrogen and Phosphorus in Channel Catfish ( Ictalurus punctatus)

    Institute of Scientific and Technical Information of China (English)

    杨雨虹; 刘行彪; 黄可; 付熊; 吴晗冰

    2011-01-01

    This study was conducted to investigate the influence of phytase on growth performance, apparent digestibility of nutrients, excretion of nitrogen and phosphorus in channel catfish (Ictalurus punctatus) and to determine the optimum additive amount of phytase. A total of 540 juvenile channel catfish with an average body weight of (1. 74 ±0. 02) g were randomly divided into 6 groups with 3 replicates per group and 30 fish per replicate. The fish in control group were fed a basal diet, and the fish in experimental groups were fed the basal diet supplemented with 300, 500, 1 000, 1 500 and 2 000 U/kg phytase, respectively. After a 90-day growth experiment, a total of 75 channel catfish [ average body weight of (16. 70 ±0.85) g ] in each group were randomly distributed into 3 replicates and 25 fish in each replicate, for measuring the nitrogen and phosphorus excretion. The nitrogen and phosphorus excretion experiment lasted for 72 h. The results of growth experiment showed as follows: compared with the control group, the weight gain rate, specific growth rate, protein efficiency ratio and apparent digestibility of nutrients in the groups with the diets supplemented with 500 to 2 000 U/kg phytase were significantly improved (P 0. 05). The results of nitrogen and phosphorus excretion experiment showed as follows: the excretion of ammonia nitrogen, total phosphorus and fecal phosphorus per kilogram of weight gain were decreased with the dietary phytase level increasing, and the above indices in the groups with the diets supplemented with 1 000 to 2 000 U/kg phytase were significantly lower compared with the control group (P <0. 05). Based on these findings, this study concludes that supplementation of 1 500 U/kg phytase in diets can improve the growth performance and apparent digestibility of nutrients in channel catfish, and decrease the nitrogen and phosphorus excretion into aquaculture water.%本试验旨在研究植酸酶对斑点叉尾鮰生长性能、营养物质

  12. Rações com níveis reduzidos de proteína bruta, cálcio e fósforo com fitase e aminoácidos para frangos de corte Diets with reduced crude protein, calcium and phosphorus levels with phytase and amino acids for broiler chickens

    Directory of Open Access Journals (Sweden)

    Elisangela Minati Gomide

    2011-11-01

    ácidos.In performance and metabolism trials, diets with reduced levels of crude protein, calcium and available phosphorus on the performance, carcass characteristics, tibia ashes, balance and retention of nitrogen and phosphorus of broiler chickens in phase from 8 to 21 and 8 to 35 days were evaluated. In each phase, 30 and 25 birds per experimental unit were used in the performance assay and five and three in the metabolism assay, respectively, in a completely randomized experimental design, with six replications. Treatments consisted of diet with 21% crude protein without phytase (control, formulated with nutritional levels recommended on the Brazilian tables, and diets with 21, 20, 19 and 18% crude protein with phytase (8 to 21 days and from 8 to 35 days, the control diet had 20% crude protein without phytase (control, and others diets with 20, 19, 18, 17% crude protein, with phytase. In the diets with phytase (80 g of Ronozyme NP(M phytase per ton of diet, the level of available phosphorus was reduced in 0.15 and the calcium in 0.30 percentage points in relation to the control diet. No effect was observed with the reduction of nutrients on feed intake, weight gain, or feed conversion in either phase or for the study of carcass characteristics to the 35 days. However, higher abdominal fat percentage was observed when the broilers were fed diet with lower protein level. The excretion of nitrogen and phosphorus decreased when birds received diets with reduced levels of these nutrients. Ash percentage in the tibia was not influenced by treatments. It is possible to reduce protein level, available phosphorus, and calcium of the diets in up to 3, 0.15 and 0.30 percentage points when the diets are supplemented with phytase and amino acids, respectively.

  13. Nicotianamine synthase overexpression positively modulates iron homeostasis-related genes in high iron rice.

    Science.gov (United States)

    Wang, Meng; Gruissem, Wilhelm; Bhullar, Navreet K

    2013-01-01

    Nearly one-third of the world population, mostly women and children, suffer from iron malnutrition and its consequences, such as anemia or impaired mental development. Biofortification of rice, which is a staple crop for nearly half of the world's population, can significantly contribute in alleviating iron deficiency. NFP rice (transgenic rice expressing nicotianamine synthase, ferritin and phytase genes) has a more than six-fold increase in iron content in polished rice grains, resulting from the synergistic action of nicotianamine synthase (NAS) and ferritin transgenes. We investigated iron homeostasis in NFP plants by analyzing the expression of 28 endogenous rice genes known to be involved in the homeostasis of iron and other metals, in iron-deficient and iron-sufficient conditions. RNA was collected from different tissues (roots, flag leaves, grains) and at three developmental stages during grain filling. NFP plants showed increased sensitivity to iron-deficiency conditions and changes in the expression of endogenous genes involved in nicotianamine (NA) metabolism, in comparison to their non-transgenic siblings (NTS). Elevated transcript levels were detected in NFP plants for several iron transporters. In contrast, expression of OsYSL2, which encodes a member of yellow stripe like protein family, and a transporter of the NA-Fe(II) complex was reduced in NFP plants under low iron conditions, indicating that expression of OsYSL2 is regulated by the endogenous iron status. Expression of the transgenes did not significantly affect overall iron homeostasis in NFP plants, which establishes the engineered push-pull mechanism as a suitable strategy to increase rice endosperm iron content.

  14. Nicotianamine synthase overexpression positively modulates iron homeostasis-related genes in high iron rice

    Directory of Open Access Journals (Sweden)

    Meng eWang

    2013-05-01

    Full Text Available Nearly one-third of the world population, mostly women and children, suffer from iron malnutrition and its consequences, such as anemia or impaired mental development. Biofortification of rice, which is a staple crop for nearly half of the world’s population, can significantly contribute in alleviating iron deficiency. NFP rice (transgenic rice expressing nicotianamine synthase, ferritin and phytase genes has a more than six-fold increase in iron content in polished rice grains, resulting from the synergistic action of nicotianamine synthase (NAS and ferritin transgenes. We investigated iron homeostasis in NFP plants by analyzing the expression of 28 endogenous rice genes known to be involved in the homeostasis of iron and other metals, in iron-deficient and iron-sufficient conditions. RNA was collected from different tissues (roots, flag leaves, grains and at three developmental stages during grain filling. NFP plants showed increased sensitivity to iron-deficiency conditions and changes in the expression of endogenous genes involved in nicotianamine (NA metabolism, in comparison to their non-transgenic siblings. Elevated transcript levels were detected in NFP plants for several iron transporters. In contrast, expression of OsYSL2, which encodes a member of Yellow Stripe-like protein family, and a transporter of the NA-Fe(II complex was reduced in NFP plants under low iron conditions, indicating that expression of OsYSL2 is regulated by the endogenous iron status. Expression of the transgenes did not significantly affect overall iron homeostasis in NFP plants, which establishes the engineered push-pull mechanism as a suitable strategy to increase rice endosperm iron content.

  15. Relação cálcio:fósforo disponível e níveis de fitase para poedeiras semipesadas no primeiro e segundo ciclos de postura Calcium:available phosphorus ratio and phytase levels for semi heavy laying hens in the first and second posture cycle

    Directory of Open Access Journals (Sweden)

    José Humberto Vilar da Silva

    2008-12-01

    ções deficientes em fósforo disponível com 600 UF melhora o peso dos ovos no segundo ciclo de postura.Two experiments were carried out to investigate the calcium (Ca:available phosphorus (aP ratio and the level of phytase in the first and second posture cycle. In the first posture cycle, 128 birds were distributed to a 3 × 2 + 1 factorial arrangement, composed of three Ca:aP ratio [14:1 (4.2% Ca and 0.30% aP, 12:1 (3.5% Ca and 0.30% aP, and 9:1 (3.5% Ca and 0.38% aP], two levels of phytase (0 and 600 Phytase Units - PU and a Ca:aP control [11:1 (4.2% Ca and 0.38% aP]. In the second posture cycle, 240 birds were distributed to a 2 × 2 × 2 factorial arrangement, composed of two Ca (3.5 and 4.2%, two aP (0.30 and 0.38% levels and two phytase levels (0 and 600 PU. In the first posture cycle, among birds fed diets without phytase, the greater eggs weight was obtained with 9:1 Ca:aP ratio and the smaller, with 12:1 Ca:aP ratio. The best eggs mass and feed conversion by eggs mass (FCEM were observed in birds fed a diet with 14:1 Ca:Pd ratio and worse, with the ration with 12:1 Ca:aP ratio. In birds fed diet with phytase, the best FCEM was obtained with 12:1 Ca:aP ratio and worse, with 9:1 Ca:aP rat