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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.

    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

  2. Localization of the Bacillus subtilis beta-propeller phytase transcripts in nodulated roots of Phaseolus vulgaris supplied with phytate.

    Science.gov (United States)

    Maougal, Rim Tinhinen; Bargaz, Adnane; Sahel, Charaf; Amenc, Laurie; Djekoun, Abdelhamid; Plassard, Claude; Drevon, Jean-Jacques

    2014-04-01

    Soil organic phosphorus (Po) such as phytate, which comprises up to 80 % of total Po, must be hydrolyzed by specific enzymes called phytases to be used by plants. In contrast to plants, bacteria, such as Bacillus subtilis, have the ability to use phytate as the sole source of P due to the excretion of a beta-propeller phytase (BPP). In order to assess whether the B. subtilis BPP could make P available from phytate for the benefit of a nodulated legume, the P-sensitive recombinant inbred line RIL147 of Phaseolus vulgaris was grown under hydroaeroponic conditions with either 12.5 μM phytate (C₆H₁₈O₂₄P₆) or 75 μmol Pi (K₂HPO₄), and inoculated with Rhizobium tropici CIAT899 alone, or co-inoculated with both B. subtilis DSM 10 and CIAT899. The in situ RT-PCR of BPP genes displayed the most intense fluorescent BPP signal on root tips. Some BPP signal was found inside the root cortex and the endorhizosphere of the root tip, suggesting endophytic bacteria expressing BPP. However, the co-inoculation with B. subtilis was associated with a decrease in plant P content, nodulation and the subsequent plant growth. Such a competitive effect of B. subtilis on P acquisition from phytate in symbiotic nitrogen fixation might be circumvented if the rate of inoculation were reasoned in order to avoid the inhibition of nodulation by excess B. subtilis proliferation. It is concluded that B. subtilis BPP gene is expressed in P. vulgaris rhizosphere.

  3. Identification of a Phytase Gene in Barley (Hordeum vulgare L.)

    Science.gov (United States)

    Dai, Fei; Qiu, Long; Ye, Lingzhen; Wu, Dezhi; Zhou, Meixue; Zhang, Guoping

    2011-01-01

    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. PMID:21533044

  4. 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.

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

    Science.gov (United States)

    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 active, extracellular phytase. The yield of total extracellular phytase activity was affected by the signal peptide and the medium composition. The expressed phytase had two pH optima (2 to 2.5 and 5 to 5.5) and a temperature optimum between 55 and 60°C, and it cross-reacted with a rabbit polyclonal antibody against the wild-type enzyme. Due to the heavy glycosylation, the expressed phytase had a molecular size of approximately 120 kDa and appeared to be more thermostable than the commercial enzyme. Deglycosylation of the phytase resulted in losses of 9% of its activity and 40% of its thermostability. The recombinant phytase was effective in hydrolyzing phytate phosphorus from corn or soybean meal in vitro. In conclusion, the phyA gene was expressed as an active, extracellular phytase in S. cerevisiae, and its thermostability was affected by glycosylation. PMID:10223979

  6. Molecular Characterization and Expression of a Phytase Gene from the Thermophilic Fungus Thermomyces lanuginosus

    Science.gov (United States)

    Berka, Randy M.; Rey, Michael W.; Brown, Kimberly M.; Byun, Tony; Klotz, Alan V.

    1998-01-01

    The phyA gene encoding an extracellular phytase from the thermophilic fungus Thermomyces lanuginosus was cloned and heterologously expressed, and the recombinant gene product was biochemically characterized. The phyA gene encodes a primary translation product (PhyA) of 475 amino acids (aa) which includes a putative signal peptide (23 aa) and propeptide (10 aa). The deduced amino acid sequence of PhyA has limited sequence identity (ca. 47%) with Aspergillus niger phytase. The phyA gene was inserted into an expression vector under transcriptional control of the Fusarium oxysporum trypsin gene promoter and used to transform a Fusarium venenatum recipient strain. The secreted recombinant phytase protein was enzymatically active between pHs 3 and 7.5, with a specific activity of 110 μmol of inorganic phosphate released per min per mg of protein at pH 6 and 37°C. The Thermomyces phytase retained activity at assay temperatures up to 75°C and demonstrated superior catalytic efficiency to any known fungal phytase at 65°C (the temperature optimum). Comparison of this new Thermomyces catalyst with the well-known Aspergillus niger phytase reveals other favorable properties for the enzyme derived from the thermophilic gene donor, including catalytic activity over an expanded pH range. PMID:9797301

  7. Cloning, Codon Optimization, and Expression of Yersinia intermedia Phytase Gene in E. coli.

    Science.gov (United States)

    Mirzaei, Maryam; Saffar, Behnaz; Shareghi, Behzad

    2016-06-01

    Phytate is an anti-nutritional factor in plants, which catches the most phosphorus contents and some vital minerals. Therefore, Phytase is added mainly as an additive to the monogastric animals' foods to hydrolyze phytate and increase absorption of phosphorus. Y. intermedia phytase is a new phytase with special characteristics such as high specific activity, pH stability, and thermostability. Our aim was to clone, express, and characterizea codon optimized Y. intermedia phytase gene in E. coli . The Y. intermedia phytase gene was optimized according to the codon usage in E. coli . The sequence was synthesized and sub-cloned in pET-22b (+) vector and transformed into E. coli Bl21 (DE3). The protein was expressed in the presence of IPTG at a final concentration of 1 mM at 30°C. The purification of recombinant protein was performed by Ni 2+ affinity chromatography. Phytase activity and stability were determined in various pH and temperatures. The codon optimized Y. intermedia phytase gene was sub-cloned successfully.The expression was confirmed by SDS-PAGE and Western blot analysis. The recombinant enzyme (approximately 45 kDa) was purified. Specific activity of enzyme was 3849 (U.mg -1 ) with optimal pH 5 and optimal temperature of 55°C. Thermostability (80°C for 15 min) and pH stability (3-6) of the enzyme were 56 and more than 80%, respectively. The results of the expression and enzyme characterization revealed that the optimized Y. intermedia phytase gene has a good potential to be produced commercially andto be applied in animals' foodsindustry.

  8. Efficacy of dietary phytase supplementation on laying performance and expression of osteopontin and calbindin genes in eggshell gland

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    Divya Shet

    2018-03-01

    Full Text Available This study was conducted to evaluate the effects of different levels of dietary phytase supplementation in the layer feed on egg production performance, egg shell quality and expression of osteopontin (OPN and calbindin (CALB1 genes. Seventy-five White Leghorn layers at 23 weeks of age were randomly divided into 5 groups consisting of a control diet with 0.33% non-phytate phosphorus (NPP and 4 low phosphorus (P diets: 2 diets (T1 and T2 with 0.24% NPP + 250 FTU/kg laboratory produced phytase or commercial phytase and another 2 diets (T3 and T4 with 0.16% NPP + 500 FTU/kg laboratory produced phytase or commercial phytase with complete replacement of inorganic P. The results indicated that there were no significant differences (P > 0.05 in egg production performance and quality of egg during the first 2 months of trial. However, in next 2 months, a significant drop in egg production and feed intake was observed in birds fed diets with low P and 500 FTU/kg supplementation of laboratory produced phytase. Osteopontin gene was up-regulated whereas the CALB1 gene was down regulated in all phytase treatment groups irrespective of the source of phytase. The current data demonstrated that 250 FTU/kg supplementation of laboratory produced phytase with 50% less NPP supplementation and 500 FTU/kg supplementation of commercial phytase even without NPP in diet can maintain the egg production. The up-regulation of OPN and down regulation of CALB1 in egg shell gland in the entire phytase treated group birds irrespective of the source of enzymes is indicative of the changes in P bio-availability at this site. Keywords: Phytase, Layer, Egg production, Gene expression, Egg shell

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

    Science.gov (United States)

    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.

  10. 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...

  11. Cartilage acidic protein 1, a new member of the beta-propeller protein family with amyloid propensity.

    Science.gov (United States)

    Anjos, Liliana; Morgado, Isabel; Guerreiro, Marta; Cardoso, João C R; Melo, Eduardo P; Power, Deborah M

    2017-02-01

    Cartilage acidic protein1 (CRTAC1) is an extracellular matrix protein of chondrogenic tissue in humans and its presence in bacteria indicate it is of ancient origin. Structural modeling of piscine CRTAC1 reveals it belongs to the large family of beta-propeller proteins that in mammals have been associated with diseases, including amyloid diseases such as Alzheimer's. In order to characterize the structure/function evolution of this new member of the beta-propeller family we exploited the unique characteristics of piscine duplicate genes Crtac1a and Crtac1b and compared their structural and biochemical modifications with human recombinant CRTAC1. We demonstrate that CRTAC1 has a beta-propeller structure that has been conserved during evolution and easily forms high molecular weight thermo-stable aggregates. We reveal for the first time the propensity of CRTAC1 to form amyloid-like structures, and hypothesize that the aggregating property of CRTAC1 may be related to its disease-association. We further contribute to the general understating of CRTAC1's and beta-propeller family evolution and function. Proteins 2017; 85:242-255. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  12. A Patient with Beta-Propeller Protein-Associated Neurodegeneration: Treatment with Iron Chelation Therapy

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    Shen-Yang Lim

    2018-05-01

    Full Text Available We present a case of beta-propeller protein-associated neurodegeneration, a form of neurodegeneration with brain iron accumulation. The patient harbored a novel mutation in the WDR45 gene. A detailed video and description of her clinical condition are provided. Her movement disorder phenomenology was characterized primarily by limb stereotypies and gait dyspraxia. The patient’s disability was advanced by the time iron-chelating therapy with deferiprone was initiated, and no clinical response in terms of cognitive function, behavior, speech, or movements were observed after one year of treatment.

  13. Ferrous Iron Up-regulation in Fibroblasts of Patients with Beta Propeller Protein-Associated Neurodegeneration (BPAN).

    OpenAIRE

    Ingrassia, Rosaria; Memo, Maurizio; Garavaglia, Barbara

    2017-01-01

    Mutations in WDR45 gene, coding for a beta-propeller protein, have been found in patients affected by Neurodegeneration with Brain Iron Accumulation, NBIA5 (also known as BPAN). BPAN is a movement disorder with Non Transferrin Bound Iron (NTBI) accumulation in the basal ganglia as common hallmark between NBIA classes (Hayflick et al., 2013). WDR45 has been predicted to have a role in autophagy, while the impairment of iron metabolism in the different NBIA subclasses has not currently been cla...

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

    Science.gov (United States)

    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. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  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

    Science.gov (United States)

    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

    2014-01-01

    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. PMID:25192284

  18. 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...

  19. Coexpression and Secretion of Endoglucanase and Phytase Genes in Lactobacillus reuteri

    Science.gov (United States)

    Wang, Lei; Yang, Yuxin; Cai, Bei; Cao, Pinghua; Yang, Mingming; Chen, Yulin

    2014-01-01

    A multifunctional transgenic Lactobacillus with probiotic characteristics and an ability to degrade β-glucan and phytic acid (phytate) was engineered to improve nutrient utilization, increase production performance and decrease digestive diseases in broiler chickens. The Bacillus subtilis WL001 endoglucanase gene (celW) and Aspergillus fumigatus WL002 phytase gene (phyW) mature peptide (phyWM) were cloned into an expression vector with the lactate dehydrogenase promoter of Lactobacillus casei and the secretion signal peptide of the Lactococcus lactis usp45 gene. This construct was then transformed into Lactobacillus reuteri XC1 that had been isolated from the gastrointestinal tract of broilers. Heterologous enzyme production and feed effectiveness of this genetically modified L. reuteri strain were investigated and evaluated. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis showed that the molecular mass of phyWM and celW was approximately 48.2 and 55 kDa, respectively, consistent with their predicted molecular weights. Endoglucanase and phytase activities in the extracellular fraction of the transformed L. reuteri culture were 0.68 and 0.42 U/mL, respectively. Transformed L. reuteri improved the feed conversion ratio of broilers from 21 to 42 days of age and over the whole feeding period. However, there was no effect on body weight gain and feed intake of chicks. Transformed L. reuteri supplementation improved levels of ash, calcium and phosphorus in tibiae at day 21 and of phosphorus at day 42. In addition, populations of Escherichia coli, Veillonella spp. and Bacteroides vulgatus were decreased, while populations of Bifidobacterium genus and Lactobacillus spp. were increased in the cecum at day 21. PMID:25050780

  20. Mining of Ruminant Microbial Phytase (RPHY1) from Metagenomic Data of Mehsani Buffalo Breed: Identification, Gene Cloning, and Characterization.

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    Mootapally, Chandra Shekar; Nathani, Neelam M; Patel, Amrutlal K; Jakhesara, Subhash J; Joshi, Chaitanya G

    2016-01-01

    Phytases have been widely used as animal feed supplements to increase the availability of digestible phosphorus, especially in monogastric animals fed cereal grains. The present study describes the identification of a full-length phytase gene of Prevotella species present in Mehsani buffalo rumen. The gene, designated as RPHY1, consists of 1,251 bp and is expressed into protein with 417 amino acids. A homology search of the deduced amino acid sequence of the RPHY1 phytase gene in a nonredundant protein database showed that it shares 92% similarity with the histidine acid phosphatase domain. Subsequently, the RPHY1 gene was expressed using a pET32a expression vector in Escherichia coli BL21 and purified using a His60 Ni-NTA gravity column. The mass of the purified RPHY1 was estimated to be approximately 63 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The optimal RPHY1 enzyme activity was observed at 55°C (pH 5) and exhibited good stability at 5°C and within the acidic pH range. Significant inhibition of RPHY1 activity was observed for Mg2+ and K+ metal ions, while Ca2+, Mn2+, and Na+ slightly inhibited enzyme activity. The RPHY1 phytase was susceptible to SDS, and it was highly stimulated in the presence of EDTA. Overall, the observed comparatively high enzyme activity levels and characteristics of the RPHY1 gene mined from rumen prove its promising candidature as a feed supplement enzyme in animal farming. © 2016 S. Karger AG, Basel.

  1. 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.

  2. 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.

  3. 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.

  4. Agrobacterium-mediated transformation of cotton (Gossypium hirsutum) shoot apex with a fungal phytase gene improves phosphorus acquisition.

    Science.gov (United States)

    Ma, Zhiying; Liu, Jianfeng; Wang, Xingfen

    2013-01-01

    Cotton is an important world economic crop plant. It is considered that cotton is recalcitrant to in vitro proliferation. Somatic embryogenesis and plant regeneration has been successful by using hypocotyl, whereas it is highly genotype dependent. Here, a genotype-independent cotton regeneration protocol from shoot apices is presented. Shoot apices from 3- to 5-day-old seedlings of cotton are infected with an Agrobacterium strain, EHA105, carrying the binary vector pC-KSA contained phytase gene (phyA) and the marker gene neomycin phosphotransferase (NPTII), and directly regenerated as shoots in vitro. Rooted shoots can be obtained within 6-8 weeks. Plants that survived by leaf painting kanamycin (kan) were -further analyzed by DNA and RNA blottings. The transgenic plants with increased the phosphorus (P) acquisition efficiency were obtained following the transformation method.

  5. Cisgenic Barley with Improved Phytase Activity

    DEFF Research Database (Denmark)

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

    2010-01-01

    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......-DNA borders in order to promote integration of the two genes at unlinked places in the plant genome. Transformed T0 plants show increases in the phytase activity of mature seeds from 1,400 in wild type to 8,950 FTU/kg in T0 plants. T1 plants of each transformant are currently screened with PCR for extra...... copies of the genomic phytase gene and the selection gene to identify segregation between the two genes. Presently, we have identified two cisgenic T1 plants without vector backbone and selection gene but with an extra copy of the genomic phytase gene....

  6. Cisgenic barley with improved phytase activity

    DEFF Research Database (Denmark)

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

    2010-01-01

    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......-DNA borders in order to promote integration of the two genes at unlinked places in the plant genome. Transformed T0 plants show increases in the phytase activity of mature seeds from 1,400 in wild type to 8,950 FTU/kg in T0 plants. T1 plants of each transformant are currently screened with PCR for extra...... copies of the genomic phytase gene and the selection gene to identify segregation between the two genes. Presently, we have identified two cisgenic T1 plants without vector backbone and selection gene but with an extra copy of the genomic phytase gene....

  7. Patient Affected by Beta-Propeller Protein-Associated Neurodegeneration: A Therapeutic Attempt with Iron Chelation Therapy

    Directory of Open Access Journals (Sweden)

    Mattia Fonderico

    2017-08-01

    Full Text Available Here, we report the case of a 36-year-old patient with a diagnosis of de novo mutation of the WDR45 gene, responsible for beta-propeller protein-associated neurodegeneration, a phenotypically distinct, X-linked dominant form of Neurodegeneration with Brain Iron Accumulation. The clinical history is characterized by a relatively stable intellectual disability and a hypo-bradykinetic and hypertonic syndrome with juvenile onset. Genetic investigations and T1 and T2-weighted MR images align with what is described in literature. The patient was also subjected to PET with 18-FDG investigation and DaT-Scan study. In reporting relevant clinical data, we want to emphasize the fact that the patient received a chelation therapy with deferiprone (treatment already used in other forms of NBIA with encouraging results, which, however, had to be interrupted because the parkinsonian symptoms worsened. Conversely, the patient has benefited from non-drug therapies and, in particular, from an adapted motor activity with assisted pedaling (method in the process of validation in treatments of parkinsonian syndromes, which started before the treatment with deferiprone and still continues.

  8. Ferrous Iron Up-regulation in Fibroblasts of Patients with Beta Propeller Protein-Associated Neurodegeneration (BPAN).

    Science.gov (United States)

    Ingrassia, Rosaria; Memo, Maurizio; Garavaglia, Barbara

    2017-01-01

    Mutations in WDR45 gene, coding for a beta-propeller protein, have been found in patients affected by Neurodegeneration with Brain Iron Accumulation, NBIA5 (also known as BPAN). BPAN is a movement disorder with Non Transferrin Bound Iron (NTBI) accumulation in the basal ganglia as common hallmark between NBIA classes (Hayflick et al., 2013). WDR45 has been predicted to have a role in autophagy, while the impairment of iron metabolism in the different NBIA subclasses has not currently been clarified. We found the up-regulation of the ferrous iron transporter (-)IRE/Divalent Metal Transporter1 and down-regulation of Transferrin receptor in the fibroblasts of two BPAN affected patients with splicing mutations 235+1G>A (BPAN1) and 517_519ΔVal 173 (BPAN2). The BPAN patients showed a concomitant increase of intracellular ferrous iron after starvation. An altered pattern of iron transporters with iron overload is highlighted in BPAN human fibroblasts, supporting for a role of DMT1 in NBIA. We here present a novel element, about iron accumulation, to the existing knowledge in field of NBIA. Attention is focused to a starvation-dependent iron overload, possibly accounting for iron accumulation in the basal ganglia. Further investigation could clarify iron regulation in BPAN.

  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. 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. Copyright © 2015 Elsevier B.V. All rights reserved.

  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. Cisgenic barley with improved phytase activity

    DEFF Research Database (Denmark)

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

    2010-01-01

    copies of the genomic phytase gene and the selection gene to identify segregation between the two genes. Presently, we have identified two cisgenic T1 plants without vector backbone and selection gene but with an extra copy of the genomic phytase gene....... 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...

  13. Overexpression of phyA and appA Genes Improves Soil Organic Phosphorus Utilisation and Seed Phytase Activity in Brassica napus

    Science.gov (United States)

    Wang, Yi; Ye, Xiangsheng; Ding, Guangda; Xu, Fangsen

    2013-01-01

    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. PMID:23573285

  14. 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.

  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.

    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.

  16. 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.

  17. Diversity of phytases in the rumen.

    Science.gov (United States)

    Nakashima, Brenda A; McAllister, Tim A; Sharma, Ranjana; Selinger, L Brent

    2007-01-01

    Examples of a new class of phytase related to protein tyrosine phosphatases (PTP) were recently isolated from several anaerobic bacteria from the rumen of cattle. In this study, the diversity of PTP-like phytase gene sequences in the rumen was surveyed by using the polymerase chain reaction (PCR). Two sets of degenerate primers were used to amplify sequences from rumen fluid total community DNA and genomic DNA from nine bacterial isolates. Four novel PTP-like phytase sequences were retrieved from rumen fluid, whereas all nine of the anaerobic bacterial isolates investigated in this work contained PTP-like phytase sequences. One isolate, Selenomonas lacticifex, contained two distinct PTP-like phytase sequences, suggesting that multiple phytate hydrolyzing enzymes are present in this bacterium. The degenerate primer and PCR conditions described here, as well as novel sequences obtained in this study, will provide a valuable resource for future studies on this new class of phytase. The observed diversity of microbial phytases in the rumen may account for the ability of ruminants to derive a significant proportion of their phosphorus requirements from phytate.

  18. Evaluation of the mature grain phytase candidate HvPAPhy_a gene in barley (Hordeum vulgare L.) using CRISPR/Cas9 and TALENs.

    Science.gov (United States)

    Holme, Inger B; Wendt, Toni; Gil-Humanes, Javier; Deleuran, Lise C; Starker, Colby G; Voytas, Daniel F; Brinch-Pedersen, Henrik

    2017-09-01

    In the present study, we utilized TALEN- and CRISPR/Cas9-induced mutations to analyze the promoter of the barley phytase gene HvPAPhy_a. The purpose of the study was dual, validation of the PAPhy_a enzyme as the main contributor of the mature grain phytase activity (MGPA), as well as validating the importance of a specific promoter region of the PAPhy_a gene which contains three overlapping cis-acting regulatory elements (GCN4, Skn1 and the RY-element) known to be involved in gene expression during grain filling. The results confirm that the barley PAPhy_a enzyme is the main contributor to the MGPA as grains of knock-out lines show very low MGPA. Additionally, the analysis of the HvPAPhy_a promoter region containing the GCN4/Skn1/RY motif highlights its importance for HvPAPhy_a expression as the MGPA in grains of plant lines with mutations within this motif is significantly reduced. Interestingly, lines with deletions located downstream of the motif show even lower MGPA levels, indicating that the GCN4/SKn1/RY motif is not the only element responsible for the level of PAPhy_a expression during grain maturation. Mutant grains with very low MPGA showed delayed germination as compared to grains of wild type barley. As grains with high levels of preformed phytases would provide more readily available phosphorous needed for a fast germination, this indicates that faster germination may be implicated in the positive selection of the ancient PAPhy gene duplication that lead to the creation of the PAPhy_a gene.

  19. Cisgenic barley with improved phytase activity

    DEFF Research Database (Denmark)

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

    2012-01-01

    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......PAPhy_a insert for further analysis. Seeds from plants homozygous for the insert showed 2.6- and 2.8-fold increases in phytase activities and the activity levels were stable over the three generations analysed. In one of the selected lines, the flanking sequences from both the left and right T-DNA borders were...

  20. C-Terminus of Progranulin Interacts with the Beta-Propeller Region of Sortilin to Regulate Progranulin Trafficking

    OpenAIRE

    Zheng, Yanqiu; Brady, Owen A.; Meng, Peter S.; Mao, Yuxin; Hu, Fenghua

    2011-01-01

    Progranulin haplo-insufficiency is a main cause of frontotemporal lobar degeneration (FTLD) with TDP-43 aggregates. Previous studies have shown that sortilin regulates progranulin trafficking and is a main determinant of progranulin level in the brain. In this study, we mapped the binding site between progranulin and sortilin. Progranulin binds to the beta-propeller region of sortilin through its C-terminal tail. The C-terminal progranulin fragment is fully sufficient for sortilin binding and...

  1. New Bacterial Phytase through Metagenomic Prospection

    Directory of Open Access Journals (Sweden)

    Nathálya Farias

    2018-02-01

    Full Text Available Alkaline phytases from uncultured microorganisms, which hydrolyze phytate to less phosphorylated myo-inositols and inorganic phosphate, have great potential as additives in agricultural industry. The development of metagenomics has stemmed from the ineluctable evidence that as-yet-uncultured microorganisms represent the vast majority of organisms in most environments on earth. In this study, a gene encoding a phytase was cloned from red rice crop residues and castor bean cake using a metagenomics strategy. The amino acid identity between this gene and its closest published counterparts is lower than 60%. The phytase was named PhyRC001 and was biochemically characterized. This recombinant protein showed activity on sodium phytate, indicating that PhyRC001 is a hydrolase enzyme. The enzymatic activity was optimal at a pH of 7.0 and at a temperature of 35 °C. β-propeller phytases possess great potential as feed additives because they are the only type of phytase with high activity at neutral pH. Therefore, to explore and exploit the underlying mechanism for β-propeller phytase functions could be of great benefit to biotechnology.

  2. Phytate addition to soil induces changes in the abundance and expression of Bacillus β-propeller phytase genes in the rhizosphere.

    Science.gov (United States)

    Jorquera, Milko A; Saavedra, Nicolás; Maruyama, Fumito; Richardson, Alan E; Crowley, David E; del C Catrilaf, Rosa; Henriquez, Evelyn J; de la Luz Mora, María

    2013-02-01

    Phytate-mineralizing rhizobacteria (PMR) perform an essential function for the mineralization of organic phosphorus but little is known about their ecology in soils and rhizosphere. In this study, PCR-based methods were developed for detection and quantification of the Bacillus β-propeller phytase (BPP) gene. Experiments were conducted to monitor the presence and persistence of a phytate-mineralizing strain, Bacillus sp. MQH19, after inoculation of soil microcosms and within the rhizosphere. The occurrence of the BPP gene in natural pasture soils from Chilean Andisols was also examined. The results showed that the Bacillus BPP gene was readily detected in sterile and nonsterile microcosms, and that the quantitative PCR (qPCR) methods could be used to monitor changes in the abundance of the BPP gene over time. Our results also show that the addition of phytate to nonsterile soils induced the expression of the BPP gene in the rhizosphere of ryegrass and the BPP gene was detected in all pasture soils sampled. This study shows that phytate addition soils induced changes in the abundance and expression of Bacillus BPP to genes in the rhizosphere and demonstrates that Bacillus BPP gene is cosmopolitan in pasture soils from Chilean Andisols. © 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  3. The structure of Rauvolfia serpentina strictosidine synthase is a novel six-bladed beta-propeller fold in plant proteins.

    Science.gov (United States)

    Ma, Xueyan; Panjikar, Santosh; Koepke, Juergen; Loris, Elke; Stöckigt, Joachim

    2006-04-01

    The enzyme strictosidine synthase (STR1) from the Indian medicinal plant Rauvolfia serpentina is of primary importance for the biosynthetic pathway of the indole alkaloid ajmaline. Moreover, STR1 initiates all biosynthetic pathways leading to the entire monoterpenoid indole alkaloid family representing an enormous structural variety of approximately 2000 compounds in higher plants. The crystal structures of STR1 in complex with its natural substrates tryptamine and secologanin provide structural understanding of the observed substrate preference and identify residues lining the active site surface that contact the substrates. STR1 catalyzes a Pictet-Spengler-type reaction and represents a novel six-bladed beta-propeller fold in plant proteins. Structure-based sequence alignment revealed a common repetitive sequence motif (three hydrophobic residues are followed by a small residue and a hydrophilic residue), indicating a possible evolutionary relationship between STR1 and several sequence-unrelated six-bladed beta-propeller structures. Structural analysis and site-directed mutagenesis experiments demonstrate the essential role of Glu-309 in catalysis. The data will aid in deciphering the details of the reaction mechanism of STR1 as well as other members of this enzyme family.

  4. 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.

  5. C-terminus of progranulin interacts with the beta-propeller region of sortilin to regulate progranulin trafficking.

    Directory of Open Access Journals (Sweden)

    Yanqiu Zheng

    Full Text Available Progranulin haplo-insufficiency is a main cause of frontotemporal lobar degeneration (FTLD with TDP-43 aggregates. Previous studies have shown that sortilin regulates progranulin trafficking and is a main determinant of progranulin level in the brain. In this study, we mapped the binding site between progranulin and sortilin. Progranulin binds to the beta-propeller region of sortilin through its C-terminal tail. The C-terminal progranulin fragment is fully sufficient for sortilin binding and progranulin C-terminal peptide displaces progranulin binding to sortilin. Deletion of the last 3 residues of progranulin (QLL abolishes its binding to sortilin and also sortilin dependent regulation of progranulin trafficking. Since progranulin haplo-insufficiency results in FTLD, these results may provide important insights into future studies of progranulin trafficking and signaling and progranulin based therapy for FTLD.

  6. C-Terminus of Progranulin Interacts with the Beta-Propeller Region of Sortilin to Regulate Progranulin Trafficking

    Science.gov (United States)

    Meng, Peter S.; Mao, Yuxin; Hu, Fenghua

    2011-01-01

    Progranulin haplo-insufficiency is a main cause of frontotemporal lobar degeneration (FTLD) with TDP-43 aggregates. Previous studies have shown that sortilin regulates progranulin trafficking and is a main determinant of progranulin level in the brain. In this study, we mapped the binding site between progranulin and sortilin. Progranulin binds to the beta-propeller region of sortilin through its C-terminal tail. The C-terminal progranulin fragment is fully sufficient for sortilin binding and progranulin C-terminal peptide displaces progranulin binding to sortilin. Deletion of the last 3 residues of progranulin (QLL) abolishes its binding to sortilin and also sortilin dependent regulation of progranulin trafficking. Since progranulin haplo-insufficiency results in FTLD, these results may provide important insights into future studies of progranulin trafficking and signaling and progranulin based therapy for FTLD. PMID:21698296

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

    Science.gov (United States)

    Zou, Li-kou; Wang, Hong-ning; Pan, Xin; Tian, Guo-bao; Xie, Zi-wen; Wu, Qi; Chen, Hui; Xie, Tao; Yang, Zhi-rong

    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 Pichia pastoris 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 extracellular 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 °C 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 °C to 95 °C 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. PMID:18600783

  8. 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.

  9. 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...

  10. Phytase for Food Application

    Directory of Open Access Journals (Sweden)

    Ursula Konietzny

    2006-01-01

    Full Text Available Phytase [myo-inositol(1,2,3,4,5,6hexakisphosphate phosphohydrolase], a phytate-specific phosphatase, is already used as a supplement in diets for monogastric animals to improvephosphate utilisation from phytate[myoinositol(1,2,3,4,5,6hexakisphosphate], the major storage form of phosphate in plant seeds. In recent years, this class of enzymes has also been found increasingly interesting for use in processing and manufacturing of food for human consumption, particularly because the decline in food phytate results in an enhancement of mineral bioavailability. Different strategies could be applied to optimise phytate degradation during food processing and digestion in the human alimentary tract such as adjustment of more favourable conditions during food processing for the phytases naturally occurring in the raw material, addition of isolated phytases to the production process, use of raw material with a high intrinsic phytate-degrading activity either naturally present or introduced by genetic engineering and the use of recombinant food-grade microorganisms as carriers for phytate-degrading activity in the human gastrointestinal tract. Furthermore, phytases may find application in the production of functional foods or food supplements with health benefits. Last but not least, technological improvements are expected to occur due to phytate degradation during processing as shown for breadmaking, production of plant protein isolates, corn wet milling and the fractionation of cereal bran.

  11. Exploratory transcriptomic analysis in muscle tissue of broilers fed a phytase-supplemented diet.

    Science.gov (United States)

    Schmeisser, J; Séon, A-A; Aureli, R; Friedel, A; Guggenbuhl, P; Duval, S; Cowieson, A J; Fru-Nji, F

    2017-06-01

    The effect of phytase on phosphorus retention, broiler (Gallus gallus) performance and bone mineralization in diets with reduced inorganic phosphate concentration is well documented. Furthermore, so-called 'extra-phosphoric' effects of phytase have been described in the literature that may be associated with changes in mineral and amino acid partitioning and requirements per se. In particular, the role of myo-inositol in phytase responses is implied but not well elucidated. It was the purpose of the experiment reported herein to explore the effect of phytase on broiler growth, nutrient digestibility, blood biochemistry and gene expression. A 5-week broiler floor pen trial was conducted to evaluate the effect of supplementation of a moderately phosphorus-deficient diet with 1000 U/kg of a 6-microbial phytase. Parameters measured were growth performance, phosphorus (P), calcium (Ca) and myo-inositol plasma concentrations, apparent ileal P digestibility, bone mineralization, breast meat weight and Pectoralis major muscle transcriptome. Supplementation of the diet with phytase improved weight gain during the starter period (18%) and the whole period (24%) compared with animals that received the control diet (p phytase. The transcriptomic analysis revealed that some differentially expressed genes (DEG) in broilers, receiving phytase in comparison with animals fed reduced phosphorus diet without phytase, were part of pathways involved in muscle development, via calmodulin/calcineurin and insulin-like growth factor. Microarray data confirmation was performed on six genes by quantitative PCR (qPCR): PI3K regulatory and catalytic subunit, Phospholipase C beta, Myocyte Enhancer Factors 2A and 2C, and calcineurin A. The results suggested that dietary supplementation with this phytase could generate low molecular weight phytate esters and indirectly myo-inositol, and could help us to understand how muscle metabolism may be affected at a gene level. Journal of Animal

  12. A Novel Phytase with Sequence Similarity to Purple Acid Phosphatases Is Expressed in Cotyledons of Germinating Soybean Seedlings 1

    Science.gov (United States)

    Hegeman, Carla E.; Grabau, Elizabeth A.

    2001-01-01

    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. PMID:11500558

  13. 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.

  14. Comparative Analysis of Peripheral Alkaline Phytase Protein Structures Expressed in E. coli

    Directory of Open Access Journals (Sweden)

    Mohammadreza Nassiri

    2015-10-01

    Full Text Available Background: Degradation of phytic acid to inorganic phosphate in domestic animals’ diets requires thermostable phytase. Although Basillus subtilis phytase shows a potential to be degraded phytate complex in high temperature, the enzyme activities and yields need to be increased to make them possible for industrial application. Methods: The phytase gene from Bacillus subtilis DR8886 was isolated from Dig Rostam hot mineral spring in Iran and cloned into pET21(+ and pET32(+. Expression was induced with 1.5 mM IPTG and the proteins were purified. Results: The recombinant protein affected by thioredoxin (Trx from pET32a-PhyC was estimated to constitute about 31% of the total soluble protein in the cells; its concentration was 3.5 μg/ml, and its maximal phytase activity was 15.9 U/ml, whereas the recombinant phytase from pET21a-PhyC was estimated to comprise about 19% of the total soluble protein; its concentration was 2.2 μg/ml, and its maximal phytase activity was 69 U/ml. The molecular masses of recombinant phytase with and without Trx were about 60 kDa and 42 kDa, respectively. Zymography confirmed that the recombinant enzymes were active. Although the concentration of the alkaline phytase expressed by pET32a was approximately 59% greater than that expressed by pET21, its phytase activity was approximately 77% less. Conclusion: This study showed that the peripheral gene (Trx encoded by the pET32a (+ vector are the principal reason for the decrease in recombinant phytase enzyme activity.

  15. 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.

  16. 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...... 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...

  17. 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......, where it serves as a storage molecule for phosphorous. Phytic acid is also associated with minerals. The minerals are bound by chelation to the negatively charged phosphate groups in phytic acid. Phytases catalyse the dephosphorylation of phytic acid, thus releasing bound minerals to make them available...... for absorption. This article presents research on phytase catalysis in gastric conditions and considers potential benefits and drawbacks for using phytases as a food supplement....

  18. Dietary microbial phytase exerts mixed effects on the gut health of tilapia: a possible reason for the null effect on growth promotion.

    Science.gov (United States)

    Hu, Jun; Ran, Chao; He, Suxu; Cao, Yanan; Yao, Bin; Ye, Yuantu; Zhang, Xuezhen; Zhou, Zhigang

    2016-06-01

    The present study evaluated the effects of dietary microbial phytase on the growth and gut health of hybrid tilapia (Oreochromis niloticus ♀×Oreochromis aureus ♂), focusing on the effect on intestinal histology, adhesive microbiota and expression of immune-related cytokine genes. Tilapia were fed either control diet or diet supplemented with microbial phytase (1000 U/kg). Each diet was randomly assigned to four groups of fish reared in cages (3×3×2 m). After 12 weeks of feeding, weight gain and feed conversion ratio of tilapia were not significantly improved by dietary microbial phytase supplementation. However, significantly higher level of P content in the scales, tighter and more regular intestinal mucosa folds were observed in the microbial phytase group and the microvilli density was significantly increased. The adhesive gut bacterial communities were strikingly altered by microbial phytase supplementation (0·41phytase, as indicated by the up-regulated intestinal expressions of the cytokine genes (tnf-α and tgf-β) and hsp70. In addition, the gut microvilli height was significantly decreased in the phytase group. These results indicate that dietary microbial phytase may exert mixed effects on hybrid tilapia, and can guide our future selection of phytases as aquafeed additives - that is, eliminating those that can stimulate intestinal inflammation.

  19. 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.

  20. 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.

  1. 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...

  2. 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.

  3. 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. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  4. A novel extracellular low-temperature active phytase from Bacillus aryabhattai RS1 with potential application in plant growth.

    Science.gov (United States)

    Pal Roy, Moushree; Datta, Subhabrata; Ghosh, Shilpi

    2017-05-01

    Bacillus aryabhattai RS1 isolated from rhizosphere produced an extracellular, low temperature active phytase. The cultural conditions for enzyme production were optimized to obtain 35 U mL -1 of activity. Purified phytase had specific activity and molecular weight of 72.97 U mg -1 and ∼40 kDa, respectively. The enzyme was optimally active at pH 6.5 and 40°C and was highly specific to phytate. It exhibited higher catalytic activity at low temperature, retaining over 40% activity at 10°C. Phytase was more thermostable in presence of Ca 2+ ion and retained 100% residual activity on preincubation at 20-50°C for 30 min. Partial phytase encoding gene, phy B (816 bp) was cloned and sequenced. The encoded amino acid sequence (272 aa) contained two conserved motifs, DA[A/T/E]DDPA[I/L/V]W and NN[V/I]D[I/L/V]R[Y/D/Q] of β-propellar phytase and had lower sequence homology with other Bacillus phytases, indicating its novelty. Phytase and the bacterial inoculum were effective in improving germination and growth of chickpea seedlings under phosphate limiting condition. Moreover, the potential applications of the enzyme with relatively high activity at lower temperatures (20-30°C) could also be extended to aquaculture and food processing. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:633-641, 2017. © 2017 American Institute of Chemical Engineers.

  5. Fingerprint motifs of phytases | Fan | African Journal of Biotechnology

    African Journals Online (AJOL)

    Among the total of potential 173 phytases gained in 11 plant genomes through MAST, PAPhys are the major phytases, and HAPhys are the minor, and other phytase groups are not found in planta. Keywords: Phytase, fingerprint motif, multiple EM for motif elicitation (MEME), MAST African Journal of Biotechnology Vol.

  6. Determining the available phosphorus release of Natuphos E 5,000 G phytase for nursery pigs.

    Science.gov (United States)

    Gourley, K M; Woodworth, J C; DeRouchey, J M; Dritz, S S; Tokach, M D; Goodband, R D

    2018-04-03

    A total of 288 pigs (PIC 327 × 1050; initially 11.1 ± 0.1 kg, and day 40 of age) were used in a 21-d growth trial to determine the available P (aP) release curve for a novel source of 6-phytase (Natuphos E 5,000 G; BASF Corporation, Florham Park, NJ). Natuphos E is a bacterial derived 6-phytase of which the phytase gene is assembled from a hybrid of phytase-producing bacteria and produced through the fermentation of Aspergillus niger. Pigs were randomly allotted to pens at weaning. From day 15 to 18 postweaning, a common corn-soybean meal diet containing 0.12% aP was fed to all pigs to acclimate them to a P-deficient diet. On day 0 of the experiment (day 19 after weaning), pens were allotted in a randomized complete block design to one of eight treatments. There were four pigs per pen and nine pens per dietary treatment. Pigs were fed a corn-soybean meal-based diet formulated to 1.25% standardized ileal digestible Lys. Experimental diets were formulated to contain 0.73% Ca and increasing aP supplied by either monocalcium P (0.12%, 0.18%, and 0.24% aP) or from increasing phytase (150, 250, 500, 750, and 1,000 phytase unit [FTU]/kg) added to the 0.12% aP diet. Analyzed phytase concentrations were 263, 397, 618, 1,100, and 1,350 FTU/kg, respectively. On day 21 of the study, one pig per pen was euthanized and the right fibula was collected for bone ash and percentage bone ash calculations. From day 0 to 21, increasing P from monocalcium P or phytase improved (linear, P ash weight and percentage bone ash increased (linear, P percentage bone ash are used as the response variables, aP release for up to 1,000 FTU/kg of Natuphos E 5,000 G phytase can be predicted by the equation: aP release = 0.000212 × FTU/kg phytase.

  7. 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.

  8. 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 CaCl 2 . 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. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. Purification, characterization of phytase enzyme from Lactobacillus ...

    African Journals Online (AJOL)

    Purification, characterization of phytase enzyme from Lactobacillus plantarum bacteria and determination of its kinetic properties. ... Many of the cereal grains, legumes and oilseeds store phosphorus in phytate form. Phytases can be produced by plants, animals and microorganisms. However, the ones with microbial origin ...

  10. Isolation and molecular characterization of phytase producing ...

    African Journals Online (AJOL)

    Isolation and molecular characterization of phytase producing bacteria from Malaysia hot springs. ... The strains were further analyzed in broth culture using sodium phytate as substrate. Among them, strain L3 was selected as the best producer (0.16 U/ml after 72 h of culture). This phytase showed optimal activity at 37 °C ...

  11. 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.

  12. Cloning and Characterization of Purple Acid Phosphatase Phytases from Wheat, Barley, Maize and Rice

    DEFF Research Database (Denmark)

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

    2011-01-01

    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......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......, it is demonstrated that wheat, barley, maize, and rice all possess purple acid phosphatase (PAP) genes that, expressed in Pichia pastoris, give fully functional phytases (PAPhys) with very similar enzyme kinetics. Preformed wheat PAPhy was localized to the protein crystalloid of the aleurone vacuole. Phylogenetic...

  13. 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.

  14. Quantitative Conversion of Phytate to Inorganic Phosphorus in Soybean Seeds Expressing a Bacterial Phytase1[OA

    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.

    2008-01-01

    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, ≥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. PMID:18162589

  15. Improved thermostability and enzyme activity of a recombinant phyA mutant phytase from Aspergillus niger N25 by directed evolution and site-directed mutagenesis.

    Science.gov (United States)

    Tang, Zizhong; Jin, Weiqiong; Sun, Rong; Liao, Yan; Zhen, Tianrun; Chen, Hui; Wu, Qi; Gou, Lin; Li, Chenlei

    2018-01-01

    We previously constructed three recombinant phyA mutant strains (PP-NP m -8, PP-NP ep -6A and I44E/T252R-PhyA), showing improved catalytic efficiency or thermostability of Aspergillus niger N25 phytase, by error-prone PCR or site-directed mutagenesis. In this study, directed evolution and site-directed mutagenesis were further applied to improve the modified phytase properties. After one-round error-prone PCR for phytase gene of PP-NP ep -6A, a single transformant, T195L/Q368E/F376Y, was obtained with the significant improvements in catalytic efficiency and thermostability. The phytase gene of T195L/Q368E/F376Y, combined with the previous mutant phytase genes of PP-NP ep -6A, PP-NP m -8 and I44E/T252R-PhyA, was then sequentially modified by DNA shuffling. Three genetically engineered strains with desirable properties were then obtained, namedQ172R, Q172R/K432R andQ368E/K432R. Among them, Q172R/K432R showed the highest thermostability with the longest half-life and the greatest remaining phytase activity after heat treatment, while Q368E/K432R showed the highest catalytic activity. Five substitutions (Q172R, T195L, Q368E, F376Y, K432R) identified from random mutagenesis were added sequentially to the phytase gene of PP-NP ep -6A to investigate how the mutant sites influence the properties of phytase. Characterization and structural analysis demonstrated that these mutations could produce cumulative or synergistic improvements in thermostability or catalytic efficiency of phytase. Copyright © 2017 Elsevier Inc. All rights reserved.

  16. A Comparative Analysis of Recombinant Expression and Solubility Screening of Two Phytases in E. coli

    Directory of Open Access Journals (Sweden)

    Ashok Pandey

    2011-01-01

    Full Text Available Microbial phytases, especially from fungal and bacterial sources, have received much attention as food additives in human nutrition and as feed supplements for monogastric animals. An effective expression screening method for recombinant production of this enzyme on a small scale is industrially desirable. An effort has been made in this work to clone and express phytase genes from Aspergillus sp. and Escherichia coli with the selected host, vector and inducer combination. Albeit the formation of insoluble inclusion bodies by fungal phytase, recombinant E. coli appA was effectively expressed in a cost-effective manner in the periplasm of BL21plysS using an inducer concentration of 0.01 mM in 4 h of growth. Enzyme was purified in three consecutive steps and functional studies were carried out.

  17. Development of phytase-expressing chlamydomonas reinhardtii for monogastric animal nutrition.

    Science.gov (United States)

    Erpel, Fernanda; Restovic, Franko; Arce-Johnson, Patricio

    2016-03-12

    In plant-derived animal feedstuffs, nearly 80 % of the total phosphorus content is stored as phytate. However, phytate is poorly digested by monogastric animals such as poultry, swine and fish, as they lack the hydrolytic enzyme phytase; hence it is regarded as a nutritionally inactive compound from a phosphate bioavailability point of view. In addition, it also chelates important dietary minerals and essential amino acids. Therefore, dietary supplementation with bioavailable phosphate and exogenous phytases are required to achieve optimal animal growth. In order to simplify the obtaining and application processes, we developed a phytase expressing cell-wall deficient Chlamydomonas reinhardtii strain. In this work, we developed a transgenic microalgae expressing a fungal phytase to be used as a food supplement for monogastric animals. A codon optimized Aspergillus niger PhyA E228K phytase (mE228K) with improved performance at pH 3.5 was transformed into the plastid genome of Chlamydomonas reinhardtii in order to achieve optimal expression. We engineered a plastid-specific construction harboring the mE228K gene, which allowed us to obtain high expression level lines with measurable in vitro phytase activity. Both wild-type and cell-wall deficient strains were selected, as the latter is a suitable model for animal digestion. The enzymatic activity of the mE228K expressing lines were approximately 5 phytase units per gram of dry biomass at pH 3.5 and 37 °C, similar to physiological conditions and economically competitive for use in commercial activities. A reference basis for the future biotechnological application of microalgae is provided in this work. A cell-wall deficient transgenic microalgae with phytase activity at gastrointestinal pH and temperature and suitable for pellet formation was developed. Moreover, the associated microalgae biomass costs of this strain would be between US$5 and US$60 per ton of feedstuff, similar to the US$2 per ton of feedstuffs

  18. Purification, characterization of phytase enzyme from Lactobacillus ...

    African Journals Online (AJOL)

    SAM

    2014-06-04

    Jun 4, 2014 ... 2Department of Food Technology, Erzurum Vocational Training School, Ataturk University, 25240, ... facultative anaerobic, catalase-negative, immobile (with ..... Partial purification of phytase from a soil isolate bacterium,.

  19. Phosphorus and phytase levels for layer hens

    OpenAIRE

    Juliana Cristina Ramos Rezende; Antonio Carlos de Laurentiz; Rosemeire da Silva Filardi; Vitor Barbosa Fascina; Daniella Aparecida Berto; Sérgio Turra Sobrane Filho

    2013-01-01

    The objective of this research was to evaluate the performance and bone quality of laying hens after peak production fed diets containing phosphorus levels and phytase. An experiment was conducted with 384 Hy-line distributed in a completely randomized in a factorial 4 x 3 with 4 levels of available phosphorus and 3 levels of phytase. The experimental period was divided into four periods of 28 days, at the end of each cycle were determined experimental feed intake, egg production, egg weight,...

  20. 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...

  1. Influence of pretreatment of agriculture residues on phytase ...

    African Journals Online (AJOL)

    phytase production. Pretreatment of agriculture residues with water to remove excess inorganic phosphate has significantly enhanced the phytase activity in case of de-oiled rice bran, wheat bran, peanut cake (low and high oil) and coconut cake. Maximum increase of 20.3 times in phytase activity was observed in case of ...

  2. 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

  3. 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

  4. 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. © 2014 The Authors. Journal of the Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

  5. Insights into phytase-containing transgenic Lemna minor (L.) as a novel feed additive.

    Science.gov (United States)

    Ghosh, Mrinmoy; Sharma, Neelesh; Gera, Meeta; Kim, Nameun; Huynh, Do; Zhang, Jiaojiao; Min, Taesun; Sodhi, Simrinder Singh; Kim, Min Bae; Rekha, V P B; Ko, Sukmin; Jeong, Dong Kee

    2018-04-01

    This study assessed the effect of supplementation of novel transgenic phytase on growth performance and bone mineralization in Korean native broiler chickens. The experiment was designed using four dietary groups: those with a diet supplemented with (A) recombinant phytase, (B) transgenic phytase from the plant Lemna minor, (C) or wild-type L. minor as well as (D) a control group that was supplemented with commercially available feed. Three hundred 1-day-old Korean native broiler chicks were used and divided into these four dietary treatment groups having three replicates of 25 birds each (n = 75). The results showed increases in growth performance and bone mineralization in Groups B and C; compared with Groups A and D. Hematological analyses revealed notable contrasts in erythrocyte sedimentation rate, red blood cell count, and hemoglobin levels among the experimental groups, whereas no impacts of dietary treatment were observed on total eosinophil, lymphocyte, heterophil, monocyte, and basophil levels. The relative expression profiling of candidate genes showed that the genes involved in growth response, meat quality, and P-Ca metabolism were significantly highly expressed in the phytase-supplemented groups. Hence, it is suggested that dietary supplementation with transgenic phytase plant L. minor for enhancing growth performance is a promising new approach in the broiler feed industry. To the best of our knowledge, we report here the most comprehensive analysis using a broiler model that provides a workable platform for further research on the cost-effective production of feed with different compositions that might be beneficial in the livestock feed industry.

  6. Clinical and Imaging Presentation of a Patient with Beta-Propeller Protein-Associated Neurodegeneration, a Rare and Sporadic form of Neurodegeneration with Brain Iron Accumulation (NBIA).

    Science.gov (United States)

    Hattingen, Elke; Handke, Nikolaus; Cremer, Kirsten; Hoffjan, Sabine; Kukuk, Guido Matthias

    2017-12-01

    Neurodegeneration with brain iron accumulation (NBIA) is a heterogeneous group of inherited neurologic disorders with iron accumulation in the basal ganglia, which share magnetic resonance (MR) imaging characteristics, histopathologic and clinical features. According to the affected basal nuclei, clinical features include extrapyramidal movement disorders and varying degrees of intellectual disability status. The most common NBIA subtype is caused by pathogenic variants in PANK2. The hallmark of MR imaging in patients with PANK2 mutations is an eye-of-the-tiger sign in the globus pallidus. We report a 33-year-old female with a rare subtype of NBIA, called beta-propeller protein-associated neurodegeneration (BPAN) with a hitherto unknown missense variant in WDR45. She presented with BPAN's particular biphasic course of neurological symptoms and with a dominant iron accumulation in the midbrain that enclosed a spotty T2-hyperintensity.

  7. Phosphorus and phytase levels for layer hens

    Directory of Open Access Journals (Sweden)

    Juliana Cristina Ramos Rezende

    2013-02-01

    Full Text Available The objective of this research was to evaluate the performance and bone quality of laying hens after peak production fed diets containing phosphorus levels and phytase. An experiment was conducted with 384 Hy-line distributed in a completely randomized in a factorial 4 x 3 with 4 levels of available phosphorus and 3 levels of phytase. The experimental period was divided into four periods of 28 days, at the end of each cycle were determined experimental feed intake, egg production, egg weight, feed conversion, mortality, and average egg weight, shell thickness, Haugh units and specific gravity. At the end of the experimental period were determined amounts of calcium and phosphorus excreted by the method of total excreta collection and a fowl per experimental unit was sacrificed for collection of bones and evaluation of width, length and level of robustness from femur and tibia. There was interaction between phosphorus levels and phytase on feed intake, feed conversion and percentage of posture. For inclusion levels of phytase all egg quality variables showed no significant differences. The treatments did not affect bone characteristics of laying hens.

  8. 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 study...... compares 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 and examines secondary protein structural changes at low pH via circular dichroism. The Peniophora...... lycii phytase was found to be the most thermostable, but the least robust enzyme in gastric conditions, whereas the Aspergillus niger and Escherichia coli phytases proved to be most resistant to gastric conditions. The phytase from Citrobacter braakii showed intermediate robustness. The extent of loss...

  9. Phytase-active lactic acid bacteria from sourdoughs

    DEFF Research Database (Denmark)

    Nuobariene, Lina; Cizeikiene, Dalia; Gradzeviciute, Egle

    2015-01-01

    Whole-grain foods play an important role in human diet as they are relatively rich in minerals, however, the absorption of those minerals in human gut can be very low due to high content of the mineral binding phytate. Therefore, the object of this study was to identify phytase-active lactic acid...... 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...... 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...

  10. Effect of nutrient components for phytase production by Aspergillus niger

    OpenAIRE

    KALIYEVA AIGUL; SULEIMENOVA ZHANARA; AKHMETSADYKOV NURLAN; SADUYEVA ZHAZIRA

    2015-01-01

    In present study the effect of carbon sources, glucose, sucrose, lactose, maltose, fructose, xylose and nitrogen sources such as ammonium sulfate, ammonium phosphate, ammonium nitrate, potassium nitrate, yeast extract, peptone on the phytase production has been studied. Maximal phytase activity of Aspergillus niger was detected in media with 1.0% sucrose as a carbon source. All other monosaccharides and disaccharides used had less effect on phytase production. Among the inorganic and organic ...

  11. Two types of phytases (histidine acid phytase and β-propeller phytase) in Serratia sp. TN49 from the gut of Batocera horsfieldi (coleoptera) larvae.

    Science.gov (United States)

    Zhang, Rui; Yang, Peilong; Huang, Huoqing; Shi, Pengjun; Yuan, Tiezheng; Yao, Bin

    2011-11-01

    Microbial phytases play a major role in the mineralization of organic phosphorous, especially in symbiotic plants and animals. In this study, we identified two types of phytases in Serratia sp. TN49 that was harbored in the gut of Batocera horsfieldi (Coleoptera) larvae. The two phytases, an acidic histidine acid phosphatase (PhyH49) and an alkaline β-propeller phytase (PhyB49), shared low identities with known phytases (61% at most). PhyH49 and PhyB49 produced in Escherichia coli exhibited maximal activities at pH 5.0 (60°C) and pH 7.5-8.0 (45°C), respectively, and are complementary in phytate degradation over the pH range 2.0-9.0. Serratia sp. TN49 harboring both PhyH49 and PhyB49 might make it more adaptive to environment change, corresponding to the evolution trend of microorganism.

  12. Phytase application in chewing gum - A technical assessment

    DEFF Research Database (Denmark)

    Nielsen, Anne Veller Friis; Meyer, Anne S.

    2016-01-01

    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....

  13. 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...

  14. 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. Copyright © 2014 Elsevier GmbH. All rights reserved.

  15. Fermentative production and application of acid phytase by ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-09-03

    Sep 3, 2008 ... the inorganic phosphate and less-phosphorylated myo- ... carbon, nitrogen, mineral sources, and culture pH, to increase the phytase ... Phytase assays were carried out using the Shimizu method. (Shimizu .... Urea. 7.27. 18.45. aControl medium: 3.0% galactose, 1.0% yeast extract, 0.6% succinic acid, pH ...

  16. Effects of supplemental microbial phytase enzyme on performance ...

    African Journals Online (AJOL)

    This experiment was conducted to investigate the effects of supplemental phytase in a corn-wheatsoybean meal basal diet on phosphorus (P) digestibility and performance of broiler chicks. 378 one-day old broiler chicks (Ross 308) were allocated to 3×3 factorial arrangements with three levels of phytase enzyme (0, 500 ...

  17. 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...

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

    DEFF Research Database (Denmark)

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

    2016-01-01

    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......BACKGROUND Enzymatic dephosphorylation of phytic acid (inositol hexakisphosphate) in cereals may improve mineral bioavailability in humans. This study quantified enzymatic dephosphorylation of phytic acid by measuring inositol tri- to hexakisphosphate (InsP3-6) degradation and iron and zinc release...... 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...

  19. Extracellular Secretion of Phytase from Transgenic Wheat Roots Allows Utilization of Phytate for Enhanced Phosphorus Uptake.

    Science.gov (United States)

    Mohsin, Samreen; Maqbool, Asma; Ashraf, Mehwish; Malik, Kauser Abdulla

    2017-08-01

    A significant portion of organic phosphorus comprises of phytates which are not available to wheat for uptake. Hence for enabling wheat to utilize organic phosphorus in form of phytate, transgenic wheat expressing phytase from Aspergillus japonicus under barley root-specific promoter was developed. Transgenic events were initially screened via selection media containing BASTA, followed by PCR and BASTA leaf paint assay after hardening. Out of 138 successfully regenerated T o events, only 12 had complete constructs and thus further analyzed. Positive T1 transgenic plants, grown in sand, exhibited 0.08-1.77, 0.02-0.67 and 0.44-2.14 fold increase in phytase activity in root extracts, intact roots and external root solution, respectively, after 4 weeks of phosphorus stress. Based on these results, T2 generation of four best transgenic events was further analyzed which showed up to 1.32, 56.89, and 15.40 fold increase in phytase activity in root extracts, intact roots and external root solution, respectively, while in case of real-time PCR, maximum fold increase of 19.8 in gene expression was observed. Transgenic lines showed 0.01-1.18 fold increase in phosphorus efficiency along with higher phosphorus content when supplied phytate or inorganic phosphorus than control plants. Thus, this transgenic wheat may aid in reducing fertilizer utilization and enhancing wheat yield.

  20. Transgenic expression of phytase in wheat endosperm increases bioavailability of iron and zinc in grains.

    Science.gov (United States)

    Abid, Nabeela; Khatoon, Asia; Maqbool, Asma; Irfan, Muhammad; Bashir, Aftab; Asif, Irsa; Shahid, Muhammad; Saeed, Asma; Brinch-Pedersen, Henrik; Malik, Kauser A

    2017-02-01

    Phytate is a major constituent of wheat seeds and chelates metal ions, thus reducing their bioavailability and so the nutritional value of grains. Transgenic plants expressing heterologous phytase are expected to enhance degradation of phytic acid stored in seeds and are proposed to increase the in vitro bioavailability of mineral nutrients. Wheat transgenic plants expressing Aspergillus japonicus phytase gene (phyA) in wheat endosperm were developed till T 3 generation. The transgenic lines exhibited 18-99 % increase in phytase activity and 12-76 % reduction of phytic acid content in seeds. The minimum phytic acid content was observed in chapatti (Asian bread) as compared to flour and dough. The transcript profiling of phyA mRNA indicated twofold to ninefold higher expression as compared to non transgenic controls. There was no significant difference in grain nutrient composition of transgenic and non-transgenic seeds. In vitro bioavailability assay for iron and zinc in dough and chapatti of transgenic lines revealed a significant increase in iron and zinc contents. The development of nutritionally enhanced cereals is a step forward to combat nutrition deficiency for iron and zinc in malnourished human population, especially women and children.

  1. 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. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. 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.

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

    Science.gov (United States)

    Osswald, Annika; Westermann, Christina; Sun, Zhongke; Riedel, Christian U.

    2015-01-01

    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. PMID:26086721

  4. Ileal microbiota of growing pigs fed different dietary calcium phosphate levels and phytase content and subjected to ileal pectin infusion.

    Science.gov (United States)

    Metzler-Zebeli, B U; Vahjen, W; Baumgärtel, T; Rodehutscord, M; Mosenthin, R

    2010-01-01

    Two experiments with growing pigs were conducted to determine the effects of dietary P and Ca levels, phytase supplementation, and ileal pectin infusion on changes in bacterial populations in the ileum and on ileal and fecal fermentation patterns. Growing pigs (BW 30.1 +/- 1.3 kg) were fitted with simple T-cannulas at the distal ileum and were fed a low-P corn-soybean meal control diet (3 g of P/kg), or the control diet supplemented with either 15 g of monocalcium phosphate (MCP)/kg (Exp. 1) or 1,000 phytase units of phytase/kg (Exp. 2). Daily infusion treatments consisted of either 60 g of pectin dissolved in 1.8 L of demineralized water or 1.8 L of demineralized water as a control infusion, infused via the ileal cannula. In each experiment, 8 barrows were assigned to 4 dietary treatments according to a double incomplete 4 x 2 Latin square design. The dietary treatments in Exp. 1 were the control diet with water infusion, the control diet with pectin infusion, the MCP diet with water infusion, or the MCP diet with pectin infusion. In Exp. 2, the pigs received the same control treatments as in Exp. 1 and the phytase diet in combination with water or pectin infusion. Gene copy numbers of total bacteria, Lactobacillus spp., Lactobacillus reuteri, Lactobacillus amylovorus/Lactobacillus sobrius, Lactobacillus mucosae, Enterococcus spp., Enterococcus faecium, Enterococcus faecalis, bifidobacteria, the Clostridium coccoides cluster, the Clostridium leptum cluster, the Bacteroides-Prevotella-Porphyrmonas group, and Enterobacteriaceae were determined by quantitative PCR in DNA extracts of ileal digesta. In Exp. 1, addition of MCP reduced ileal gene copy numbers of Enterococcus spp. (P = 0.048), E. faecium (P = 0.015), and the C. leptum cluster (P = 0.028), whereas pectin infusion enhanced (P = 0.008) ileal d-lactate concentration. In Exp. 2, supplemental phytase led to greater ileal gene copy numbers of the C. coccoides (P = 0.041) and C. leptum (P = 0.048) clusters and

  5. 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...

  6. Microbial and biochemical studies on phytase enzyme in some microorganisms

    International Nuclear Information System (INIS)

    Abdelbary, N.A.

    1997-01-01

    Mixed calcium and magnesium salts of phytic acid myoinositol hexa phosphoric acid are widely distributed in food stuffs of plant origin, they may bind essential proteins, phospholipids and microelements to form indigestible compounds. In this concern, destruction of phytic acid and its salts by different methods is very important, one of them is by using microbial phytase. This study aims to produce phytase enzyme from microorganisms and study the best conditions of production and purification and also the properties of the partially purified phytase. 22 figs., 29 tabs., 61 refs

  7. Phytase activity, phytic acid, zinc, phosphorus and protein contents ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-09-15

    Sep 15, 2009 ... Key words: Zinc, nitrogen, phosphorus, phytic acid, phytase. .... speculated that the synthesizing metabolism of grain PA probably was closely ..... Efficiency when Grown in the Chelate-Buffered Nutrient Solution. II. Nutrient ...

  8. Consecutive treatment with phytase and arazyme influence protein ...

    African Journals Online (AJOL)

    Jane

    2011-08-01

    Aug 1, 2011 ... accelerated livestock growth. Key words: Soybean meal, phytase, arazyme, hydrolysis. .... Fast protein liquid chromatography (FPLC) analysis. The FPLC of ..... A study on the effect of substituting soybean oil meal for imported ...

  9. Aspergillus ficuum phytase activity is inhibited by cereal grain components.

    Science.gov (United States)

    Bekalu, Zelalem Eshetu; Madsen, Claus Krogh; Dionisio, Giuseppe; Brinch-Pedersen, Henrik

    2017-01-01

    In the current study, we report for the first time that grain components of barley, rice, wheat and maize can inhibit the activity of Aspergillus ficuum phytase. The phytase inhibition is dose dependent and varies significantly between cereal species, between cultivars of barley and cultivars of wheat and between Fusarium graminearum infected and non-infected wheat grains. The highest endpoint level of phytase activity inhibition was 90%, observed with grain protein extracts (GPE) from F. graminearum infected wheat. Wheat GPE from grains infected with F. graminearum inhibits phytase activity significantly more than GPE from non-infected grains. For four barley cultivars studied, the IC50 value ranged from 0.978 ± 0.271 to 3.616 ± 0.087 mg×ml-1. For two non-infected wheat cultivars investigated, the IC50 values were varying from 2.478 ± 0.114 to 3.038 ± 0.097 mg×ml-1. The maize and rice cultivars tested gaveIC50 values on 0.983 ± 0.205 and 1.972 ± 0.019 mg×ml-1, respectively. After purifying the inhibitor from barley grains via Superdex G200, an approximately 30-35 kDa protein was identified. No clear trend for the mechanism of inhibition could be identified via Michaelis-Menten kinetics and Lineweaver-Burk plots. However, testing of the purified phytase inhibitor together with the A. ficuum phytase and the specific protease inhibitors pepstatin A, E64, EDTA and PMSF revealed that pepstatin A repealed the phytase inhibition. This indicates that the observed inhibition of A. ficuum phytase by cereal grain extracts is caused by protease activity of the aspartic proteinase type.

  10. Phosphorus digestibility is highly influenced by phytase but slightly by calcium in growing pigs

    DEFF Research Database (Denmark)

    Poulsen, Hanne Damgaard; Carlson, Dorthe; Nørgaard, Jan Værum

    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...... phytases: one fungal 3-phytase from Aspergillus ficuum and two formulations of a bacterial 6-phytase from E. coli. The basal diet was composed of wheat, barley, soybean and rapeseed meal supplemented with vitamins and minerals without any added inorganic phosphate. Calcium was adjusted to 6.3 g....../kg in the basal diet which was heat-treated at 90 °C. Phytase was then added at 250, 500 and 750 FTU/kg diet. A negative control without added phytase was included (plant phytase

  11. Phytase in broiler diets with reduced nonphytate phosphorus levels

    Directory of Open Access Journals (Sweden)

    Vera Maria Barbosa de Moraes

    2009-12-01

    Full Text Available This work evaluated the inclusion of phytase in diets with reduced nonphytate phosphorus (NPP levels in relation to the performance and carcass yield of broilers and litter characteristics. The experimental design was completely randomized in a 2 x 3 factorial arrangement – two phytase levels (0 and 25U/kg and three NPP levels (100, 85 and 70% of the requirement, in a total of six treatments with fi ve replicates. The reduction of the NPP to 70% of the requirement decreased the weight gain and feed intake and increased the feed:gain ratio. Phytase inclusion increased the weight gain and the feed intake at 21 days. The NPP x phytase interaction was signifi cant with regard to the weight gain at 42 days. The reduction of NPP to 70% diminished the feed intake at 42 days and increased the liver and heart relative weight. Nitrogen content was lower in the litter of broilers fed diets with phytase and decreased phosphorus content, due to the NPP reduction in the diets. It was concluded that diets with 85% of NPP, supplemented with 25U/kg of phytase, can be used with no negative effect on performance or carcass and litter characteristics.

  12. 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

    2017-04-01

    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.

  13. Screening of mutant strains producing phytase from A. niger by 60Co γ-ray irradiation

    International Nuclear Information System (INIS)

    Yang Pingping; Wang Yan; Tao Wenyi

    2004-01-01

    60 Co γ-ray was used to irradiate Aspergillus niger 447-92 for screening the mutant strain of producing phytase, and the effects of mutation induction were determined and analyzed. A mutant strain A. niger 496-1 with high level of phytase was selected, the phytase properties of A. niger 496-1 were analyzed

  14. 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 ...

  15. Co-feeding strategy to enhance phytase production in Pichia pastoris

    African Journals Online (AJOL)

    The experimental designs, the methanol/L-alanine co-feeding strategy and optimization of phytase production by P. pastoris supported by the optimum levels of variables and lower temperature expression produced high level of phytase activity which could be scaled up to produce phytase for food additives at industrial ...

  16. Differential influence of phytase supplementation on the balance of phosphorus and other elements in laying hens' feed

    OpenAIRE

    Sobolewska, Sylwia; Orda, Janusz; Budny-Walczak, Anna

    2015-01-01

    The aim of this study was to evaluate the impact of microbial phytase addition to laying hen diet on the balance of phosphorus, calcium, manganese and zinc. Phytase supplementation caused a decrease in phosphorus excretion by 8% in the group with low-phytase diet and 21% in the group with phytase addition of 1000 FTU (one unit of phytase activity) to a diet. Phytase supplementation increased the retention of phosphorus by 31% in laying hens receiving 500 FTU and 57% in the group with phytase ...

  17. 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.

  18. 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.

  19. “In Silico” Characterization of 3-Phytase A and 3-Phytase B from Aspergillus niger

    Directory of Open Access Journals (Sweden)

    Doris C. Niño-Gómez

    2017-01-01

    Full Text Available Phytases are used for feeding monogastric animals, because they hydrolyze phytic acid generating inorganic phosphate. Aspergillus niger 3-phytase A (PDB: 3K4Q and 3-phytase B (PDB: 1QFX were characterized using bioinformatic tools. Results showed that both enzymes have highly conserved catalytic pockets, supporting their classification as histidine acid phosphatases. 2D structures consist of 43% alpha-helix, 12% beta-sheet, and 45% others and 38% alpha-helix, 12% beta-sheet, and 50% others, respectively, and pI 4.94 and 4.60, aliphatic index 72.25 and 70.26 and average hydrophobicity of −0,304 and −0.330, respectively, suggesting aqueous media interaction. Glycosylation and glycation sites allowed detecting zones that can affect folding and biological activity, suggesting fragmentation. Docking showed that H59 and H63 act as nucleophiles and that D339 and D319 are proton donor residues. MW of 3K4Q (48.84 kDa and 1QFX (50.78 kDa is similar; 1QFX forms homodimers which will originate homotetramers with several catalytic center accessible to the ligand. 3K4Q is less stable (instability index 45.41 than 1QFX (instability index 33.66, but the estimated lifespan for 3K4Q is superior. Van der Waals interactions generate hydrogen bonds between the active center and O2 or H of the phytic acid phosphate groups, providing greater stability to these temporal molecular interactions.

  20. Phytase-mediated enzymatic mineralization of chitosan-enriched hydrogels

    DEFF Research Database (Denmark)

    Lišková, Jana; Douglas, Timothy E.L.; Wijnants, Robbe

    2018-01-01

    Hydrogels mineralized with calcium phosphate (CaP) are increasingly popular bone regeneration biomaterials. Mineralization can be achieved by phosphatase enzyme incorporation and incubation in calcium glycerophosphate (CaGP). Gellan gum (GG) hydrogels containing the enzyme phytase and chitosan...... oligomer were mineralized in CaGP solution and characterized with human osteoblast-like MG63 cells and adipose tissue-derived stem cells (ADSC). Phytase induced CaP formation. Chitosan concentration determined mineralization extent and hydrogel mechanical reinforcement. Phytase-induced mineralization...... promoted MG63 adhesion and proliferation, especially in the presence of chitosan, and was non-toxic to MG63 cells (with and without chitosan). ADSC adhesion and proliferation were poor without mineralization. Chitosan did not affect ADSC osteogenic differentiation....

  1. Waste vinegar residue as substrate for phytase production.

    Science.gov (United States)

    Wang, Zhi-Hong; Dong, Xiao-Fang; Zhang, Guo-Qing; Tong, Jian-Ming; Zhang, Qi; Xu, Shang-Zhong

    2011-12-01

    Waste vinegar residue, the by-product of vinegar processing, was used as substrate for phytase production from Aspergillus ficuum NTG-23 in solid-state fermentation to investigate the potential for the efficient re-utilization or recycling of waste vinegar residue. Statistical designs were applied in the processing of phytase production. First, a Plackett-Burman (PB) design was used to evaluate eleven parameters: glucose, starch, wheat bran, (NH(4))(2)SO(4), NH(4)NO(3), tryptone, soybean meal, MgSO(4)·7H(2)O, CaCl(2)·7H(2)O, FeSO(4)·7H(2)O, incubation time. The PB experiments showed that there were three significant factors: glucose, soybean meal and incubation time. The closest values to the optimum point were then derived by steepest ascent path. Finally, a mathematical model was created and validated to explain the behavioural process after these three significant factors were optimized using response surface methodology (RSM). The best phytase activity was attained using the following conditions: glucose (7.2%), soybean meal (5.1%), and incubation time (271 h). The phytase activity was 7.34-fold higher due to optimization by PB design, steepest ascent path design and RSM. The phytase activity was enhanced 0.26-fold in comparison with the results by the second step of steepest ascent path design. The results indicate that with waste vinegar residue as a substrate higher production of phytase from Aspergillus ficuum NTG-23 could be obtained through an optimization process and that this method might be applied to an integrated system for recycling of the waste vinegar residue.

  2. ECONOMIC RETURNS FROM REDUCING POULTRY LITTER PHOSPHORUS WITH MICROBIAL PHYTASE

    OpenAIRE

    Bosch, Darrell J.; Zhu, Minkang; Kornegay, Ervin T.

    1997-01-01

    Requiring that crop applications of manure be based on phosphorus content (P-standard) could increase poultry litter disposal costs. Microbial phytase reduces litter P content and could reduce litter disposal costs under a P-standard. For a representative Virginia turkey farm, phytase costs $2,500 and could increase value of litter used for fertilizer on the turkey farm by $390 and reduce supplemental P feed costs by $1,431. Based on assumed litter demand and supply, estimated litter export p...

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

    Directory of Open Access Journals (Sweden)

    Aline Puspita Kusumadjaja

    2010-06-01

    Full Text Available 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 affect the enzyme activity. Thermostabile phytase can be isolated from microorganism live in hot spring water or volcano crater. In this study, the screening of thermophylic microorganism having thermostabile phytase activity in Ijen Crater, Banyuwangi, has been done. From this process, it was obtained 33 isolates that produce phytase enzyme. Isolate was code by AP-17 yields highest phytase activity, that is 0.0296 U/mL, so this isolate was choosen for further study. The activity of crude phytase enzyme was measured based on the amount of anorganic phosphate that was produced in enzymatic reaction using UV-VIS spectrophotometer at 392 nm. Based on morphology test to identify the gram type of microorganism, isolate AP-17 has a bacill cell type and identified as positive gram bacteria. This isolate was assumed as Bacillus type.   Keywords: Phytase, thermophilic microorganism, phytase activity

  4. Secretion of non-cell-bound phytase by the yeast Pichia kudriavzevii TY13.

    Science.gov (United States)

    Hellström, A; Qvirist, L; Svanberg, U; Veide Vilg, J; Andlid, T

    2015-05-01

    Mineral deficiencies cause several health problems in the world, especially for populations consuming cereal-based diets rich in the anti-nutrient phytate. Our aim was to characterize the phytate-degrading capacity of the yeast Pichia kudriavzevii TY13 and its secretion of phytase. The phytase activity in cell-free supernatants from cultures with 100% intact cells was 35-190 mU ml(-1) depending on the media. The Km was 0.28 mmol l(-1) and the specific phytase activity 0.32 U mg(-1) total protein. The phytase activity and secretion of extracellular non-cell-bound phytase was affected by the medium phosphate concentrations. Further, addition of yeast extract had a clearly inducing effect, resulting in over 60% of the cultures total phytase activity as non-cell-bound. Our study reveals that it is possible to achieve high extracellular phytase activity from the yeast P. kudriavzevii TY13 by proper composition of the growth medium. TY13 could be a promising future starter culture for fermented foods with improved mineral bioavailability. Using strains that secrete phytase to the food matrix may significantly improve the phytate degradation by facilitating the enzyme-to-substrate interaction. The secreted non-cell-bound phytase activities by TY13 could further be advantageous in industrial production of phytase. © 2015 The Society for Applied Microbiology.

  5. 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. © 2015 Poultry Science Association Inc.

  6. Increased iron level in phytase-supplemented diets reduces performance and nutrient utilisation in broiler chickens.

    Science.gov (United States)

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

    2017-08-01

    1. The effect of different levels of dietary iron on phytase activity and its subsequent effect on broiler performance were investigated in a 3 × 2 factorial arrangement. A total of 360 day-old Ross 308 male broiler chicks were distributed to 6 experimental diets, formulated with three levels of Fe (60, 80 and 100 mg/kg) and two levels of phytase (0 and 500 FTU/kg). 2. Phytase supplemented to mid-Fe diets increased feed consumption more than the non-supplemented diet at d 24. From hatch to d 35, Fe × phytase interaction significantly influenced the feed intake (FI), body weight gain (BWG) and feed conversion ratio (FCR). The high-Fe diet supplemented with phytase significantly reduced FI and BWG of broilers than those supplemented with low- or mid-Fe diets. The overall FCR was significantly better in birds fed on the mid-Fe diets with phytase supplementation. 3. A significant improvement in ileal digestibility of N, P, Mg and Fe was observed in birds feed diets containing 60 mg Fe/kg, with significant interaction between Fe and phytase. 4. Phytase improved the bone breaking strength when supplemented to low- or mid-Fe diets, compared to the non-supplemented diets. There was a significant Fe × phytase interaction effect. Tibia Fe content was higher in birds fed on phytase-free diets with high Fe but the reverse was the case when phytase was added and their interaction was significant. High dietary Fe significantly increased the accumulation of Fe in liver. 5. Phytase improved Ca-Mg-ATPase, Ca-ATPase and Mg-ATPase activities in jejunum when supplemented to the diet containing 80 mg Fe/kg. 6. This study indicates that high (100 mg/kg) dietary Fe inhibited phytase efficacy and subsequently reduced the overall performance and nutrient utilisation of broilers.

  7. 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...

  8. Effect of combination of citric acid and microbial phytase on ...

    African Journals Online (AJOL)

    CA) and microbial phytase (MP) on digestibility of calcium, phosphorus and mineralization parameters of tibia bone in broilers chicks. A total of 360 Ross-308 male broiler chicks were used in a completely randomized design with a 3 × 2 factorial ...

  9. Increasing zinc levels in phytase-supplemented diets improves the ...

    African Journals Online (AJOL)

    Three hundred and thirty-six day-old Ross-308 male broiler chicks were used in a 35-day trial to investigate the effect of different concentrations of dietary Zn and phytase on broiler performance and energy utilization. Twelve day-old birds were used for the initial slaughter group to provide baseline body compositional data, ...

  10. Biosynthesis of phytases and phosphatases by Aspergillus niger 551

    African Journals Online (AJOL)

    ARyw

    2012-02-16

    Feb 16, 2012 ... The culture was stored on potato dextrose agar (PDA) slants at 4°C. Medium. The medium consisted of: starch (40 g/L), glucose (5 g/L), KCl (0.5 ..... Production, purification and properties of microbial phytases. Bioresour.

  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

  12. Effect of phosphorus limiting on phytase activity, proton efflux and ...

    African Journals Online (AJOL)

    This work intended to measure the nodulated-roots oxygen consumption, proton efflux and phytase activity in 2 lines of common bean (Phaseolus vulgaris) (115, 147) at 2 levels of P supply. Rooted seedlings were inoculated with Rhizobium tropici CIAT 899 in hydroaeroponic cultivation under glasshouse. Phosphorus was ...

  13. 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.

  14. 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...

  15. Enhancing the Thermal Resistance of a Novel Acidobacteria-Derived Phytase by Engineering of Disulfide Bridges.

    Science.gov (United States)

    Tan, Hao; Miao, Renyun; Liu, Tianhai; Cao, Xuelian; Wu, Xiang; Xie, Liyuan; Huang, Zhongqian; Peng, Weihong; Gan, Bingcheng

    2016-10-28

    A novel phytase of Acidobacteria was identified from a soil metagenome, cloned, overexpressed, and purified. It has low sequence similarity (phytases. At the optimum pH (2.5), the phytase shows an activity level of 1,792 μmol/min/mg at physiological temperature (37°C) and could retain 92% residual activity after 30 min, indicating the phytase is acidophilic and acidostable. However the phytase shows poor stability at high temperatures. To improve its thermal resistance, the enzyme was redesigned using Disulfide by Design 2.0, introducing four additional disulfide bridges. The half-life time of the engineered phytase at 60°C and 80°C, respectively, is 3.0× and 2.8× longer than the wild-type, and its activity and acidostability are not significantly affected.

  16. 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....

  17. Co-feeding strategy to enhance phytase production in Pichia pastoris

    African Journals Online (AJOL)

    SAM

    2014-05-21

    May 21, 2014 ... phytase was induced by methanol/L-Alanine (0.5% for first day and 1%, 0.3925 and 0.7185 g/L/h, respectively for subsequent days till 7 days). Due to the heavy glycol- sylation, the expressed phytase was found to have mole- cular sizes of around 120, 116, and 66 kDa and this showed that phytase is a ...

  18. 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...

  19. Phytase-active yeasts from grain-based food and beer.

    Science.gov (United States)

    Nuobariene, L; Hansen, A S; Jespersen, L; Arneborg, N

    2011-06-01

    To screen yeast strains isolated from grain-based food and beer for phytase activity to identify high phytase-active strains. The screening of phytase-positive strains was carried out at conditions optimal for leavening of bread dough (pH 5·5 and 30°C), in order to identify strains that could be used for the baking industry. Two growth-based tests were used for the initial testing of phytase-active strains. Tested strains belonged to six species: Saccharomyces cerevisiae, Saccharomyces pastorianus, Saccharomyces bayanus, Kazachstania exigua (former name Saccharomyces exiguus), Candida krusei (teleomorph Issachenkia orientalis) and Arxula adeninivorans. On the basis of initial testing results, 14 strains were selected for the further determination of extracellular and intracellular (cytoplasmic and/or cell-wall bound) phytase activities. The most prominent strains for extracellular phytase production were found to be S. pastorianus KVL008 (a lager beer strain), followed by S. cerevisiae KVL015 (an ale beer strain) and C. krusei P2 (isolated from sorghum beer). Intracellular phytase activities were relatively low in all tested strains. Herein, for the first time, beer-related strains of S. pastorianus and S. cerevisiae are reported as phytase-positive strains. The high level of extracellular phytase activity by the strains mentioned previously suggests them to be strains for the production of wholemeal bread with high content of bioavailable minerals. © 2011 The Authors. Journal of Applied Microbiology © 2011 The Society for Applied Microbiology.

  20. 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

  1. Isolation and identification of phytase-producing strains from soil samples and optimization of production parameters

    Directory of Open Access Journals (Sweden)

    Masoud Mohammadi

    2017-09-01

    Discussion and conclusion: Penicillium sp. isolated from a soil sample near Qazvin, was able to produce highly active phytase in optimized environmental conditions, which could be a suitable candidate for commercial production of phytase to be used as complement in poultry feeding industries.

  2. Effect of citric acid and microbial phytase on serum enzyme activities ...

    African Journals Online (AJOL)

    Effect of citric acid and microbial phytase on serum enzyme activities and plasma minerals retention in broiler chicks. ... African Journal of Biotechnology ... An experiment was conducted to study the effect of microbial phytase supplementation and citric acid in broiler chicks fed corn-soybean meal base diets on enzyme ...

  3. Phytase activity of fungi from oil polluted soils and their ability to ...

    African Journals Online (AJOL)

    Mrs. Ekundayo Esther

    2013-09-04

    Sep 4, 2013 ... Full Length Research Paper. Phytase activity of ... Key words: Fungi, biodegradation, bonny light crude oil, phytase. ... bon compounds bind to different soil components and these are ... tium of microorganisms and more than 200 species of ..... ducts contain hydrocarbon that are toxic to microorga- nisms.

  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. 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 in samples of the TMR revealed that the exogenous phytase started degrading the inositol phosphate when feeds and phytase were mixed, and thus the InsP6 phosphorus (InsP6-P) content in the TMR was found to decrease with higher doses of phytase (1.69, 1.51, 1.39, and 1.25 g/kg of DM for the none, low, medium...

  6. 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

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

    DEFF Research Database (Denmark)

    Dionisio, Giuseppe; Brinch-Pedersen, Henrik; 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...

  8. 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

  9. 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

  10. 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

  11. 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.

  12. Increasing zinc levels in phytase-supplemented diets improves the ...

    African Journals Online (AJOL)

    Marjina Akter

    2017-09-02

    Sep 2, 2017 ... This result indicate that the Zn concentrations used in this study were not ... levels of phytase, the concentrations of Zn in the experimental diets of these studies ... combustion technique, as described by Sweeney (1989) using a LECO® ... Premix A. 2.0. 2.0. 2.0. 2.0. 2.0. 2.0. Choline Cl. 0.9. 0.9. 0.9. 1.1. 0.9.

  13. 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...... reduced APS by 48% in maize, 30% in SBM and 26% in maize–SBM. No interaction between screen size and microbial phytase on phytate degradation was observed, but the interaction between microbial phytase and incubation time was significant (P... of screen size and feed on microbial phytase efficacy on phytate degradation. The interaction between screen size and feed affected the relative phytate degradation rate (Rd) of microbial phytase as well as the time to decrease 50% of the phytate P (t) (P

  14. 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

    The present paper summarizes the current state of knowledge on cereal phytase that are particular relevant for improving mineral and phosphate bio-availability. Phytases can initiate the hydrolysis of phytate, the main storage form of phosphate in cereals and the major anti-nutritional factor...... 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...... wheat, barley and rye whereas non-Triticeae cereals such as maize and rice have very little MGPA. Recent studies have determined the evolutionary relationships of phytases in Triticeae and non-Triticeae and highlighted the importance of the purple acid phosphatase phytases (PAPhys). In the Triticeae...

  15. 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 phytase purple acid phosphatase (HvPAPhy_a) expressed during barley seed development was evaluated as transgene for overexpression in barley. The phytase was expressed constitutively driven by the cauliflower mosaic virus 35S-promoter, and the phytase activity was measured in the mature grains......, 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...... activities were increased significantly by 110-fold (52 000 FTU/kg) and 57-fold (51 000 FTU/kg), respectively. The HvPAPhy_a-transformed barley plants with high phytase activities possess triple potential utilities for the improvement of phosphate bioavailability. First of all, the utilization of the mature...

  16. 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.

  17. 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. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  18. 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

  19. 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).

  20. Fungal phytases: characteristics and amelioration of nutritional quality and growth of non-ruminants.

    Science.gov (United States)

    Singh, B; Satyanarayana, T

    2015-08-01

    Fungal phytases are histidine acid phosphatases, a subclass of acid phosphatases, which catalyse the hydrolysis of phytic acid resulting in the release of phosphate moieties and thus mitigate its antinutritional properties. The supplementation of feed with phytases increases the bioavailability of phosphorus and minerals in non-ruminant animals and reduces the phosphorus pollution due to phosphorus excretion in the areas of intensive livestock production. Although phytases are reported in plants, animals and micro-organisms, fungal sources are used extensively for the production of phytases on a commercial scale. Phytases have been produced by fungi in both solid-state fermentation (SSF) and submerged fermentation (SmF). The fungal phytases are high molecular weight proteins ranging from 35 to 500 kDa. They are optimally active within pH and temperature ranges between 4.5 and 6.0, and 45 and 70 °C respectively. Phytate degradation leads to amelioration in the nutritional status of foods and feeds by improving the availability of minerals, phosphorus and proteins in non-ruminant animals and human beings and thus mitigates the environmental phosphorus pollution. Our article focuses on the role of fungal phytases in improving nutritional value of foods and feeds with concomitant increase in growth of non-ruminant animals and mitigating environmental phosphorus pollution. Journal of Animal Physiology and Animal Nutrition © 2014 Blackwell Verlag GmbH.

  1. 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.

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

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    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.

  3. 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.

  4. Effect of two microbial phytase preparations on phosphorus utilisation in broilers fed maize-soybean meal based diets

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    J. VALAJA

    2008-12-01

    Full Text Available The present study was carried out to determine the effect of two microbial phytases, Aspergillus niger (FINASEâ FP-500, 291 PU (phytase units/g and Trichoderma reesei phytase (FINASEâ P, 5880 PU/g on phosphorus (P and calcium (Ca utilisation and ileal P and Ca digestibility in broiler chickens fed diets based on maize and soybean meal. A total of 96 Ross broiler chickens housed four birds to a cage were used. Four dietary treatments consisted of a positive control supplemented with dicalcium phosphate (17 g/kg, a negative control without inorganic P, basal diet without inorganic P supplemented with Aspergillus niger phytase (2.6 g/kg and basal diet without inorganic P supplemented with Trichoderma reesei phytase (0.13 g/kg. Both phytases provided 750 PU/kg feed. P retention per unit intake was lowest and P excretion highest in birds fed the positive control diet with inorganic P (P

  5. Effect of phytase in laying hen diets with different phosphorus sources

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    EM Casartelli

    2005-06-01

    Full Text Available An experiment was conducted to evaluate the effects of the enzyme phytase in diets formulated with different phosphorus sources on performance, eggshell quality and excretion of commercial laying hens. Two hundred and eighty-eight commercial Hyssex Brown laying hens were evaluated during two production phases, which included eight twenty-eight-day cycles, using a completely randomized design in a 3x2 factorial with six replicates of eight birds per treatment. Three phosphorus sources (calcium and sodium phosphate, micro-granulated dicalcium phosphate and triple super phosphate and two phytase levels (0 or 1000 FTU/kg diet were tested in the composition of the diets. After the post-peak period, triple super phosphate decreased bird performance and eggshell quality. It was possible to reduce the levels of phosphorus supplementation when phytase was added to the diet. Besides, phytase supplementation reduced phosphorus, calcium and nitrogen excretions, but affected mean egg weight at production peak.

  6. Purification and identification of a phytase from fruity bodies of the ...

    African Journals Online (AJOL)

    use

    2011-12-05

    Dec 5, 2011 ... The chromatographic procedure used for isolation of the phytase included ion exchange ... phosphorus in plant foods such as oil seeds, cereal grains and ... anhydrase (30 kDa), soybean trypsin inhibitor (20 kDa) and.

  7. Candida albicans orf19.3727 encodes phytase activity and is essential for human tissue damage

    Science.gov (United States)

    Fong, Wing-Ping; Samaranayake, Lakshman Perera

    2017-01-01

    Candida albicans is a clinically important human fungal pathogen. We previously identified the presence of cell-associated phytase activity in C. albicans. Here, we reveal for the first time, that orf19.3727 contributes to phytase activity in C. albicans and ultimately to its virulence potency. Compared with its wild type counterpart, disruption of C. albicans orf19.3727 led to decreased phytase activity, reduced ability to form hyphae, attenuated in vitro adhesion, and reduced ability to penetrate human epithelium, which are the major virulence attributes of this yeast. Thus, orf19.3727 of C. albicans plays a key role in fungal pathogenesis. Further, our data uncover a putative novel strategy for anti-Candidal drug design through inhibition of phytase activity of this common pathogen. PMID:29216308

  8. Standardized total tract digestibility of phosphorus in flaxseed meal fed to growing and finishing pigs without or with phytase supplementation.

    Science.gov (United States)

    Kim, J W; Ndou, S P; Mejicanos, G A; Nyachoti, C M

    2017-02-01

    Two experiments were conducted to determine the apparent total tract digestibility (ATTD) and standardized total tract digestibility (STTD) of P in flaxseed meal (FM) and the effect of dietary microbial phytase on the digestibility of P in FM fed to growing and finishing pigs. In Exp. 1, eighteen growing barrows (26.6 ± 1.8 kg BW) were allotted to 1 of 3 experimental diets consisting of a diet containing 32% FM that was fed with or without phytase at 500 phytase units (FTU/kg and a P-free diet in a completely randomized design to give 6 replicates per diet. The experimental period lasted 12 d including first 7 d for adaptation and 5 d for total collection of feces. Pigs were fed their assigned diets at 4% of BW at the beginning of the experiment. The daily feed allowance was offered in 2 equal portions at 0800 and 1600 h. All experimental diets were provided in mash form. Results indicated that pigs fed the diets containing FM with dietary phytase had less ( phytase supplementation. Also, phytase supplementation increased ( phytase supplementation had less ( phytase supplementation. Also, daily P output was reduced ( = 0.08) when pigs were fed the FM diets with phytase compared to those fed the FM diets without phytase. The ATTD of P in FM diets was increased ( phytase supplementation. The basal EPL was calculated at 164 ± 19 mg/kg of DMI in finishing pigs fed the P-free diet. In conclusion, the ATTD and STTD of P in FM fed to growing pigs were 37.3% and 43.2%, respectively, whereas respective values for finishing pigs were 31.4%, and 37.8%, respectively. Also, dietary phytase supplementation improved both ATTD and STTD of P in FM for both stages of pigs by an average of 33%.

  9. Phytase supplementation improved growth performance and bone characteristics in broilers fed varying levels of dietary calcium.

    Science.gov (United States)

    Powell, S; Bidner, T D; Southern, L L

    2011-03-01

    An experiment was conducted to investigate the effect of dietary Ca level on the efficacy of phytase. A total of 288 male Ross × Ross 708 broilers with initial and final BW of 37 and 705 g, respectively, were used in brooder batteries from 0 to 21 d posthatch. Each treatment had 8 replications with 6 broilers/replicate pen. All diets were corn-soybean meal based and formulated to contain 1.26% total Lys. The treatments were positive control with 0.45% nonphytate P and 1% Ca and a negative control with 0.20% nonphytate P with 0.67, 1.00, or 1.33% Ca fed with or without 500 phytase units of Optiphos (Escherichia coli-derived phytase; JBS United Inc., Sheridan, IN). Increasing Ca from 0.67 to 1.33% linearly decreased (P ≤ 0.003) ADG, ADFI, bone breaking strength, bone weight, tibia ash weight, and percentage tibia ash; however, quadratic effects were found for ADFI, G:F, percentage tibia ash, and mortality (P ≤ 0.09). Phytase supplementation increased (P ash weight, and percentage tibia ash and decreased (P = 0.054) mortality. The increase in ADG, ADFI, bone weight, ash weight, and percentage tibia ash (P ≤ 0.026) and decrease in mortality (phytase × Ca linear; P = 0.058) from phytase supplementation was greater in broilers fed the higher levels of Ca. Calcium utilization was linearly decreased (P < 0.002) with increasing Ca. Phosphorus digestibility and utilization were increased with increasing levels of Ca (P ≤ 0.002); however, P utilization decreased at 1% Ca and increased at 1.33% (quadratic; P < 0.070). Phytase supplementation increased Ca utilization (P < 0.024), P digestibility (P < 0.001), and P utilization (P < 0.029). However, the increase in P digestibility (phytase × Ca; P < 0.021) was greater at the lower levels of Ca whereas P utilization (phytase × Ca; P < 0.001) was greater at 1.33% Ca with phytase supplementation. The results of this research indicate that dietary Ca level, within the ranges used in this experiment, does not negatively

  10. 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

  11. INFLUENCE OF DIETARY PHYTIC ACID AND PHYTASE ON GROWTH, DIGESTIBILITY, AND VERTEBRAL PHOSPHORUS OF JUVENILE JAPANESE FLOUNDER, Paralichthys olivaceus

    Directory of Open Access Journals (Sweden)

    Asda Laining

    2013-06-01

    Full Text Available Triplicate groups of juvenile Japanese flounder (0.56 g were fed with six experimental diets to examine the effects of phytic acid, with or without phytase on growth performances, nutrient digestibility, and vertebral phosphorus (P content. Diet without both phytic acid (PA and phytase supplementation was used as control. One diet was added with 10 g PA/kg without phytase supplement. Four diets were formulated to contain two levels of phytase (1,000 FTU or 2,000 FTU phytase/kg diet combined with 2 levels of PA (10 and 20 g/kg diet. All diets were added with 10 g/kg in-organic P to meet flounder requirement. After 50 days culture, fish fed 10PA/2,000P grew significantly (P0.05 effects on feed intake and FCR. However, fish fed 10 g PA/kg combined with phytase had significant (P<0.05 higher whole body lipid, ash, and P than other groups. Moreover, P digestibility and vertebral P content were significantly increased by dietary phytase. This finding suggested that dietary phytase had potential to enhance the growth and nutrient utilization in juvenile Japanese flounder fed diet containing phytic acid. Specifically, inclusion of 2,000 FTU phytase/kg diet gave better performances when diet containing PA at level of 10 g/kg diet.

  12. Computational-based structural, functional and phylogenetic analysis of Enterobacter phytases.

    Science.gov (United States)

    Pramanik, Krishnendu; Kundu, Shreyasi; Banerjee, Sandipan; Ghosh, Pallab Kumar; Maiti, Tushar Kanti

    2018-06-01

    Myo-inositol hexakisphosphate phosphohydrolases (i.e., phytases) are known to be a very important enzyme responsible for solubilization of insoluble phosphates. In the present study, Enterobacter phytases have characterized by different phylogenetic, structural and functional parameters using some standard bio-computational tools. Results showed that majority of the Enterobacter phytases are acidic in nature as most of the isoelectric points were under 7.0. The aliphatic indices predicted for the selected proteins were below 40 indicating their thermostable nature. The average molecular weight of the proteins was 48 kDa. The lower values of GRAVY of the said proteins implied that they have better interactions with water. Secondary structure prediction revealed that alpha-helical content was highest among the other forms such as sheets, coils, etc. Moreover, the predicted 3D structure of Enterobacter phytases divulged that the proteins consisted of four monomeric polypeptide chains i.e., it was a tetrameric protein. The predicted tertiary model of E. aerogenes (A0A0M3HCJ2) was deposited in Protein Model Database (Acc. No.: PM0080561) for further utilization after a thorough quality check from QMEAN and SAVES server. Functional analysis supported their classification as histidine acid phosphatases. Besides, multiple sequence alignment revealed that "DG-DP-LG" was the most highly conserved residues within the Enterobacter phytases. Thus, the present study will be useful in selecting suitable phytase-producing microbe exclusively for using in the animal food industry as a food additive.

  13. Citrobacter farmeri phas32, an isolate from bean (Phaseolus vulgaris farm soil with high phytase production

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    Maryam Ebrahimian

    2017-12-01

    Full Text Available Introduction: Phytase hydrolyzes phytic acid and enhances bioavailability of phosphorus and other nutritive minerals for monogastric animals, so it is commonly used as an important food additive. Materials and methods: The aim of this study was isolation of phytase producing bacteria from one of Shushtar's bean farms, Southwest of Iran by phytase screening medium (PSM and optimization of the growth and enzyme productive conditions by the best isolate. Results: The best isolate was identified as Citrobacter farmeri strain phas32. Optimized conditions for phytase production by this isolate were 30˚C, pH 7, 0.25% phytic acid and 48 h incubation and phytase enzyme of phas32 had the best activity at 65˚C and pH 8.5. Enzyme unit and its molecular weight were 31 U/ml and 40 KD, respectively. Discussion and conclusion: Finally, based on these results it can be concluded that the Citrobacter farmeri strain phas32 is potent phytase producer that can be used for large scale enzyme production.

  14. Manipulation of factors affecting phytate hydrolysis in enhancing phytase efficacy in poultry: A review

    Directory of Open Access Journals (Sweden)

    Noraini*, S.

    2017-06-01

    Full Text Available Phosphorus in phytate is largely unavailable to chickens unless they are provided with dietary phytase. Phytase was shown to increase phytate degradation in the crop and proventriculus-gizzard and very little phytate degradation occurred in the duodenum-jejunum or ileum. These previous investigations were conducted on chickens fed corn based diet but not with wheat based diet. Increase in digesta passage or mean retention time (MRT along the gastrointestinal tract could enhance phytase efficacy as the prolonged reaction time between substrates and phytase may further facilitate phytate dephosphorylation. Dietary fat and fibre supplementation have been shown to influence intestinal MRT in chickens therefore it is expected that inclusion of both dietary fat and fibre could be manipulated to further improve phytase efficacy in broiler chickens. This paper provides a brief review of in vitro phytate hydrolysis, phytate hydrolysis in the gastrointestinal tract of broilers and factors that affect phytate hydrolysis that can be manipulated to enhance the efficacy of phytase in poultry diets.

  15. Phosphate Uptake from Phytate Due to Hyphae-Mediated Phytase Activity by Arbuscular Mycorrhizal Maize.

    Science.gov (United States)

    Wang, Xin-Xin; Hoffland, Ellis; Feng, Gu; Kuyper, Thomas W

    2017-01-01

    Phytate is the most abundant form of soil organic phosphorus (P). Increased P nutrition of arbuscular mycorrhizal plants derived from phytate has been repeatedly reported. Earlier studies assessed acid phosphatase rather than phytase as an indication of mycorrhizal fungi-mediated phytate use. We investigated the effect of mycorrhizal hyphae-mediated phytase activity on P uptake by maize. Two maize ( Zea mays L.) cultivars, non-inoculated or inoculated with the arbuscular mycorrhizal fungi Funneliformis mosseae or Claroideoglomus etunicatum , were grown for 45 days in two-compartment rhizoboxes, containing a root compartment and a hyphal compartment. The soil in the hyphal compartment was supplemented with 20, 100, and 200 mg P kg -1 soil as calcium phytate. We measured activity of phytase and acid phosphatase in the hyphal compartment, hyphal length density, P uptake, and plant biomass. Our results showed: (1) phytate addition increased phytase and acid phosphatase activity, and resulted in larger P uptake and plant biomass; (2) increases in P uptake and biomass were correlated with phytase activity but not with acid phosphatase activity; (3) lower phytate addition rate increased, but higher addition rate decreased hyphal length density. We conclude that P from phytate can be taken up by arbuscular mycorrhizal plants and that phytase plays a more important role in mineralizing phytate than acid phosphatase.

  16. Molecular Characterization and Comparative Phylogenetic Analysis of Phytases from Fungi with Their Prospective Applications

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    Sharad Tiwari

    2013-01-01

    Full Text Available Plant seeds that have high phytate content are used as animal feed. Phytases, enzymes that catalyze the breakdown of phytate into inorganic phosphorus and myoinositol phosphate derivatives, have been intensively studied in recent years and gained immense attention because of their application in reducing phytate content in animal feed and food for human consumption, thus indirectly lowering environmental pollution caused by undigested phytate. This review is focused on summarising the current knowledge on recent developments of fungal and yeast phytases. Comparative account on diverse sources and physiological roles, molecular characteristics and regulation mechanisms of phytases are discussed. Phylogenetic relationship of phytases from different classes of fungi is studied in details. It is inferred on the basis of phylogeny that phytases from Ascomycetes and Basidiomycetes differ in the amino acid sequences, therefore they fall in separate clade in the tree. The prospective biotechnological applications of microbial phytases such as animal feed additives, probiotics, pharmaceuticals, as well as in aquaculture, food industry, paper manufacturing, development of transgenic plants and animals with special reference to its use as biofertilizers are also emphasised in this review.

  17. 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 ...

  18. 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 ...

  19. 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. ©2014 Poultry Science Association Inc.

  20. Phosphorus utilization response of pigs and broiler chickens to diets supplemented with antimicrobials and phytase

    Directory of Open Access Journals (Sweden)

    Katherine McCormick

    2017-03-01

    Full Text Available Three experiments were conducted to evaluate the phosphorus (P utilization responses of pigs and broiler chickens to dietary supplementation with antimicrobials and phytase and to determine if P digestibility response to phytase is affected by supplementation with antimicrobials. Experiment 1 used 4 diets (a basal negative control formulated to contain 0.41% total P and 0.71% calcium [Ca] without added antimicrobials, basal negative control with added carbadox, basal negative control with added tylosin, or basal negative control with added virginiamycin and six 18-kg barrows in individual metabolism crates per diet. There was no effect of antimicrobials on P and Ca digestibility or retention. Carbadox supplementation increased (P < 0.05 digestibility and retention of gross energy (GE and supplementation with tylosin increased (P < 0.05 N retention relative to the basal negative control diet. Experiment 2 used eight 19-kg barrows in individual metabolism crates per treatment and 9 dietary treatments arranged in a 3 × 3 factorial of antimicrobials (none, tylosin, or virginiamycin and phytase (0, 500, or 1,500 FTU/kg. Phytase addition to the diets linearly increased (P < 0.05 apparent total tract digestibility or retention of P, Ca, nitrogen (N and GE. Supplementation with antimicrobials did not affect apparent total tract digestibility or retention of P, Ca, N or GE. There were linear effects (P < 0.01 of phytase on Ca utilization in diets that were not supplemented with antimicrobials but only tendencies (P < 0.10 in diets supplemented with tylosin or virginiamycin. Phytase linearly improved (P < 0.05 N utilization in diets supplemented with tylosin or virginiamycin but not in diets without added antimicrobials. Experiment 3 was a broiler chicken experiment with the same experimental design as Exp. 2 but feeding 8 birds per cage and 10 replicate cages per diet. Antimicrobial supplementation improved (P < 0.05 feed efficiency and

  1. 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.

  2. 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. © 2015 Poultry Science Association Inc.

  3. 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.

  4. Effect of Exogenous Phytase Addition on Soil Phosphatase Activities: a Fluorescence Spectroscopy Study.

    Science.gov (United States)

    Yang, Xiao-zhu; Chen, Zhen-hua; Zhang, Yu-lan; Chen, Li-jun

    2015-05-01

    The utilization of organic phosphorus (P) has directly or indirectly improved after exogenous phytase was added to soil. However, the mechanism by which exogenous phytase affected the soil phosphatases (phosphomonoesterase and phosphodiesterase) activities was not clear. The present work was aimed to study red soil, brown soil and cinnamon soil phosphomonoesterase (acid and alkaline) (AcP and AlP) and phosphodiesterase (PD) activities responding to the addition of exogenous phytase (1 g phytase/50 g air dry soil sample) based on the measurements performed via a fluorescence detection method combined with 96 microplates using a TECAN Infinite 200 Multi-Mode Microplate Reader. The results indicated that the acid phosphomonoesterase activity was significantly enhanced in red soil (p≤0. 01), while it was significantly reduced in cinnamon soil; alkaline phosphomonoesterase activity was significantly enhanced in cinnamon soil (p≤ 0. 01), while it was significantly reduced in red soil; phosphodiesterase activity was increased in three soils but it was significantly increased in brown soil (p≤0. 01) after the addition of exogenous phytase. The activities still remained strong after eight days in different soils, which indicated that exogenous phytase addition could be enhance soil phosphatases activities effectively. This effect was not only related to soil properties, such as pH and phosphorus forms, but might also be related to the excreted enzyme amount of the stimulating microorganism. Using fluorescence spectroscopy to study exogenous phytase addition influence on soil phosphatase activities was the first time at home and abroad. Compared with the conventional spectrophotometric method, the fluorescence microplate method is an accurate, fast and simple to use method to determine the relationships among the soil phosphatases activities.

  5. 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.

  6. 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

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

    Science.gov (United States)

    Holme, Inger Baeksted; Dionisio, Giuseppe; Madsen, Claus Krogh; Brinch-Pedersen, Henrik

    2017-04-01

    The phytase purple acid phosphatase (HvPAPhy_a) expressed during barley seed development was evaluated as transgene for overexpression in barley. The phytase was expressed constitutively driven by the cauliflower mosaic virus 35S-promoter, and the phytase activity was measured in the mature grains, the green leaves and in the dry mature vegetative plant parts left after harvest of the grains. The T 2 -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 activities were increased significantly by 110-fold (52 000 FTU/kg) and 57-fold (51 000 FTU/kg), respectively. The HvPAPhy_a-transformed barley plants with high phytase activities possess triple potential utilities for the improvement of phosphate bioavailability. First of all, the utilization of the mature grains as feed to increase the release of bio-available phosphate and minerals bound to the phytate of the grains; secondly, the utilization of the powdered straw either directly or phytase extracted hereof as a supplement to high phytate feed or food; and finally, the use of the stubble to be ploughed into the soil for mobilizing phytate-bound phosphate for plant growth. © 2016 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

  8. 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.

  9. 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

  10. 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...

  11. Heterologous Expression of Secreted Bacterial BPP and HAP Phytases in Plants Stimulates Arabidopsis thaliana Growth on Phytate

    Directory of Open Access Journals (Sweden)

    Lia R. Valeeva

    2018-02-01

    Full Text Available Phytases are specialized phosphatases capable of releasing inorganic phosphate from myo-inositol hexakisphosphate (phytate, which is highly abundant in many soils. As inorganic phosphorus reserves decrease over time in many agricultural soils, genetic manipulation of plants to enable secretion of potent phytases into the rhizosphere has been proposed as a promising approach to improve plant phosphorus nutrition. Several families of biotechnologically important phytases have been discovered and characterized, but little data are available on which phytase families can offer the most benefits toward improving plant phosphorus intake. We have developed transgenic Arabidopsis thaliana plants expressing bacterial phytases PaPhyC (HAP family of phytases and 168phyA (BPP family under the control of root-specific inducible promoter Pht1;2. The effects of each phytase expression on growth, morphology and inorganic phosphorus accumulation in plants grown on phytate hydroponically or in perlite as the only source of phosphorus were investigated. The most enzymatic activity for both phytases was detected in cell wall-bound fractions of roots, indicating that these enzymes were efficiently secreted. Expression of both bacterial phytases in roots improved plant growth on phytate and resulted in larger rosette leaf area and diameter, higher phosphorus content and increased shoot dry weight, implying that these plants were indeed capable of utilizing phytate as the source of phosphorus for growth and development. When grown on phytate the HAP-type phytase outperformed its BPP-type counterpart for plant biomass production, though this effect was only observed in hydroponic conditions and not in perlite. Furthermore, we found no evidence of adverse side effects of microbial phytase expression in A. thaliana on plant physiology and seed germination. Our data highlight important functional differences between these members of bacterial phytase families and indicate

  12. Heterologous Expression of Secreted Bacterial BPP and HAP Phytases in Plants Stimulates Arabidopsis thaliana Growth on Phytate.

    Science.gov (United States)

    Valeeva, Lia R; Nyamsuren, Chuluuntsetseg; Sharipova, Margarita R; Shakirov, Eugene V

    2018-01-01

    Phytases are specialized phosphatases capable of releasing inorganic phosphate from myo -inositol hexakisphosphate (phytate), which is highly abundant in many soils. As inorganic phosphorus reserves decrease over time in many agricultural soils, genetic manipulation of plants to enable secretion of potent phytases into the rhizosphere has been proposed as a promising approach to improve plant phosphorus nutrition. Several families of biotechnologically important phytases have been discovered and characterized, but little data are available on which phytase families can offer the most benefits toward improving plant phosphorus intake. We have developed transgenic Arabidopsis thaliana plants expressing bacterial phytases PaPhyC (HAP family of phytases) and 168phyA (BPP family) under the control of root-specific inducible promoter Pht1;2 . The effects of each phytase expression on growth, morphology and inorganic phosphorus accumulation in plants grown on phytate hydroponically or in perlite as the only source of phosphorus were investigated. The most enzymatic activity for both phytases was detected in cell wall-bound fractions of roots, indicating that these enzymes were efficiently secreted. Expression of both bacterial phytases in roots improved plant growth on phytate and resulted in larger rosette leaf area and diameter, higher phosphorus content and increased shoot dry weight, implying that these plants were indeed capable of utilizing phytate as the source of phosphorus for growth and development. When grown on phytate the HAP-type phytase outperformed its BPP-type counterpart for plant biomass production, though this effect was only observed in hydroponic conditions and not in perlite. Furthermore, we found no evidence of adverse side effects of microbial phytase expression in A. thaliana on plant physiology and seed germination. Our data highlight important functional differences between these members of bacterial phytase families and indicate that future

  13. Heterologous Expression of Secreted Bacterial BPP and HAP Phytases in Plants Stimulates Arabidopsis thaliana Growth on Phytate

    Science.gov (United States)

    Valeeva, Lia R.; Nyamsuren, Chuluuntsetseg; Sharipova, Margarita R.; Shakirov, Eugene V.

    2018-01-01

    Phytases are specialized phosphatases capable of releasing inorganic phosphate from myo-inositol hexakisphosphate (phytate), which is highly abundant in many soils. As inorganic phosphorus reserves decrease over time in many agricultural soils, genetic manipulation of plants to enable secretion of potent phytases into the rhizosphere has been proposed as a promising approach to improve plant phosphorus nutrition. Several families of biotechnologically important phytases have been discovered and characterized, but little data are available on which phytase families can offer the most benefits toward improving plant phosphorus intake. We have developed transgenic Arabidopsis thaliana plants expressing bacterial phytases PaPhyC (HAP family of phytases) and 168phyA (BPP family) under the control of root-specific inducible promoter Pht1;2. The effects of each phytase expression on growth, morphology and inorganic phosphorus accumulation in plants grown on phytate hydroponically or in perlite as the only source of phosphorus were investigated. The most enzymatic activity for both phytases was detected in cell wall-bound fractions of roots, indicating that these enzymes were efficiently secreted. Expression of both bacterial phytases in roots improved plant growth on phytate and resulted in larger rosette leaf area and diameter, higher phosphorus content and increased shoot dry weight, implying that these plants were indeed capable of utilizing phytate as the source of phosphorus for growth and development. When grown on phytate the HAP-type phytase outperformed its BPP-type counterpart for plant biomass production, though this effect was only observed in hydroponic conditions and not in perlite. Furthermore, we found no evidence of adverse side effects of microbial phytase expression in A. thaliana on plant physiology and seed germination. Our data highlight important functional differences between these members of bacterial phytase families and indicate that future crop

  14. 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.

  15. Interaction of phytase RONOZYME?P(L and citric acid on the utilization of phosphorus by common carp (Cyprinus carpio

    Directory of Open Access Journals (Sweden)

    Jacques Gabaudan

    2010-12-01

    Full Text Available A feeding trial was conducted for 60 days to study the effects of the combination of microbial phytase and citric acidon phosphorus utilization in Cyprinus carpio fingerlings. Four diets designated as diet without phytase or inorganic phosphorussupplementation (T1, with 1.1% MSP (T2, with 0.55% MSP and phytase (T3 and with 0.55% MSP, phytase andcitric acid (T4. Four replicate groups of 20 fish were fed two times daily until satiation. Phosphorus digestibility and retentionwere measured as well as the fish growth performance. It is concluded from the results that the addition of microbial phytaseand citric acid enhances the availability of phosphorus from plant sources, improves bone mineralization, growth and feedefficiency. Combining a low dose of citric acid to the phytase significantly increased the positive effects of the enzyme.

  16. 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

  17. Activity of Escherichia coli, Aspergillus niger, and Rye Phytase toward Partially Phosphorylated myo-Inositol Phosphates.

    Science.gov (United States)

    Greiner, Ralf

    2017-11-08

    Kinetic parameters for the dephosphorylation of sodium phytate and a series of partially phosphorylated myo-inositol phosphates were determined at pH 3.0 and pH 5.0 for three phytase preparations (Aspergillus niger, Escherichia coli, rye). The enzymes showed lower affinity and turnover numbers at pH 3 compared to pH 5 toward all myo-inositol phosphates included in the study. The number and distribution of phosphate groups on the myo-inositol ring affected the kinetic parameters. Representatives of the individual phytate dephosphorylation pathways were identified as the best substrates of the phytases. Within the individual phytate dephosphorylation pathways, the pentakisphosphates were better substrates compared to the tetrakisphosphates or phytate itself. E. coli and rye phytase showed comparable activities at both pH values toward the tetrakis- and trisphosphate, whereas A. niger phytase exhibited a higher activity toward the tetrakisphosphate. A myo-inositol phosphate with alternate phosphate groups was shown to be not significantly dephosphorylated by the phytases.

  18. Digestibility, Determination of Metabolizable Energy and Bone Mineralization of Broilers Fed with Nutritionally Valued Phytase

    Directory of Open Access Journals (Sweden)

    FH Litz

    Full Text Available ABSTRACT The objective of this study was to evaluate the effect of using exoenzyme phytase in broiler's diets on digestibility of nutrients, feed energy and tibia bone mineralization. A completely randomized design was used, with the following treatments: sorghum with dicalcium phosphate (SDP, corn with dicalcium phosphate (CDP, sorghum with meat and bone meal (SMBM, sorghum with valued phytase (SVP and sorghum with phytase without valued (SPWV. For digestibility analysis, eighty 15 day old broilers were used, a total of 1400 male Hubbard Flex chickens, which were submitted to total excreta collection to obtain the percentages of food digestibility, crude protein, ether extract, apparent metabolizable energy, calcium and phosphorus while for tibias mineralization. Six birds per treatment were used, where determination of mineral matter, calcium and phosphorus were performed. Metabolizable energy (ME and apparent metabolizable energy corrected for nitrogen (AMEn of the feed were also calculated. Data were subjected to variation analysis and the average compared by 5% Tukey test. There was no difference between treatments for the digestibility at 15-20 day old as well as for the feed energy values, but the diets with phytase had higher phosphorous percentage values for tibia bone mineralization, demonstrating that exogenous phytase enzyme is able to hydrolyze phytate origininated from plant and release the phosphorus for assimilation by animals, acting as a substitute for phosphorus plant sources.

  19. 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.

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

    Science.gov (United States)

    Nielsen, Anne V. F.; 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-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. PMID:23917170

  1. Purification and characterization of a novel neutral and heat-tolerant phytase from a newly isolated strain Bacillus nealsonii ZJ0702

    Science.gov (United States)

    2013-01-01

    Background Phytic acid and phytates can interact with biomolecules, such as proteins and carbohydrates, and are anti-nutritional factors found in food and feed. Therefore, it is necessary to remove these compounds in food and feed processing. Phytase can hydrolyze phytic acid and phytates to release a series of lower phosphate esters of myoinositol and orthophosphate. Thus, the purification and characterization of novel phytases that can be used in food and feed processing is of particular interest to the food and feed industries. Results A novel neutral and heat-tolerant phytase from a newly isolated strain Bacillus nealsonii ZJ0702 was purified to homogeneity with a yield of 5.7% and a purification fold of 44. The molecular weight of the purified phytase obtained by SDS-PAGE was 43 kDa. The homology analysis based on N-terminal amino acid and DNA sequencing indicated that the purified phytase was different from other known phytases. The optimal thermal and pH activity of the phytase was observed at 55°C and 7.5, respectively. Seventy-three percent of the original activity of the phytase was maintained following incubation at 90°C for 10 min. The phytase was stable within a pH range of 6.0 − 8.0 and showed high substrate specificity for sodium phytate. Cu2+, Co2+, Zn2+, Mn2+, Ba2+ and Ni2+ ions were found to inhibit the activity of the phytase. Conclusions A novel phytase purified from B. nealsonii ZJ0702 was identified. The phytase was found to be thermally stable over a wide temperature range at neutral pH. These properties suggest that this phytase is a suitable alternative to fungal phytases for the hydrolysis of phytic acid and phytates in food and feed processing industries. PMID:24073799

  2. 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...... 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....

  3. Uranium incorporation biokinetics in poultry bones as function of phytase doses

    International Nuclear Information System (INIS)

    Arruda-Neto, J.D.T.; Cestari, A.C.; Zamboni, C.B.; Saiki, M.; Nogueira, G.P.; Fonseca, L.E.C.; Manso-Guevara, M.V.; Deppman, A.; Likhachev, V.P.; Mesa, J.

    2005-01-01

    Neutron activation analysis has been used to study uranium incorporation in poultry bones as function of chow doped with: (a) uranium (20 ppm); (b) U-doped food (20 ppm) plus phytase (120 ppm) and (c) U-doped food (20 ppm) plus phytase (180 ppm). To investigate this situation experiments involving several groups of Cobb broilers was performed. Two animals per group were sacrificed weekly up to their adultness and uranium concentration in the tibia was measured. It was observed that the concentration of uranium (μg U/g bone) is decreasing all along the animal life spanning period of 14-42 days. This behavior suggests that the skeleton mass is growing faster than the corresponding accumulation of uranium. The administration of phytase seems not to alter this scenario. (author)

  4. Phytase production by Aspergillus niger NCIM 563 for a novel application to degrade organophosphorus pesticides.

    Science.gov (United States)

    Shah, Parin C; Kumar, V Ravi; Dastager, Syed G; Khire, Jayant M

    2017-12-01

    The production of phytase using Aspergillus niger NCIM 563 under submerged fermentation conditions was studied using protein rich chickpea flour as substrate. Employing a hybrid statistical media optimization strategy of Plackett-Burman and Box-Behnken experimental designs in shake-flasks gave an increased phytase activity from an initial 66 IU/mL in 216 h to 160 IU/mL in a reduced time of 132 h. Productivity, thus increased by 3.97 times from 7.3 to 29 IU/mL/day. Using the optimized media, the production was successfully scaled-up further and improved up to 164 IU/mL in 96 h by studies carried out employing 2 and 10-L fermenters. The enzyme supernatant was recovered using centrifugal separation of biomass and the stability of the produced phytase was tested for animal feed applications under gastric conditions. In vitro degradation studies of water soluble monocrotophos, methyl parathion and water insoluble chlorpyrifos, pesticides used extensively in agriculture was carried out. It was observed by HPLC analysis that phytase could degrade 72% of chlorpyrifos at pH 7.0, 35 °C. Comparable results were obtained with monocrotophos and methyl parathion. With chlorpyrifos at higher temperature 50 °C as much as 91% degradation could be obtained. The degradation of chlorpyrifos was further validated by spraying phytase on harvested green chilli (Capsicum annuum L) under normal conditions of pH 7.0, 35 °C and the degradation products obtained analyzed by LCMS. Thus, the present study brings out a potentially novel application of phytase for biodegradation of organophosphorus pesticides.

  5. Xylanase, protease and superdosing phytase interactions in broiler performance, carcass yield and digesta transit time

    Directory of Open Access Journals (Sweden)

    Tiago T. dos Santos

    2017-06-01

    Full Text Available The interaction of xylanase, protease and superdosing (1,500 FTU/kg phytase in a 2 × 2 × 2 factorial arrangement was studied in broilers fed sorghum-based diets. A total of 2,800 one-day-old unsexed Ross 308 chicks were housed in 56 pens with 50 birds per pen, with or without inclusion of xylanase, protease and phytase, totaling 8 treatments and 7 replicates per treatment. Body weight (BW and feed intake (FI were measured at 21 and 42 days of age, and mortality corrected feed conversion ratio (FCR was calculated for each period and cumulatively. Tibia ash and carcass yield were determined in 2 birds per replicate at 21 and 42 days of age, respectively. Digesta transit time was determined at 21, 28, 35 and 42 days of age using 5 birds per replicate. Results showed that superdosing phytase increased BW and FI at 42 days of age (P < 0.05 and xylanase improved FCR (P < 0.05. Xylanase and phytase also positively influenced carcass yield and breast weight, respectively. Overall, inclusion of superdosing phytase increased transit time when included in a diet containing xylanase, and no change with protease inclusion. In conclusion, the beneficial effects of xylanase, protease and superdosing phytase in broiler performance were not additive. This limitation is likely not related to the lack of efficacy of any one of the individual enzymes but to a limitation of the bird to respond additively to successive additions of enzymes.

  6. Tibia mineralization of chickens determined to meat production using a microbial phytase

    Directory of Open Access Journals (Sweden)

    Mária Angelovičová

    2018-02-01

    Full Text Available The target of the research was 6-phytase of microbial origin. It was used in feed mixtures for chickens determined to meat production. Its effect has been studied in relation to the tibia mineralization by calcium, phosphorus and magnesium. 6-phytase is a product of Aspergillus oryzae. That was obtained by means of biotechnological processes of production of commercially available enzymes. It was incorporated in the feed mixtures 0.1%. In a 38-day feeding trial, 300 one-day-old, as hatched, Cobb 500 chickens determined to meat production (100 birds per group were fed on one concentrations of dietary non-phytate phosphorus (2.32, 2.31 g.kg-1, respectively and supplemental microbial phytase (0 and 500   FTU.kg-1 feed mixtures. Control group was used to compare the results and control feed mixtures contained 4.5 g.kg-1 without microbial phytase. At days 40 it was selected 6 birds in every group, which were slaughter in accordance with the principles of welfare. Left tibias of every bird were used to determination of calcium, phosphorus and magnesium contents. According to in vivo, it was found that the addition of microbial phytase to reduced dietary non-phytate phosphorus increased concentrations of calcium (Ca, phosphorus (P and magnesium (Mg in tibia. The differences among groups were statistically significant (p <0.05. It was concluded that reducing of dietary non-phytate phosphorus on the 2.32, 2.31 g.kg-1, respectively, by monocalcium phosphate and microbial phytase supplementation in feed mixtures facilitated tibia mineralization at chicken determined to meat production. Normal 0 21 false false false EN-GB X-NONE X-NONE

  7. Fabrication of phytic acid sensor based on mixed phytase-lipid Langmuir-Blodgett films.

    Science.gov (United States)

    Caseli, Luciano; Moraes, Marli L; Zucolotto, Valtencir; Ferreira, Marystela; Nobre, Thatyane M; Zaniquelli, Maria Elisabete D; Rodrigues Filho, Ubirajara P; Oliveira, Osvaldo N

    2006-09-26

    This paper reports the surface activity of phytase at the air-water interface, its interaction with lipid monolayers, and the construction of a new phytic acid biosensor on the basis of the Langmuir-Blodgett (LB) technique. Phytase was inserted in the subphase solution of dipalmitoylphosphatidylglycerol (DPPG) Langmuir monolayers, and its incorporation to the air-water interface was monitored with surface pressure measurements. Phytase was able to incorporate into DPPG monolayers even at high surface pressures, ca. 30 mN/m, under controlled ionic strength, pH, and temperature. Mixed Langmuir monolayers of phytase and DPPG were characterized by surface pressure-area and surface potential-area isotherms, and the presence of the enzyme provided an expansion in the monolayers (when compared to the pure lipid at the interface). The enzyme incorporation also led to significant changes in the equilibrium surface compressibility (in-plane elasticity), especially in liquid-expanded and liquid-condensed regions. The dynamic surface elasticity for phytase-containing interfaces was investigated using harmonic oscillation and axisymmetric drop shape analysis. The insertion of the enzyme at DPPG monolayers caused an increase in the dynamic surface elasticity at 30 mN m(-)(1), indicating a strong interaction between the enzyme and lipid molecules at a high-surface packing. Langmuir-Blodgett (LB) films containing 35 layers of mixed phytase-DPPG were characterized by ultraviolet-visible and fluorescence spectroscopy and crystal quartz microbalance nanogravimetry. The ability in detecting phytic acid was studied with voltammetric measurements.

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

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    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

  9. 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. © 2015 Poultry Science Association Inc.

  10. A Novel Phytase Derived from an Acidic Peat-Soil Microbiome Showing High Stability under Acidic Plus Pepsin Conditions.

    Science.gov (United States)

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

    2016-01-01

    Four novel phytases of the histidine acid phosphatase family were identified in two publicly available metagenomic datasets of an acidic peat-soil microbiome in northeastern Bavaria, Germany. These enzymes have low similarity to all the reported phytases. They were overexpressed in Escherichia coli and purified. Catalytic efficacy in simulated gastric fluid was measured and compared among the four candidates. The phytase named rPhyPt4 was selected for its high activity. It is the first phytase identified from unculturable Acidobacteria. The phytase showed a longer half-life than all the gastric-stable phytases that have been reported to date, suggesting a strong resistance to low pH and pepsin. A wide pH profile was observed between pH 1.5 and 5.0. At the optimum pH (2.5) the activity was 2,790 μmol/min/mg at the physiological temperature of 37°C and 3,989 μmol/min/mg at the optimum temperature of 60°C. Due to the competent activity level as well as the high gastric stability, the phytase could be a potential candidate for practical use in livestock and poultry feeding. © 2016 S. Karger AG, Basel.

  11. 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.

  12. 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. ©2014 Poultry Science Association Inc.

  13. Genetics Home Reference: beta-propeller protein-associated neurodegeneration

    Science.gov (United States)

    ... signs and symptoms, affected individuals can live into middle age. Death may result from complications of dementia or ... What is precision medicine? What is newborn screening? New Pages RAB18 deficiency Depression Pelizaeus-Merzbacher-like disease ...

  14. 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

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

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    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.

  16. 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.

  17. 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...

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

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    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.

  19. Phytase inclusion in pig diets improves zinc status but its effect on copper availability is inconsistent

    NARCIS (Netherlands)

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

    2012-01-01

    Complexation of dietary phytate with cations is a major cause of reduced bioavailability of Zn and possibly Cu in pig diets. We conducted 2 studies with 2 treatments in young growing pigs (8 to 40 kg) to estimate potential contributions of phytase to availability and supply of Zn and Cu,

  20. Phosphate uptake from phytate due to hyphae-mediated phytase activity by arbuscular mycorrhizal maize

    NARCIS (Netherlands)

    Wang, Xinxin; Hoffland, Ellis; Feng, Gu; Kuijper, Thomas

    2017-01-01

    Phytate is the most abundant form of soil organic phosphorus (P). Increased P nutrition of arbuscular mycorrhizal plants derived from phytate has been repeatedly reported. Earlier studies assessed acid phosphatase rather than phytase as an indication of mycorrhizal fungi-mediated phytate use. We

  1. Dose-dependent effects of a microbial phytase on phosphorus digestibility of common feedstuffs in pigs

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    Ferdinando N. Almeida

    2017-07-01

    Full Text Available Objective The objective of this study was to evaluate increasing doses of a novel microbial phytase (Cibenza Phytaverse, Novus International, St. Charles, MO, USA on standardized total tract digestibility (STTD of P in canola meal (CM, corn, corn-derived distiller’s dried grains with solubles (DDGS, rice bran (RB, sorghum, soybean meal (SBM, sunflower meal (SFM, and wheat. Methods Two cohorts of 36 pigs each (initial body weight = 78.5±3.7 kg were randomly assigned to 2 rooms, each housing 36 pigs, and then allotted to 6 diets with 6 replicates per diet in a randomized complete block design. Test ingredient was the only dietary source of P and diets contained 6 concentrations of phytase (0, 125, 250, 500, 1,000, or 2,000 phytase units [FTU]/kg with 0.4% of TiO2 as a digestibility marker. Feeding schedule for each ingredient was 5 d acclimation, 5 d fecal collection, and 4 d washout. The STTD of P increased (linear or exponential p≤0.001 with the inclusion of phytase for all ingredients. Results Basal STTD of P was 37.6% for CM, 37.6% for corn, 68.6% for DDGS, 10.3% for RB, 41.2% for sorghum, 36.7% for SBM, 26.2% for SFM, and 55.1% for wheat. The efficiency of this novel phytase to hydrolyze phytate is best described with a broken-line model for corn, an exponential model for CM, RB, SBM, SFM, and wheat, and a linear model for DDGS and sorghum. Based on best-fit model the phytase dose (FTU/kg needed for highest STTD of P (%, respectively, was 735 for 64.3% in CM, 550 for 69.4% in corn, 160 for 55.5% in SBM, 1,219 for 57.8% in SFM, and 881 for 64.0% in wheat, whereas a maximum response was not obtained for sorghum, DDGS and RB within the evaluated phytase range of 0 to 2,000 FTU/kg. These differences in the phytase concentration needed to maximize the STTD of P clearly indicate that the enzyme does not have the same hydrolysis efficiency among the evaluated ingredients. Conclusion Variations in enzyme efficacy to release P from phytate in

  2. 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

  3. Statistical Optimization of the Induction of Phytase Production by Arabinose in a recombinant E. coli using Response Surface Methodology

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    Abd-El Aziem Farouk

    2017-11-01

    Full Text Available The production of phytase in a recombinant E.coli using the pBAD expression  system was optimized using response surface methodology with full-factorial faced centered central composite design. The ampicilin and arabinose concentration in the cultivation media and the incubation temperature were optimized in order to maximize phytase production using 2 3  central composite experimental design. With this design the number of actual experiment performed could be reduced while allowing eludidation of possible interactions among these factors. The most significant parameter was shown to be the linear and quadratic effect of the incubation temperature.  Optimal conditions for phytase production were determined to be 100 µg/ml ampicilin, 0.2 % arabinose and an incubation temperature of 37ºC. The production of phytase in the recombinant E. coli was scaled up to 100 ml and 1000 ml.

  4. Performance, litter quality and gaseous odour emissions of broilers fed phytase supplemented diets.

    Science.gov (United States)

    Sharma, Nishchal K; Choct, Mingan; Wu, Shu-Biao; Smillie, Robert; Morgan, Natalie; Omar, Amal S; Sharma, Nisha; Swick, Robert A

    2016-12-01

    The effect of graded levels of phytase on performance, bone characteristics, excreta/litter quality and odorant emissions was examined using 720 Ross 308 male d-old broilers. A 2 × 4 factorial arrangement of treatments was employed with 6 replicates of 15 birds per pen. Factors were: diets-positive and negative control (PC, NC); phytase - 0, 500, 1,000, 1,500 FTU/kg. The PC was formulated to meet the 2014 Ross 308 nutrient specifications, whereas the NC was formulated with lower Ca (-1.4 g/kg), available P (-1.5 g/kg), Na (-0.3 g/kg), dLys (-0.2 g/kg) and MEn (-0.28 MJ/kg) equivalent to nutrient matrix values for 500 FTU/kg phytase in the starter, grower and finisher periods (i.e., downspec diet). On d 24, phytase decreased FCR by 1.6, 4.3 and 4.6 points at inclusion levels of 500, 1,000 and 1,500 FTU/kg, respectively ( P  litter quality and water to feed intake ratio ( P   0.05). Solid phase micro-extraction gas chromatography-mass spectrometry (SPME-GC-MS) analysis of gaseous emissions on d 39 indicated no difference in the emission of alcohols, aldehydes, ketones, volatile fatty acids and phenols between treatments ( P  > 0.05). The results indicate that phytase has greater benefits when formulated using nutrient matrix values as compared with adding it over the top in an already nutrient sufficient diet. The later method would be expected to increase feed costs without concomitant performance benefits.

  5. Performance, litter quality and gaseous odour emissions of broilers fed phytase supplemented diets

    Directory of Open Access Journals (Sweden)

    Nishchal K. Sharma

    2016-12-01

    Full Text Available The effect of graded levels of phytase on performance, bone characteristics, excreta/litter quality and odorant emissions was examined using 720 Ross 308 male d-old broilers. A 2 × 4 factorial arrangement of treatments was employed with 6 replicates of 15 birds per pen. Factors were: diets-positive and negative control (PC, NC; phytase – 0, 500, 1,000, 1,500 FTU/kg. The PC was formulated to meet the 2014 Ross 308 nutrient specifications, whereas the NC was formulated with lower Ca (−1.4 g/kg, available P (−1.5 g/kg, Na (−0.3 g/kg, dLys (−0.2 g/kg and MEn (−0.28 MJ/kg equivalent to nutrient matrix values for 500 FTU/kg phytase in the starter, grower and finisher periods (i.e., downspec diet. On d 24, phytase decreased FCR by 1.6, 4.3 and 4.6 points at inclusion levels of 500, 1,000 and 1,500 FTU/kg, respectively (P  0.05. Solid phase micro-extraction gas chromatography-mass spectrometry (SPME-GC-MS analysis of gaseous emissions on d 39 indicated no difference in the emission of alcohols, aldehydes, ketones, volatile fatty acids and phenols between treatments (P > 0.05. The results indicate that phytase has greater benefits when formulated using nutrient matrix values as compared with adding it over the top in an already nutrient sufficient diet. The later method would be expected to increase feed costs without concomitant performance benefits.

  6. Digestibility marker and ileal amino acid digestibility in phytase-supplemented soybean or canola meals for growing pigs.

    Science.gov (United States)

    Favero, A; Ragland, D; Vieira, S L; Owusu-Asiedu, A; Adeola, O

    2014-12-01

    Two experiments using soybean meal (SBM) or canola meal (CM) were conducted to investigate whether the choice of digestibility marker influenced the apparent ileal digestibility (AID) or standardized ileal digestibility (SID) of N and AA in diets supplemented with phytase. In each experiment, 18 barrows fitted with T-cannulas at the ileocecal junction were assigned to 3 diets consisting of a N-free diet to determine endogenous losses of N and AA, a semipurified diet (SBM in Exp. 1 or CM in Exp. 2), and the semipurified diet supplemented with phytase at 1,000 phytase units/kg. Three digestibility markers including acid-insoluble ash (AIA), chromic oxide (Cr2O3), and titanium dioxide (TiO2) were added to each diet at 3 g/kg. Each diet was fed for 7 d, consisting of a 5-d adjustment and a 2-d collection of ileal digesta. In both studies, basal ileal endogenous losses determined with Cr2O3 as a digestibility marker were lower (Pdigestibility markers. Using SBM as the protein source in Exp. 1, there was no interaction between phytase and digestibility marker on AID or SID of AA. The AID of N and AA in SBM using AIA as a digestibility marker tended to be lower (Pdigestibility markers. Phytase supplementation increased (Pdigestibility marker tended to be associated with lower (Pdigestibility marker and phytase. Phytase supplementation had effects (Pdigestibility marker. With Cr2O3 or TiO2 as the digestibility marker in the CM diets, phytase supplementation increased (Pdigestibility marker. In contrast, there were no clear improvements in AA digestibility from phytase supplementation for SBM. Phytase effects on AID or SID of AA were dependent on the digestibility marker used in diets when CM was used as the protein source but not when SBM was used as the protein source. Therefore, AA digestibility response to phytase supplementation may depend on the protein being evaluated as well as the choice of digestibility marker.

  7. P and Ca digestibility is increased in broiler diets supplemented with the high-phytase HIGHPHY wheat.

    Science.gov (United States)

    Scholey, D; Burton, E; Morgan, N; Sanni, C; Madsen, C K; Dionisio, G; Brinch-Pedersen, H

    2017-09-01

    Around 70% of total seed phosphorus is represented by phytate which must be hydrolysed to be bioavailable in non-ruminant diets. The limited endogenous phytase activity in non-ruminant animals make it common practice to add an exogenous phytase source to most poultry and pig feeds. The mature grain phytase activity (MGPA) of cereal seeds provides a route for the seeds themselves to contribute to phytate digestion, but MGPA varies considerably between species and most varieties in current use make negligible contributions. Currently, all phytases used for feed supplementation and transgenic improvement of MGPA are derived from microbial enzymes belonging to the group of histidine acid phosphatases (HAP). Cereals contain HAP phytases, but the bulk of MGPA can be attributed to phytases belonging to a completely different group of phosphatases, the purple acid phosphatases (PAPhy). In recent years, increased MGPAs were achieved in cisgenic barley holding extra copies of barley PAPhy and in the wheat HIGHPHY mutant, where MGPA was increased to ~6200 FTU/kg. In the present study, the effect of replacing 33%, 66% and 100% of a standard wheat with HIGHPHY wheat was compared with a control diet with and without 500 FTU of supplemental phytase. Diets were compared by evaluating broiler performance, ileal Ca and P digestibility and tibia development, using nine replicate pens of four birds per diet over 3 weeks from hatch. There were no differences between treatments in any tibia or bird performance parameters, indicating the control diet did not contain sufficiently low levels of phosphorus to distinguish effect of phytase addition. However, in a comparison of the two wheats, the ileal Ca and P digestibility coefficients for the 100% HIGHPHY wheat diets are 22.9% and 35.6% higher, respectively, than for the control diet, indicating the wheat PAPhy is functional in the broiler digestive tract. Furthermore, 33% HIGHPHY replacement of conventional wheat, significantly improved

  8. What aspect of dietary modification in broilers controls litter water-soluble phosphorus: dietary phosphorus, phytase, or calcium?

    Science.gov (United States)

    Leytem, A B; Plumstead, P W; Maguire, R O; Kwanyuen, P; Brake, J

    2007-01-01

    Environmental concerns about phosphorus (P) losses from animal agriculture have led to interest in dietary strategies to reduce the concentration and solubility of P in manures and litters. To address the effects of dietary available phosphorus (AvP), calcium (Ca), and phytase on P excretion in broilers, 18 dietary treatments were applied in a randomized complete block design to each of four replicate pens of 28 broilers from 18 to 42 d of age. Treatments consisted of three levels of AvP (3.5, 3.0, and 2.5 g kg(-1)) combined with three levels of Ca (8.0, 6.9, and 5.7 g kg(-1)) and two levels of phytase (0 and 600 phytase units [FTU]). Phytase was added at the expense of 1.0 g kg(-1) P from dicalcium phosphate. Fresh litter was collected from pens when the broilers were 41 d of age and analyzed for total P, soluble P, and phytate P as well as P composition by (31)P nuclear magnetic resonance (NMR) spectroscopy. Results indicated that the inclusion of phytase at the expense of inorganic P or reductions in AvP decreased litter total P by 28 to 43%. Litter water-soluble P (WSP) decreased by up to 73% with an increasing dietary Ca/AvP ratio, irrespective of phytase addition. The ratio of WSP/total P in litter decreased as the dietary Ca/AvP ratio increased and was greater in the phytase-amended diets. This study indicated that while feeding reduced AvP diets with phytase decreased litter total P, the ratio of Ca/AvP in the diet was primarily responsible for effects on WSP. This is important from an environmental perspective as the amount of WSP in litter could be related to potential for off-site P losses following land application of litter.

  9. Time-series responses of swine plasma metabolites to ingestion of diets containing myo-inositol or phytase.

    Science.gov (United States)

    Cowieson, Aaron J; Roos, Franz F; Ruckebusch, Jean-Paul; Wilson, Jonathan W; Guggenbuhl, Patrick; Lu, Hang; Ajuwon, Kolapo M; Adeola, Olayiwola

    2017-12-01

    The effect of the ingestion of diets containing either myo-inositol or exogenous phytase on plasma metabolites was examined using 29 kg barrows. The diets were: control (maize, soya, rapeseed, rice bran), control plus 2 g/kg myo-inositol, control plus 1000 phytase units (FYT)/kg or 3000 FYT/kg exogenous phytase. Pigs were housed in a PigTurn device and blood was collected, from jugular catheters, via an automated system at -30, (30 min before feeding), 0, 15, 30, 45, 60, 90, 120, 150, 180, 240, 300 and 360 min post-feeding. The addition of 2 g/kg myo-inositol to the basal diet resulted in an increase in plasma myo-inositol concentration that was evident 45-60 min after diet introduction and persisted to 360 min post-feeding. Similarly, supplementation of the basal diet with either 1000 or 3000 FYT/kg exogenous phytase resulted in an increase in plasma myo-inositol concentration that was still rising 360 min post-feeding. Plasma P concentration was increased over time by the addition of 1000 and 3000 FYT/kg phytase, but not by the addition of myo-inositol. Other plasma metabolites examined were not affected by dietary treatment. It can be concluded that oral delivery of myo-inositol results in rapid increase in plasma myo-inositol concentrations that peak approximately 45-60 min after feeding. Use of supplemental phytase achieves similar increases in myo-inositol concentration in plasma but the appearance is more gradual. Furthermore, supplementation of pig diets with exogenous phytase results in rapid appearance of P in plasma that may be sustained over time relative to diets with no added phytase.

  10. Effect of diet containing phytate and phytase on the activity and messenger ribonucleic acid expression of carbohydrase and transporter in chickens.

    Science.gov (United States)

    Liu, N; Ru, Y J; Li, F D; Cowieson, A J

    2008-12-01

    The effect of dietary phytate and phytase on carbohydrase activity and hexose transport was investigated in broiler chickens. Diets containing phytate P (2.2 or 4.4 g/kg) with different phytase dose rates (0, 500, or 1,000 phytase units/kg) were fed to 504 female Cobb chicks for 3 wk. Diets containing high phytate concentrations depressed (P diets containing phytate also decreased (P diets containing increased phytate upregulated (P competence, and phytase can ameliorate these effects for chickens.

  11. Evaluation of increasing levels of a microbial phytase in phosphorus deficient broiler diets via live broiler performance, tibia bone ash, apparent metabolizable energy, and amino acid digestibility.

    Science.gov (United States)

    Pieniazek, J; Smith, K A; Williams, M P; Manangi, M K; Vazquez-Anon, M; Solbak, A; Miller, M; Lee, J T

    2017-02-01

    The objective was to investigate increasing concentrations of an evolved microbial phytase on male broiler performance, tibia bone ash, AME, and amino acid digestibility when fed diets deficient in available phosphorus (aP). Experiment 1 evaluated the effects of phytase during a 21 d battery cage study and Experiment 2 was a 42 d grow-out. Experiment 1 included six treatments; negative control (NC) with an aP level of 0.23% (starter) and 0.19% (grower), two positive controls (PC) consisting of an additional 0.12% and 0.22% aP (PC 1 and PC 2), and the NC supplemented with three levels of phytase (250, 500, and 2,000 U/kg). The NC diet reduced (P Phytase increased (P phytase yielding similar results to the PC2, and improved FCR and increased bone ash was observed at all phytase levels. Amino acid digestibility coefficients were increased (P phytase at 250 U/kg. Phytase at all rates increased (P phytase inclusion of 250, 500, and 2000 U/kg, respectively. Experiment 2 included a PC consisting of 0.45%, 0.41%, and 0.38% aP for the starter, grower, and finisher, respectively; NC with reduced aP of 0.17%; and phytase at 500 and 2,000 U/kg. Phytase increased BW (P phytase resulted in further BW increases compared to the PC (starter and grower). Phytase improved FCR to levels comparable to the PC, with supplementation at 2,000 U/kg resulting in improvements beyond the PC in the starter phase. Amino acid digestibility coefficients were increased with phytase at 2,000 U/kg to levels comparable to that of the PC. These data confirm that the inclusion of phytase improves broiler performance and bone mineralization in aP reduced diets and levels beyond the traditional 500 U/kg can result in further improvements. © 2016 Poultry Science Association Inc.

  12. 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. Journal of Animal Physiology and Animal Nutrition © 2014 Blackwell Verlag GmbH.

  13. Optimization of phytase production by Penicillium purpurogenum GE1 under solid state fermentation by using Box-Behnken design.

    Science.gov (United States)

    Awad, Ghada E A; Helal, Mohamed M I; Danial, Enas N; Esawy, Mona A

    2014-01-01

    Phytase production by Penicillium purpurogenum GE1 isolated from soil around bean root nodules was investigated by solid state fermentation (SSF) using mixed substrates consisted of corn cob and corn bran. The SSF conditions were optimized by using one-variable-at-a-time strategy. The optimum conditions for phytase production were at 27 °C, pH 8 and 66% moisture content. The study of different carbon and nitrogen sources revealed that glucose and peptone registered the highest enzyme productivity (92 ± 5.6 U/g ds, 125 ± 4.9 U/g ds). Among different surfactants, maximum phytase productivity was observed with Tween 80 at 0.001 concentrations (170 ± 4.2 U/g ds). A Box-Behnken design was employed to investigate the optimization of the most significant variables affecting the enzyme production. Maximal phytase production was detected after the addition of (g/5 g ds): 0.75 glucose, 0.375 peptone and 0, 01 tween 80. This result represented an improvement in phytase production of 2.6 folds when compared to that previously obtained using the basal medium under the same cultivation conditions. The generated model was found to be very adequate for phytase production (90% accuracy) as the experimental value was 444 ± 3.5 U/g ds compared to 401 U/g ds for the predicted value. In brief, the production of phytase using corn cob and corn bran is a novel and cheap way for the production of this important enzyme and opens a new way for researchers to discover and explore this arena.

  14. Phosphorus acquisition by citrate- and phytase-exuding Nicotiana tabacum plant mixtures depends on soil phosphorus availability and root intermingling.

    Science.gov (United States)

    Giles, Courtney D; Richardson, Alan E; Cade-Menun, Barbara J; Mezeli, Malika M; Brown, Lawrie K; Menezes-Blackburn, Daniel; Darch, Tegan; Blackwell, Martin Sa; Shand, Charles A; Stutter, Marc I; Wendler, Renate; Cooper, Patricia; Lumsdon, David G; Wearing, Catherine; Zhang, Hao; Haygarth, Philip M; George, Timothy S

    2018-03-02

    Citrate and phytase root exudates contribute to improved phosphorus (P) acquisition efficiency in Nicotiana tabacum (tobacco) when both exudates are produced in a P deficient soil. To test the importance of root intermingling in the interaction of citrate and phytase exudates, Nicotiana tabacum plant-lines with constitutive expression of heterologous citrate (Cit) or fungal phytase (Phy) exudation traits were grown under two root treatments (roots separated or intermingled) and in two soils with contrasting soil P availability. Complementarity of plant mixtures varying in citrate efflux rate and mobility of the expressed phytase in soil was determined based on plant biomass and P accumulation. Soil P composition was evaluated using solution 31 P NMR spectroscopy. In the soil with limited available P, positive complementarity occurred in Cit+Phy mixtures with roots intermingled. Root separation eliminated positive interactions in mixtures expressing the less mobile phytase (Aspergillus niger PhyA) whereas positive complementarity persisted in mixtures that expressed the more mobile phytase (Peniophora lycii PhyA). Soils from Cit+Phy mixtures contained less inorganic P and more organic P compared to monocultures. Exudate-specific strategies for the acquisition of soil P were most effective in P-limited soil and depended on citrate efflux rate and the relative mobility of the expressed phytase in soil. Plant growth and soil P utilization in plant systems with complementary exudation strategies are expected to be greatest where exudates persist in soil and are expressed synchronously in space and time. This article is protected by copyright. All rights reserved.

  15. Optimization of phytase production by Penicillium purpurogenum GE1 under solid state fermentation by using Box–Behnken design

    Science.gov (United States)

    Awad, Ghada E.A.; Helal, Mohamed M.I.; Danial, Enas N.; Esawy, Mona A.

    2013-01-01

    Phytase production by Penicillium purpurogenum GE1 isolated from soil around bean root nodules was investigated by solid state fermentation (SSF) using mixed substrates consisted of corn cob and corn bran. The SSF conditions were optimized by using one-variable–at-a-time strategy. The optimum conditions for phytase production were at 27 °C, pH 8 and 66% moisture content. The study of different carbon and nitrogen sources revealed that glucose and peptone registered the highest enzyme productivity (92 ± 5.6 U/g ds, 125 ± 4.9 U/g ds). Among different surfactants, maximum phytase productivity was observed with Tween 80 at 0.001 concentrations (170 ± 4.2 U/g ds). A Box–Behnken design was employed to investigate the optimization of the most significant variables affecting the enzyme production. Maximal phytase production was detected after the addition of (g/5 g ds): 0.75 glucose, 0.375 peptone and 0, 01 tween 80. This result represented an improvement in phytase production of 2.6 folds when compared to that previously obtained using the basal medium under the same cultivation conditions. The generated model was found to be very adequate for phytase production (90% accuracy) as the experimental value was 444 ± 3.5 U/g ds compared to 401 U/g ds for the predicted value. In brief, the production of phytase using corn cob and corn bran is a novel and cheap way for the production of this important enzyme and opens a new way for researchers to discover and explore this arena. PMID:24596503

  16. Effects of administration of four different doses of Escherichia coli phytase on femur properties of 16-week-old turkeys.

    Science.gov (United States)

    Tatara, Marcin R; Krupski, Witold; Kozłowski, Krzysztof; Drażbo, Aleksandra; Jankowski, Jan

    2015-03-18

    The enzyme phytase is able to initiate the release of phosphates from phytic acid, making it available for absorption within gastrointestinal tract and following utilization. The aim of the study was to determine effects of Escherichia coli phytase administration on morphological, densitometric and mechanical properties of femur in 16-week-old turkeys. One-day-old BUT Big-6 males were assigned to six weight-matched groups. Turkeys receiving diet with standard phosphorus (P) and calcium (Ca) content belonged to the positive control group (Group I). Negative control group (Group II) consisted of birds fed diet with lowered P and Ca content. Turkeys belonging to the remaining groups have received the same diet as group II but enriched with graded levels of Escherichia coli phytase: 125 (Group III), 250 (Group IV), 500 (Group V) and 1000 (Group VI) FTU/kg. At the age of 112 days of life, the final body weights were determined and the turkeys were sacrificed to obtain right femur for analyses. Geometric and densitometric properties of femur were determined using quantitative computed tomography (QCT) technique, while mechanical evaluation was performed in three-point bending test. Phytase administration increased cross-sectional area, second moment of inertia, mean relative wall thickness, cortical bone mineral density and maximum elastic strength decreasing cortical bone area of femur (P phytase administration on geometric, densitometric and mechanical properties of femur were observed in turkeys receiving 125 and 250 FTU/kg of the diet. Phytase administration at the dosages of 500 and 1000 FTU/kg of the diet improved the final body weight in turkeys. The results obtained in this study indicate a possible practical application of Escherichia coli phytase in turkey feeding to improve skeletal system properties and function.

  17. Phytase Activity of Lactic Acid Bacteria Isolated from Dairy and Pharmaceutical Probiotic Products

    Directory of Open Access Journals (Sweden)

    Zohreh Khodaii

    2013-01-01

    Full Text Available Phytate, the major storage form of phosphorus in plant seeds, can form insoluble complexes with minerals such as iron, zinc and calcium thus reducing their bioavailability. Phytase enzymes are often used to upgrade the nutritional quality of phytate-rich foods and feeds such as grains. The phytate-degrading activity of 43 lactic acid bacteria including isolates from commercial probiotic preparations, dairy products and type strains were measured. The phytate-degrading activity of bifidobacteria and lactobacillus isolates from pharmaceutical probiotics, dairy products and type strains were determined. The enzyme activity of probotic bacteria ranged between 1.1-5.4 mU and was strain not species specific. Phytase activity may thus be a useful additional attribute of probiotics to be used as food supplements.

  18. Pond-raised hybrid catfish, male Ictalurus punctatus X female Ictalurus furcatus, do not respond to microbial phytase “super-dosing”

    Science.gov (United States)

    Two experiments were conducted in consecutive years to evaluate responses of hybrid catfish, male Ictalurus punctatus X female Ictalurus furcatus, to “super-dosing” of 6-phytase added to existing commercial catfish feeds. In each experiment, two diets with or without a phytase super-dose (2,500 and ...

  19. 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

  20. Impacts of age and calcium on Phytase efficacy in broiler chickens

    Science.gov (United States)

    A total of 648 straight-run hatchling Heritage 56M × fast feathering Cobb 500F broiler birds were used to determine the effects of Ca concentration and age on phytase efficacy. Corn and SBM based diets with 0.19%non-phytate P were prepared with three Ca (6.5, 8.0 and 9.5 g/kg) concentrations. A 6-ph...

  1. 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.

  2. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  4. Extracellular Phytase Production by the Wine Yeast S. cerevisiae (Finarome Strain) during Submerged Fermentation.

    Science.gov (United States)

    Kłosowski, Grzegorz; Mikulski, Dawid; Jankowiak, Oliwia

    2018-04-08

    One of the key steps in the production of phytases of microbial origin is selection of culture parameters, followed by isolation of the enzyme and evaluation of its catalytic activity. It was found that conditions for S. cerevisiae yeast culture, strain Finarome, giving the reduction in phytic acid concentration of more than 98% within 24 h of incubation were as follows: pH 5.5, 32 °C, continuous stirring at 80 rpm, the use of mannose as a carbon source and aspartic acid as a source of nitrogen. The highest catalytic activity of the isolated phytase was observed at 37 °C, pH 4.0 and using phytate as substrate at concentration of 5.0 mM. The presence of ethanol in the medium at a concentration of 12% v / v reduces the catalytic activity to above 60%. Properties of phytase derived from S. cerevisiae yeast culture, strain Finarome, indicate the possibility of its application in the form of a cell's free crude protein isolate for the hydrolysis of phytic acid to improve the efficiency of alcoholic fermentation processes. Our results also suggest a possibility to use the strain under study to obtain a fusant derived with specialized distillery strains, capable of carrying out a highly efficient fermentation process combined with the utilization of phytates.

  5. Characterization of phytase enzymes as feed additive for poultry and feed

    Science.gov (United States)

    Lamid, M.; Al-Arif, A.; Asmarani, O.; Warsito, S. H.

    2018-04-01

    One of the obstacles to utilizing rice bran as feed is the presence of antinutrition in the form of phytic acid which binds in minerals to form complex compounds with P, Mg, Mn, Fe, Zn, Ca. Phytic acid and its salts are the main forms of P, Mg, Mn, Fe, Zn, Ca deposits contained in cereals, legume and grains, about 60-90% of total minerals P, Mg, Mn, Fe, Zn, Ca in the form of phytic acid or phytate salts. Phytate is one of the enzymes belonging to the phosphatase group capable of hydrolyzing phytate compounds of myo-inositol (1,2,3,4,5,6) hexsa phosphatase into myo-inositol and organic phosphat. The aim of this study was to obtain characterization of phytase enzymes from isolate Actinobacillus sp., Bacillus pumilus, Bacillus vallimortis and IBR-1. Determination of phytase activity and the absorbance was measured using a UV-Vis spectrophotometer at a wavelength of 392 nm. The result of Actinobacillus sp, Bacillus pumilus, Bacillus vallimortis, IBR-1 each having optimum temperature were 50°C, 40°C, 45°C, 45°C, and optimum pH were 4, 4, 5.5. Bacteria especially Actinobacillus sp, Bacillus pumilus, Bacillus vallimortis, IBR-1 are proven capable of producing the high enough phytase enzymes required for mineral availability for livestock and fish.

  6. 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.

  7. 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.

  8. Improving specific activity and thermostability of Escherichia coli phytase by structure-based rational design.

    Science.gov (United States)

    Wu, Tzu-Hui; Chen, Chun-Chi; Cheng, Ya-Shan; Ko, Tzu-Ping; Lin, Cheng-Yen; Lai, Hui-Lin; Huang, Ting-Yung; Liu, Je-Ruei; Guo, Rey-Ting

    2014-04-10

    Escherichia coli phytase (EcAppA) which hydrolyzes phytate has been widely applied in the feed industry, but the need to improve the enzyme activity and thermostability remains. Here, we conduct rational design with two strategies to enhance the EcAppA performance. First, residues near the substrate binding pocket of EcAppA were modified according to the consensus sequence of two highly active Citrobacter phytases. One out of the eleven mutants, V89T, exhibited 17.5% increase in catalytic activity, which might be a result of stabilized protein folding. Second, the EcAppA glycosylation pattern was modified in accordance with the Citrobacter phytases. An N-glycosylation motif near the substrate binding site was disrupted to remove spatial hindrance for phytate entry and product departure. The de-glycosylated mutants showed 9.6% increase in specific activity. On the other hand, the EcAppA mutants that adopt N-glycosylation motifs from CbAppA showed improved thermostability that three mutants carrying single N-glycosylation motif exhibited 5.6-9.5% residual activity after treatment at 80°C (1.8% for wild type). Furthermore, the mutant carrying all three glycosylation motifs exhibited 27% residual activity. In conclusion, a successful rational design was performed to obtain several useful EcAppA mutants with better properties for further applications. Copyright © 2014 Elsevier B.V. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    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.

  10. 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...... with microbial phytase (750 FTU/kg) fed dry; diet 3, diet 2 fed in liquid form (fermented 17.5 h, 20 °C, 50% residual in the tank). InsP6-P was not present in gastric or ileal digesta in pigs fed diet 3 due to complete InsP6 degradation before feeding. In pigs fed diet 2 the amount of gastric InsP6-P...... was considerably smaller compared with pigs fed diet 1 due to phytase addition (P ≤ 0.001). On the other hand, the amount of ileal InsP6-P was only slightly less in pigs fed diet 2 compared with diet 1 indicating that InsP6 is greatly degraded in the small intestine. Furthermore, the amounts of gastric or ileal...

  11. Bioavailability of Iron, Zinc, Phytate and Phytase Activity during Soaking and Germination of White Sorghum Varieties

    Science.gov (United States)

    Afify, Abd El-Moneim M. R.; El-Beltagi, Hossam S.; Abd El-Salam, Samiha M.; Omran, Azza A.

    2011-01-01

    The changes in phytate, phytase activity and in vitro bioavailability of iron and zinc during soaking and germination of three white sorghum varieties (Sorghum bicolor L. Moench), named Dorado, Shandweel-6, and Giza-15 were investigated. Sorghum varieties were soaked for 20 h and germinated for 72 h after soaking for 20 h to reduce phytate content and increase iron and zinc in vitro bioavailability. The results revealed that iron and zinc content was significantly reduced from 28.16 to 32.16% and 13.78 to 26.69% for soaking treatment and 38.43 to 39.18% and 21.80 to 31.27% for germination treatments, respectively. Phytate content was significantly reduced from 23.59 to 32.40% for soaking treatment and 24.92 to 35.27% for germination treatments, respectively. Phytase enzymes will be activated during drying in equal form in all varieties. The results proved that the main distinct point is the change of phytase activity as well as specific activity during different treatment which showed no significant differences between the varieties used. The in vitro bioavailability of iron and zinc were significantly improved as a result of soaking and germination treatments. PMID:22003395

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

    Directory of Open Access Journals (Sweden)

    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.

  13. 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

    Wheat (Triticum aestivum L.) possesses preformed phytase activity in the grain that is essential to make phosphate available to cell metabolism and in food and feed (Brejnholt S. et al., 2011). Cereals contain the purple acid phosphatase type of phytases, PAPhy (Dionisio G. et al., 2011a). Mature......., 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......) 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.). Plant Physiol. [in press, Jan 10, Epub ahead of print] Dionisio G., Brinch-Pedersen H., Welinder K.G., Jørgensen M. (2011b...

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

    Directory of Open Access Journals (Sweden)

    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.

  15. 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 Ca2+, but inhibited by Al3+, Mn2+, Zn2+, and Cu2+ at a tested concentration of 5 mM. PMID:23586045

  16. The effect of phytase levels in the diet on the digestibility and growth performance of white shrimp Litopenaeus vannamei

    Directory of Open Access Journals (Sweden)

    Muhammmad Agus Suprayudi

    2013-11-01

    Full Text Available This experiment was conducted to evaluate the effect of phytase levels in the diet on the digestibility and growth performance of white shrimp Litopenaeus vanname. White shrimp were used in the trial with initial body weight of 3.8±0.01 g were reared in  aquarium (60×40×50 cm3 and filled with 60L treated sea water at a density of ten individu. Experimental diet were formulated in isonitrogeneous (36% protein and isocaloric (3960 kcal/kg to contain four levels of phytase: 0 iu/kg (diet A, 500 iu/kg (diet B, 750 iu/kg (diet C, and 1000 iu/kg(diet D. All diets were supplied with 0.5% of Cr2O3 as an indicator for digestibility measurement shrimp fed three times daily at satiation levelfor how many days of rearing. A factorial completely randomized experimental design was selected consisted of four treatments and triplicates. The result showed that the phytase addition improved the digestibility of protein, calcium, and phosphor as well as improve growth performance of white shrimp. Based on evaluation of those parameters, it is concluded that phytase level of500 IU support the best growth performance ofjuvenile white shrimp. Keywords: phytase, feed digestibility, growth,Litopenaeus vannamei

  17. Effect of zinc bacitracin and phytase on growth performance, nutrient digestibility, carcass and meat traits of broilers.

    Science.gov (United States)

    Attia, Y A; Bovera, F; Abd El-Hamid, A E; Tag El-Din, A E; Al-Harthi, M A; El-Shafy, A S

    2016-06-01

    A total of 336 one-day-old Hubbard broiler chickens were randomly distributed among 8 groups, each containing six replicates (7 chickens/replicate). From 1 to 40 days of age, the groups fed the same starter, grower and finisher diets. The control group was unsupplemented; zinc bacitracin (ZnB) group received the antibiotic at 0.5 g/kg; fungal phytase (FP) groups received 250, 500 and 1000 U/kg diet of Aspergillus niger phytase (FP_250, FP_500 and FP_1000 groups), respectively; bacterial phyatse (BP) groups received 250, 500 and 1000 U/kg diet of Escherichia coli phytase (BP_250EP, BP_500EP and BP_1000EP groups) respectively. Considering the whole experimental period, body weight gain was unaffected by ZnB and different concentrations of bacterial and fungal phytase; however, the feed conversion ratio of the group fed a diet supplemented with 500 U of BP was better (p phytase at 500 U may enhance performance of broiler chickens fed during days 1-40 of age and yield similar growth performance and economic efficiency to those of eB-supplemented groups. Journal of Animal Physiology and Animal Nutrition © 2015 Blackwell Verlag GmbH.

  18. Effect of Dietary Phytase Supplementation on Bone and Hyaline Cartilage Development of Broilers Fed with Organically Complexed Copper in a Cu-Deficient Diet.

    Science.gov (United States)

    Muszyński, Siemowit; Tomaszewska, Ewa; Kwiecień, Małgorzata; Dobrowolski, Piotr; Tomczyk, Agnieszka

    2018-04-01

    Tibial mechanical, chemical, and histomorphometrical traits were investigated for growing male Ross 308 broiler chickens fed diets that had copper (Cu) from organic source at a lowered level of 25% of the daily requirement (4 mg kg -1 of a premix) with or without phytase. Dietary treatments were control non-copper, non-phytase group (0 Suppl); 4 mg kg -1 Cu non-phytase group (25%Cu); and 4 mg kg -1 Cu + 500 FTU kg -1 phytase group (25%Cu + phyt). The results show that birds fed with the addition of phytase exhibited improved weight gain and final body weight and had increased serum IGF-1 and osteocalcin concentrations. The serum concentration of Cu and P did not differ between groups; however, Ca concentration decreased in the 25%Cu + phyt group when compared to the 25%Cu group. Added Cu increased bone Ca, P, Cu, and ash content in Cu-supplemented groups, but bone weight and length increased only by the addition of phytase. Bone geometry, yield, and ultimate strengths were affected by Cu and phytase addition. A decrease of the elastic stress and ultimate stress of the tibia in Cu-supplemented groups was observed. The histomorphometric analysis showed a positive effect of Cu supplementation on real bone volume and trabecular thickness in the tibia metaphyseal trabeculae; additionally, phytase increased the trabeculea number. The supplementation with Cu significantly increased the total articular cartilage and growth plate cartilage thickness; however, the changes in thickness of particular zones were dependent upon phytase addition. In summary, dietary Cu supplements given to growing broilers with Cu in their diet restricted to 25% of the daily requirement had a positive effect on bone metabolism, and phytase supplementation additionally improved cartilage development.

  19. 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.

  20. 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

    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......, and Phy4 led to an undegraded InsP6 content of 56, 49, 70, and 18%, respectively, when incubated with rapeseed cake for 6 h, indicating that Phy2 and Phy4 were the most effective phytases. However, Phy2 had a higher specific activity than Phy4, as 60% of the original InsP6 content was remaining after 3 h...

  1. 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 calcareum Ca, or sugar beet co-product. Inclusion of microbial phytase increased (P calcareum Ca, or the sugar beet co-product, but pigs fed calcium carbonate diets had greater ( P< 0.05) ATTD of P than pigs fed L. calcareumCa or the sugar beet co-product. Regardless of Ca source, inclusion of microbial phytase increased (P < 0.001) the ATTD of P. In conclusion, MCP has the greatest ATTD and STTD of Ca among the calcium 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.

  2. Water-soluble phosphorus in fresh broiler litter is dependent upon phosphorus concentration fed but not on fungal phytase supplementation.

    Science.gov (United States)

    Applegate, T J; Joern, B C; Nussbaum-Wagler, D L; Angel, R

    2003-06-01

    This experiment determined the effects of different phosphorus (P) feeding programs on total and water-soluble P excretion by broilers. Ross 308, male broilers were fed an industry (IND) diet (0.48, 0.35, 0.31, and 0.30% nonphytate P; NPP), an industry diet with reduced NPP and supplemental phytase [IND + PT; 600 phytase units (FTU)/kg; 0.36, 0.26, 0.20, and 0.19% NPP], a diet to more closely meet the birds' NPP requirements in which NPP was reduced further with supplemental phytase (REQ + P; 600 FTU/kg; 0.36, 0.26, 0.19, and 0.09% NPP), or a diet with low-phytate(lpa 1-1) corn with supplemental phytase (LPA + P; 600 FTU/kg; 0.37, 0.29, 0.19, 0.19% NPP). These diets were fed from hatch to 17 d, 17 to 31 d, 31 to 42 d, and 42 to 49 d of age, respectively. Fungal phytase was analyzed prior to diet formulation. Diets were fed to six replicate pens of 39 birds per pen. Litter samples were collected at 49 d of age and frozen prior to analyses. Diet did not significantly affect broiler performance (average BW at 49 d = 3.03 kg), tibia, or toe ash throughout the study (P > 0.05). Litter from broilers fed the IND diet was significantly higher (P litter from broilers fed IND + PT (0.84 and 0.14% of DM, respectively), REQ + P (0.78 and 0.11% of DM, respectively), or LPA + PT (0.64 and 0.12% of DM, respectively). Litter total and water-soluble P were not significantly different among broilers fed IND + PT, REQ + PT, or LPA + PT. In conclusion, phytase supplementation did not affect the solubility of P in the litter regardless of P feeding program.

  3. 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. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. Effect of immobilized fungal phytase on growth performance and bone traits of broilers fed with low dietary calcium and phosphorus

    Directory of Open Access Journals (Sweden)

    Sreeja Ajith

    2018-06-01

    Full Text Available Aim: The aim of this study was to investigate the effects of phytase which was laboratory produced by Aspergillus foetidus on the growth performance, mineral retention, and bone traits of broilers fed with low dietary calcium and phosphorus. Materials and Methods: The extracellular phytase enzyme secreted into the crude filtrate was concentrated by ammonium sulfate precipitation to obtain an activity of 500 phytase units (FTU. A total of 90 1-day-old chicks (Cobb 500 were randomly divided into three treatment groups with five replicates having six birds each. Dietary treatment, T1, was with 0.45% non-phytate P (NPP during starter and 0.40% during finisher phase with 1% Ca. Dietary treatment, T2, had 0.37% NPP during starter and 0.32% in finisher phase with 1% Ca and supplemental lab phytase at 500 FTU/kg. Dietary treatment, T3, was similar to T2 with a lower Ca of 0.8%. Results: There was no significant difference among the dietary treatments with regard to body weight gain, feed intake, feed conversion ratio, and Ca retention (p>0.05. However, a significant improvement in retention of P by birds was observed in phytase supplemental groups T2 and T3 (p<0.05. Dry weight of tibia (2.58-2.78 g/kg live weight and ash content (39.7- 41.8% was comparable among treatments. A similar trend was observed for bone Ca, P, and Mn content. Conclusion: The study indicated that 500 FTU/kg phytase can be effectively supplemented in a broiler diet with low phosphorus (0.37% in starter and 0.32% NPP in finisher diet and low calcium (0.8% in diet for better growth performance and with successful replacement of dietary P by 0.08 % and reduced P excretion into the environment in broiler chicken.

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

    Directory of Open Access Journals (Sweden)

    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.

  6. FLUCTUATION OF POST-PRANDIAL PLASMA MINERAL LEVEL OF JUVENILE JAPANESE FLOUNDER, Paralichthys olivaceus FED DIETARY PHOSPHORUS AND PHYTASE SUPPLEMENTATION

    Directory of Open Access Journals (Sweden)

    Asda Laining

    2010-12-01

    Full Text Available In order to investigate the phytic acid degradation in the gut of post juvenile Japanese flounder, indirect method was carried out by measuring the pre-prandial and postprandial plasma mineral and alkaline phosphatase (ALP level as well as liver phosphorus content. The experiment was designed into a Randomized Block in which experiment units were grouped according to sampling days at 10, 20 and 30 days of feeding time. Experimental diets contained three levels of dietary inorganic phosphorus at 0.0; 0.25 and 0.5% combined with two levels of dietary phytase at 0 and 2,000 FTU/kg diet. Juvenile Japanese flounder (IBW = 36.2 g were randomly distributed into 6 tanks of a 200 L capacity with density of 15 fish/tank. Blood sampling was carried out at 0 hour (before feeding or pre-prandial and at 1, 3, 6 and 12 hour post feeding (post-prandial time in three sampling days, respectively. Plasma was measured for mineral and ALP levels, while liver was analyzed for P content. The observation showed that fish fed without both dietary IP and phytase supplements had the lowest postprandial plasma IP, Mg and ALP levels during 12-h postprandial period. Plasma IP level at 6-h post-feeding in groups fed dietary 0.25 and 0.5% IP were significant higher when diet supplemented with phytase than those without phytase supplement. Peak level of plasma IP in fish fed 0.25% IP was similar to fish fed 0.5% with the presence of dietary phytase. At 1 and 3-h post-feeding, plasma Ca level increased in all groups, but significant difference was only observed between group fed diet without both dietary IP and phytase and other groups. Similar to plasma IP level, peak of plasma Mg and ALP concentration occurred in fish fed 0.25% IP together with phytase, and did not significantly differ from fish fed with 0.5% IP even when phytase was included in diet.

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

    OpenAIRE

    Wutiporn Phromkunthong; Jacques Gabaudan

    2006-01-01

    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 d...

  8. 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...

  9. Hydrolysis of phytate to its lower esters can influence the growth performance and nutrient utilization of broilers with regular or super doses of phytase.

    Science.gov (United States)

    Beeson, L A; Walk, C L; Bedford, M R; Olukosi, O A

    2017-07-01

    The aim of the study was to observe the effects of dietary available phosphorus (aP) and calcium (Ca), with regular or super doses of phytase, on phytate hydrolysis and subsequent influences on broiler growth performance and nutrient utilization. In a 2 × 3 factorial design, 384 Ross-308 broilers were allocated to one of 6 dietary treatments with 8 replicates in a randomized complete block design for 21 days. Diets were nutritionally adequate (positive control, PC) or marginally deficient in aP and Ca (negative control, NC), with 0, 500 or 1,500 FTU/kg phytase. Bird and feed weights were recorded on d 0 and 21, excreta were collected on d 19 and 20, and gizzard and ileal contents were collected on d 21. Body weight gain (P phytase in the PC and quadratically in the NC. There was an interactive effect on ileal DM, N, and P utilization, increasing quadratically with phytase supplementation in the NC, but there was no phytase influence in the PC (P Phytase linearly increased copper (P Phytase decreased ileal (IPx, inositol x-phosphate) IP6 and IP5 and increased inositol (quadratic, P phytase was added. Phytate hydrolysis improves the growth potential of birds fed NC diets, allowing them to match the growth performance of birds fed PC diets and improve nutrient utilization. These results indicate that dietary Ca and aP concentrations can be reduced when phytase is supplemented. It also may be beneficial to apply the enzyme nutrient matrix to other nutrients in the diet to maintain an optimal balance of nutrients in the digesta. © The Author 2017. Published by Oxford University Press on behalf of Poultry Science Association.

  10. 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

  11. Effect of superdosing phytase on productive performance and egg quality in laying hens

    Directory of Open Access Journals (Sweden)

    Jong Hyuk Kim

    2017-07-01

    Full Text Available Objective An experiment was conducted to determine the effect of superdosing phytase on productive performance and egg quality in laying hens. Methods A total of 200 42-wk-old Hy-Line Brown laying hens were allotted into 1 of 5 dietary treatments with 5 replicates consisting of 8 hens per replicate. The positive control (PC and negative control diets (NC were prepared based on the recommended P levels in layer diets. Supplemental phytase was added to the negative control diet at 10,000 (SD10, 20,000 (SD20, or 30,000 (SD30 fytase units (FTU/kg. Productive performance was summarized for 6 weeks from 42 weeks to 47 weeks of age. Egg quality was assessed from 4 eggs per replicate randomly collected at the conclusion of the experiment. Results The SD20 treatment had greater (p<0.05 hen-day egg production than PC, NC, and SD10 treatment groups. There was no difference in hen-day egg production between SD20 and SD30 treatment groups. However, SD30 treatment had greater (p<0.05 hen-day egg production than PC treatment, but showed no difference in hen-day egg production as compared to NC and SD10 treatment groups. However, egg weight, egg mass, feed intake, and feed conversion ratio were not affected by dietary treatments. Egg quality including eggshell strength, eggshell color, egg yolk color, and haugh unit was not influenced by dietary treatments. Conclusion Superdosing level of 20,000 FTU/kg phytase in diets has a positive effect on egg production rate, but no beneficial effect on egg quality in laying hens.

  12. 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.

  13. Effects of neutral phytase on growth performance and phosphorus utilization in crucian carp (Carassius auratus).

    Science.gov (United States)

    Nie, Xin-Zheng; Chen, Sha; Zhang, Xiao-Xu; Dai, Bin-Yang; Qian, Li-Chun

    A feeding trial was conducted for nine weeks to investigate the effects of partially replacing Ca(H 2 PO 4 ) 2 with neutral phytase on the growth performance, phosphorus utilization, nutrient digestibility, serum biochemical parameters, bone and carcass mineral composition, and digestive-enzyme-specific activity in crucian carp (Carassius auratus). The diets prepared with 0.8%, 0%, and 1.8% Ca(H 2 PO 4 ) 2 (1%=1 g/100 g) supplements were regarded as the P 1 E 0 , negative control (NC), and positive control (PC) groups, respectively; the other three experimental diets were prepared with the addition of 200, 300, and 500 U/kg of neutral phytase, respectively, based on the P 1 E 0 group. Three hundred and eighty-four fish ((1.50±0.01) g) were randomly distributed in the six treatments with four replicates each. The fish were initially fed with 2%-3% diets of their body weight per day, with feeding twice daily (08:00 and 16:00), under a 12-h light/12-h dark cycle at the temperature of (27.56±0.89) °C. The results showed that supplemental phytase at different levels in the diet improved the final body weight, average daily gain, feed conversion ratio, phosphorus utilization, and protein efficiency ratio of crucian carp (PPhytase supplementation increased the mineral content in serum (P), bone (P, Ca), and carcass (P, Ca, Zn, Na, and Mg) (Pphytase-supplemented diets (Pphytase improved the growth performance, phosphorus utilization as well as nutrient utilization in crucian carp, and it can be considered an important nutritional replacement for Ca(H 2 PO 4 ) 2 .

  14. 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.

  15. Radioisotope and mathematical modeling in the assessment of supplementation of Phytase in diets for growing pigs

    International Nuclear Information System (INIS)

    Mota, L.C.; Moreira, J.A.; Oliveira, R.L.R.; Vitti, D.M.S.S.; Patino, R.M.

    2014-01-01

    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 10 ), the P excreted in urine (F 02 ) and the output flow of P in the bones (F 32 e F 23 ) and soft tissue (F 42 e F 24 ), however, there was a reduction in P excreted in feces (F 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 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)

  16. Effects of copper source and concentration on in vitro phytate phosphorus hydrolysis by phytase.

    Science.gov (United States)

    Pang, Yanfang; Applegate, Todd J

    2006-03-08

    Five copper (Cu) sources were studied at pH 2.5, 5.5, and 6.5 to determine how Cu affects phytate phosphorus (PP) hydrolysis by phytase at concentrations up to 500 mg/kg diet (60 min, 40-41 degrees C). Subsequently, Cu solubility with and without sodium phytate was measured. Adding Cu inhibited PP hydrolysis at pH 5.5 and pH 6.5 (P copper chloride and copper lysinate inhibited PP hydrolysis much less than copper sulfate pentahydrate, copper chloride, and copper citrate (P copper-phytin complexes.

  17. Effect of combined xylanase and phytase on growth performance, apparent total tract digestibility, and carcass characteristics in growing pigs fed corn-based diets containing high-fiber coproducts.

    Science.gov (United States)

    Jang, Y D; Wilcock, P; Boyd, R D; Lindemann, M D

    2017-09-01

    Phytate has been shown to be an antinutrient, and the feeding of high levels of phytase can break down phytate to improve nutrient utilization and pig performance. Dietary xylanase targets arabinoxylan breakdown, thereby improving energy utilization in pigs. However, the effects of simultaneous supplementation have not been clearly determined. Crossbred pigs ( = 45; mean initial weight, 26.4 ± 0.2 kg) were allotted to 1 of 9 treatments to evaluate the effects of both xylanase (endo-1,4-β xylanase [EC 3.2.1.8]) and phytase (6-phytase [EC 3.1.3.26]) supplementation as follows: 1) positive control (PC), a corn-soybean meal-based diet with 15% corn distillers dried grains with solubles, 15% wheat middlings, and 13% corn germ meal; 2) negative control (NC), ME was reduced by 103 kcal/kg from the PC diet by replacement of fat with corn starch; 3) NC + phytase (500 phytase units (FTU)/kg diet); 4) NC + phytase (1,000 FTU/kg diet); 5) NC + phytase (2,000 FTU/kg diet); 6) NC + xylanase (24,000 xylanase units [BXU]/kg diet); 7) NC + phytase (500 FTU/kg diet) + xylanase (24,000 BXU/kg diet); 8) NC + phytase (1,000 FTU/kg diet) + xylanase (24,000 BXU/kg diet); and 9) NC + phytase (2,000 FTU/kg diet) + xylanase (24,000 BXU/kg diet). All diets were formulated to meet nutrient requirements before phytase and xylanase addition to the diets. There were no significant interactions between xylanase and phytase supplementation on growth performance, carcass characteristics, and apparent total tract digestibility (ATTD). The ADG ( phytase level increased. The ATTD of P increased as phytase supplementation level increased ( phytase level increased. Estimated carcass lean percentage and lean gain increased ( phytase level increased. Xylanase supplementation had no effect on growth performance, ATTD, and carcass characteristics. The results demonstrated an improved nutrient digestibility, performance, and carcass response to phytase supplementation beyond P provision because all diets

  18. Degradation of Phytate Pentamagnesium Salt by Bacillus sp. T4 Phytase as a Potential Eco-friendly Feed Additive

    Science.gov (United States)

    Park, Inkyung; Lee, Jaekoo; Cho, Jaiesoon

    2012-01-01

    A bacterial isolate derived from soil samples near a cattle farm was found to display extracellular phytase activity. Based on 16S rRNA sequence analysis, the strain was named Bacillus sp. T4. The optimum temperature for the phytase activity toward magnesium phytate (Mg-InsP6) was 40°C without 5 mM Ca2+ and 50°C with 5 mM Ca2+. T4 phytase had a characteristic bi-hump two pH optima of 6.0 to 6.5 and 7.4 for Mg-InsP6. The enzyme showed higher specificity for Mg-InsP6 than sodium phytate (Na-InsP6). Its activity was fairly inhibited by EDTA, Cu2+, Mn2+, Co2+, Ba2+ and Zn2+. T4 phytase may have great potential for use as an eco-friendly feed additive to enhance the nutritive quality of phytate and reduce phosphorus pollution. PMID:25049504

  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. 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 ...

  1. 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

  2. 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.)

  3. Purification and biochemical characterization of an Aspergillus niger phytase produced by solid-state fermentation using triticale residues as substrate.

    Science.gov (United States)

    Neira-Vielma, Alberto A; Aguilar, Cristóbal N; Ilyina, Anna; Contreras-Esquivel, Juan C; Carneiro-da-Cunha, María das Graça; Michelena-Álvarez, Georgina; Martínez-Hernández, José L

    2018-03-01

    In this study, an extracellular phytase produced by Aspergillus niger 7A-1, was biochemically characterized for possible industrial application. The enzyme was purified from a crude extract obtained by solid-state fermentation (SSF) of triticale waste. The extract was obtained by microfiltration, ultrafiltration (300, 100 and 30 kDa) and DEAE-Sepharose column chromatography. The molecular weight of the purified enzyme was estimated to be 89 kDa by SDS-PAGE. The purified enzyme was most active at pH 5.3 and 56 °C, and retained 50% activity over a wide pH range of 4 to 7. The enzymatic thermostability assay showed that the enzyme retained more than 70% activity at 80 °C for 60 s, 40% activity for 120 s and 9% after 300 s. The phytase showed broad substrate specificity, a K m value of 220 μM and V max of 25 μM/min. The purified phytase retained 50% of its activity with phosphorylated compounds such as phenyl phosphate, 1-Naphthyl phosphate, 2-Naphthyl phosphate, p-Nitrophenyl phosphate and Glycerol-2-phosphate. The inhibition of phytase activity by metal ions was observed to be drastically inhibited (50%) by Ca ++ and was slightly inhibited (10%) by Ni ++ , K + , and Na + , at 10 and 20 mM concentrations. A positive effect was obtained with Mg ++ , Mn ++ , Cu ++ , Cd ++ and Ba ++ at 25 and 35% with stimulatory effect on the phytase activity.

  4. Purification and biochemical characterization of an Aspergillus niger phytase produced by solid-state fermentation using triticale residues as substrate

    Directory of Open Access Journals (Sweden)

    Alberto A. Neira-Vielma

    2018-03-01

    Full Text Available In this study, an extracellular phytase produced by Aspergillus niger 7A-1, was biochemically characterized for possible industrial application. The enzyme was purified from a crude extract obtained by solid-state fermentation (SSF of triticale waste. The extract was obtained by microfiltration, ultrafiltration (300, 100 and 30 kDa and DEAE-Sepharose column chromatography. The molecular weight of the purified enzyme was estimated to be 89 kDa by SDS-PAGE. The purified enzyme was most active at pH 5.3 and 56 °C, and retained 50% activity over a wide pH range of 4 to 7. The enzymatic thermostability assay showed that the enzyme retained more than 70% activity at 80 °C for 60 s, 40% activity for 120 s and 9% after 300 s. The phytase showed broad substrate specificity, a Km value of 220 μM and Vmax of 25 μM/min. The purified phytase retained 50% of its activity with phosphorylated compounds such as phenyl phosphate, 1-Naphthyl phosphate, 2-Naphthyl phosphate, p-Nitrophenyl phosphate and Glycerol-2-phosphate. The inhibition of phytase activity by metal ions was observed to be drastically inhibited (50% by Ca++ and was slightly inhibited (10% by Ni++, K+, and Na+, at 10 and 20 mM concentrations. A positive effect was obtained with Mg++, Mn++, Cu++, Cd++ and Ba++ at 25 and 35% with stimulatory effect on the phytase activity.

  5. Engineering Technology Of Fish Farming Floating Nets Cages On Polka Dot Grouper (Cromileptes Altivelis) Used Artificial Feed Enriched Phytase Enzyme

    Science.gov (United States)

    Samidjan, Istiyanto; Rachmawati, Diana

    2018-02-01

    One solution is to utilize engineering technology cultivation floating cage net polka dot grouper (ducker grouper), which is given artificial feed enriched with phytase enzymes. The objectives of this study was to examine the use of technology engineering floating net on ducker grouper on artificial feed that is enriched with different dose phytase enzymes to accelerate growth and survival. The research method used ducker grouper fish size 15,5 ± 0,5 cm in the net cages unit (1 m x 1 m x 1 m), 250 fish per cage, using 12 cages. Each net-cages was made of polyethylens netting, mesh size 12.5 mm. with complete randomized design (CRD) 4 treatment and 3 replication were feed Artificial enriched of phytase enzyme with the doses of A (0 FTU · kg-1 diet), B (200 FTU · kg-1 diet), C (500 FTU · kg-1 diet), and D (800 FTU · kg-1 diet) phytase enzyme. Feed was given 2 times a day in the morning and afternoon with 5% biomass per day. Data includes the growth of absolute weight polka dot grouper, FCR, and survival rate analyzed variety and Test Tukey.The result of the research showed that the difference of artificial feeding enriched phytase enzyme significantly (P <0,05) to growth, food conversion ratio (FCR), survival rete of polka dot grouper. The best treatment at C (500 mg / kg of feed) increase growth of absolute weight of 128.75 g, 1.75 (FCR), and a survival rate of 93.5%.

  6. 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 broilers fed the LCaP diet. Total P, soluble P (SP), and reactive soluble P (RSP) were decreased (P litter of broilers fed the LCaP diets. Total P, SP, and RSP were decreased (P 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.

  7. 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

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

    International Nuclear Information System (INIS)

    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.; Nogueira, G.P.; Fonseca, L.E.C.; Jorge, S.A.C.; Garcia, F.

    2002-01-01

    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 (μg-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)

  9. An in-silico insight into the characteristics of β-propeller phytase.

    Science.gov (United States)

    Mathew, Akash; Verma, Anukriti; Gaur, Smriti

    2014-06-01

    Phytase is an enzyme that is found extensively in the plant kingdom and in some species of bacteria and fungi. This paper identifies and analyses the available full length sequences of β-propeller phytases (BPP). BPP was chosen due to its potential applicability in the field of aquaculture. The sequences were obtained from the Uniprot database and subject to various online bioinformatics tools to elucidate the physio-chemical characteristics, secondary structures and active site compositions of BPP. Protparam and SOPMA were used to analyse the physiochemical and secondary structure characteristics, while the Expasy online modelling tool and CASTp were used to model the 3-D structure and identify the active sites of the BPP sequences. The amino acid compositions of the four sequences were compared and composed in a graphical format to identify similarities and highlight the potentially important amino acids that form the active site of BPP. This study aims to analyse BPP and contribute to the clarification of the molecular mechanism involved in the enzyme activity of BPP and contribute in part to the possibility of constructing a synthetic version of BPP.

  10. 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)

  11. Impacts of dietary calcium, phytate, and phytase on inositol hexakisphosphate degradation and inositol phosphate release in different segments of digestive tract of broilers.

    Science.gov (United States)

    Li, W; Angel, R; Kim, S-W; Brady, K; Yu, S; Plumstead, P W

    2017-10-01

    A total of 720 straight-run Heritage 56 M × fast feathering Cobb 500F broiler chickens was fed from 11 to 13 d of age to determine the impacts of dietary calcium (Ca), phytate phosphorus (PP), and phytase concentrations on inositol phosphate (IP3-6) profile in different digestive tract (GI) segments. The experiment was a 2 × 2 × 3 randomized block design with 2 Ca (0.7 and 1.0%) and 2 PP (0.23 and 0.34%) concentrations and 3 doses of Buttiauxella sp. phytase (0, 500, and 1,000 FTU/kg). The experiment was replicated in time (block) with 3 replicates per treatment (Trt) of 10 birds per block. Concentrations of IP3-6 in the crop, proventriculus (Prov) plus (+) gizzard (Giz), and distal ileum, as well as the ileal IP6 and P disappearance were determined at 13 d of age. The detrimental impact of Ca on IP6 and P disappearance was observed only in the ileum, where 11% reduction in both IP6 and P disappearance was seen when Ca increased from 0.7 to 1.0% (P phytase (P  0.05). Inclusion of phytase, at both 500 and 1,000 FTU/kg, resulted in lower IP6 and the accumulation of lower IP ester (IP3-5) concentrations in all GI segments (P phytase inclusion, despite the degree of improvement affected by PP (P phytase/kg inclusion, respectively, resulting in 41 and 64% greater P digestibility, respectively. In conclusion, phytase can effectively degrade IP6 to lower esters and increase P utilization. However, the efficacy of phytase can be affected by diet Ca and PP concentrations. © The Author 2017. Published by Oxford University Press on behalf of Poultry Science Association.

  12. Impacts of dietary calcium, phytate, and phytase on inositol hexakisphosphate degradation and inositol phosphate release in different segments of digestive tract of broilers

    Science.gov (United States)

    Li, W.; Angel, R.; Kim, S.-W.; Brady, K.; Yu, S.; Plumstead, P. W.

    2017-01-01

    Abstract A total of 720 straight-run Heritage 56 M × fast feathering Cobb 500F broiler chickens was fed from 11 to 13 d of age to determine the impacts of dietary calcium (Ca), phytate phosphorus (PP), and phytase concentrations on inositol phosphate (IP3–6) profile in different digestive tract (GI) segments. The experiment was a 2 × 2 × 3 randomized block design with 2 Ca (0.7 and 1.0%) and 2 PP (0.23 and 0.34%) concentrations and 3 doses of Buttiauxella sp. phytase (0, 500, and 1,000 FTU/kg). The experiment was replicated in time (block) with 3 replicates per treatment (Trt) of 10 birds per block. Concentrations of IP3–6 in the crop, proventriculus (Prov) plus (+) gizzard (Giz), and distal ileum, as well as the ileal IP6 and P disappearance were determined at 13 d of age. The detrimental impact of Ca on IP6 and P disappearance was observed only in the ileum, where 11% reduction in both IP6 and P disappearance was seen when Ca increased from 0.7 to 1.0% (P phytase (P  0.05). Inclusion of phytase, at both 500 and 1,000 FTU/kg, resulted in lower IP6 and the accumulation of lower IP ester (IP3–5) concentrations in all GI segments (P phytase inclusion, despite the degree of improvement affected by PP (P phytase/kg inclusion, respectively, resulting in 41 and 64% greater P digestibility, respectively. In conclusion, phytase can effectively degrade IP6 to lower esters and increase P utilization. However, the efficacy of phytase can be affected by diet Ca and PP concentrations. PMID:28938789

  13. Phytases Improve Myo-Inositol Bioaccessibility in Rye Bread: A Study Using an In Vitro Method of Digestion and a Caco-2 Cell Culture Model.

    Science.gov (United States)

    Duliński, Robert; Cielecka, Emilia Katarzyna; Pierzchalska, Małgorzata; Żyła, Krzysztof

    2015-03-01

    Preparations of 6-phytase A (EC 3.1.3.26) and phytase B (acid phosphatase, EC 3.1.3.2) were applied alone and combined in the preparation of dough to estimate their catalytic potential for myo- inositol liberation from rye flour in the breadmaking technology. The experimental bread samples were ground after baking and subjected to determination of myo- inositol bioavailability by an in vitro method that simulated digestion in a human alimentary tract, followed by measurements of myo- inositol transport through enterocyte- -like differentiated Caco-2 cells to determine its bioaccessibility. Myo- inositol content was measured by a high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) technique. The concentration of myo- inositol in the dialysates of control bread was 25.3 µg/mL, whereas in the dialysates of bread sample baked with 6-phytase A, the concentration increased to 35.4 µg/mL, and in the bread baked with phytase B to 64.98 µg/mL. Simultaneous application of both enzymes resulted in myo- inositol release of 64.04 µg/mL. The highest bioaccessibility of myo- inositol, assessed by the measurement of the passage through the Caco-2 monolayer was determined in the bread baked with the addition of 6-phytase A. Enzymatically modified rye bread, particularly by the addition of 6-phytase A, may be therefore a rich source of a highly bioaccessible myo - -inositol.

  14. 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.

  15. 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.

  16. Purification and characterization of a protease-resistant phytase of Aspergillus oryzae SBS50 whose properties make it exceptionally useful as a feed supplement.

    Science.gov (United States)

    Sapna; Singh, Bijender

    2017-10-01

    An extracellular phytase of Aspergillus oryzae SBS50 was purified to homogeneity using ammonium sulphate precipitation, ion-exchange and gel filtration chromatography. Purified phytase has a monomeric molecular mass of ∼80kDa exhibiting its optimal activity at pH 5.0 and 50°C with a T 1/2 of 300min at 50°C. Phytase of A. oryzae displayed broad substrate specificity with V max and K m values of 58.82μmol/ml/min and 1.14mM, respectively, for calcium phytate. Purity and homogeneity of the phytase was confirmed by high performance liquid chromatography and MALDI-TOF analysis revealed the identification of a peptide showing homology with acid phosphatase of Aspergillus oryzae RIB40. Among the inhibitors, 2,3-butanedione and sodium molybdate significantly inhibited the enzyme activity. Phytase of A. oryzae showed protease-resistance and was more stable during storage at 4°C and -20°C as compared to room temperature. Among all the feed samples, mustard oil cake was dephytinized more efficiently than other feed samples. These unique properties suggested that the phytase has the potential to be useful as an animal feed supplement. Copyright © 2017 Elsevier B.V. All rights reserved.

  17. Effects of high inclusion of soybean meal and a phytase superdose on growth performance of weaned pigs housed under the rigors of commercial conditions.

    Science.gov (United States)

    Moran, K; Boyd, R D; Zier-Rush, C; Wilcock, P; Bajjalieh, N; van Heugten, E

    2017-12-01

    Two studies were conducted to determine whether soybean meal (SBM) use in nursery pig diets can be increased by superdosing with phytase. In Exp. 1, 2,550 pigs (BW of 5.54 ± 0.09 kg) were used to evaluate the optimal level of phytase in low- or high-SBM diets. Two SBM levels (low and high) and 4 phytase doses (0, 1,250, 2,500, and 3,750 phytase units [FTU]/kg) were combined to create 8 dietary treatments in a 2 × 4 factorial arrangement. Pigs were fed a 3-phase feeding program, with each period being 10, 10, and 22 d, respectively. Inclusion of low and high SBM was 15.0 and 25.0%, respectively, for Phase 1; 19.0 and 29.0%, respectively, for Phase 2; and 32.5% for the common Phase 3 diet. Pigs fed diets with high SBM had improved G:F for Phase 1 and 2 and overall ( Phytase quadratically improved G:F during Phase 3 and overall ( phytase dose being 2,500 FTU/kg. High-SBM diets tended ( = 0.09) to decrease stool firmness (determined daily from d 1 to 10) only on d 2. In Exp. 2, 2,112 pigs (BW of 5.99 ± 0.10 kg) were used to evaluate the impact of high levels of SBM and phytase on performance, stool firmness, mortality, and morbidity in weaned pigs originating from a porcine reproductive and respiratory syndrome (PRRS) virus-positive sow farm. Pigs were fed a 3-phase feeding program as in Exp. 1. Three levels of SBM (low, medium, or high) and 2 phytase levels (600 or 2,600 FTU) were combined to create 6 dietary treatments in a 3 × 2 factorial arrangement. Inclusion of SBM was 15.0, 22.5, and 30.0% for Phase 1 and 20.0, 27.5, and 35.0% for Phase 2 for low, medium, and high SBM, respectively, and 29.0% for the common Phase 3 diet. Inclusion of SBM did not affect growth performance. The percentage of pigs removed for medical treatment linearly declined with increasing SBM levels ( = 0.04). High-SBM diets tended ( phytase tended ( phytase at superdose levels can improve growth performance independently from the level of SBM in the diet.

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

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    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.

  19. 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.

  20. Valorisation of untreated cane molasses for enhanced phytase production by Bacillus subtilis K46b and its potential role in dephytinisation.

    Science.gov (United States)

    Rocky-Salimi, Karim; Hashemi, Maryam; Safari, Mohammad; Mousivand, Maryam

    2017-01-01

    The high cost of phytase production is the most limiting factor in its application in animal feeds. The present study aimed to develop a low-cost medium for production of a novel phytase in submerged fermentation using inexpensive agro-industrial by-products. The applicability of phytase in dephytinisation of commonly used food/feed ingredients, i.e. soybean meal and wheat bran, was also investigated. Using a one-factor-at-a-time approach, soybean meal and cane molasses were identified as significant agro-industrial by-products and these factors were subsequently optimised using response surface methodology (RSM). A central composite design was employed to further enhance phytase yield. Under optimum conditions of soybean meal 22.3 g L -1 , cane molasses 100 g L -1 and 39 h fermentation, phytase production increased to 56.562 U mL -1 , indicating more than 28-fold enhancement. The enzyme efficiently dephytinised wheat bran and soybean meal after 24 h incubation at 56.5 °C and increased inorganic phosphate content by 240% and 155%, respectively. Soybean meal and cane molasses were successfully used for enhancement of phytase production as economical carbon, nitrogen and phytic acid sources using RSM. The phytase showed a good capability to dephytinise wheat bran and soybean meal, demonstrating that the enzyme can be considered as a potential candidate for industrial food and feed applications. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  1. Effects of rapeseed meal fiber content on phosphorus and calcium digestibility in growing pigs fed diets without or with microbial phytase.

    Science.gov (United States)

    Bournazel, M; Lessire, M; Duclos, M J; Magnin, M; Même, N; Peyronnet, C; Recoules, E; Quinsac, A; Labussière, E; Narcy, A

    2018-01-01

    The optimization of dietary phosphorus (P) and calcium (Ca) supply requires a better understanding of the effect of dietary fiber content of co-products on the digestive utilization of minerals. This study was designed to evaluate the effects of dietary fiber content from 00-rapeseed meal (RSM) on P and Ca digestibility throughout the gastrointestinal tract in growing pigs fed diets without or with microbial phytase. In total, 48 castrated male pigs (initial BW=36.1±0.4 kg) were housed in metabolic crates for 29 days. After an 8-day adaptation period, pigs were allocated to one of the eight treatments. The impact of dietary fiber was modulated by adding whole RSM (wRSM), dehulled RSM (dRSM) or dRSM supplemented with 4.5% or 9.0% rapeseed hulls (dRSMh1 and dRSMh2). Diets contained 0 or 500 phytase unit of microbial phytase per kg. From day 14 to day 23, feces and urine were collected separately to determine apparent total tract digestibility (ATTD) and apparent retention (AR) of P and Ca. At the end of the experiment, femurs and digestive contents were sampled. No effect of variables of interest was observed on growth performance. Microbial phytase increased ATTD and AR of P (Pphytase (Pphytase which increased AR of Ca and femur characteristics (Pphytase but cecal recovery was considerably reduced by microbial phytase (Pphytase and R 2=0.24, P=0.026 with microbial phytase). The inclusion of hulls improved the solubility of iP (Pphytase in releasing phosphate in the stomach. Moreover, dietary fiber may affect solubilization process in the cecum which potentiates the effect of microbial phytase on P digestibility.

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

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    E. Kiarie

    2015-10-01

    Full Text Available 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 (p<0.05 for NC vs PC birds. Phytase response in NC birds was linear (p<0.05 with 2,000 FTU showing the greatest improvement on body weight gain (20%, feed conversion (7.4%, tibia ash (18%, AR of Ca (38%, AR of P (51% and apparent metabolizable energy corrected for nitrogen (5.1% relative to NC. Furthermore, phytase at ≥750 FTU resulted in AID of total AA commensurate to that of PC fed birds and at ≥1,000 FTU improved (p<0.05 AR of P, dry matter, and N beyond that of the lower doses of phytase 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.

  3. 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.

  4. 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.

    2015-01-01

    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. PMID:26221114

  5. Strain improvement of Pichia kudriavzevii TY13 for raised phytase production and reduced phosphate repression.

    Science.gov (United States)

    Qvirist, Linnea; Vorontsov, Egor; Veide Vilg, Jenny; Andlid, Thomas

    2017-03-01

    In this work, we present the development and characterization of a strain of Pichia kudriavzevii (TY1322), with highly improved phytate-degrading capacity. The mutant strain TY1322 shows a biomass-specific phytate degradation of 1.26 mmol g -1  h -1 after 8 h of cultivation in a high-phosphate medium, which is about 8 times higher compared with the wild-type strain. Strain TY1322 was able to grow at low pH (pH 2), at high temperature (46°C) and in the presence of ox bile (2% w/v), indicating this strain's ability to survive passage through the gastrointestinal tract. The purified phytase showed two pH optima, at pH 3.5 and 5.5, and one temperature optimum at 55°C. The lower pH optimum of 3.5 matches the reported pH of the pig stomach, meaning that TY1322 and/or its phytase is highly suitable for use in feed production. Furthermore, P. kudriavzevii TY1322 tolerates ethanol up to 6% (v/v) and shows high osmotic stress tolerance. Owing to the phenotypic characteristics and non-genetically modified organisms nature of TY1322, this strain show great potential for future uses in (i) cereal fermentations for increased mineral bioavailability, and (ii) feed production to increase the phosphate bioavailability for monogastric animals to reduce the need for artificial phosphate fortification. © 2016 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

  6. Effect of High Phytase Inclusion Rates on Performance of Broilers Fed Diets Not Severely Limited in Available Phosphorus

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    T. T. dos Santos

    2013-02-01

    Full Text Available Phytate is not only an unavailable source of phosphorus (P for broilers but it also acts as an anti-nutrient, reducing protein and mineral absorption, increasing endogenous losses and reducing broiler performance. The objective of this study was to investigate the anti-nutritional effects of phytate by including high levels of phytase in diets not severely limited in available P. A total of 768 male Arbor Acres broilers were distributed in six treatments of eight replicate pens of 16 birds each consisting of a positive control diet (PC, positive control with 500 FTU/kg phytase, negative control (NC diet with lower available P and calcium (Ca levels and the same NC diet with 500, 1,000 or 1,500 FTU/kg phytase. Body weight gain (BWG, feed intake (FI, feed conversion ratio (FCR and mortality were determined at 21 and 35 d of age while foot ash was determined in four birds per pen at 21 d of age. FI, FCR and foot ash where not affected by the lower mineral diets at 21 d of age nor by the enzyme inclusion but broilers fed lower Ca and available P diets had lower BWG. At 35 d of age no difference was observed between broilers fed the positive or NC diets but broilers fed 500, 1,000 and 1,500 FTU/kg on top of the NC diet had better FCR than broilers fed the positive control diet. When compared to birds fed a diet adequate in P, birds fed the same diet included with 500, 1,000 and 1,500 FTU/kg of phytase in marginally deficient available P and Ca diets had an improvement of performance. These results support the concept that hydrolysing phytate and reducing the anti-nutritional effects of phytate improves bird performance on marginally deficient diets that were not covering the P requirement of birds.

  7. 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 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 lean yield in finisher CA gilts, although no change in these parameters was observed in CA finisher boars. The increase in lean yield in CA gilts by selective breeding without affecting the level of salivary phytase activity documents the value of conventional genetic selection in conjunction with genetic modification.

  8. [Effect of a supplemental Aspergillus niger phytase on the utilization of plant phosphorus by rainbow trout (Oncorhynchus mykiss)].

    Science.gov (United States)

    Rodehutscord, M; Becker, A; Pfeffer, E

    1995-01-01

    Effects of a supplemental Aspergillus niger-phytase on digestibility and utilization of dietary phosphorus (P) were studied in three experiments with rainbow trout. P concentration in the diets was 4.8 and 5.8 g/kg DM, respectively. The P contained in the diet originated solely from plants, mainly soy-products. Digestibility of P was studied using the stripping method and hydrochloride insoluble ash as marker. Utilization was studied in growth trials by use of the comparative body analysis. At a water temperature of 15 degrees C, both digestibility and utilization of P were increased from 25 to 57% and from 17 to 49%, respectively when 1000 U/kg phytase were supplemented. Feed consumption and gain of trout were significantly increased. At a water temperature of 10 degrees C, utilization of P was also increased from 6 to 25%. However, feed consumption and gain of trout were very low at this water temperature and not influenced by the supplemental phytase.

  9. SpyRing interrogation: analyzing how enzyme resilience can be achieved with phytase and distinct cyclization chemistries

    Science.gov (United States)

    Schoene, Christopher; Bennett, S. Paul; Howarth, Mark

    2016-01-01

    Enzymes catalyze reactions with exceptional selectivity and rate acceleration but are often limited by instability. Towards a generic route to thermo-resilience, we established the SpyRing approach, cyclizing enzymes by sandwiching between SpyTag and SpyCatcher (peptide and protein partners which lock together via a spontaneous isopeptide bond). Here we first investigated the basis for this resilience, comparing alternative reactive peptide/protein pairs we engineered from Gram-positive bacteria. Both SnoopRing and PilinRing cyclization gave dramatic enzyme resilience, but SpyRing cyclization was the best. Differential scanning calorimetry for each ring showed that cyclization did not inhibit unfolding of the inserted β-lactamase. Cyclization conferred resilience even at 100 °C, where the cyclizing domains themselves were unfolded. Phytases hydrolyze phytic acid and improve dietary absorption of phosphate and essential metal ions, important for agriculture and with potential against human malnutrition. SpyRing phytase (PhyC) resisted aggregation and retained catalytic activity even following heating at 100 °C. In addition, SpyRing cyclization made it possible to purify phytase simply by heating the cell lysate, to drive aggregation of non-cyclized proteins. Cyclization via domains forming spontaneous isopeptide bonds is a general strategy to generate resilient enzymes and may extend the range of conditions for isolation and application of enzymes. PMID:26861173

  10. Bioprocess for the production of recombinant HAP phytase of the thermophilic mold Sporotrichum thermophile and its structural and biochemical characteristics.

    Science.gov (United States)

    Maurya, Anay Kumar; Parashar, Deepak; Satyanarayana, T

    2017-01-01

    Thermophilc mold Sporotrichum thermophile secretes an acidstable and thermostable phytase, which finds application as a food and feed additive because of its adequate thermostability, acid stability, protease insensitivity and broad substrate spectrum. Low extracellular phytase production by the mold is a major bottleneck for its application on a commercial scale. We have successfully overcome this problem by constitutive secretary expression of codon optimized rStPhy under glyceraldehyde phosphate dehydrogenase (GAP) promoter in Pichia pastoris. A ∼41-fold improvement in rStPhy production has been achieved. Circular Dichroism (CD) spectra revealed that rStPhy is composed of 26.65% α-helices, 5.26% β-sheets and 68.09% random coils at pH 5.0 and 60°C, the optima for the enzyme activity. The melting temperature (T m ) of the enzyme is ∼73°C. The 3D structure of rStPhy displayed characteristic signature sequences (RHGXRXP and HD) of HAP phytase. The catalytically important amino acids (Arg74, His75, Arg78, His368 and Asp369) were identified by docking and site directed mutagenesis. Fluorescence quenching by N-bromosuccinimide (NBS) and CsCl exposed tryptophan residues surrounded by negative charges, which play a key role in maintaining structural integrity of rStPhy. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. 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. PMID:24688383

  12. 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

    Directory of Open Access Journals (Sweden)

    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.

  13. Effects of Adding Super Dose Phytase to the Phosphorus-deficient Diets of Young Pigs on Growth Performance, Bone Quality, Minerals and Amino Acids Digestibilities

    Directory of Open Access Journals (Sweden)

    Z. K. Zeng

    2014-02-01

    Full Text Available Two experiments were conducted to evaluate the efficacy of feeding an Escherichia coli (E. coli derived phytase to pigs fed P deficient, corn-soybean meal diets. In Exp. 1, 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, 0.46% total P and 0.24% non-phytate P supplemented with 0, 500, 1,000, or 20,000 FTU/kg E. coli phytase as well as a positive control formulated to be adequate in all nutrients (0.77% Ca, 0.62% total P and 0.42% non-phytate P. The treatments were applied to six pens with four pigs per pen for 28 days. In Exp. 2, ten crossbred pigs (19.66±1.16 kg fitted with ileal T-cannula were used in a nutrient balance study. The pigs were assigned to treatments similar to those used in Exp. 1 in a doubly replicated 5×4 incomplete Latin square design (5 diets with 4 periods. Each period consisted of a 5-d adjustment period followed by a 3-d total collection of feces and urine and then a 2-d collection of ileal digesta. Supplementation with phytase linearly increased (p<0.05 weight gain, feed intake, feed efficiency, bone breaking strength and fat-free dry and ash bone weight. There were linear increases (p<0.01 in the apparent ileal digestibility (AID of DM, GE, CP, Ca, total P, inositol hexaphosphate (IP6 and some AA with increasing dose of E. coli phytase. Pigs fed 20,000 FTU/kg had a greater (p<0.05 AID of IP6 (80% vs 59% or 64%, respectively than pigs fed diets with 500 or 1,000 FTU/kg phytase. There were linear increases (p<0.05 in the total tract digestibility of Ca, total P, Na, K, Mg, and Zn as well as in the retention of Mg and Zn with increased phytase dose. The retention and utilization of Cu, and the total tract digestibility of CP and Cu quadratic increased (p<0.05 with increased phytase dose. In conclusion, supplementation of 500 FTU of phytase/kg and above effectively hydrolyzed phytate in low-P corn-soybean diets for pigs. In

  14. 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).

  15. 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.

  16. 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.

    2015-01-01

    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. PMID:26323404

  17. 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,...

  18. 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

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

    Directory of Open Access Journals (Sweden)

    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.

  20. 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

    be used as an alternative to inorganic phosphorus supplementation to feed and the study addresses the environmental implications of substituting inorganic phosphorus with Ronozyme Phytase in intensive pig production in Denmark. Methods. Life cycle assessment is used as an analytical tool, and modelling...... pig production is justified by major advantages in terms of avoided contributions to global warming, acidification, photochemical ozone formation and particularly nutrient enrichment and by significant energy savings and particularly phosphate savings. A single trade-off in terms of agricultural land......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...

  1. Phytases Improve Myo-Inositol Bioaccessibility in Rye Bread: A Study Using an In Vitro Method of Digestion and a Caco-2 Cell Culture Model

    Directory of Open Access Journals (Sweden)

    Emilia Katarzyna Cielecka

    2015-01-01

    Full Text Available Preparations of 6-phytase A (EC 3.1.3.26 and phytase B (acid phosphatase, EC 3.1.3.2 were applied alone and combined in the preparation of dough to estimate their catalytic potential for myo-inositol liberation from rye flour in the breadmaking technology. The experimental bread samples were ground after baking and subjected to determination of myo-inositol bioavailability by an in vitro method that simulated digestion in a human alimentary tract, followed by measurements of myo-inositol transport through enterocyte-like differentiated Caco-2 cells to determine its bioaccessibility. Myo-inositol content was measured by a high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD technique. The concentration of myo-inositol in the dialysates of control bread was 25.3 μg/mL, whereas in the dialysates of bread sample baked with 6-phytase A, the concentration increased to 35.4 μg/mL, and in the bread baked with phytase B to 64.98 μg/mL. Simultaneous application of both enzymes resulted in myo-inositol release of 64.04 μg/mL. The highest bioaccessibility of myo-inositol, assessed by the measurement of the passage through the Caco-2 monolayer was determined in the bread baked with the addition of 6-phytase A. Enzymatically modifi ed rye bread, particularly by the addition of 6-phytase A, may be therefore a rich source of a highly bioaccessible myo-inositol.

  2. Phytase supplementation increases bone mineral density, lean body mass and voluntary physical activity in rats fed a low-zinc diet.

    Science.gov (United States)

    Scrimgeour, Angus G; Marchitelli, Louis J; Whicker, Jered S; Song, Yang; Ho, Emily; Young, Andrew J

    2010-07-01

    Phytic acid forms insoluble complexes with nutritionally essential minerals, including zinc (Zn). Animal studies show that addition of microbial phytase (P) to low-Zn diets improves Zn status and bone strength. The present study determined the effects of phytase supplementation on bone mineral density (BMD), body composition and voluntary running activity of male rats fed a high phytic acid, low-Zn diet. In a factorial design, rats were assigned to ZnLO (5 mg/kg diet), ZnLO+P (ZnLO diet with 1500 U phytase/kg) or ZnAD (30 mg/kg diet) groups and were divided into voluntary exercise (EX) or sedentary (SED) groups, for 9 weeks. SED rats were significantly heavier from the second week, and no catch-up growth occurred in EX rats. Feed intakes were not different between groups throughout the study. ZnLO animals had decreased food efficiency ratios compared to both phytase-supplemented (ZnLO+P) and Zn-adequate (ZnAD) animals (Pbone mineral content (BMC), bone area (BA) and BMD than rats fed ZnLO diets; and in rats fed ZnAD diets these indices were the highest. The dietary effects on BMC, BA and BMD were independent of activity level. We conclude that consuming supplemental dietary phytase or dietary Zn additively enhances Zn status to increase BMD, LBM and voluntary physical activity in rats fed a low-Zn diet. While the findings confirm that bone health is vulnerable to disruption by moderate Zn deficiency in rats, this new data suggests that if dietary Zn is limiting, supplemental phytase may have beneficial effects on LBM and performance activity. (c) 2010 Elsevier Inc. All rights reserved.

  3. Subsequent somatic axis and bone tissue metabolism responses to a low-zinc diet with or without phytase inclusion in broiler chickens.

    Directory of Open Access Journals (Sweden)

    Siemowit Muszyński

    Full Text Available Zinc is required for normal bone development and cartilage formation. The purpose of this study was to assess the effect of with adding organic Zn (alone or phytase inclusion at the reduced dose to growing male Ross 308 chickens on somatic axis and bone tissue metabolism. 200 one-day old broilers were divided into the negative control group fed diet without Zn or phytase inclusion, positive control group receiving Zn in the 100% of daily recommended dose from ZnO, and two experimental groups fed diet introduced Zn in 25% of daily recommendation as a glycine chelate (Zn-Gly with or without phytase inclusion (500 FTU·kg-1. Supplemental organic Zn increased bone Zn and Mg content, serum IGF-1, growth hormone and leptin concentration. Additional phytase inclusion increased body weight gain, blood plasma Ca, Fe, Zn and osteocalcin concentration and tibia ash percentage when compared to the Zn-deprived control. Bone geometry, yield and ultimate strengths were enhanced in both organic Zn supplemented groups, and the overall mechanical strength parameters of bone were better in these groups than in the positive control group supplemented with standard dose of inorganic Zn. Also marked improvements in the thickness of articular and the growth plate cartilages as well as real bone volume and thickness of metaphyseal trabeculae were achieved in all broilers fed Zn-supplemented diet irrespective of phytase inclusion, however, the highest cancellous bone mass and the best trabecular structure were noted after ZnO supplementation. In concludion, although dietary organic Zn given to growing broilers in 25% of daily recommended dose improved general bone properties and mechanical strength, the obtained results do not allow to unambiguously state that organic Zn supplementation at this level, even after phytase inclusion, is sufficient for proper bone development.

  4. 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. Journal of Animal Physiology and Animal Nutrition © 2015 Blackwell Verlag GmbH.

  5. Subsequent somatic axis and bone tissue metabolism responses to a low-zinc diet with or without phytase inclusion in broiler chickens.

    Science.gov (United States)

    Muszyński, Siemowit; Tomaszewska, Ewa; Kwiecień, Małgorzata; Dobrowolski, Piotr; Tomczyk-Warunek, Agnieszka

    2018-01-01

    Zinc is required for normal bone development and cartilage formation. The purpose of this study was to assess the effect of with adding organic Zn (alone or phytase inclusion) at the reduced dose to growing male Ross 308 chickens on somatic axis and bone tissue metabolism. 200 one-day old broilers were divided into the negative control group fed diet without Zn or phytase inclusion, positive control group receiving Zn in the 100% of daily recommended dose from ZnO, and two experimental groups fed diet introduced Zn in 25% of daily recommendation as a glycine chelate (Zn-Gly) with or without phytase inclusion (500 FTU·kg-1). Supplemental organic Zn increased bone Zn and Mg content, serum IGF-1, growth hormone and leptin concentration. Additional phytase inclusion increased body weight gain, blood plasma Ca, Fe, Zn and osteocalcin concentration and tibia ash percentage when compared to the Zn-deprived control. Bone geometry, yield and ultimate strengths were enhanced in both organic Zn supplemented groups, and the overall mechanical strength parameters of bone were better in these groups than in the positive control group supplemented with standard dose of inorganic Zn. Also marked improvements in the thickness of articular and the growth plate cartilages as well as real bone volume and thickness of metaphyseal trabeculae were achieved in all broilers fed Zn-supplemented diet irrespective of phytase inclusion, however, the highest cancellous bone mass and the best trabecular structure were noted after ZnO supplementation. In concludion, although dietary organic Zn given to growing broilers in 25% of daily recommended dose improved general bone properties and mechanical strength, the obtained results do not allow to unambiguously state that organic Zn supplementation at this level, even after phytase inclusion, is sufficient for proper bone development.

  6. Phytases Improve Myo-Inositol Bioaccessibility in Rye Bread: A Study Using an In Vitro Method of Digestion and a Caco-2 Cell Culture Model

    Science.gov (United States)

    Cielecka, Emilia Katarzyna; Pierzchalska, Małgorzata; Żyła, Krzysztof

    2015-01-01

    Summary Preparations of 6-phytase A (EC 3.1.3.26) and phytase B (acid phosphatase, EC 3.1.3.2) were applied alone and combined in the preparation of dough to estimate their catalytic potential for myo-inositol liberation from rye flour in the breadmaking technology. The experimental bread samples were ground after baking and subjected to determination of myo-inositol bioavailability by an in vitro method that simulated digestion in a human alimentary tract, followed by measurements of myo-inositol transport through enterocyte- -like differentiated Caco-2 cells to determine its bioaccessibility. Myo-inositol content was measured by a high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) technique. The concentration of myo-inositol in the dialysates of control bread was 25.3 µg/mL, whereas in the dialysates of bread sample baked with 6-phytase A, the concentration increased to 35.4 µg/mL, and in the bread baked with phytase B to 64.98 µg/mL. Simultaneous application of both enzymes resulted in myo-inositol release of 64.04 µg/mL. The highest bioaccessibility of myo-inositol, assessed by the measurement of the passage through the Caco-2 monolayer was determined in the bread baked with the addition of 6-phytase A. Enzymatically modified rye bread, particularly by the addition of 6-phytase A, may be therefore a rich source of a highly bioaccessible myo- -inositol. PMID:27904333

  7. Effects of zinc oxide and microbial phytase on digestibility of calcium and phosphorus in maize-based diets fed to growing pigs.

    Science.gov (United States)

    Blavi, L; Sola-Oriol, D; Perez, J F; Stein, H H

    2017-02-01

    An experiment was conducted to test the hypothesis that inclusion of Zn at a pharmacological level in diets fed to pigs affects apparent total tract digestibility (ATTD) of Ca and P and standardized total tract digestibility (STTD) of Ca. The second hypothesis was that inclusion of microbial phytase increases the ATTD of Ca and P and the STTD of Ca regardless of the concentration of Zn in the diet. Fifty-six growing barrows (15.4 ± 1.9 kg average BW) were allotted to a randomized complete block design with 7 dietary treatments and 8 pigs per treatment. A maize-based basal diet was formulated with either 0 or 2,400 mg/kg Zn from ZnO and 0, 1,000, or 3,000 units of phytase (FTU) per kilogram. A Ca-free diet was used to determine basal endogenous losses of Ca. Experimental diets were fed for 13 d, and feces were collected from the feed provided from d 6 to 11 using the marker-to-marker approach; urine was also collected from d 6 to 11. Retention of Ca, ATTD of Ca, and STTD of Ca increased ( phytase in the diet increased and were less ( phytase increased in the diet, but the increase was greater if ZnO was not added than if ZnO was added to the diet (interaction, phytase in the diets. Inclusion of microbial phytase increased the ATTD and STTD of Ca in diets and also the ATTD of P.

  8. 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

  9. INTERACTION BETWEEN DIETARY MINERAL AND PHYTASE ON BIOLOGICAL PERFORMANCES OF JAPANESE FLOUNDER, Paralichthys olivaceus. PART II. MINERAL DIGESTIBILITY AND VERTEBRAL MINERAL CONTENT

    Directory of Open Access Journals (Sweden)

    Asda Laining

    2015-06-01

    Full Text Available Interactive effects between dietary inorganic phosphorus (IP and phytase (P on mineral digestibility and vertebral mineral content were investigated in a 30 days feeding trial followed by three weeks digestibility trial with Japanese flounder, Paralichthys olivaceus. Eight experimental diets were formulated based on two levels of dietary Ca at 0% and 0.2% combined with either 0% or 0.25% of dietary IP and either with 0 and 2,000 fytase unit (FTU/kg of phytase in diet, respectively. Result indicated that digestibility of total phosphorus significantly increased by three dietary compounds where the highest was observed in fish fed diet contained 0.25% IP and 2,000 FTU phytase/kg and dietary Ca also included in diet. Significant interaction was only detected between dietary IP and P on this parameter. Supplementation of IP and Ca not phytase significantly improved Ca digestibility. Ca digestibility was very poor when dietary IP and Ca were not supplemented in diet even with when phytase supplemented in diet. There was significant interaction between dietary IP and Ca on Ca digestibility. Vertebral total phosphorus, Ca, and Mg content as well as Ca:P ratio were significantly enhanced by dietary IP and phytase. Dietary Ca has significant effect only on vertebral total phosphorus. Interaction between dietary IP and Ca was significantly found on vertebral Ca content and Ca:P ratio. No significant second-order interaction was observed among the three dietary mineral on overall parameters. Based on total phosphorus and Ca digestibility as well vertebral phosphorus content found in this study, dietary IP, Ca, and phytase at rate of 0.25%, 0.2%, and 2,000 FTU phytase/kg diet, respectively are needed to supplement in diet for a better mineral absorption and bone mineralization.

  10. INTERACTION BETWEEN DIETARY MINERAL AND PHYTASE ON BIOLOGICAL PERFORMANCES OF JAPANESE FLOUNDER, Paralichthys olivaceus. PART I. GROWTH, FEED INTAKE, AND WHOLE BODY MINERAL CONTENT

    Directory of Open Access Journals (Sweden)

    Asda Laining

    2013-06-01

    Full Text Available In order to determine the effect of dietary calcium (Ca, inorganic phosphorus (IP, and phytase (P supplementation in marine fish, a 2 x 2 x 2 factorial design was arranged to observe the interrelationship between dietary Ca and IP with the presence of dietary phytase. Two levels of dietary Ca at 0% and 0.2% combined with either 0% or 0.25% of dietary IP and either with 0 and 2,000 FTU phytase/kg diet, respectively to formulate eight experimental diets. SPI-based diet was used as basal diet and the sources of Ca, IP, and phytase were similar to those used in the previous experiment. Juvenile Japanese flounder with initial body weight around 0.6 g was fed the test diets. After 30 days of feeding trial, the results showed that both dietary IP and phytase, but not dietary Ca significantly enhanced the growth and feed intake. The highest growth was achieved in fish fed a diet containing the Ca, IP, and phytase supplement among groups. Fish fed diet without the three dietary supplements had the lowest SGR and did not significantly improve by supplementing dietary Ca. Feed intake (FI and was significantly influenced by dietary IP and phytase, not dietary Ca while feed conversion ratio (FCR was significantly affected by all dietary treatments. Interaction effect was detected between dietary Ca and IP, between dietary IP and P on FCR. Total phosphorus content in whole body was significantly increased by supplementing all dietary treatments which was highest in fish fed 0.25 IP/0.2 Ca/P. Significant interaction was observed between dietary IP and P on this parameter. Whole body Ca content was significantly improved by either dietary IP or Ca and not dietary P. As conclusion even without inorganic Ca supplement, dietary IP at level of 0.25% or 2,000 FTU phytase/kg diet could enhance growth and FI of fish as well as whole body phosphorus content of juvenile Japanese flounder when diet basal contained organic Ca around 1.2%.

  11. 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.

  12. 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.

  13. Purification and characterization of two distinct acidic phytases with broad pH stability from Aspergillus niger NCIM 563.

    Science.gov (United States)

    Soni, S K; Magdum, A; Khire, J M

    2010-11-01

    Aspergillus niger NCIM 563 produced two different extracellular phytases (Phy I and Phy II) under submerged fermentation conditions at 30°C in medium containing dextrin-glucose-sodium nitrate-salts. Both the enzymes were purified to homogeneity using Rotavapor concentration, Phenyl-Sepharose column chromatography and Sephacryl S-200 gel filtration. The molecular mass of Phy I and II as determined by SDS-PAGE and gel filtration were 66, 264, 150 and 148 kDa respectively, indicating that Phy I consists of four identical subunits and Phy II is a monomer. The pI values of Phy I and II were 3.55 and 3.91, respectively. Phy I was highly acidic with optimum pH of 2.5 and was stable over a broad pH range (1.5-9.0) while Phy II showed a pH optimum of 5.0 with stability in the range of pH 3.5-9.0. Phy I exhibited very broad substrate specificity while Phy II was more specific for sodium phytate. Similarly Phy II was strongly inhibited by Ag(+), Hg(2+) (1 mM) metal ions and Phy I was partially inhibited. Peptide analysis by Mass Spectrometry (MS) MALDI-TOF also indicated that both the proteins were totally different. The K(m) for Phy I and II for sodium phytate was 2.01 and 0.145 mM while V(max) was 5,018 and 1,671 μmol min(-1) mg(-1), respectively. The N-terminal amino acid sequences of Phy I and Phy II were FSYGAAIPQQ and GVDERFPYTG, respectively. Phy II showed no homology with Phy I and any other known phytases from the literature suggesting its unique nature. This, according to us, is the first report of two distinct novel phytases from Aspergillus niger.

  14. 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. © 2015 Poultry Science Association Inc.

  15. 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,...

  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

    DEFF Research Database (Denmark)

    Abdollahi, M.R.; Duangnumsawang, Y.; Kwakkel, R.P.

    2016-01-01

    was investigated. The effect of phytase on the CAID and TTR of P in birds with access to a separate Ca source was of particular interest. Five dietary Ca concentrations (1.3, 4.3, 7.3, 10.3 and 13.3 g/kg) and two inclusion levels (0 or 1000 phytase units [FYT/kg] of feed) of a 6-phytase were evaluated in a 5 × 2...... factorial arrangement of treatments. All diets, based on wheat and soybean meal, were formulated to be equivalent in respect of total and non-phytate P contents (5.7 and 3.0 g/kg, respectively). A total of 480, one-day-old male broilers were allocated to 48 cages (8 birds/cage), and cages were randomly...... 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 g Ca/kg, whereas in the phytase...

  17. Effect of Quantum phytase on nutrient digestibility and bone ash in White Leghorn laying hens fed corn-soybean meal-based diets.

    Science.gov (United States)

    Hughes, A L; Dahiya, J P; Wyatt, C L; Classen, H L

    2009-06-01

    The efficacy of an Escherichia coli 6-phytase supplementation (Quantum) on nutrient digestibility-retention and bone ash in laying hens fed corn-soybean meal (CSM) diets was investigated. White Leghorn hens (Shaver and Bovan strains) were fed CSM diets containing 0.35% (positive control, PC), 0.25% (negative control 1, NC1), or 0.15% (negative control 2, NC2) nonphytate P from 21 to 61 wk of age. Six more diets were manufactured by supplementing the negative control diets with 200, 400, and 600 units per kilogram of exogenous phytase resulting in a total of 9 treatments. Each dietary treatment x strain subclass was replicated twice with 6 hens per replication. Fecal and ileal digesta samples were collected at 42 wk of age to determine apparent nutrient digestibility or retention. Left tibiae were collected at 42 and 61 wk of age to determine bone ash. The coefficients for ileal digestibility and fecal retention for protein were higher (P ash percentage was higher (P < 0.05) in 61-wk-old hens fed 200 or 400 units per kilogram of phytase-supplemented NC2 diets. Significantly higher diet AME and fecal protein retention were demonstrated for Shaver hens in comparison to the Bovan hens. Overall, the Quantum phytase was not efficacious at improving nutrient digestibility-retention in laying hens fed CSM diets deficient in nonphytate P.

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

    OpenAIRE

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

    2015-01-01

    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 (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 ...

  19. Effects of phytase, cellulase, and dehulling treatments on iron and zinc in vitro solubility in faba bean (Vicia faba L.) Flour and Legume Fractions.

    Science.gov (United States)

    Luo, Yu-Wei; Xie, Wei-Hua; Cui, Qun-Xiang

    2010-02-24

    Simulations of gastrointestinal digestion were used to try to identify the nature of the complexes between antinutritional factors and iron and zinc in faba bean and legume fractions. In digestible residue of raw faba bean flour, simultaneous action of cellulase and phytases made it possible to release about 28% units more iron than that released with the treatment without enzymes. About 49.8% of iron in raw faba bean flour was solubilized after in vitro digestion and simultaneous action of cellulase and phytase. In the hull fraction, the action of phytases and the simultaneous action of cellulase and phytase allowed about 7 and 35% units of additional zinc to be solubilized, respectively. Single enzymatic degradation of phytates from dehulled faba bean allowed solubilization from 65 to 93% of zinc, depending upon the treatment. In dehulled faba bean, iron was chelated by phytates and by fibers, whereas zinc was almost exclusively chelated by phytates. In the hull of faba bean, a high proportion of iron was chelated by iron-tannins, while the rest of iron as well as the majority of zinc were chelated in complexes between phytates and fibers.

  20. 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.

  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, H.; Tauson, Anne-Helene

    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...... × 4 Latin square with four pigs fed four diets. A basal wheat/barley-based diet was fed either as non-heat-treated or heat-treated (steam-pelleted at 90°C). The heat-treatment resulted in an inactivation of plant phytase below detectable level. Diet 1 (non-heat-treated basal diet fed dry); diet 2...... (heat-treated basal diet fed dry); diet 3 (as diet 2 but with microbial phytase (750 FTU/kg as fed) fed dry); diet 4 (as diet 3 fed liquid (fermented for 17.5 h nighttime and 6.5 h daytime at 20°C with 50% residue in the tank)). Chromic oxide (Cr2O3) was included as marker and ATTD was determined both...

  2. 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...

  3. Performance efficiency of feed utilization, relative growth rate, and survival rate of common carp (Cyprinus carpio) through the addition of phytase in the feed

    Science.gov (United States)

    Rachmawati, D.; Samidjan, I.

    2018-04-01

    The purpose of this study was to determine the effect of adding phytase enzyme in the feed on digestibility of feed, efficiency of feed utilization, relative growth rate and survival rate of Common carp (Cyprinus carpio). Fish samples in this research were Common carp with an average - weight of 3.34 ± 0,16 g/fish. The treatments were adding the phytase enzyme in the feed with the different level of doses. Those were A (0 U kg-1 feed), B (500 U kg-1 feed), C (1.000 U kg-1 feed g) and D (1.500 U kg-1 feed). Observation was conducted on digestibility of protein (ADCP), digestibility of phosphor (ADCF), efficiency of feed utilization (EFU), relative growth rate (RGR), protein efficiency ratio (PER), feed conversion ratio (FCR), survival rate (SR) and water quality parameters. The results show that the addition of phytase enzyme significantly (P0.05) affected on SR of common carp. Based on results, it was concluded that optimum doses of phytase enzyme feed in terms of digestibility of feed, efficiency utilization of Feed and growth rate of Common carp ranges from 943 to 1100 U kg-1 feed

  4. 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

  5. The efficacy of Aspergillus niger phytase in rendering phytate phosphorus available for absorption in pigs is influenced by pig physiological status

    NARCIS (Netherlands)

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

    1997-01-01

    We performed an experiment with 112 piglets, 32 growing-finishing pigs, and 12 sows during pregnancy and lactation to test the hypotheses that apparent total tract digestibilities of P and Ca as well as the efficacy of Aspergillus niger phytase in rendering phytate P available in pigs depend on pig

  6. Effect of a microbial phytase on growth performance, plasma parameters and apparent ileal amino acid digestibility in Youxian Sheldrake fed a low-phosphorus corn-soybean diet

    Directory of Open Access Journals (Sweden)

    Shaoping He

    2017-10-01

    Full Text Available Objective This study investigated the effect of microbial phytase supplementation on growth performance, tibia ash, plasma parameters, apparent ileal digestibility (AID of amino acid (AA and apparent digestibility of nutrients in Youxian Sheldrakes fed with low-phosphorus (P corn-soybean diets. Methods A total of 350 Youxian Sheldrakes (7d old were randomly divided into 5 treatment groups: positive control (PC group has adequate available P diet (0.42% and 0.38%, starter and grower, negative control (NC group were deficient in available P (0.32% and 0.28%, starter and grower and NC diet was supplemented with 3 levels of microbial phytase (500, 750, and 1,000 U/kg. Results Dietary supplementation of phytase in NC diet improved the average daily gain, increased the levels of serum calcium (Ca, tibia Ca and P, AID of AA and apparent digestibility of energy and Ca in starter stage (p<0.05. There was an increased (p<0.001 in the utilization of P from 17.3% to 23.9%. Phytase supplementation (1,000 U/kg has shown that the AID of His, Thr, Val, indispensable AA, Glu, Pro, and dispensable AA was higher (p<0.05 than that of NC. Moreover, phytase supplementation improved (p<0.05 serum and tibia Ca and P, AID of AA and apparent digestibility of dry matter, crude protein, energy, P and Ca, and reduced (p<0.05 feed to gain ratio (F/G and the levels of serum alkaline phosphatase in grower stage. Likewise, an increase (p<0.001 in the utilization of P was noticed from 12.6% to 17.2%. Supplement phytase at 750 U/kg improved the AID of His, Thr, Asp, Cys, Pro, and Ser (p<0.05. Conclusion The microbial phytase supplement could improve growth performance, AID of some AA and apparent utilization of other nutrients in Youxian Sheldrakes, and reduce excreta P load to environment.

  7. 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

    The effect of a supplemental fungal phytase on performance and phosphorus availability by juvenile rainbow trout fed diets with a high inclusion of plant based protein and on the magnitude and composition of the waste phosphorus production was tested in a 2 × 3 factorial design at a temperature....../suspended phosphorus waste output from fish fed the phytase supplemented diet containing 0.71% available phosphorus, suggesting that the phosphorus requirement was reached at this phosphorus level. Consistent with this, there was a substantial increase in the dissolved/suspended phosphorus waste output from fish fed...... the phytase supplemented diet containing 0.81% available phosphorus, suggesting that the phosphorus requirement was exceeded in this group. This study demonstrated that phytase supplementation will be advantageous to the fish and the environment if supplemented to low-phosphorus diets containing a large share...

  8. 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.

  9. 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.

  10. Effects of graded levels of microbial phytase on apparent total tract digestibility of calcium and phosphorus and standardized total tract digestibility of phosphorus in four sources of canola meal and in soybean meal fed to growing pigs.

    Science.gov (United States)

    She, Y; Liu, Y; Stein, H H

    2017-05-01

    One hundred twenty pigs were used to determine effects of graded levels of microbial phytase on the apparent total tract digestibility (ATTD) of P and Ca and the standardized total tract digestibility (STTD) of P in 4 sources of canola meal and in 1 source of soybean meal (SBM) fed to growing pigs. The 4 sources of canola meal were produced from 1 source of high-protein canola seeds and 2 sources of conventional canola seeds with 1 of the conventional canola seeds being divided into 2 separate batches before crushing. Pigs (16.2 ± 5.3 kg initial BW) were individually housed in metabolism crates and were randomly allotted to 1 of 20 diets in a 5 × 4 factorial arrangement of treatments with 5 ingredients and 4 levels of phytase. There were 6 replicate pigs per diet. Five basal diets based on high-protein canola meal (CM-HP), high-temperature processed canola meal (CM-HT), low-temperature processed canola meal (CM-LT), conventional canola meal (CM-CV), or SBM were formulated. The basal diets contained no phytase. Fifteen additional diets were prepared by adding approximately 500, 1,500, or 2,500 phytase units/kg to each of the 5 basal diets. Feces were quantitatively collected for 5 d based on the marker-to-marker approach after a 7-d adaptation period. Results indicated that supplementation of microbial phytase increased (linear, phytase also increased (linear and quadratic, phytase was added, but no differences were observed in the ATTD and STTD of P in SBM, CM-HP, CM-HT, or CM-CV if the highest amount of phytase were added (interaction, phytase on the STTD of P in CM-HP, CM-HT, CM-LT, CM-CV, and SBM. In conclusion, inclusion of graded levels of microbial phytase increased the ATTD and STTD of P in CM-HP, CM-HT, CM-LT, CM-CV, and SBM and the response to microbial phytase added to each ingredient can be predicted by regression equations.

  11. 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.

  12. Conservation of the human integrin-type beta-propeller domain in bacteria.

    Directory of Open Access Journals (Sweden)

    Bhanupratap Chouhan

    Full Text Available Integrins are heterodimeric cell-surface receptors with key functions in cell-cell and cell-matrix adhesion. Integrin α and β subunits are present throughout the metazoans, but it is unclear whether the subunits predate the origin of multicellular organisms. Several component domains have been detected in bacteria, one of which, a specific 7-bladed β-propeller domain, is a unique feature of the integrin α subunits. Here, we describe a structure-derived motif, which incorporates key features of each blade from the X-ray structures of human αIIbβ3 and αVβ3, includes elements of the FG-GAP/Cage and Ca(2+-binding motifs, and is specific only for the metazoan integrin domains. Separately, we searched for the metazoan integrin type β-propeller domains among all available sequences from bacteria and unicellular eukaryotic organisms, which must incorporate seven repeats, corresponding to the seven blades of the β-propeller domain, and so that the newly found structure-derived motif would exist in every repeat. As the result, among 47 available genomes of unicellular eukaryotes we could not find a single instance of seven repeats with the motif. Several sequences contained three repeats, a predicted transmembrane segment, and a short cytoplasmic motif associated with some integrins, but otherwise differ from the metazoan integrin α subunits. Among the available bacterial sequences, we found five examples containing seven sequential metazoan integrin-specific motifs within the seven repeats. The motifs differ in having one Ca(2+-binding site per repeat, whereas metazoan integrins have three or four sites. The bacterial sequences are more conserved in terms of motif conservation and loop length, suggesting that the structure is more regular and compact than those example structures from human integrins. Although the bacterial examples are not full-length integrins, the full-length metazoan-type 7-bladed β-propeller domains are present, and sometimes two tandem copies are found.

  13. 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.

  14. Effect of dietary calcium level and source on mineral utilisation by piglets fed diets containing exogenous phytase.

    Science.gov (United States)

    Schlegel, P; Gutzwiller, A

    2017-10-01

    Calcium and phosphorus are essential minerals, closely linked in digestive processes and metabolism. With widespread use of low P diets containing exogenous phytase, the optimal dietary Ca level was verified. The 40-day study evaluated the effects of Ca level (4, 7 and 10 g/kg diet) and Ca source (Ca from CaCO 3 and from Lithothamnium calcareum) on mineral utilisation in 72 piglets (7.9 ± 1.0 kg BW) fed an exogenous phytase containing diet with 2.9 g digestible P/kg. Measured parameters were growth performance, stomach mineral solubility, bone breaking strength and urinary, serum and bone mineral concentration. The apparent total tract digestibility of minerals was also assessed in the two diets with 7 g Ca/kg, using 12 additional pigs. Regardless of Ca source, increasing dietary Ca impaired feed conversion ratio, increased urinary pH, increased serum and urinary Ca, decreased serum and urinary P, decreased serum Mg and increased urinary Mg, increased serum AP activity, decreased bone Mg increased bone Zn. Bone breaking strength was improved with 7 compared to 4 g Ca/kg. Compared to CaCO 3 , Ca from Lithothamnium calcareum increased serum Mg and with, 10 g Ca/kg, it limited body weight gain. The dose response of Ca in a diet with 2.9 g digestible P/kg and including exogenous phytase indicated that: (i) a low dietary Ca was beneficial for piglet growth, but was limiting the metabolic use of P; (ii) a high dietary Ca level impaired P utilisation; (iii) the optimal P utilisation and bone breaking strength was obtained with a dietary Ca-to-digestible P ratio of 2.1 to 2.4:1; (iv). Increasing dietary Ca reduced Mg utilisation, but not Zn status, when fed at adequate level. Finally, Ca from Lithothamnium calcareum had similar effects on Ca and P metabolism as CaCO 3 , but impaired growth when fed at the highest inclusion level. Journal of Animal Physiology and Animal Nutrition © 2016 Blackwell Verlag GmbH.

  15. 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.

    2016-01-01

    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. PMID:26740131

  16. 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.

  17. Detoxification of castor bean residues and the simultaneous production of tannase and phytase by solid-state fermentation using Paecilomyces variotii.

    Science.gov (United States)

    Madeira, Jose Valdo; Macedo, Juliana Alves; Macedo, Gabriela Alves

    2011-08-01

    In this work, we introduce a biological detoxification method that converts toxic waste from castor beans into animal feed material. This method simultaneously induces the production of tannase and phytase by Paecilomyces variotii; both enzymes have high levels of activity and have the potential to be used in feedstuffs because they decrease overall anti-nutritional factors. The maximum tannase and phytase activities obtained were 2600 and 260 U/g after 48 and 72 h, respectively. SDS-PAGE electrophoresis of the fermented castor cake extracts revealed a reduction in ricin bands during fermentation, and the bands were no longer visible after 48 h. The cytotoxicity of the extracts was evaluated by MTT testing on RAW cells, and a progressive increase in cellular viability was obtained, reaching almost 100% after 72 h of fermentation. Copyright © 2011 Elsevier Ltd. All rights reserved.

  18. Energy and protein levels in diets containing phytase for broilers from 22 to 42 days of age: performance and nutrient excretion

    Directory of Open Access Journals (Sweden)

    Adriano Kaneo Nagata

    2011-08-01

    Full Text Available This study was conducted in order to evaluate the influence of different levels of metabolizable energy and crude protein in diets formulated according to the ideal protein concept with phytase supplementation on performance and nutrient excretion of broilers from 22 to 42 days age. It was used 1,500 Coob lineage broilers at 22 days of age and with initial weight of 833 ± 7g, distributed in completely randomized design in a 3 × 3 + 1 factorial scheme composed of three levels of correct apparent metabolizable energy (2,950; 3,100 and 3,250 kcal/kg, three levels of crude protein (14, 16 and 18% and a control treatment, totaling ten treatments with six repetitions of 25 birds each. All diets, with the exception of the control, were supplemented with phytase. For determination of excretion of pollutants, it was used 180 broilers from the same lineage at 35 days of age,placed in metabolic cages, with ten treatments each one with six repetitions and three birds per experimental unit. The protein and energy levels in diets containing phytase influenced feed intake, weight gain, feed conversion and excretion of nitrogen, phosphorus, calcium, potassium, copper and zinc by the birds. The corrected apparent metabolizable energy level in the diets for broilers in the studied period must be increased up to 3,250 kcal/kg of metabolizable energy and the levels of crude protein, calcium and phosphorus must be reduced down to 18, 0.70 and 0.31%, respectively, provided that supplemented with amino acids and phytase to improve the performance and to reduce excretion of pollutants by birds.

  19. A new process for simultaneous production of tannase and phytase by Paecilomyces variotii in solid-state fermentation of orange pomace.

    Science.gov (United States)

    Madeira, Jose Valdo; Macedo, Juliana Alves; Macedo, Gabriela Alves

    2012-03-01

    The production of enzymes such as tannases and phytases by solid-state fermentation and their use in animal feed have become a subject of great interest. In the present work, Paecilomyces variotii was used to produce tannase and phytase simultaneously. Solid-state fermentation, a process initially designed for tannase production, was implemented here using orange pomace as substrate. Orange pomace is the waste product of the large orange juice industry in Brazil, and it has also been used as an ingredient in animal feed. In addition to enzymatic production, biotransformation of the phenolic content and antioxidant capacity of the orange pomace were analyzed after fermentation. Fermentation conditions, namely moisture level and tannic acid concentration rate, were studied using CCD methodology. The response surface obtained indicated that the highest tannase activity was 5,000 U/gds after 96 h at 59% (v/w) and 3% (w/w) and that of phytase was 350 U/gds after 72 h at 66% (v/w) and 5.8% (w/w) of moisture level and tannic acid concentration, respectively. The amount of tannase production was similar to the levels achieved in previous studies, but this was accomplished with a 7% (w/w) reduction in the amount of supplemental tannic acid required. These results are the first to show that P. variotii is capable of producing phytase at significant levels. Moreover, the antioxidant capacity of orange pomace when tested against the free radical ABTS was increased by approximately tenfold as a result of the fermentation process.

  20. Effect of diet phase change, dietary Ca and P level and phytase on bird performance and real-time gizzard pH measurements.

    Science.gov (United States)

    Lee, S A; Dunne, J; Mottram, T; Bedford, M R

    2017-06-01

    In this study, a novel capsule technique was used to capture real-time pH readings from the gizzard over several hours, in response to different dietary treatments. 1. The first experiment was a preliminary study into capsule administration and pH recordings using 9 male Ross 308 broilers from 20 d. In the second experiment, broilers (576) were fed in two phases (0-21 and 21-42 d) with 4 treatment groups; low and adequate Ca and AvP diets with and without Quantum Blue phytase (1500 FTU/kg). Capsules were administered to 8 birds from each treatment group, pre and post diet phase change, with readings captured over a 2.5 h period. 2. Phytase addition improved body weight gain (BWG) and feed conversion ratio (FCR) of birds fed low dietary Ca, while having no significant effect on birds fed adequate Ca diets. Unexpectantly, diets with higher Ca levels gave a lower average gizzard pH compared to the low Ca diet. Phytase addition, irrespective of Ca level, increased average gizzard pH. Fluctuations in gizzard pH (0.6-3.8) were observed across all treatment groups. Higher frequencies of pH readings below pH 1.0 were seen in birds fed an adequate Ca diet and with phytase supplementation of a low Ca diet. 3. These results signify the potential use of capsule techniques to monitor real-time pH changes. The implication on gastric protein and fibre hydrolysis as a result of large fluctuations in pH should be considered.

  1. 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)

    Moretti, Diego; Schuth, Stephan; 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 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 (Pphytase increased fractional iron absorption 3.2 times, suggesting it could be a possible strategy to decrease PA in cereal-based porridges. PMID:24124445

  2. EFFECTS OF THE COMBINATION OF NON-PHYTATE PHOSPHORUS, PHYTASE AND 25-HYDROXYCHOLECALCIFEROL ON THE PERFORMANCE AND MEAT QUALITY OF BROILER CHICKENS

    Directory of Open Access Journals (Sweden)

    L Ren

    2015-09-01

    Full Text Available ABSTRACTThis experiment was conducted to evaluate the combination effect of low dietary non-phytate phosphorus (NPP concentrations, phytase (PHY levels, and 25-hydroxycholecalciferol (25-OH-D3 levels on the growth performance and meat quality of broilers. Two levels of NPP, two levels of PHY, and two levels of 25-OH-D3 resulted in a 2�2�2 factorial arrangements, with eight treatments (TRT. The birds on TRT 1-4 were fed diet 1 (NRC NPP was reduced by 0.1 and the birds on TRT 5-8 were fed with diet 2 (NRC NPP was reduced by 0.2. Each diet was mixed with different levels PHY and 25-OH-D3. Performance and meat quality parameters were measured. Results showed that during entire experiment the most advantageous effects were obtained with TRT 3 (NRC NPP reduced by 0.1 + 600 U/kg phytase + 34.5�g/kg 25-OH-D3 and TRT 4 (NRC NPP reduced by 0.1 + 600 U/kg phytase + 69�g/kg 25-OH-D3. The lowest body weight gain (BWG and feed intake(FI were observed with TRT 5 (NRC NPP reduced by 0.2 + 300 U/kg phytase + 34.5�g/kg 25-OH-D3. Lowering NRC NPP by 0.1 to 0.2 significantly reduced weight gain (WG (p0.05 and produced small decrease in FI. BWG, FI and feed conversion ratio were not influenced (p>0.05 by different PHY or 25-OH-D3 levels. In addition, the meat color, pH, and shear force were not affected by the different NPP, PHY or 25-OH-D3levels.

  3. Supplementation of broiler diets with high levels of microbial protease and phytase enables partial replacement of commercial soybean meal with raw, full-fat soybean.

    Science.gov (United States)

    Erdaw, M M; Perez-Maldonado, R A; Iji, P A

    2018-02-22

    A 3 × 3 + 1 factorial, involving three levels of protease (0, 15,000 or 30,000 PROT/kg) and three levels of phytase (1,000, 2,000 or 3,000 FYT/kg), was used to evaluate the effect of replacing commercial soybean meal (SBM) with raw, full-fat soybean (RFSB) at 75 g/kg of diet for broilers. A control diet was used for comparison. Each treatment was replicated six times, with nine birds per replicate. The concentration of trypsin inhibitors (TIs) in the test diets was approximately 10,193.4 TIU/kg. Regardless of enzyme supplementation, feed intake (FI) and body weight gain (BWG) of birds in the control group were superior to those on the test diets. Birds that received the protease-free test diets had reduced FI and BWG, but when supplemented with protease, were similar to the control diet in BWG, FI (except 0-35 days) and feed conversion ratio (FCR). When the test diet was supplemented with elevated levels (extradose) of protease and phytase, the BWG was improved during 0-10 days (p = .05) and 0-24 days (p replace SBM in broiler diets, provided the diets are supplemented with elevated levels of protease and phytase. © 2018 Blackwell Verlag GmbH.

  4. Effect of heat-treatment, phytase, xylanase and soaking time on inositol phosphate degradation in vitro in wheat, soybean meal and rapeseed cake

    DEFF Research Database (Denmark)

    Blaabjerg, Karoline; Carlsson, N G; Hansen-Møller, Jens

    2010-01-01

    An in vitro method was used to evaluate the degradation of myo-inositol hexakisphosphate (InsP6) in non-heat-treated wheat (NHW), heat-treated wheat (HW), soybean meal (SBM) or rapeseed cake (RSC) soaked separately or in combination. The feedstuffs were soaked in water (20 °C) and samples were...... heat-treatment and soaking time (P≤0.001). This was mainly due to a smaller proportion of non-degraded InsP6-P at 24 h in HW compared with NHW (0.13 vs. 0.47) (P≤0.001) possibly caused by structural changes imposed by the heat-treatment. In SBM, RSC, SBM/NHW or RSC/NHW, the InsP6 degradation...... was affected by the interaction between phytase addition and soaking time (P≤0.001) as phytase reduced the proportion of non-degraded InsP6-P at 2, 4, 8 or 24 h. Soaking of NHW, SBM or RSC (without phytase) separately resulted in a limited InsP6 degradation, whereas a pronounced InsP6 degradation occurred when...

  5. Effects of a high dose of microbial phytase and myo-inositol supplementation on growth performance, tibia mineralization, nutrient digestibility, litter moisture content, and foot problems in broiler chickens fed phosphorus-deficient diets.

    Science.gov (United States)

    Farhadi, D; Karimi, A; Sadeghi, Gh; Rostamzadeh, J; Bedford, M R

    2017-10-01

    A total of 660 one-day-old Ross 308 broiler chicks were randomly distributed into eleven dietary treatments. Treatments included a maize-soybean meal-based diet with recommended calcium (Ca) and non-phytate phosphorus (nPP) (positive control; PC), an nPP-deficient diet (negative control; NC), NC diets supplemented with different levels of phytase (0, 500, 1,000, 2,000, 3,000, 4,000, 5,000, and 6,000 FTU/kg), a NC diet plus 0.15% myo-inositol, and a NC diet with reduced Ca level (Ca to nPP ratio same as PC). Feeding the NC diet had no effects on birds' body weight (BW), weight gain (WG), feed intake (FI), and feed conversion ratio (FCR), but decreased (P Phytase supplementation at ≥4,000 FTU/kg improved (P phytase returned (P phytase in a dose-dependent manner, especially at ≥4,000 FTU/kg levels, was effective in overcoming the negative consequences of NC diets, primarily due to the ability to improve nutrient utilization. In addition, reducing the Ca level or supplementation of inositol of NC diet can correct some the negative effects of feeding a NC diet confirming the negative effect of a wide Ca: P ratio in a P-deficient diet and suggesting that inositol may play a role in the response to phytase addition. © 2017 Poultry Science Association Inc.

  6. Response of broilers to feeding low-calcium and phosphorus diets plus phytase under different environmental conditions: body weight and tibiotarsus mineralization.

    Science.gov (United States)

    Catalá-Gregori, P; García, V; Hernández, F; Madrid, J; Cerón, J J

    2006-11-01

    Three experiments on Ross broiler chickens were conducted in 3 locations: cages (Experiment 1), floor pens (Experiment 2), and commercial farms (Experiment 3). The effect of low-total P (TP) wheat-soybean based diets plus microbial phytase (Natuphos) was evaluated. Four experimental starter and finisher diets were used in a 2-phase feeding program, as follows: control diet (SC until 21 d, FC from 22 to 42 d); 2 diets (SL400 and SL600 until 21 d, FL400 and FL600 from 22 to 42 d) with low TP (0.61% for starter and 0.54% for finisher), including 400 and 600 U/kg of phytase, respectively; and a very low-TP (0.52% for starter and 0.44% for finisher) diet (SVL600 until 21 d, FVL600 from 22 to 42 d) with 600 U/kg of phytase. In Experiment 1 (broilers in cages had movement limitation and no access to litter), no differences in BW, tibiotarsus mineralization, or mineral metabolism were observed among diets. In Experiment 2 (broilers in floor pens had movement limitation and access to litter), at 21 d of age, the lowest tibiotarsus ash percentage and BW were shown by birds fed the SVL600 diet. At 42 d of age, broilers fed the FC diet were the lightest. For the rest of the parameters of tibiotarsus mineralization and mineral metabolism measured in Experiment 2, no differences were shown. In Experiment 3 (broilers in commercial farms had access to litter without movement limitation), the BW of broilers fed the SC diet was the highest at 21 d of age. At 42 d of age, the broilers fed FL400 and FL600 diets were the heaviest. At the end of Experiment 3, broilers fed the FC diet had the highest dry litter Ca and P, whereas broilers fed the FVL600 diet had the lowest values. In conclusion, the very low-TP wheat-soybean based diet supplemented with 600 U/kg of phytase was sufficient to optimize all the parameters measured in Experiment 1 but not in Experiments 2 and 3. Therefore, when evaluating Ca and P in phytase-supplemented diets for broilers, it is necessary to bear in mind the

  7. 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.

  8. 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

  9. Phosphate, phytate and phytases in plants: from fundamental knowledge gained in Arabidopsis to potential biotechnological applications in wheat.

    Science.gov (United States)

    Secco, David; Bouain, Nadia; Rouached, Aida; Prom-U-Thai, Chanakan; Hanin, Moez; Pandey, Ajay K; Rouached, Hatem

    2017-11-01

    Phosphorus (P) is an essential macronutrient for all living organisms. In plants, P is taken up from the rhizosphere by the roots mainly as inorganic phosphate (Pi), which is required in large and sufficient quantities to maximize crop yields. In today's agricultural society, crop yield is mostly ensured by the excessive use of Pi fertilizers, a costly practice neither eco-friendly or sustainable. Therefore, generating plants with improved P use efficiency (PUE) is of major interest. Among the various strategies employed to date, attempts to engineer genetically modified crops with improved capacity to utilize phytate (PA), the largest soil P form and unfortunately not taken up by plants, remains a key challenge. To meet these challenges, we need a better understanding of the mechanisms regulating Pi sensing, signaling, transport and storage in plants. In this review, we summarize the current knowledge on these aspects, which are mainly gained from investigations conducted in Arabidopsis thaliana, and we extended it to those available on an economically important crop, wheat. Strategies to enhance the PA use, through the use of bacterial or fungal phytases and other attempts of reducing seed PA levels, are also discussed. We critically review these data in terms of their potential for use as a technology for genetic manipulation of PUE in wheat, which would be both economically and environmentally beneficial.

  10. 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

  11. Femur morphometry, densitometry, geometry and mechanical properties in young pigs fed a diet free of inorganic phosphorus and supplemented with phytase.

    Science.gov (United States)

    Skiba, Grzegorz; Sobol, Monika; Raj, Stanisława

    2017-02-01

    The study investigated in piglets the effect of replacing dietary inorganic P by addition of microbial phytase and its impact on performance, nutrient digestibility and on the geometrical characteristics and mineralisation of the femur. Sixteen pigs on day 58 of age were divided into two groups and fed either a diet free of additional inorganic phosphorus (P) and supplemented with phytase (Diet LP, 4.23 g total P/kg diet) or a diet with a mineral source of P and not supplemented with phytase (Diet SP, 5.38 g total P/kg diet). Performance data and the apparent total tract digestibility of nutrients were estimated between days 58 and 114, and 72 and 86 of age, respectively. On day 114 of age, the pigs were slaughtered, the femur was dissected and the mineral content and mineral density, maximum strength and maximum elastic strength, cortical wall thickness, cross-sectional area and cortical index were analysed. The growth performance and digestibility of nutrient fractions (with exception of P) did not differ between treatment groups. The P-digestibility was significantly higher in Group LP. The femur of pigs in Group LP had significantly greater cortical wall thickness, cortical index, bone mineral content, bone mineral density, maximum strength and maximum elastic strength than Group SP. Femur maximum strength and maximum elastic strength were correlated with cortical wall thickness and cortical index. Resulting from the different supply of digestible P, the femur geometrical, densitometric and mechanical properties of Group LP were better than those of Group SP. The mechanical properties of the femur of pigs depended more on its geometrical characteristics than on the degree of its mineralisation.

  12. Shifting the pH Profile of Aspergillus niger PhyA Phytase To Match the Stomach pH Enhances Its Effectiveness as an Animal Feed Additive

    OpenAIRE

    Kim, Taewan; Mullaney, Edward J.; Porres, Jesus M.; Roneker, Karl R.; Crowe, Sarah; Rice, Sarah; Ko, Taegu; Ullah, Abul H. J.; Daly, Catherine B.; Welch, Ross; Lei, Xin Gen

    2006-01-01

    Environmental pollution by phosphorus from animal waste is a major problem in agriculture because simple-stomached animals, such as swine, poultry, and fish, cannot digest phosphorus (as phytate) present in plant feeds. To alleviate this problem, a phytase from Aspergillus niger PhyA is widely used as a feed additive to hydrolyze phytate-phosphorus. However, it has the lowest relative activity at the pH of the stomach (3.5), where the hydrolysis occurs. Our objective was to shift the pH optim...

  13. Prediction of ingredient quality and the effect of a combination of xylanase, amylase, protease and phytase in the diets of broiler chicks. 2. Energy and nutrient utilisation.

    Science.gov (United States)

    Cowieson, A J; Singh, D N; Adeola, O

    2006-08-01

    1. In order to investigate the effects of xylanase, amylase, protease and phytase in the diets of broiler chickens containing graded concentrations of metabolisable energy (ME), two 42-d experiments were conducted using a total of 2208 broiler chicks (8 treatments with 12 replicate pens in each experiment). 2. Four diets including one positive and three negative control diets were used. Three maize/soybean meal-based negative control (NC) diets were formulated to be identical in available phosphorus (P), calcium (Ca) and amino acids but NC1 contained approximately 0.17 MJ/kg less ME than NC2 and approximately 0.34 MJ/kg less ME than NC3. A positive control (PC) was fed for comparison and was formulated to be adequate in all nutrients, providing approximately 0.63 MJ/kg ME, 0.13% available P, 0.12% Ca and 1 to 2% amino acids more than NC1. 3. The reduction in nutrient density between NC1 and PC was determined using ingredient quality models Avichecktrade mark Corn and Phychecktrade mark that can predict the response to exogenous enzymes in maize/soybean meal-based broiler diets. Supplementation of each diet with or without a cocktail of xylanase, amylase, protease and phytase gave a total of 8 dietary treatments in a 4 x 2 factorial arrangement. The same treatments and diet designs were used in both experiments but conducted in different locations using different batches of maize, soybean meal and minor ingredients. 4. In both experiments, digestibility was improved by the addition of exogenous enzymes, particularly those for P, Ca and certain amino acids. In addition, the supplementation of the PC with enzymes elicited a positive response indicating that over-the-top addition of xylanase, amylase, protease and phytase may offer a nutritionally and economically viable alternative to feed cost reduction. 5. It can be concluded that the digestibility of nutrients by broilers fed on maize/soybean meal-based diets can be improved by the use of a combination of xylanase

  14. 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

    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 a...

  15. Effect of The Phytase Enzyme Addition in The Artificial Feed on Digestibility of Feed, Feed Conversion Ratio and Growth of Gift Tilapia Saline Fish (Oreochromis niloticus) Nursery Stadia I

    Science.gov (United States)

    Rachmawati, Diana; Samidjan, Istiyanto; Elfitasari, Tita

    2018-02-01

    The purpose of this study was to determine the effect of adding the phytase enzyme in the artificial feed on digestibility of feed, feed conversion ratio and growth of gift tilapia saline fish (Oreochromis niloticus) nursery stadia I. The fish samples in this study used gift tilapia saline fish (O. niloticus) with an average weight of 0,62 ± 0,008 g/fish and the stocking density of 1 fish1 L. Experimental method used in this study was completely randomized design with 4 treatments and 3 repetitions. The treatments were by adding phytase enzyme in artificial feed with the different level of doses those were A (0 FTU kg1 feed), B (500 FTU kg1 feed), C (1000 FTU kg1 feed) and D (1500 FTU kg1 feed). The results show that the addition of phytase enzyme was significantly (P0.05) affected on Survival Rate (SR) of gift tilapia saline fish. The optimum doses of phytase enzyme on RGR, FCR, PER, ADCP and ADCF of gift tilapia saline fish ranged from 1060 to 1100 FTU kg-1 feed.

  16. 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...

  17. 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...

  18. 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 ...

  19. 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...

  20. The effect of lipids, a lipid-rich ready-to-use therapeutic food, or a phytase on iron absorption from maize-based meals fortified with micronutrient powders.

    Science.gov (United States)

    Monnard, Arnaud; Moretti, Diego; Zeder, Christophe; Steingötter, Andreas; Zimmermann, Michael B

    2017-06-01

    Background: Ready-to-use-therapeutic foods (RUTFs) high in lipid, protein, and iron are used to treat malnutrition. Lipids increase gastric residence time, which could increase iron absorption, particularly from poorly soluble iron compounds and in combination with phytase. Objectives: The objectives were to 1 ) assess the effect on iron absorption of a lipid emulsion given 20 min before or together with an iron-fortified maize meal and 2 ) assess iron absorption from a micronutrient powder (MNP) given with a nutrient-dense RUTF and/or a microbial phytase. Design: A total of 41 women participated in 3 studies. They consumed a maize meal fortified with isotopically labeled ferrous sulfate (FeSO 4 ; study 1) or ferric pyrophosphate (FePP; study 2). In studies 1 and 2, a lipid emulsion was given with or 20 min before the meal. In study 3, with the use of a 2 × 2 factorial design, subjects consumed a maize meal fortified with an MNP containing labeled FeSO 4 (MNP) given with an RUTF (MNP+RUTF), with a phytase (MNP+phytase), or both (MNP+RUTF+phytase). Iron absorption was assessed by isotope incorporation in erythrocytes 14 d after the test meals. Results: The lipid emulsion given either before or with the meal significantly increased iron absorption from FePP by 2.55-fold (95% CI: 1.48-, 4.37-fold; P = 0.001) but not from FeSO 4 There was a trend to increase iron absorption with the MNP+RUTF meal, which did not reach significance (1.21-fold; 95% CI: 0.92-, 1.61-fold; P = 0.060). The addition of phytase to MNP and MNP+RUTF significantly increased iron absorption by 1.85-fold (95% CI: 1.49-, 2.29-fold; P phytase and RUTF. Conclusions: In iron-fortified maize-based meals, the addition of lipids more than doubles iron absorption from FePP. Our results suggest the possibility of an enhancing effect on iron absorption of lipid-rich RUTFs, but more research is needed to determine this. This trial was registered at clinicaltrials.gov as NCT01991626. © 2017 American Society

  1. Evaluation of normal yellow dent corn and high available phosphorus corn in combination with reduced dietary phosphorus and phytase supplementation for broilers grown to market weights in litter pens.

    Science.gov (United States)

    Yan, F; Kersey, J H; Fritts, C A; Waldroup, P W; Stilborn, H L; Crum, R C; Rice, D W; Raboy, V

    2000-09-01

    A study was conducted to determine the extent fecal P levels could be reduced while maintaining performance. Various strategies were employed including the use of a high available phosphorus hybrid of corn (HAPC), supplementation with phytase enzyme, and reduced dietary P levels. The use of HAPC resulted in a 50% reduction in phytate-bound dietary P as compared with a normal yellow dent corn (YDC) diet. Dietary nonphytate P was maintained at either NRC (1994) recommendations for appropriate age periods or reduced by 0.075 or 0.15%. Portions of the diets were supplemented with 1,000 units of phytase/kg. Male chicks of a commercial strain were grown to 56 d on the test diets. Broilers fed diets with HAPC had BW, feed conversion, livability, and tibia ash that were equal to or superior to those fed diets with YDC with considerably reduced fecal P content at any dietary level of nonphytate P. Phytase supplementation enabled birds to maintain live performance at lower levels of nonphytate P, further reducing the fecal P output. One of the greatest contributions of phytase was a reduction in mortality at the lower levels of nonphytate P. Dietary P levels could be reduced by 0.075% under NRC (1994) recommendations without adversely affecting live performance; a reduction of 0.15% in conjunction with phytase supplementation maintained BW, feed conversion, and livability but reduced tibia ash. The extent to which dietary P levels can be reduced over the entire feeding program is subject to further research.

  2. 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.

  3. 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.

  4. 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. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. Engineering of Yersinia Phytases to Improve Pepsin and Trypsin Resistance and Thermostability and Application Potential in the Food and Feed Industry.

    Science.gov (United States)

    Niu, Canfang; Yang, Peilong; Luo, Huiying; Huang, Huoqing; Wang, Yaru; Yao, Bin

    2017-08-30

    Susceptibility to proteases usually limits the application of phytase. We sought to improve the pepsin and trypsin resistance of YeAPPA from Yersinia enterocolitica and YkAPPA from Y. kristensenii by optimizing amino acid polarity and charge. The predicted pepsin/trypsin cleavage sites F89/K226 in pepsin/trypsin-sensitive YeAPPA and the corresponding sites (F89/E226) in pepsin-sensitive but trypsin-resistant YkAPPA were substituted with S and H, respectively. Six variants were produced in Pichia pastoris for catalytic and biochemical characterization. F89S, E226H, and F89S/E226H elevated pepsin resistance and thermostability and K226H and F89S/K226H improved pepsin and trypsin resistance and stability at 60 °C and low pH. All the variants increased the ability of the proteins to hydrolyze phytate in corn meal by 2.6-14.9-fold in the presence of pepsin at 37 °C and low pH. This study developed a genetic manipulation strategy specific for pepsin/trypsin-sensitive phytases that can improve enzyme tolerance against proteases and heat and benefit the food and feed industry in a cost-effective way.

  6. 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.

  7. 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.

  8. Ileal Amino Acid Digestibility of Broken Rice Fed to Postweaned Piglets with or without Multicarbohydrase and Phytase Supplementation.

    Science.gov (United States)

    Dadalt, J C; Gallardo, C; Polycarpo, G V; Budiño, F E L; Rogiewicz, A; Berto, D A; Trindade Neto, M A

    2016-10-01

    Most of amino acid (AA) digestibility values for feed ingredients are obtained using pigs cannulated in the distal ileum. The ileal-cannulated pig model uses pigs older than six weeks due to difficulties related to implanting the T-cannula in distal ileum of younger pigs and complications during the post-surgical recovery. However, to properly formulate the diet of weaned pigs, the nutritive value of feed ingredients should be determined with younger pigs. Thus, 25 weaned pigs were used to determine the apparent total tract digestibility (ATTD) of nutrients, energy, and apparent ileal digestibility (AID) and standardized ileal digestibility (SID) ileal AA digestibility of broken rice (BR), with or without multicarbohydrase (MC) and phytase (Phy) supplementation. Piglets were weaned at 23 d of age and individually housed in digestibility cages until 45 d of age. The trial consisted of 7 d of adaptation to the experimental diets and 3 d of excreta (feces and urine) collection. Ileal digesta was collected at slaughter (about 6 weeks of age). A completely randomized experimental design was used to determine the effects of MC and Phy. Reference diets (RD, 5% casein) was replaced by 30% of BR with or without MC, Phy, or MC+Phy. The RD was used to quantify endogenous AA losses. BR with Phy supplied had increased the ATTD of dry matter (pdigestible energy and protein and SID for histidine (pdigestibilities. Standardized amino acid digestibilities of BR, without enzymes, were lower than those values reported in the literature. The MC and Phy improved the digestibility of some nutrients and energy of BR in post-weaned piglet diets.

  9. Ileal Amino Acid Digestibility of Broken Rice Fed to Postweaned Piglets with or without Multicarbohydrase and Phytase Supplementation

    Directory of Open Access Journals (Sweden)

    J. C. Dadalt

    2016-10-01

    Full Text Available Most of amino acid (AA digestibility values for feed ingredients are obtained using pigs cannulated in the distal ileum. The ileal-cannulated pig model uses pigs older than six weeks due to difficulties related to implanting the T-cannula in distal ileum of younger pigs and complications during the post-surgical recovery. However, to properly formulate the diet of weaned pigs, the nutritive value of feed ingredients should be determined with younger pigs. Thus, 25 weaned pigs were used to determine the apparent total tract digestibility (ATTD of nutrients, energy, and apparent ileal digestibility (AID and standardized ileal digestibility (SID ileal AA digestibility of broken rice (BR, with or without multicarbohydrase (MC and phytase (Phy supplementation. Piglets were weaned at 23 d of age and individually housed in digestibility cages until 45 d of age. The trial consisted of 7 d of adaptation to the experimental diets and 3 d of excreta (feces and urine collection. Ileal digesta was collected at slaughter (about 6 weeks of age. A completely randomized experimental design was used to determine the effects of MC and Phy. Reference diets (RD, 5% casein was replaced by 30% of BR with or without MC, Phy, or MC+Phy. The RD was used to quantify endogenous AA losses. BR with Phy supplied had increased the ATTD of dry matter (p<0.05 and SID of histidine (p = 0.05, arginine, leucine, lysine, valine, alanine, and proline (p<0.05. BR with MC had been increased digestible energy and protein and SID for histidine (p<0.05. There was no interaction between Phy and MC on the BR nutrient digestibilities. Standardized amino acid digestibilities of BR, without enzymes, were lower than those values reported in the literature. The MC and Phy improved the digestibility of some nutrients and energy of BR in post-weaned piglet diets.

  10. Suplementação dietética de fitase sobre o metabolismo de nutrientes de frangos de corte Phytase dietetic supplementation on nutrients metabolism of broilers

    Directory of Open Access Journals (Sweden)

    Guilherme Rodrigues Lelis

    2010-08-01

    Full Text Available O experimento foi conduzido com os objetivos de avaliar o efeito da suplementação de fitase em dietas sobre a energia metabolizável (EMAn e sobre os coeficientes de digestibilidade ileal aparente de matéria seca, proteína bruta, cálcio e fósforo e determinar a deposição de cinzas, cálcio e fósforo na tíbia de frangos de corte. Foram utilizados 350 frangos de corte machos Ross de 16 a 25 dias de idade, distribuídos num delineamento inteiramente ao acaso, com cinco dietas e dez repetições de sete aves por unidade experimental. As dietas, à base de milho e de farelo de soja, foram formuladas considerando a disponibilidade de energia metabolizável, proteína bruta, lisina, cálcio e fósforo disponível, de acordo com a matriz nutricional da enzima fitase, e suplementadas com 0,5% de oxido crômico. As dietas foram: controle positivo; controle negativo 1; controle negativo 2; controle negativo 1 + 250 uft; controle negativo 2 + 500 uft. A suplementação de enzima fitase nos níveis de 250 e de 500 uft/kg de dieta melhorou os valores energéticos das dietas, que aumentaram, em média, 36 e 54 kcal/kg de MS, respectivamente. A utilização de fitase melhorou os coeficientes de digestibilidade da proteína bruta e do fósforo, cujos maiores valores foram obtidos com a suplementação de 500 uft/kg de ração. A suplementação de fitase melhorou o coeficiente de digestibilidade e a retenção de fósforo, reduzindo o fósforo excretado, e aumentando a composição de fósforo na tíbia das aves, principalmente nos frangos de corte alimentados com a dieta suplementada com fitase (500 uft/kg. A suplementação de fitase (500 uft/kg melhora o coeficiente de digestibilidade da proteína bruta e do fósforo, melhora a retenção de fósforo e diminui a excreção de fósforo de frangos de corte.The experiment was carried out to evaluate the effect of phytase enzyme supplementation in diets on the metabolizable energy (ME and on the

  11. Effects of exogenous phytase and xylanase, individually or in combination, and pelleting on nutrient digestibility, available energy content of wheat and performance of growing pigs fed wheat-based diets.

    Science.gov (United States)

    Yang, Y Y; Fan, Y F; Cao, Y H; Guo, P P; Dong, B; Ma, Y X

    2017-01-01

    Two experiments were conducted to determine the effects of adding exogenous phytase and xylanase, individually or in combination, as well as pelleting on nutrient digestibility, available energy content of wheat and the performance of growing pigs fed wheat-based diets. In Experiment 1, forty-eight barrows with an initial body weight of 35.9±0.6 kg were randomly assigned to a 2×4 factorial experiment with the main effects being feed form (pellet vs meal) and enzyme supplementation (none, 10,000 U/kg phytase, 4,000 U/kg xylanase or 10,000 U/kg phytase plus 4,000 U/kg xylanase). The basal diet contained 97.8% wheat. Pigs were placed in metabolic cages for a 7-d adaptation period followed by a 5-d total collection of feces and urine. Nutrient digestibility and available energy content were determined. Experiment 2 was conducted to evaluate the effects of pelleting and enzymes on performance of wheat for growing pigs. In this experiment, 180 growing pigs (35.2±9.0 kg BW) were allocated to 1 of 6 treatments according to a 2×3 factorial treatment arrangement with the main effects being feed form (meal vs pellet) and enzyme supplementation (0, 2,500 or 5,000 U/kg xylanase). In Experiment 1, there were no interactions between feed form and enzyme supplementation. Pelleting reduced the digestibility of acid detergent fiber (ADF) by 6.4 percentage units (pdigestibility of energy by 0.6 percentage units (pdigestibility of crude protein by 0.5 percentage units (p = 0.07) compared with diets in mash form. The addition of phytase improved the digestibility of phosphorus (pdigestibility of crude protein by 1.0 percentage units (p = 0.09) and increased the digestibility of neutral detergent fiber (NDF) (pdigestibility of phosphorus (pdigestibility (pdigestibility but decreased ADF digestibility. Adding xylanase increased crude protein digestibility and pig performance. Phytase increased the apparent total tract digestibility of phosphorus and calcium. The combination of

  12. 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

  13. Effect of dietary phosphorus, phytase, and 25-hydroxycholecalciferol on broiler chicken bone mineralization, litter phosphorus, and processing yields.

    Science.gov (United States)

    Angel, R; Saylor, W W; Mitchell, A D; Powers, W; Applegate, T J

    2006-07-01

    Three floor pen experiments (Exp) were conducted to evaluate low nonphytin P (NPP) concentrations and the NPP sparing effect of phytase (PHY) and 25-hydroxycholecalciferol (25D) on bone mineralization, bone breaking during commercial processing, litter P, and water-soluble P (WSP) concentrations. Tested treatments (TRT) were control, National Research Council NPP; University of Maryland (UMD) NPP; UMD + PHY, UMD NPP reduced by 0.064% NPP + 600 U of PHY/kg; UMD + PHY + 25D, UMD NPP reduced by 0.090% NPP + 600 U of PHY and 70 microg of 25D/kg; control + PHY mimicked the industry practice of diets by 0.1% when PHY is added; and negative control with 90% UMD NPP concentrations. UMD + PHY and control + PHY diets contained 600 U of PHY/kg, and UMD + PHY + 25D contained 600 U of PHY + 70 microg of 25D/kg. Performance results were presented separately. After each Exp, litter P and WSP were determined, and bone measurements were obtained on 8 or 10 broilers per pen. Tested TRT did not affect broiler BW. Femur ash weight of broilers fed the UMD and UMD + PHY + 25D was lower in all Exp compared with that of broilers fed the control diet. Femur ash was similar for control and UMD + PHY broilers, yet averaged over all Exp, UMD + PHY broilers consumed 39% less NPP and required less NPP per gram of femur ash than those on the control (4.87 and 7.77 g of NPP/g of ash, Exp 3). At the end of Exp 3, broilers were processed in a commercial facility. Despite reductions in NPP intake and bone mineralization, no differences were observed in measurements of economic importance (parts lost, carcass yield, and incidence of broken bones). The P excretion per bird was lowest for birds fed the UMD + PHY + 25D diet followed by those fed the UMD + PHY and negative control diets (10.44, 12.00, and 13.78 g of P/bird, respectively) and were highest for those fed the control diet (19.55 g of P/bird). These results suggest that feeding diets low in P together with PHY and 25D will not affect

  14. Ractopamina e fitase em dietas para suínos na fase de terminação Ractopamine and phytase in diets for finishing pigs

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    Anderson Corassa

    2009-11-01

    ractopamine and enzyme phytase on the performance and carcass characteristics of finishing pigs. Three hundred and fifty-four commercial type barrows were used from a single source with 94.1 ± 5.7 kg initial body weight in a randomized complete block design, with six treatments and five replications with 11 or 12 animals per experimental unit evaluated. The following feeds were: control diet based on tables of nutritional requirements; (A diet control with nutritional adjustment for ractopamine; (FIT diet A with phytase; (RAC diet A with ractopamine; (RAC+FIT diet A with ractopamine and phytase; and (RAC+FITe diet A with ractopamine and phytase with part of the demands for calcium and available phosphorus met by the enzyme. The experiment lasted 21 days and at the end the animals were slaughtered for carcass evaluation. The results were compared by orthogonal contrasts. Feed intake was not influenced by the diets. The diet with adjusted protein and amino acid levels and without ractopamine did not improve performance compared with the control diet. There was effect of ractopamine on daily gain and feed conversion at the end of the 21-day period, and also on the pre-slaughter weight, hot carcass weight, carcass dressing, total fat-free lean and payment index. Phytase inclusion in diets containing ractopamin for calcium and phosphorus available levels above the finishing pigs´ requirements did not effect the weight gain, feed conversion and carcass traits. Ractopamine improved performance and carcass traits. The use of phytase in diets containing ractopamine in partial substitution of inorganic mineral sources made possibly to maintain performance and carcass traits.

  15. Scientific skepticism and new discoveries: an analysis of a report of zinc/phytase supplementation and the efficacy of botulinum toxins in treating cosmetic facial rhytides, hemifacial spasm and benign essential blepharospasm.

    Science.gov (United States)

    Cohen, Joel L

    2014-10-01

    A recent paper in the Journal of Drugs in Dermatology by Koshy and colleagues (2012, 11( 4 ):507-512) report on "Effect of Dietary Zinc and Phytase Supplementation on Botulinum Toxin Treatments" and conclude by claiming the discovery of "a potentially meaningful role for zinc and/or phytase supplementation in increasing the degree and duration of botulinum toxin effect in the treatment of cosmetic facial rhytids, benign essential blepharospasm, and hemifacial spasm". The purpose of this paper is to examine these published claims for possible methodological and design errors and potential sources of bias. The authors evaluated the published results in comparison to the published literature on zinc deficiency, the role of phytase, prior reports of an effect of zinc on activity of botulinum toxin, issues of study design and execution and if the reported results of the study supported the study's conclusions. Multiple issues are present in the reported study, which appear to invalidate its conclusions. These areas include lack of direct evidence for the presence of clinical or subclinical zinc deficiency in the study population or for the level of phytate in the study population sufficient to interfere with zinc absorption in these subjects. Additionally, there is ambiguity as to the actual dose of zinc used as well as in the study design itself. Also there is a failure of the study through the "unmasking" of the crossover design. There is potential financial conflict of interest in the study execution that may have biased the reported results. Finally there is inadequate data presented to evaluate the claims made of a "new discovery" as to the three disease entities reported on and the various botulinum toxins used in each of the three treatment arms of the study. Based on this evaluation, it appears that a high level of clinical and scientific skepticism is warranted concerning any claim of a beneficial effect of zinc and phytase supplementation on the efficacy or

  16. 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 ...

  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. Effect of dietary calcium concentrations in low non-phytate phosphorus diets containing phytase on growth performance, bone mineralization, litter quality, and footpad dermatitis incidence in growing broiler chickens

    Directory of Open Access Journals (Sweden)

    Jong Hyuk Kim

    2017-07-01

    Full Text Available Objective An experiment was conducted to investigate the effect of dietary Ca concentrations in low non-phytate phosphorus (NPP diets containing phytase on growth performance, bone mineralization, litter quality, and footpad dermatitis (FPD incidence in growing broiler chickens. Methods A total of 1,800 21-day-old Ross 308 growing broiler chickens were allotted to 1 of 6 dietary treatments with 6 replicated cages. Six diets were formulated to provide increasing Ca concentrations of 4.0, 5.0, 6.0, 7.0, 8.0, or 9.0 g/kg in diets. The concentrations of NPP in all diets were maintained at 3.0 g/kg, and phytase was supplemented to all diets at the level of 1,000 fytase units (FTU/kg. At the end of the 14-d feeding trial, birds were euthanized for tibia sampling, and litter samples were collected from 3 areas in the cage. The FPD incidence was measured based on a 6-point scoring system. Results Dietary Ca concentrations had no effect on growth performance of growing broiler chickens. However, a tendency (linear, p = 0.05 for decreased feed efficiency was observed as dietary Ca concentrations were increased. The concentrations of Ca and P in the tibia of broiler chickens increased (linear and quadratic, p<0.01 with increasing Ca concentrations in low NPP diets containing phytase. Litter pH, moisture, and N contents were not affected by increasing Ca concentrations in low NPP diets containing phytase. However, a tendency (quadratic, p = 0.10 for increased FPD incidence with increasing dietary Ca concentrations was observed. Conclusion Dietary Ca concentrations from 4.0 to 9.0 g/kg in low NPP diets containing phytase have little effects on growth performance of growing broiler chickens. However, Ca and P concentrations in the tibia are decreased if dietary Ca concentrations are less than 5.0 g/kg. The FPD incidence for growing broiler chickens may be decreased if less than 9.0 g/kg of Ca is included in diets.

  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. Avaliação da matriz nutricional da enzima fitase em rações contendo sorgo para poedeiras comerciais Evaluation of the nutritional matrix values for phytase enzyme in laying hens diets with sorghum

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    Elaine Cristina Ligeiro

    2009-10-01

    Full Text Available Avaliaram-se os efeitos da inclusão de fitase e de sua matriz nutricional em rações contendo sorgo sobre o desempenho das aves, a qualidade dos ovos, a ingestão e a excreção e retenção de fósforo e nitrogênio em poedeiras comerciais. Utilizaram-se 180 poedeiras comerciais, distribuídas em delineamento inteiramente ao acaso em esquema fatorial 2 × 2 + 1, com dois níveis de fitase (0 e 500 FTU/kg de ração e dois níveis de substituição do milho pelo sorgo (50 e 100% e uma ração testemunha (isenta de sorgo e fitase, constituindo cinco tratamentos com seis repetições de seis aves. As rações foram à base de milho e farelo de soja, sem fitase e sorgo, considerando a matriz nutricional da fitase. O desempenho e a qualidade dos ovos foram avaliados em quatro períodos de 28 dias. Ao final do experimento, um ensaio de metabolismo foi realizado para quantificar a ingestão, excreção e retenção aparente de fósforo e nitrogênio e avaliar a viabilidade econômica das rações. Ao considerar a matriz nutricional da fitase, as exigências em energia, cálcio, fósforo, proteína e aminoácidos foram atendidas, mesmo com a redução dos níveis nutricionais da dieta, e o desempenho e a qualidade dos ovos não foram comprometidos. A adição de fitase nas rações possibilitou reduzir todos os parâmetros econômicos avaliados. O sorgo pode substituir totalmente o milho e ser o único grão energético da dieta.The effects were assessed of including phytase and its nutritional matrix in diets containing sorghum on the performance of the birds, egg quality, intake and phosphorus excretion and retention in laying hens. One hundred and eighty laying hens were arranged in a randomized complete design in a 2 × 2 + 1 factorial arrangement with two phytase levels (0 and 500 FTU/kg diet, two levels of corn replacement for sorghum (50 and 100% and a control diet (without phytase and sorghum forming five treatments and six replications of

  1. FITASE EM DIETAS PARA SUÍNOS EM CRESCIMENTO: (I IMPACTO AMBIENTAL PHYTASE IN DIETS FOR GROWING PIGS: (I ENVIRONMENTAL IMPACT

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    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

  2. 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.

  3. 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.

  4. A simple and accurate two-step long DNA sequences synthesis strategy to improve heterologous gene expression in pichia.

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    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.

  5. A novel protein tyrosine phosphatase like phytase from Lactobacillus fermentum NKN51: Cloning, characterization and application in mineral release for food technology applications.

    Science.gov (United States)

    Sharma, Rekha; Kumar, Piyush; Kaushal, Vandana; Das, Rahul; Kumar Navani, Naveen

    2018-02-01

    A novel protein tyrosine phosphatase like phytase (PTPLP), designated as PhyLf from probiotic bacterium Lactobacillus fermentum NKN51 was identified, cloned, expressed and characterized. The recombinant PhyLf showed specific activity of 174.5 U/mg. PhyLf exhibited strict specificity towards phytate and optimum temperature at 60 °C, pH 5.0 and ionic strength of 100 mM. K m and K cat of PhyLf for phytate were 0.773 mM and 84.31 s -1 , respectively. PhyLf exhibited high resistance against oxidative inactivation. PhyLf shares no homology, sans the active site with reported PTLPs, warranting classification as a new subclass. Dephytinization of durum wheat and finger millet under in vitro gastrointestinal conditions using PhyLf enhanced the bioaccessibility of mineral ions. Probiotic origin, phytate specificity, resistance to oxidative environment and gastric milieu coupled with ability to release micronutrients are unique properties of PhyLf which present a strong case for its use in ameliorating nutritional value of cereals and animal feed. Copyright © 2017 Elsevier Ltd. All rights reserved.

  6. 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.

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    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.

  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)

    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. PMID:26599444

  8. Nutrient reduction in rations with phytase for growing pigs Redução de nutrientes em rações com fitase para suínos em crescimento

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    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

  9. 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)

  10. N-Glycosylation Improves the Pepsin Resistance of Histidine Acid Phosphatase Phytases by Enhancing Their Stability at Acidic pHs and Reducing Pepsin's Accessibility to Its Cleavage Sites

    Science.gov (United States)

    Niu, Canfang; Luo, Huiying; Shi, Pengjun; Huang, Huoqing; Wang, Yaru; Yang, Peilong

    2015-01-01

    N-Glycosylation can modulate enzyme structure and function. In this study, we identified two pepsin-resistant histidine acid phosphatase (HAP) phytases from Yersinia kristensenii (YkAPPA) and Yersinia rohdei (YrAPPA), each having an N-glycosylation motif, and one pepsin-sensitive HAP phytase from Yersinia enterocolitica (YeAPPA) that lacked an N-glycosylation site. Site-directed mutagenesis was employed to construct mutants by altering the N-glycosylation status of each enzyme, and the mutant and wild-type enzymes were expressed in Pichia pastoris for biochemical characterization. Compared with those of the N-glycosylation site deletion mutants and N-deglycosylated enzymes, all N-glycosylated counterparts exhibited enhanced pepsin resistance. Introduction of the N-glycosylation site into YeAPPA as YkAPPA and YrAPPA conferred pepsin resistance, shifted the pH optimum (0.5 and 1.5 pH units downward, respectively) and improved stability at acidic pH (83.2 and 98.8% residual activities at pH 2.0 for 1 h). Replacing the pepsin cleavage sites L197 and L396 in the immediate vicinity of the N-glycosylation motifs of YkAPPA and YrAPPA with V promoted their resistance to pepsin digestion when produced in Escherichia coli but had no effect on the pepsin resistance of N-glycosylated enzymes produced in P. pastoris. Thus, N-glycosylation may improve pepsin resistance by enhancing the stability at acidic pH and reducing pepsin's accessibility to peptic cleavage sites. This study provides a strategy, namely, the manipulation of N-glycosylation, for improvement of phytase properties for use in animal feed. PMID:26637601

  11. The influence of meat-and-bone meal and exogenous phytase on growth performance, bone mineralisation and digestibility coefficients of protein (N), amino acids and starch in broiler chickens.

    Science.gov (United States)

    Liu, Sonia Y; Cowieson, Aaron J; Selle, Peter H

    2016-06-01

    The objective of this study was to examine the influence of meat-and-bone meal (MBM) and phytase inclusion on growth performance, bone mineralisation and apparent digestibility coefficients of nutrients in broiler chickens offered wheat-based diets. The feeding study comprised 7 dietary treatments: positive control (PC, 9.0% Ca and 4.5% available phosphorous [AvP] in starter, 7.0% Ca and 3.5% AvP in finisher); negative control (NC, 7.2% Ca and 3.0% AvP in starter, 5.2% Ca and 2.0% AvP in finisher) diets incorporating a 3 × 2 factorial array of 3 MBM inclusions (0, 60, 120 g/kg) and 2 levels of phytase supplementation (0 and 1,000 FYT/kg). Each treatment was allocated to 6 replicated pens with 30 birds per pen in an environmentally-controlled deep litter facility. A total of 1,260 one-day-old male Ross 308 chicks were offered starter diets from 1 to 14 days post-hatch and finisher diets from 15 to 36 days post-hatch. There were significant ( P  digestibility coefficients of starch and protein except that diets without MBM had significantly ( P  digestibility and diets with 120 g/kg MBM had significantly ( P  digestibility. No dietary influence on ileal fat digestibility was observed. There were dietary interactions on ileal digestibilities of isoleucine, valine and glycine. Phytase significantly increased glycine digestibility in diets with 60 and 120 g/kg MBM but not in diets without MBM. Including 120 g/kg MBM significantly ( P  digestibility coefficients of 13 ex 16 amino acids in the distal ileum. This study demonstrated the negative impacts of MBM on amino acid digestibility and growth performance. Also, responses to phytase were more pronounced in diets without MBM, which may have been due to their relatively lower available P and higher phytate concentrations in comparison to diets containing MBM.

  12. Effect of dietary calcium concentrations in low non-phytate phosphorus diets containing phytase on growth performance, bone mineralization, litter quality, and footpad dermatitis incidence in growing broiler chickens.

    Science.gov (United States)

    Kim, Jong Hyuk; Jung, Hyunjung; Pitargue, Franco Martinez; Han, Gi Ppeum; Choi, Hyeon Seok; Kil, Dong Yong

    2017-07-01

    An experiment was conducted to investigate the effect of dietary Ca concentrations in low non-phytate phosphorus (NPP) diets containing phytase on growth performance, bone mineralization, litter quality, and footpad dermatitis (FPD) incidence in growing broiler chickens. A total of 1,800 21-day-old Ross 308 growing broiler chickens were allotted to 1 of 6 dietary treatments with 6 replicated cages. Six diets were formulated to provide increasing Ca concentrations of 4.0, 5.0, 6.0, 7.0, 8.0, or 9.0 g/kg in diets. The concentrations of NPP in all diets were maintained at 3.0 g/kg, and phytase was supplemented to all diets at the level of 1,000 fytase units (FTU)/kg. At the end of the 14-d feeding trial, birds were euthanized for tibia sampling, and litter samples were collected from 3 areas in the cage. The FPD incidence was measured based on a 6-point scoring system. Dietary Ca concentrations had no effect on growth performance of growing broiler chickens. However, a tendency (linear, p = 0.05) for decreased feed efficiency was observed as dietary Ca concentrations were increased. The concentrations of Ca and P in the tibia of broiler chickens increased (linear and quadratic, pLitter pH, moisture, and N contents were not affected by increasing Ca concentrations in low NPP diets containing phytase. However, a tendency (quadratic, p = 0.10) for increased FPD incidence with increasing dietary Ca concentrations was observed. Dietary Ca concentrations from 4.0 to 9.0 g/kg in low NPP diets containing phytase have little effects on growth performance of growing broiler chickens. However, Ca and P concentrations in the tibia are decreased if dietary Ca concentrations are less than 5.0 g/kg. The FPD incidence for growing broiler chickens may be decreased if less than 9.0 g/kg of Ca is included in diets.

  13. Isolation of phytase-producing bacteria from Himalayan soils and their effect on growth and phosphorus uptake of Indian mustard (Brassica juncea).

    Science.gov (United States)

    Kumar, Vinod; Singh, Prashant; Jorquera, Milko A; Sangwan, Punesh; Kumar, Piyush; Verma, A K; Agrawal, Sanjeev

    2013-08-01

    Phytase-producing bacteria (PPB) is being investigated as plant growth promoting rhizobacteria (PGPR) to improve the phosphorus (P) nutrition and growth of plants grown in soil with high phytate content. Phytate is dominant organic P forms in many soils and must be hydrolyzed to be available for plants. Indian mustard (Brassica juncea) is a plant with economic importance in agriculture and phytoremediation, therefore biotechnological tools to improve growth and environmental stress tolerance are needed. In this study, we isolated and characterized PPB from Himalayan soils and evaluated their effect on growth and P uptake by B. juncea under greenhouse conditions. Sixty five PPB were isolated and based on phytate hydrolysis, three efficient PPB were chosen and identified as Acromobacter sp. PB-01, Tetrathiobacter sp. PB-03 and Bacillus sp. PB-13. Selected PPB showed ability to grow at wide range of pH, temperature and salt concentrations as well as to harbour diverse PGPR activities, such as: solubilization of insoluble Ca-phosphate (193-642 μg ml(-1)), production of phytohormone indole acetic acid (5-39 μg ml(-1)) and siderophore. Tetrathiobacter sp. PB-03 and Bacillus sp. PB-13 showed 50 and 70 % inhibition of phytopathogen Rhizoctonia solani, respectively. Greenhouse potting assay also showed that the bacterization of B. juncea seeds with Tetrathiobacter sp. PB-03 and Bacillus sp. PB-13 significantly increased the biomass and P content in 30 days old seedlings. This study reveals the potential of PPB as PGPR to improve the growth of B. juncea.

  14. 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

  15. Gene Therapy

    Science.gov (United States)

    Gene therapy Overview Gene therapy involves altering the genes inside your body's cells in an effort to treat or stop disease. Genes contain your ... that don't work properly can cause disease. Gene therapy replaces a faulty gene or adds a new ...

  16. Effects of multi-carbohydrase and phytase on standardized ileal digestibility of amino acids and apparent metabolizable energy in canola meal fed to broiler chicks.

    Science.gov (United States)

    Gallardo, C; Dadalt, J C; Kiarie, E; Trindade Neto, M A

    2017-09-01

    Two assays were conducted to evaluate nutritive value of canola meal (CM) fed to broiler chicks without or with a multi-carbohydrase (MC) preparation (700 U α-galactosidase, 2,200 U galactomannanase, 30,000 U xylanase, and 22,000 U β-glucanase per kg of diet) and phytase (Phy, 500 FTU per kg of diet). Assay 1 determined apparent total tract digestibility (ATTD) of nutrients and metabolizable energy (AME) by the difference method. Assay 2 determined apparent ileal digestibility (AID) and standardized ileal digestibility (SID) of amino acids by the index method. Two reference diets (RD) - an 85% corn based and a 5% casein-cornstarch diet fortified with vitamins and minerals - were made for assays 1 and 2, respectively. For each assay, the test diets were made by mixing RD and CM 7:3 wt/wt basis and fed without or with MC or Phy or combination. A total of 245 day-old male broilers (Cobb 500) was allocated to 5 treatments to give 7 replicates (7 birds/cage). The birds were fed a commercial diet from day zero to 10 followed by assay 1 fed from d 11 to 18 and assay 2 fed from d 19 to 21. Excreta samples were collected on d 15 to18, and all birds were slaughtered on d 21 for ileal digesta. There was an interaction (P  0.05) between MC and Phy on AMEn; however, MC and Phy individually improved AMEn retention. Enzymes interacted (P < 0.05) on SID of Arg, His, Leu, Met, Thr, Ala, Asp, Gln, and Gly. In this context, feeding a combination of MC and Phy resulted in higher (P < 0.05) SID of Arg, His, Met, and Thr relative to single activity or control. Both enzymes improved (P < 0.05) SID of Lys independently. The combination of carbohydrase and Phy may be an effective strategy to improve amino acid utilization in CM for poultry. © 2017 Poultry Science Association Inc.

  17. 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

  18. 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

  19. Avaliação técnica e econômica da matriz nutricional da enzima fitase em rações contendo farelo de girassol para poedeiras comerciais Technical and economical evaluation of the nutritional matrix of phytase enzyme diets with sunflower meal for laying hens

    Directory of Open Access Journals (Sweden)

    Otto Mack Junqueira

    2010-10-01

    Full Text Available Cento e oitenta poedeiras comerciais Isa Brown, com 60 semanas de idade, foram distribuídas em delineamento inteiramente ao acaso, em esquema fatorial 2 õ 2, com dois níveis de fitase (0 e 500 ftu/kg de ração e dois de farelo de girassol (4 e 8%, totalizando cinco dietas, avaliadas com seis repetições de seis aves. As dietas foram formuladas a partir de uma ração controle, à base de milho e farelo de soja, isenta de fitase e farelo de girassol, valorizando na formulação das dietas a matriz nutricional da fitase. O período experimental foi dividido em quatro ciclos de 28 dias cada. A matriz nutricional preconizada para fitase permitiu o atendimento pleno das exigências das aves, mesmo quando as dietas foram formuladas com níveis nutricionais reduzidos, isso quando desconsiderados os nutrientes presentes na matriz nutricional da enzima, de acordo com recomendação da empresa produtora. A adição de fitase na dieta reduz a excreção de fósforo pelas aves e melhora os parâmetros econômicos. A inclusão de farelo de girassol no nível de 8% prejudica a produção de ovos, mas não afeta os demais parâmetros de desempenho e qualidade dos ovos.One hundred and eighty Isa Brown laying hens at 60 weeks of age were arranged in a completely randomized design in a 2 õ 2 factorial scheme, with two levels of phytase (0 and 500 ftu/kg ration and two levels of sunflower meal (4 and 8% totalizing five diets, evaluated with six replicates with six birds. Diets were formulated from a control ration based on corn and soybean meal free from phytase and sunflower meal, valuing the nutritional phytase matrix in diets formulation. The experimental period was divided in four 28-day cycles. The nutritional matrix suggested for phytase completely met the requirement by hens, even when diets were formulated with reduced nutritional levels when the nutrients present in the enzyme nutritional matrix were not considered accordingly to recommendations by

  20. Digestibilidade de nutrientes e balanço de Ca e P em suínos recebendo dietas com ácido butírico, fitase e diferentes níveis de cálcio Nutrient digestibility and Ca and P balance in pigs receiving butyric acid, phytase and different calcium levels

    Directory of Open Access Journals (Sweden)

    Taiane Golfetto Machinsky

    2010-11-01

    Full Text Available Estudou-se o uso do ácido butírico (AB e da fitase em dietas de suínos na fase de crescimento, variando no nível de cálcio. O experimento foi subdivido no tempo em dois períodos de 17 dias, sendo três de adaptação e 14 de mensurações. Em cada período, foram utilizados 16 suínos machos castrados, com peso de 24,6±0,7kg no primeiro e 43,2±1,77kg no segundo período. As dietas diferiam no nível de cálcio (0,5 ou 0,72%, de AB (0 ou 0,3% de butirato de sódio 84% e de fitase (0 ou 500 FTU kg-1, fitase de origem bacteriana derivada de Escherichia coli. O delineamento foi em blocos casualizados (períodos, em decomposição fatorial 2x2x2, com quatro repetições. Foram avaliadas a digestibilidade aparente dos nutrientes e da energia bruta e o balanço de Ca e P. O AB melhorou a digestibilidade da proteína bruta, mas, de forma individual ou em combinação com a fitase, não aumentou a retenção de minerais. A fitase aumentou a retenção de P, reduzindo sua excreção fecal e urinária. O menor nível de Ca na dieta proporcionou maior retenção de Ca e menor retenção de P, em decorrência do aumento da excreção de P na urina (PIt was studied the use of butyric acid (BA and phytase in growing pigs diets, varying calcium level. The experiment was divided into two periods of time, of 17 days, with 3 days of adaptation and 14 of measurements. In each period, it was used 16 barrows weighing 24.6±0.7kg in the first and 43.2±1.77kg in the second. Diets were different in calcium level (0.5 or 0.72%, AB (0 or 0.3% sodium butyrate 84% and phytase (0 or 500FTU kg-1 phytase of bacterial origin derived from Escherichia coli. The experimental design was in randomized blocks (periods, decomposed in 2x2x2 factorial, with four replications. It was evaluated the apparent digestibility of nutrients, gross energy and balance of Ca and P. AB improved crude protein digestibility, but individually or in combination with phytase did not increase

  1. Digestibilidade de dietas suplementadas com fitase para suínos sob diferentes ambientes térmicos Digestibility of diets supplemented with phytase for pigs under different thermal environments

    Directory of Open Access Journals (Sweden)

    Charles Kiefer

    2012-08-01

    Full Text Available Realizou-se um ensaio de digestibilidade com o objetivo de avaliar níveis de fitase sobre a digestibilidade e excreção de nutrientes para suínos submetidos a condições de conforto e estresse térmico. Foram utilizados 24 suínos machos castrados, alojados em gaiolas metabólicas, em câmara climatizada, distribuídos em delineamento de blocos ao acaso em esquema fatorial 2x4, com dois ambientes (23 e 31°C e quatro níveis de fitase (0, 300, 600 e 900UF kg-1 de ração, com três repetições, sendo cada unidade experimental constituída por um animal. Concluiu-se que as dietas fornecidas aos suínos submetidos ao estresse térmico (31°C apresentam menores coeficientes de digestibilidade da proteína, fósforo, cálcio, zinco, ferro e magnésio e, portanto, apresentam maior excreção percentual desses nutrientes nas fezes, quando comparadas às dietas fornecidas aos suínos submetidos à termoneutralidade (23°C. A suplementação de fitase não altera o valor energético da dieta e o coeficiente de digestibilidade da proteína, mas aumenta o coeficiente de digestibilidade de fósforo, cálcio e magnésio e reduz a excreção percentual de fósforo e cálcio nas fezes dos suínos. Recomenda-se o nível de inclusão de 300UF kg-1 de ração, independentemente da condição térmica ambiental.We conducted a digestibility trial with the aim of evaluate phytase levels on digestibility and nutrient excretion for pigs submitted to thermoneutral conditions and heat stress. We used 24 barrows housed in metabolic cages in climatic chamber, distributed in randomized blocks in a 2x4 factorial design, two environments (23 and 31°C and four phytase levels (0, 300, 600 and 900UF kg-1 diet, with three replicates and each experimental unit consists of one animal. It was concluded that pigs submitted to high ambient temperatures (31°C have worse digestibility of protein, phosphorus, calcium, zinc, iron and magnesium and therefore have a higher

  2. 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.

  3. 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

  4. Gene expression

    International Nuclear Information System (INIS)

    Hildebrand, C.E.; Crawford, B.D.; Walters, R.A.; Enger, M.D.

    1983-01-01

    We prepared probes for isolating functional pieces of the metallothionein locus. The probes enabled a variety of experiments, eventually revealing two mechanisms for metallothionein gene expression, the order of the DNA coding units at the locus, and the location of the gene site in its chromosome. Once the switch regulating metallothionein synthesis was located, it could be joined by recombinant DNA methods to other, unrelated genes, then reintroduced into cells by gene-transfer techniques. The expression of these recombinant genes could then be induced by exposing the cells to Zn 2+ or Cd 2+ . We would thus take advantage of the clearly defined switching properties of the metallothionein gene to manipulate the expression of other, perhaps normally constitutive, genes. Already, despite an incomplete understanding of how the regulatory switch of the metallothionein locus operates, such experiments have been performed successfully

  5. Feed efficiency in Mangalarga Marchador foals fed diet supplemented with probiotics or phytase Eficiência alimentar de potros da raça Mangalarga Marchador alimentados com dieta suplementada com probióticos ou fitase

    Directory of Open Access Journals (Sweden)

    Raquel Silva de Moura

    2009-06-01

    Full Text Available The objective was to assess the effect of probiotic or phytase supplementation on dry matter intake (DMI, average daily gain (ADG and feed conversion (FC in 24 Mangalarga Marchador foals (16 males, 8 females, with 241.6 ± 23.2 days of age, and 216.7 ± 27.8 kg of body weight (BW. The study lasted 28 days, the first 23 days for diet and handling adaptation, and the last five days for feces collection. The basal diet consisted of Cynodon pasture and pelleted concentrate (1.2 kg/100 kg BW, as well as water and mineral salt ad libitum. Treatments groups were assigned as follows: basal diet (control; basal diet + 5 g/day of live yeast culture (10(9 CFU/g of S. cerevisiae; basal diet + 2 g/day of live bacteria (10(8 CFU/g of B. bifidum, E. faecium, L. acidophilus, L. plantarum and yeast culture (10(5 CFU/g of S. cerevisiae; and basal diet + 1250 FTU/day of phytase. Pasture DMI was estimated with an external marker (LIPE™ associated with cecal in situ digestibility of dry matter. All foals were randomized, by replicated, using a 4 × 2 factorial, and results were compared using Student t-test, taking into consideration the effect of initial BW as covariate for the percentage DMI per BW. Among foals, DMI varied from 2.4 to 2.8% BW, with foals supplemented with live bacteria and yeast having lower DMI (total diet and forage. All animals had an ADG of 0.33 kg/day, and foals supplemented with live bacteria and yeast had FC 32.42 to 43.83% lower than foals supplemented with phytase or fed only basal diet. The phytase dose of 205 FTU/kg DM in the basal diet had no effects. Probiotic composed of live bacteria and yeast culture improved feed efficiency of forage and can be used as a nutritional strategy to maintain body condition in foals during the dry season.O trabalho foi realizado visando avaliar a influência da suplementação de probióticos ou fitase sobre o consumo diário de matéria seca (CMS, o ganho de peso diário (GPD e a conversão alimentar

  6. 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.

  7. 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.

  8. 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.

  9. Ageing genes

    DEFF Research Database (Denmark)

    Rattan, Suresh

    2018-01-01

    The idea of gerontogenes is in line with the evolutionary explanation of ageing as being an emergent phenomenon as a result of the imperfect maintenance and repair systems. Although evolutionary processes did not select for any specific ageing genes that restrict and determine the lifespan...... of an individual, the term ‘gerontogenes’ primarily refers to any genes that may seem to influence ageing and longevity, without being specifically selected for that role. Such genes can also be called ‘virtual gerontogenes’ by virtue of their indirect influence on the rate and process of ageing. More than 1000...... virtual gerontogenes have been associated with ageing and longevity in model organisms and humans. The ‘real’ genes, which do influence the essential lifespan of a species, and have been selected for in accordance with the evolutionary life history of the species, are known as the longevity assurance...

  10. 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

    Directory of Open Access Journals (Sweden)

    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

  11. 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.

  12. Effects of Protease, Phytase and a Bacillus sp. Direct-Fed Microbial on Nutrient and Energy Digestibility, Ileal Brush Border Digestive Enzyme Activity and Cecal Short-Chain Fatty Acid Concentration in Broiler Chickens

    Science.gov (United States)

    Murugesan, Ganapathi R.; Romero, Luis F.; Persia, Michael E.

    2014-01-01

    Two experiments were conducted to determine the effects of protease and phytase (PP) and a Bacillus sp. direct-fed microbial (DFM) on dietary energy and nutrient utilization in broiler chickens. In the first experiment, Ross 308 broiler chicks were fed diets supplemented with PP and DFM in a 2×2 factorial arrangement. The 4 diets (control (CON), CON + PP, CON + DFM, and CON + PP + DFM) were fed from 15–21 days of age. In Experiment 1, significant interaction (P≤0.01) between PP and DFM on the apparent ileal digestibility coefficient for starch, crude protein, and amino acid indicated that both additives increased the digestibility. Both additives increased the nitrogen retention coefficient with a significant interaction (P≤0.01). Although no interaction was observed, significant main effects (P≤0.01) for nitrogen-corrected apparent ME (AMEn) for PP or DFM indicated an additive response. In a follow-up experiment, Ross 308 broiler chicks were fed the same experimental diets from 1–21 days of age. Activities of ileal brush border maltase, sucrase, and L-alanine aminopeptidase were increased (P≤0.01) by PP addition, while a trend (P = 0.07) for increased sucrase activity was observed in chickens fed DFM, in Experiment 2. The proportion of cecal butyrate was increased (P≤0.01) by DFM addition. Increased nutrient utilization and nitrogen retention appear to involve separate but complementary mechanisms for PP and DFM, however AMEn responses appear to have separate and additive mechanisms. PMID:25013936

  13. 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.

  14. Níveis de cálcio em rações de frangos de corte na fase inicial suplementadas com fitase Levels of calcium in broiler diets supplemented with phytase in the initial phase

    Directory of Open Access Journals (Sweden)

    Neudi Artemio Schoulten

    2003-10-01

    levels of calcium in diets supplemented with the enzyme phytase for chickens from 1 to 21 days of age, an experiment was conducted using 600 1-d old Hubbard-MPK strain chicks. The broilers were allotted to a completely randomized design, factorial arrangement five x two (levels of calcium x sexes, three replicates and 20 birds per plot. The basal ration with 0.54% total phosphorus and 600 units of phytase/kg, was formulated to meet the nutritional requirements of the broilers, according to NRC (1994, except for calcium, whose levels were 0.46; 0.67; 0.88; 1.09 and 1.30%. At 14 days, two chicks of each treatment were transferred to metabolism cages for total collection of the excreta during four consecutive days. At 21 days of age, two broilers from each treatment were slaughtered for removal of the left tibia for subsequent analyses. The males presented higher fed intake and weight gain, and shown a linear reduction as the level of calcium was increased. The levels of 0.67 and 0.71% presented better digestibility of dry matter and higher nitrogen retention, respectively. The females presented content of ashes and calcium in the tibia higher than those of males (1.68 and 2.34% respectively. The level of 0.55% of calcium in the diets provided the maximum deposition of ashes (51.5% while the level of 0.59% was enabled to reach the maximum calcification (17.8%. Phosphorus deposition showed quadratic effect in relation to increasing levels of calcium in the diets, and 0.75% of calcium provided to reach the largest deposition (10.8%, while deposition of manganese was reduced linearly with the increasing levels of calcium in the diets. It was concluded that high levels of calcium in the diets supplemented with phytase decreased weight gain and shown negative effect on the phosphorus and manganese deposition in the broilers tibia from one to 21 days of age. Low levels of calcium impaired the digestibility of dry matter and nitrogen retention of diets of broilers one to 21 day

  15. Desempenho, composição da cama, das tíbias, do fígado e das excretas de frangos de corte alimentados com rações contendo fitase e baixos níveis de fósforo Performance, litter composition, tibia, liver and excreta of broilers fed diets containing reduced levels of phosphorus and phytase enzyme

    Directory of Open Access Journals (Sweden)

    Antonio Carlos de Laurentiz

    2009-10-01

    Full Text Available Um experimento foi realizado para avaliar o desempenho e a composição da cama, das tíbias, do fígado e das excretas de frangos de corte em diversas fases utilizando-se diferentes níveis de fósforo disponível (Pd e da enzima fitase. Foram utilizados 1.200 pintos machos de 1 dia de idade, da marca comercial Cobb, distribuídos em delineamento inteiramente casualizado, em esquema fatorial 4 × 3, composto de quatro níveis de fósforo disponível e três níveis de fitase: 0, 500 e 1.000 ftu/kg de ração, cada um com quatro repetições de 25 aves. Os níveis de fósforo disponível foram obtidos considerando quatro disponibilidades do fósforo total (Pt do milho e do farelo de soja (34, 56, 78 e 100% de disponibilidade. Foram avaliados os índices de desempenho no final de cada fase de criação e aos 42 dias de idade. Os parâmetros de desempenho foram afetados pela redução dos níveis de fósforo disponível das rações e a redução desse elemento para níveis inferiores a 19% piorou o desempenho das aves, enquanto a utilização de fitase no nível de Pd reduzido em 39% melhorou o desempenho. Nas fases de criação, a redução do teor de fósforo disponível com a utilização da enzima fitase influencia o desempenho e as características da cama e dos ossos.An experiment was conducted to evaluate the performance and composition of litter, tibia, liver and excreta of broiler chicks at different phases using different levels of available phosphorous (Pd and the enzyme phytase. One thousand and two hundred one-day-Cobb-male chicks were used, distributed in a randomized complete design, using a 4 × 3 factorial arrangement, consisting of four available phosphorus levels and three phytase levels - 0, 500 and 1,000 FTU phytase/kg of diet with four replications of 25 birds each. The levels of available phosphorus were obtained considering four different availabilities of total phosphorus (Pt, corn and soybean meal (34, 56, 78 and 100

  16. 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

    Directory of Open Access Journals (Sweden)

    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

  17. 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

    Directory of Open Access Journals (Sweden)

    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

  18. Gene doping.

    Science.gov (United States)

    Haisma, H J; de Hon, O

    2006-04-01

    Together with the rapidly increasing knowledge on genetic therapies as a promising new branch of regular medicine, the issue has arisen whether these techniques might be abused in the field of sports. Previous experiences have shown that drugs that are still in the experimental phases of research may find their way into the athletic world. Both the World Anti-Doping Agency (WADA) and the International Olympic Committee (IOC) have expressed concerns about this possibility. As a result, the method of gene doping has been included in the list of prohibited classes of substances and prohibited methods. This review addresses the possible ways in which knowledge gained in the field of genetic therapies may be misused in elite sports. Many genes are readily available which may potentially have an effect on athletic performance. The sporting world will eventually be faced with the phenomena of gene doping to improve athletic performance. A combination of developing detection methods based on gene arrays or proteomics and a clear education program on the associated risks seems to be the most promising preventive method to counteract the possible application of gene doping.

  19. Gene Locater

    DEFF Research Database (Denmark)

    Anwar, Muhammad Zohaib; Sehar, Anoosha; Rehman, Inayat-Ur

    2012-01-01

    software's for calculating recombination frequency is mostly limited to the range and flexibility of this type of analysis. GENE LOCATER is a fully customizable program for calculating recombination frequency, written in JAVA. Through an easy-to-use interface, GENE LOCATOR allows users a high degree...... of flexibility in calculating genetic linkage and displaying linkage group. Among other features, this software enables user to identify linkage groups with output visualized graphically. The program calculates interference and coefficient of coincidence with elevated accuracy in sample datasets. AVAILABILITY...

  20. 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 ratio. In the second posture cycle, the diet consumption decreased with increasing levels of calcium (3.5 to 4.2%, available phosphorus (0.30 to 0.38% and phytase (0 to 600 PU. Feed conversion by eggs mass or per dozen eggs improved, respectively, with increasing levels of available phosphorus and phytase, while specific gravity improved with the increase of dietary calcium level. For laying hens in the first posture cycle, it is recommended 14:1 (4.2% Ca:0.30% Pd ratio without phytase and 12:1 (3.5% Ca:0.30% Pd ratio with 600 PU and, for birds in the second posture cycle, it is recommended 14:1 (4.2% Ca and 0.38% Pd ratio. The supplementation of diets deficient in available phosphorus with 600 PU improves egg weight in the second posture cycle.

  1. Efeitos da suplementação de fitase sobre a disponibilidade aparente de Mg, Ca, Zn, Cu, Mn e Fe em alimentos vegetais para a tilápia-do-nilo Effects of phytase supplementation on apparent availability of Mg, Ca, Zn, Cu, Mn, and Fe of plant feedstuffs for nile tilapia

    Directory of Open Access Journals (Sweden)

    Giovani Sampaio Gonçalves

    2005-12-01

    Full Text Available Cem juvenis de tilápia-do-nilo (Oreochromis niloticus; PV = 100.0 ± 5.0 g foram distribuídos em 10 tanques-rede com o objetivo de avaliar o efeito da suplementação da enzima fitase (0, 1.000 e 2.000 UFA/kg sobre a disponibilidade de minerais em alguns alimentos energéticos (milho, milho extrusado, farelo de trigo, farelo de arroz e farelo de sorgo e protéicos (farelo de soja extrusado, farelo de soja, farelo de girassol, farelo de algodão e glúten de milho utilizados na alimentação de tilápia-do-nilo (Oreochromis niloticus. Para determinação dos coeficientes de disponibilidade aparente (CDA do cálcio (Ca, magnésio (Mg, zinco (Zn, cobre (Cu, ferro (Fe e manganês (Mn, foram confeccionadas 31 rações, marcadas com 0,10% de óxido de crômio III uma referência (ração purificada e 30 contendo os dez alimentos e os diferentes níveis de suplementação da enzima fitase. O CDA dos nutrientes foi calculado com base no teor de crômio da ração e das fezes. A fitase aumenta, nos vegetais, a disponibilidade do Mg, Cu, Zn e Mn, os quais apresentam tendência diferenciada, em razão do seu valor biológico e do nível de suplementação de enzima.One hundred Nile tilapia juveniles (Oreochromis niloticus; BW= 100.0 ± 5.0 g were assigned to 10 experimental cages to evaluate the effects of phytase supplementation (0, 1,000, and 2,000 FTU/kg on calcium (Ca, magnesium (Mg, zinc (Zn, copper (Cu, iron (Fe and manganese (Mn availability of ten feedstuffs: five energetic (corn, extruded corn, wheat meal, rice meal and low-tannin sorghum and five protein (extruded soybean, soybean meal, sunflower meal, cottonseed meal and corn gluten meal. As reference, an albumin and gelatin-based diet [with 0.10 % chromic oxide (III as external marker] was fed to the juveniles. Thirty-one diets (one reference and 30 based on all feedstuffs and increasing levels of phytase were formulated to determine the coefficients of apparent availability of minerals (Mg

  2. Gene Ontology

    Directory of Open Access Journals (Sweden)

    Gaston K. Mazandu

    2012-01-01

    Full Text Available The wide coverage and biological relevance of the Gene Ontology (GO, confirmed through its successful use in protein function prediction, have led to the growth in its popularity. In order to exploit the extent of biological knowledge that GO offers in describing genes or groups of genes, there is a need for an efficient, scalable similarity measure for GO terms and GO-annotated proteins. While several GO similarity measures exist, none adequately addresses all issues surrounding the design and usage of the ontology. We introduce a new metric for measuring the distance between two GO terms using the intrinsic topology of the GO-DAG, thus enabling the measurement of functional similarities between proteins based on their GO annotations. We assess the performance of this metric using a ROC analysis on human protein-protein interaction datasets and correlation coefficient analysis on the selected set of protein pairs from the CESSM online tool. This metric achieves good performance compared to the existing annotation-based GO measures. We used this new metric to assess functional similarity between orthologues, and show that it is effective at determining whether orthologues are annotated with similar functions and identifying cases where annotation is inconsistent between orthologues.

  3. Gene doping: gene delivery for olympic victory

    OpenAIRE

    Gould, David

    2012-01-01

    With one recently recommended gene therapy in Europe and a number of other gene therapy treatments now proving effective in clinical trials it is feasible that the same technologies will soon be adopted in the world of sport by unscrupulous athletes and their trainers in so called ‘gene doping’. In this article an overview of the successful gene therapy clinical trials is provided and the potential targets for gene doping are highlighted. Depending on whether a doping gene product is secreted...

  4. Metabolismo do ferro em suínos recebendo dietas contendo fitase, níveis reduzidos de fósforo inorgânico e sem suplemento micromineral e vitamínico Iron metabolism in swine fed phytase-added diets without mineral vitamin supplement and reduced inorganic phosphorus

    Directory of Open Access Journals (Sweden)

    Renzo Freire de Almeida

    2007-08-01

    Full Text Available O presente estudo foi desenvolvido com o objetivo de avaliar o metabolismo do ferro por meio da determinação do eritrograma, contagem de reticulócitos, dosagem de ferro sérico, ferritina sérica e transferrina sérica de suínos em fase de terminação alimentados com dietas contendo fitase, sem suplemento micromineral/vitamínico e redução dos níveis de fósforo inorgânico (Pi. Foram utilizadas 48 fêmeas suínas de linhagem comercial, com peso inicial de 60kg, distribuídas em seis tratamentos com oito animais em cada grupo. A colheita de sangue foi feita em um grupo de 24 animais com 100kg e em outro grupo de 24 animais com 120kg. Não foram observadas diferenças (P>0,05 nos valores obtidos do eritrograma, da contagem de reticulócitos, de ferro sérico e de transferrina para os animais nos tratamentos testados. Com relação à ferritina, verificou-se que os animais até os 100kg de peso vivo que receberam ração sem suplemento micromineral/vitamínico, sem fósforo inorgânico e com fitase apresentaram valores superiores (PThis research was aimed at evaluating the effect of mineral-vitamin supplement withdrawal associated to reduction of inorganic phosphorus level and addition of phytase in feed on iron metabolism of finishing-phase pigs. Erythrocyte and reticulocyte count, serum iron, ferritin and transferrin quantification was performed. Forty eight hybrid swine females with initial average weight of 60kg were allotted to a completely randomized experimental design with six with four replications of two animals each. Blood was drawn from a group of twenty four 100kg animals and from a second group of twenty four 120kg animals. No differences (P>0.05 were observed in erythrocyte and reticulocyte count or serum iron and transferrin quantification. However, ferritin levels were increased in 100kg animals fed basal feed without mineral/vitamin supplement and inorganic phosphorus with phytase when compared to animals fed basal feed

  5. Genes and Hearing Loss

    Science.gov (United States)

    ... ENTCareers Marketplace Find an ENT Doctor Near You Genes and Hearing Loss Genes and Hearing Loss Patient ... mutation may only have dystopia canthorum. How Do Genes Work? Genes are a road map for the ...

  6. Efeito da fitase e xilanase sobre o desempenho e as características ósseas de frangos de corte alimentados com dietas contendo farelo de arroz Effect of phytase and xilanase on the performance and bone characteristics of broiler chicks fed diets with rice bran

    Directory of Open Access Journals (Sweden)

    Ademir José Conte

    2003-10-01

    Full Text Available O presente trabalho foi conduzido com o objetivo de verificar o efeito do uso da fitase e xilanase no desempenho e na deposição óssea de minerais em frangos de corte alimentados com dietas contendo 15% de farelo de arroz, baixo fósforo disponível e sem suplementação de ferro, cobre, zinco e manganês. A fitase utilizada foi da marca comercial Natuphos 5000 e a xilanase, da marca Avizyme 1300. O delineamento experimental foi inteiramente casualizado, em esquema fatorial 4x2+1. A dieta controle foi composta por níveis adequados de nutrientes, sem farelo de arroz e sem enzimas. As dietas dos demais tratamentos foram formuladas com 15% de farelo de arroz, contendo baixo fósforo disponível (40% do recomendado e sem suplementação inorgânica de Fe, Cu, Zn e Mn, com quatro níveis de fitase (0, 400, 800 e 1.200 FTU/kg, com ou sem xilanase (1 kg/t. As variáveis estudadas aos 21 e 42 dias de idade foram desempenho e cinzas, P, Zn, Fe, Mn e Cu na tíbia. O peso vivo e consumo de ração aumentaram de maneira significativa com a adição de fitase, enquanto que a conversão alimentar não foi afetada, tanto aos 21 como aos 42 dias. O melhor nível de fitase foi de 1.105 e 1.023 FTU/kg para melhor desempenho, aos 21 e 42 dias de idade, respectivamente. A xilanase não afetou significativamente o peso vivo e o consumo de ração, porém melhorou significativamente a conversão alimentar nas duas idades avaliadas, em função de menor consumo de ração nas aves que receberam xilanase. Os níveis de fitase promoveram aumento linear nos teores de cinzas e fósforo da tíbia e não afetaram a deposição de Zn, Fe, Mn e Cu, nas duas idades estudadas. A xilanase não afetou os parâmetros avaliados na tíbia.The present work was designed to verify the effect of the use of phytase and xylanase upon the performance and bone deposition of minerals in broiler diets containing 15% of rice bran, low available phosphorus and without supplementation of iron

  7. Gene expression and gene therapy imaging

    International Nuclear Information System (INIS)

    Rome, Claire; Couillaud, Franck; Moonen, Chrit T.W.

    2007-01-01

    The fast growing field of molecular imaging has achieved major advances in imaging gene expression, an important element of gene therapy. Gene expression imaging is based on specific probes or contrast agents that allow either direct or indirect spatio-temporal evaluation of gene expression. Direct evaluation is possible with, for example, contrast agents that bind directly to a specific target (e.g., receptor). Indirect evaluation may be achieved by using specific substrate probes for a target enzyme. The use of marker genes, also called reporter genes, is an essential element of MI approaches for gene expression in gene therapy. The marker gene may not have a therapeutic role itself, but by coupling the marker gene to a therapeutic gene, expression of the marker gene reports on the expression of the therapeutic gene. Nuclear medicine and optical approaches are highly sensitive (detection of probes in the picomolar range), whereas MRI and ultrasound imaging are less sensitive and require amplification techniques and/or accumulation of contrast agents in enlarged contrast particles. Recently developed MI techniques are particularly relevant for gene therapy. Amongst these are the possibility to track gene therapy vectors such as stem cells, and the techniques that allow spatiotemporal control of gene expression by non-invasive heating (with MRI guided focused ultrasound) and the use of temperature sensitive promoters. (orig.)

  8. Imaging reporter gene for monitoring gene therapy

    International Nuclear Information System (INIS)

    Beco, V. de; Baillet, G.; Tamgac, F.; Tofighi, M.; Weinmann, P.; Vergote, J.; Moretti, J.L.; Tamgac, G.

    2002-01-01

    Scintigraphic images can be obtained to document gene function at cellular level. This approach is presented here and the use of a reporter gene to monitor gene therapy is described. Two main ways are presented: either the use of a reporter gene coding for an enzyme the action of which will be monitored by radiolabeled pro-drug, or a cellular receptor gene, the action of which is documented by a radio labeled cognate receptor ligand. (author)

  9. Redução de proteína e fósforo em rações com fitase para frangos de corte no período de 1 a 21 dias de idade: desempenho e teores de minerais na cama Decreasing dietary levels of protein and phosphorum supplemented with phytase for broilers from 1 to 21 old: performance and mineral levels in the litter

    Directory of Open Access Journals (Sweden)

    Yolanda Lopes da Silva

    2006-06-01

    Full Text Available O experimento foi realizado para avaliar o desempenho e o teor de minerais na cama de frangos no período de 1 a 21 dias de idade, alimentados com rações contendo níveis reduzidos de proteína bruta (PB e fósforo disponível (Pd suplementadas com aminoácidos e fitase. Foram utilizados 1.500 pintos de corte machos de um dia de idade, da marca comercial Cobb, distribuídos em delineamento de blocos casualizados, em esquema fatorial 3 x 3 + 1, com três níveis de PB (15, 17 e 19%, três de Pd (0,25; 0,34 e 0,45% e uma ração controle formulada de acordo com os níveis nutricionais recomendados por pesquisadores brasileiros, totalizando dez tratamentos e seis repetições de 25 aves. Nas rações com 0,25 e 0,34% de Pd, adicionaram-se 500 FTU de fitase, reduzindo em 17% o teor de cálcio. O ganho de peso das aves que consumiram as rações com 17% de PB, 0,34 e 0,45 de Pd ou com 19% de PB e diferentes de níveis de Pd foi similar ao daquelas alimentadas com a ração controle. O consumo das aves que receberam as rações com 15 e 17% de PB e 0,25% de Pd + fitase foi menor que o daquelas que consumiram a ração controle. Não houve diferenças entre as rações para a conversão alimentar. Com exceção do nitrogênio, houve redução nos teores de minerais na cama quando as aves foram alimentadas com a ração contendo 17% de PB e 0,34% de Pd. Reduções para 17% nos teores de PB e para 0,34% nos teores de Pd das rações para frangos na fase inicial não prejudicam o desempenho das aves, desde que as rações sejam suplementadas com aminoácidos e fitase. Nesses níveis de PB e Pd, observou-se redução no impacto ambiental, como conseqüência da redução nos teores de P, Ca, Cu e Zn na cama.The trial was carried out to evaluate the performance and mineral levels in the litter of broilers from 1 to 21 days old, fed diets with low-crude protein (CP and low available phosphorus (AP supplemented with synthetic amino acids and phytase. A

  10. Nitrogen, calcium and phosphorus balance of broilers fed diets with phytase and crystalline amino acids Balanço de nitrogênio, cálcio e fósforo em frangos de corte recebendo dietas com fitase e aminoácidos cristalinos

    Directory of Open Access Journals (Sweden)

    Elisangela Minati Gomide

    2011-06-01

    Full Text Available The objective of this study was to evaluate the effect of reduced-crude protein (CP and available phosphorus (aP diets, added with phytase and amino acids for broilers in the growing (14 to 21 days old and retirement (35 to 42 days phases on nitrogen, potassium and phosphorus balance. Ninety Cobb broilers were distributed in a completely randomized design with three treatments and six replications with five and three birds per experimental unit in the growing and retirement phases, respectively. Diets were elaborated to contain reduced CP and aP levels (19 and 17% CP in the growing phase, with 0.34% aP; and 18 and 16% CP in the retirement phase, with 0.30% aP, supplemented with crystalline amino acids and 500 FTU of phytase/kg diet. These were compared to a control diet (21% CP and 0.46% aP for growing phase and 19% CP and 0.40% aP for retirement phase. The reduction of CP in four percentual units reduced the N excretion and increased the retention of this element, showing that the phytase had no influence on N balance when used the crystalline amino acids in the diets. The phytase reduced the P excretion and increased the retention ones. The potassium excretion decreased in reduced-CP diets, but this excessive reduction decreased the retention of this element, except in the retirement phase. It was concluded that the reduced-CP and aP diets, supplemented with phytase and amino acids, are efficient in reducing the pollutant power of the broilers excrements in the growing and retirement phases of production.Objetivou-se, com este trabalho, avaliar o efeito de rações com reduzidos teores de proteína bruta (PB e fósforo disponível (Pd, suplementadas com fitase e aminoácidos, para frangos de corte na fase de crescimento (14 a 21 dias e terminação (35 a 42 dias de idade sobre o balanço de nitrogênio, potássio e fósforo. Foram utilizados 90 pintos Cobb, distribuídos em um delineamento inteiramente casualizado com três tratamentos e seis

  11. Phylotype dynamics of bacterial P utilization genes in microbialites and bacterioplankton of a monomictic endorheic lake

    Czech Academy of Sciences Publication Activity Database

    Valdespino-Castillo, P.M.; Alcantara-Hernandez, R.J.; Merino-Ibarra, M.; Alcocer, J.; Macek, Miroslav; Moreno-Guillen, O.A.; Falcon, L.I.

    2017-01-01

    Roč. 73, č. 2 (2017), s. 296-309 ISSN 0095-3628 Institutional support: RVO:60077344 Keywords : extracellular enzymes * DOP utilization * phytase * P turnover Subject RIV: EE - Microbiology, Virology OBOR OECD: Microbiology Impact factor: 3.630, year: 2016

  12. Imaging gene expression in gene therapy

    International Nuclear Information System (INIS)

    Wiebe, Leonard I.

    1997-01-01

    Full text. Gene therapy can be used to introduce new genes, or to supplement the function of indigenous genes. At the present time, however, there is non-invasive test to demonstrate efficacy of the gene transfer and expression processes. It has been postulated that scintigraphic imaging can offer unique information on both the site at which the transferred gene is expressed, and the degree of expression, both of which are critical issue for safety and clinical efficacy. Many current studies are based on 'suicide gene therapy' of cancer. Cells modified to express these genes commit metabolic suicide in the presence of an enzyme encoded by the transferred gene and a specifically-convertible pro drug. Pro drug metabolism can lead to selective metabolic trapping, required for scintigraphy. Herpes simplex virus type-1 thymidine kinase (H S V-1 t k + ) has been use for 'suicide' in vivo tumor gene therapy. It has been proposed that radiolabelled nucleosides can be used as radiopharmaceuticals to detect H S V-1 t k + gene expression where the H S V-1 t k + gene serves a reporter or therapeutic function. Animal gene therapy models have been studied using purine-([ 18 F]F H P G; [ 18 F]-A C V), and pyrimidine- ([ 123 / 131 I]I V R F U; [ 124 / 131I ]) antiviral nucleosides. Principles of gene therapy and gene therapy imaging will be reviewed and experimental data for [ 123 / 131I ]I V R F U imaging with the H S V-1 t k + reporter gene will be presented

  13. Imaging gene expression in gene therapy

    Energy Technology Data Exchange (ETDEWEB)

    Wiebe, Leonard I. [Alberta Univ., Edmonton (Canada). Noujaim Institute for Pharmaceutical Oncology Research

    1997-12-31

    Full text. Gene therapy can be used to introduce new genes, or to supplement the function of indigenous genes. At the present time, however, there is non-invasive test to demonstrate efficacy of the gene transfer and expression processes. It has been postulated that scintigraphic imaging can offer unique information on both the site at which the transferred gene is expressed, and the degree of expression, both of which are critical issue for safety and clinical efficacy. Many current studies are based on `suicide gene therapy` of cancer. Cells modified to express these genes commit metabolic suicide in the presence of an enzyme encoded by the transferred gene and a specifically-convertible pro drug. Pro drug metabolism can lead to selective metabolic trapping, required for scintigraphy. Herpes simplex virus type-1 thymidine kinase (H S V-1 t k{sup +}) has been use for `suicide` in vivo tumor gene therapy. It has been proposed that radiolabelled nucleosides can be used as radiopharmaceuticals to detect H S V-1 t k{sup +} gene expression where the H S V-1 t k{sup +} gene serves a reporter or therapeutic function. Animal gene therapy models have been studied using purine-([{sup 18} F]F H P G; [{sup 18} F]-A C V), and pyrimidine- ([{sup 123}/{sup 131} I]I V R F U; [{sup 124}/{sup 131I}]) antiviral nucleosides. Principles of gene therapy and gene therapy imaging will be reviewed and experimental data for [{sup 123}/{sup 131I}]I V R F U imaging with the H S V-1 t k{sup +} reporter gene will be presented

  14. Gene doping: gene delivery for olympic victory.

    Science.gov (United States)

    Gould, David

    2013-08-01

    With one recently recommended gene therapy in Europe and a number of other gene therapy treatments now proving effective in clinical trials it is feasible that the same technologies will soon be adopted in the world of sport by unscrupulous athletes and their trainers in so called 'gene doping'. In this article an overview of the successful gene therapy clinical trials is provided and the potential targets for gene doping are highlighted. Depending on whether a doping gene product is secreted from the engineered cells or is retained locally to, or inside engineered cells will, to some extent, determine the likelihood of detection. It is clear that effective gene delivery technologies now exist and it is important that detection and prevention plans are in place. © 2012 The Author. British Journal of Clinical Pharmacology © 2012 The British Pharmacological Society.

  15. Disponibilidade aparente do fósforo em alimentos vegetais e suplementação da enzima fitase para tilápia-do-nilo Apparent phosphorus availability in vegetable feedstuffs and supplementation of phytase enzyme for Nile tilapia Oreochromis niloticus

    Directory of Open Access Journals (Sweden)

    Giovani Sampaio Gonçalves

    2007-10-01

    Full Text Available Este estudo foi realizado com o objetivo de avaliar o efeito da fitase (0, 1.000 e 2.000 uf/kg na disponibilidade aparente de P de cinco alimentos energéticos (milho, milho extrusado, farelo de trigo, farelo de arroz e sorgo baixo tanino e cinco alimentos protéicos (soja extrusada, farelo de soja, farelo de girassol, farelo de algodão e glúten de milho utilizando-se uma ração purificada como controle. Foram utilizados 100 juvenis (100,00 ± 5,00 g de tilápia-do-nilo (Oreochromis niloticus alojados em dez gaiolas para manejo de alimentação e coleta de fezes. Para cada alimento e cada nível de fitase avaliado, foram coletadas amostras em cinco gaiolas, totalizando cinco repetições por tratamento. Os resultados obtidos indicaram que a suplementação de até 2.000 uf/kg de fitase não aumentou a disponibilidade aparente de P do farelo de trigo e do farelo de algodão. Entretanto, o nível de 1.000 uf/kg aumentou a disponibilidade aparente de P da soja extrusada e do farelo de girassol, enquanto o nível de 2.000 uf/kg proporcionou essa resposta para o farelo de milho, o milho extrusado, o sorgo baixo tanino, o farelo de arroz, o farelo de soja e o glúten de milho.This study was carried out to evaluate effects of phytase (0, 1,000, and 2,000 uf/kg on apparent P availability of five energetic feeds (corn, extruded corn, wheat meal, rice meal and low-tannin sorghum and five protein feeds (extruded soy, soybean meal, sunflower meal, cottonseed meal and corn gluten using a purified diet as control. One hundred (100.00 ± 5.00 g Nile tilapia juveniles (Oreochromis niloticus were allotted to ten cages to allow feeding and fecal collection. For each feed and each phytase level evaluated samples were collected from five cages, in a total of five replications per treatment. The obtained results showed that the phytase supplementation up to 2,000 uf/kg did not increase apparent P availability present in wheat meal or cottonseed meal. However, the

  16. Evolution of homeobox genes.

    Science.gov (United States)

    Holland, Peter W H

    2013-01-01

    Many homeobox genes encode transcription factors with regulatory roles in animal and plant development. Homeobox genes are found in almost all eukaryotes, and have diversified into 11 gene classes and over 100 gene families in animal evolution, and 10 to 14 gene classes in plants. The largest group in animals is the ANTP class which includes the well-known Hox genes, plus other genes implicated in development including ParaHox (Cdx, Xlox, Gsx), Evx, Dlx, En, NK4, NK3, Msx, and Nanog. Genomic data suggest that the ANTP class diversified by extensive tandem duplication to generate a large array of genes, including an NK gene cluster and a hypothetical ProtoHox gene cluster that duplicated to generate Hox and ParaHox genes. Expression and functional data suggest that NK, Hox, and ParaHox gene clusters acquired distinct roles in patterning the mesoderm, nervous system, and gut. The PRD class is also diverse and includes Pax2/5/8, Pax3/7, Pax4/6, Gsc, Hesx, Otx, Otp, and Pitx genes. PRD genes are not generally arranged in ancient genomic clusters, although the Dux, Obox, and Rhox gene clusters arose in mammalian evolution as did several non-clustered PRD genes. Tandem duplication and genome duplication expanded the number of homeobox genes, possibly contributing to the evolution of developmental complexity, but homeobox gene loss must not be ignored. Evolutionary changes to homeobox gene expression have also been documented, including Hox gene expression patterns shifting in concert with segmental diversification in vertebrates and crustaceans, and deletion of a Pitx1 gene enhancer in pelvic-reduced sticklebacks. WIREs Dev Biol 2013, 2:31-45. doi: 10.1002/wdev.78 For further resources related to this article, please visit the WIREs website. The author declares that he has no conflicts of interest. Copyright © 2012 Wiley Periodicals, Inc.

  17. Gene cluster statistics with gene families.

    Science.gov (United States)

    Raghupathy, Narayanan; Durand, Dannie

    2009-05-01

    Identifying genomic regions that descended from a common ancestor is important for understanding the function and evolution of genomes. In distantly related genomes, clusters of homologous gene pairs are evidence of candidate homologous regions. Demonstrating the statistical significance of such "gene clusters" is an essential component of comparative genomic analyses. However, currently there are no practical statistical tests for gene clusters that model the influence of the number of homologs in each gene family on cluster significance. In this work, we demonstrate empirically that failure to incorporate gene family size in gene cluster statistics results in overestimation of significance, leading to incorrect conclusions. We further present novel analytical methods for estimating gene cluster significance that take gene family size into account. Our methods do not require complete genome data and are suitable for testing individual clusters found in local regions, such as contigs in an unfinished assembly. We consider pairs of regions drawn from the same genome (paralogous clusters), as well as regions drawn from two different genomes (orthologous clusters). Determining cluster significance under general models of gene family size is computationally intractable. By assuming that all gene families are of equal size, we obtain analytical expressions that allow fast approximation of cluster probabilities. We evaluate the accuracy of this approximation by comparing the resulting gene cluster probabilities with cluster probabilities obtained by simulating a realistic, power-law distributed model of gene family size, with parameters inferred from genomic data. Surprisingly, despite the simplicity of the underlying assumption, our method accurately approximates the true cluster probabilities. It slightly overestimates these probabilities, yielding a conservative test. We present additional simulation results indicating the best choice of parameter values for data

  18. Carboxylesterase 1 genes

    DEFF Research Database (Denmark)

    Rasmussen, Henrik Berg; Madsen, Majbritt Busk

    2018-01-01

    The carboxylesterase 1 gene (CES1) encodes a hydrolase that metabolizes commonly used drugs. The CES1-related pseudogene, carboxylesterase 1 pseudogene 1 (CES1P1), has been implicated in gene exchange with CES1 and in the formation of hybrid genes including the carboxylesterase 1A2 gene (CES1A2...

  19. Gene doping in sports.

    Science.gov (United States)

    Unal, Mehmet; Ozer Unal, Durisehvar

    2004-01-01

    Gene or cell doping is defined by the World Anti-Doping Agency (WADA) as "the non-therapeutic use of genes, genetic elements and/or cells that have the capacity to enhance athletic performance". New research in genetics and genomics will be used not only to diagnose and treat disease, but also to attempt to enhance human performance. In recent years, gene therapy has shown progress and positive results that have highlighted the potential misuse of this technology and the debate of 'gene doping'. Gene therapies developed for the treatment of diseases such as anaemia (the gene for erythropoietin), muscular dystrophy (the gene for insulin-like growth factor-1) and peripheral vascular diseases (the gene for vascular endothelial growth factor) are potential doping methods. With progress in gene technology, many other genes with this potential will be discovered. For this reason, it is important to develop timely legal regulations and to research the field of gene doping in order to develop methods of detection. To protect the health of athletes and to ensure equal competitive conditions, the International Olympic Committee, WADA and International Sports Federations have accepted performance-enhancing substances and methods as being doping, and have forbidden them. Nevertheless, the desire to win causes athletes to misuse these drugs and methods. This paper reviews the current status of gene doping and candidate performance enhancement genes, and also the use of gene therapy in sports medicine and ethics of genetic enhancement. Copyright 2004 Adis Data Information BV

  20. Human Gene Therapy: Genes without Frontiers?

    Science.gov (United States)

    Simon, Eric J.

    2002-01-01

    Describes the latest advancements and setbacks in human gene therapy to provide reference material for biology teachers to use in their science classes. Focuses on basic concepts such as recombinant DNA technology, and provides examples of human gene therapy such as severe combined immunodeficiency syndrome, familial hypercholesterolemia, and…

  1. Níveis de energia metabolizável em rações formuladas com base no conceito de proteína ideal e suplementadas com fitase para leitões dos 15 aos 35 kg Metabolizable energy levels in diets formulated according to the ideal protein concept and supplemented with phytase for piglets from 15 to 35 kg

    Directory of Open Access Journals (Sweden)

    Marcelo José Milagres de Almeida

    2008-05-01

    15 to 35 kg to evaluate the effect of different dietary levels of metabolizable energy (ME and crude protein (CP formulated according to the ideal protein concept and phytase supplementation on performance and carcass characteristics. The experiment was analyzed as a complete randomized block design, in a factorial arrangement of treatments 3 × 2 + 1, with three ME levels (3,080, 3,230, and 3,380 kcal/kg, two of CP (14% and 16%, supplemented with synthetic amino acids and 1000 FTU/kg of phytase, with calcium level reduced by 25% and phosphorus by 30%. Additional control treatment was formulated with 18% CP and without phytase, to attend the nutrient requirements of pigs according to Brazilian Feedstuffs Tables recommendations. Therefore, seven treatments, six replications and two animals per experimental unit were used. No treatment effect on daily weight gain, feed conversion, daily ME intake and weight of organs (absolute and relative of pigs was observed.Feed intake was higher and fat deposition rate, lower in pigs fed diet with 3,380 kcal ME/kg. Decreasing nitrogen intake, increasing efficiency of nitrogen utilization for gain and lower blood urea content were observed for pigs fed 14% CP diets compared to those with 18%. The reduction of levels of ME, CP, available phosphorus and calcium to 3,080 kcal/kg, 14%, 0.54% and 0.28%, respectively, in rations for pigs, formulated based on ideal protein concept and phytase, do not affect performance of pigs from 15 to 35 kg.

  2. Tumor targeted gene therapy

    International Nuclear Information System (INIS)

    Kang, Joo Hyun

    2006-01-01

    Knowledge of molecular mechanisms governing malignant transformation brings new opportunities for therapeutic intervention against cancer using novel approaches. One of them is gene therapy based on the transfer of genetic material to an organism with the aim of correcting a disease. The application of gene therapy to the cancer treatment had led to the development of new experimental approaches such as suicidal gene therapy, inhibition of oncogenes and restoration of tumor-suppressor genes. Suicidal gene therapy is based on the expression in tumor cells of a gene encoding an enzyme that converts a prodrug into a toxic product. Representative suicidal genes are Herpes simplex virus type 1 thymidine kinase (HSV1-tk) and cytosine deaminase (CD). Especially, physicians and scientists of nuclear medicine field take an interest in suicidal gene therapy because they can monitor the location and magnitude, and duration of expression of HSV1-tk and CD by PET scanner

  3. Essential Bacillus subtilis genes

    DEFF Research Database (Denmark)

    Kobayashi, K.; Ehrlich, S.D.; Albertini, A.

    2003-01-01

    To estimate the minimal gene set required to sustain bacterial life in nutritious conditions, we carried out a systematic inactivation of Bacillus subtilis genes. Among approximate to4,100 genes of the organism, only 192 were shown to be indispensable by this or previous work. Another 79 genes were...... predicted to be essential. The vast majority of essential genes were categorized in relatively few domains of cell metabolism, with about half involved in information processing, one-fifth involved in the synthesis of cell envelope and the determination of cell shape and division, and one-tenth related...... to cell energetics. Only 4% of essential genes encode unknown functions. Most essential genes are present throughout a wide range of Bacteria, and almost 70% can also be found in Archaea and Eucarya. However, essential genes related to cell envelope, shape, division, and respiration tend to be lost from...

  4. Biodisponibilidade e perdas endógenas mínimas de P em dietas com níveis crescentes de fitase para suínos em crescimento pela técnica de diluição isotópica Bioavailability and minimum endogenous losses of P in diets with phytase increasing levels for growing swine by using the isotopic dilution technique

    Directory of Open Access Journals (Sweden)

    J.A. Moreira

    2003-06-01

    Full Text Available O objetivo do trabalho foi avaliar rações à base de milho, farelo de soja e farelo de arroz desengordurado contendo níveis crescentes de fitase (253, 759, 1265 e 1748 UF/kg de ração, por meio da biodisponibilidade e das perdas endógenas mínimas de P com o uso da técnica de diluição isotópica. Foram utilizados 20 suínos mestiços (machos castrados, com peso médio de 32,19+2,17 kg, em delineamento experimental de blocos ao acaso, contendo cinco tratamentos e quatro repetições. Os leitões foram mantidos em gaiolas metabólicas durante 10 dias para adaptação e sete dias para coletas de sangue, fezes e urina. No primeiro dia de coleta foram injetados em cada animal, por via endovenosa, 7,4 MBq de 32P. No final do experimento os animais foram abatidos e os tecidos do músculo, coração, fígado, rins e ossos coletados para estudos posteriores. Os níveis de enzima fitase não afetaram o P retido, o P urinário, o P plasma e a biodisponibilidade. Observou-se efeito quadrático dos níveis de fitase sobre o P nas fezes e endógeno. A variável perda endógena de fósforo foi mais precisa do que a variável biodisponibilidade para avaliar os efeitos da enzima fitase. O melhor aproveitamento do fósforo fítico ocorreu no nível 759 UF.The objective of this study was to evaluate diets containing increasing levels of phytase (253, 759, 1265 and 1748 PU/kg of diets, by mean of bioavailability and minimum endogenous P losses. Twenty crossbred barrow weighting 32.19+2.17 kg were used in a randomized block design experiment, with five treatments and four replicates. The animals were kept in metabolic cages for a ten-day adaptation period and seven days for total collection of feces and urine. Blood samples were taken for seven days. At the first day of the collection period, each animal was injected intravenously with 7.4 MBq of 32P. At the end of the experimental period the animals were slaughtered and tissues of the muscle, heart, liver

  5. Radiotechnologies and gene therapy

    International Nuclear Information System (INIS)

    Xia Jinsong

    2001-01-01

    Gene therapy is an exciting frontier in medicine today. Radiologist will make an uniquely contribution to these exciting new technologies at every level by choosing sites for targeting therapy, perfecting and establishing routes of delivery, developing imaging strategies to monitor therapy and assess gene expression, developing radiotherapeutic used of gene therapy

  6. Discovering genes underlying QTL

    Energy Technology Data Exchange (ETDEWEB)

    Vanavichit, Apichart [Kasetsart University, Kamphaengsaen, Nakorn Pathom (Thailand)

    2002-02-01

    A map-based approach has allowed scientists to discover few genes at a time. In addition, the reproductive barrier between cultivated rice and wild relatives has prevented us from utilizing the germ plasm by a map-based approach. Most genetic traits important to agriculture or human diseases are manifested as observable, quantitative phenotypes called Quantitative Trait Loci (QTL). In many instances, the complexity of the phenotype/genotype interaction and the general lack of clearly identifiable gene products render the direct molecular cloning approach ineffective, thus additional strategies like genome mapping are required to identify the QTL in question. Genome mapping requires no prior knowledge of the gene function, but utilizes statistical methods to identify the most likely gene location. To completely characterize genes of interest, the initially mapped region of a gene location will have to be narrowed down to a size that is suitable for cloning and sequencing. Strategies for gene identification within the critical region have to be applied after the sequencing of a potentially large clone or set of clones that contains this gene(s). Tremendous success of positional cloning has been shown for cloning many genes responsible for human diseases, including cystic fibrosis and muscular dystrophy as well as plant disease resistance genes. Genome and QTL mapping, positional cloning: the pre-genomics era, comparative approaches to gene identification, and positional cloning: the genomics era are discussed in the report. (M. Suetake)

  7. Gene therapy: An overview

    Directory of Open Access Journals (Sweden)

    Sudip Indu

    2013-01-01

    Full Text Available Gene therapy "the use of genes as medicine" involves the transfer of a therapeutic or working copy of a gene into specific cells of an individual in order to repair a faulty gene copy. The technique may be used to replace a faulty gene, or to introduce a new gene whose function is to cure or to favorably modify the clinical course of a condition. The objective of gene therapy is to introduce new genetic material into target cells while causing no damage to the surrounding healthy cells and tissues, hence the treatment related morbidity is decreased. The delivery system includes a vector that delivers a therapeutic gene into the patient′s target cell. Functional proteins are created from the therapeutic gene causing the cell to return to a normal stage. The vectors used in gene therapy can be viral and non-viral. Gene therapy, an emerging field of biomedicine, is still at infancy and much research remains to be done before this approach to the treatment of condition will realize its full potential.

  8. Gene therapy in periodontics.

    Science.gov (United States)

    Chatterjee, Anirban; Singh, Nidhi; Saluja, Mini

    2013-03-01

    GENES are made of DNA - the code of life. They are made up of two types of base pair from different number of hydrogen bonds AT, GC which can be turned into instruction. Everyone inherits genes from their parents and passes them on in turn to their children. Every person's genes are different, and the changes in sequence determine the inherited differences between each of us. Some changes, usually in a single gene, may cause serious diseases. Gene therapy is 'the use of genes as medicine'. It involves the transfer of a therapeutic or working gene copy into specific cells of an individual in order to repair a faulty gene copy. Thus it may be used to replace a faulty gene, or to introduce a new gene whose function is to cure or to favorably modify the clinical course of a condition. It has a promising era in the field of periodontics. Gene therapy has been used as a mode of tissue engineering in periodontics. The tissue engineering approach reconstructs the natural target tissue by combining four elements namely: Scaffold, signaling molecules, cells and blood supply and thus can help in the reconstruction of damaged periodontium including cementum, gingival, periodontal ligament and bone.

  9. Primetime for Learning Genes.

    Science.gov (United States)

    Keifer, Joyce

    2017-02-11

    Learning genes in mature neurons are uniquely suited to respond rapidly to specific environmental stimuli. Expression of individual learning genes, therefore, requires regulatory mechanisms that have the flexibility to respond with transcriptional activation or repression to select appropriate physiological and behavioral responses. Among the mechanisms that equip genes to respond adaptively are bivalent domains. These are specific histone modifications localized to gene promoters that are characteristic of both gene activation and repression, and have been studied primarily for developmental genes in embryonic stem cells. In this review, studies of the epigenetic regulation of learning genes in neurons, particularly the brain-derived neurotrophic factor gene ( BDNF ), by methylation/demethylation and chromatin modifications in the context of learning and memory will be highlighted. Because of the unique function of learning genes in the mature brain, it is proposed that bivalent domains are a characteristic feature of the chromatin landscape surrounding their promoters. This allows them to be "poised" for rapid response to activate or repress gene expression depending on environmental stimuli.

  10. Genes and Social Behavior

    OpenAIRE

    Robinson, Gene E.; Fernald, Russell D.; Clayton, David F.

    2008-01-01

    What specific genes and regulatory sequences contribute to the organization and functioning of brain circuits that support social behavior? How does social experience interact with information in the genome to modulate these brain circuits? Here we address these questions by highlighting progress that has been made in identifying and understanding two key “vectors of influence” that link genes, brain, and social behavior: 1) social information alters gene readout in the brain to influence beh...

  11. History of gene therapy.

    Science.gov (United States)

    Wirth, Thomas; Parker, Nigel; Ylä-Herttuala, Seppo

    2013-08-10

    Two decades after the initial gene therapy trials and more than 1700 approved clinical trials worldwide we not only have gained much new information and knowledge regarding gene therapy in general, but also learned to understand the concern that has persisted in society. Despite the setbacks gene therapy has faced, success stories have increasingly emerged. Examples for these are the positive recommendation for a gene therapy product (Glybera) by the EMA for approval in the European Union and the positive trials for the treatment of ADA deficiency, SCID-X1 and adrenoleukodystrophy. Nevertheless, our knowledge continues to grow and during the course of time more safety data has become available that helps us to develop better gene therapy approaches. Also, with the increased understanding of molecular medicine, we have been able to develop more specific and efficient gene transfer vectors which are now producing clinical results. In this review, we will take a historical view and highlight some of the milestones that had an important impact on the development of gene therapy. We will also discuss briefly the safety and ethical aspects of gene therapy and address some concerns that have been connected with gene therapy as an important therapeutic modality. Copyright © 2013 Elsevier B.V. All rights reserved.

  12. Refining discordant gene trees.

    Science.gov (United States)

    Górecki, Pawel; Eulenstein, Oliver

    2014-01-01

    Evolutionary studies are complicated by discordance between gene trees and the species tree in which they evolved. Dealing with discordant trees often relies on comparison costs between gene and species trees, including the well-established Robinson-Foulds, gene duplication, and deep coalescence costs. While these costs have provided credible results for binary rooted gene trees, corresponding cost definitions for non-binary unrooted gene trees, which are frequently occurring in practice, are challenged by biological realism. We propose a natural extension of the well-established costs for comparing unrooted and non-binary gene trees with rooted binary species trees using a binary refinement model. For the duplication cost we describe an efficient algorithm that is based on a linear time reduction and also computes an optimal rooted binary refinement of the given gene tree. Finally, we show that similar reductions lead to solutions for computing the deep coalescence and the Robinson-Foulds costs. Our binary refinement of Robinson-Foulds, gene duplication, and deep coalescence costs for unrooted and non-binary gene trees together with the linear time reductions provided here for computing these costs significantly extends the range of trees that can be incorporated into approaches dealing with discordance.

  13. Digestibilidade aparente e suplementação de fitase em alimentos vegetais para tilápia do Nilo - DOI: 10.4025/actascianimsci.v26i3.1795 Apparent digestibility and supplementation of phytase in vegetable feedstuff for Nile tilapia - DOI: 10.4025/actascianimsci.v26i3.1795

    Directory of Open Access Journals (Sweden)

    Hamilton Hisano

    2004-04-01

    Full Text Available Alimentos de origem vegetal utilizados em rações comerciais para peixes contêm fatores antinutricionais tais como o fitato, molécula capaz de complexar com minerais, aminoácidos, amido e que pode, também, atuar de forma negativa nas reações enzimáticas digestivas. Desse modo, esse estudo teve por objetivo avaliar o efeito da enzima fitase (0, 1.000 e 2.000 UF/kg na digestibilidade aparente de 5 alimentos energéticos (milho, milho extrusado, farelos de trigo, de arroz e de sorgo e 5 protéicos (soja extrusada, farelos de soja, de girassol, de algodão e glúten de milho utilizando uma ração à base de albumina e gelatina e 0,10% de óxido de crômio (Cr2O3 como referência. Foram determinados os coeficientes de digestibilidade aparente (CDA da matéria seca, a proteína bruta e energia bruta. Foram utilizados 100 juvenis de tilápia do Nilo com peso médio de 100,0 g os quais foram alojados em 10 gaiolas para o manejo de alimentação e coleta de fezes. Alimentos de origem vegetal respondem à presença da enzima fitase de forma diferenciada. Os valores digestíveis da matéria seca, da proteína bruta e energia bruta dependem do valor biológico dos alimentos, da concentração e da natureza do fitato e do nível de suplementação de enzima.Vegetable feedstuff used in commercial fish diets contains antinutrients, such as phytate, molecule able to complex minerals, amino acids, starch and, which can also acts in a negative way on the digestive enzymatic reactions. Thus, this study aimed to evaluate the effect of phytase enzyme (0; 1,000; 2,000 UF/kg on the apparent digestibility of five energetic feedstuff (corn, extruded corn, wheat middling, rice meal and sorghum and five protein feedstuff (soybean meal, extruded soybean meal, sunflower meal, cottonseed meal and gluten meal using an albumin-gelatin diet and 0.10% of chromic oxide (Cr2O3, as indicator. Apparent digestibility coefficient (ADC of dry matter, crude protein and gross

  14. Efeito de diferentes níveis de inclusão de farelo de arroz em dietas suplementadas com fitase para frangos de corte = Effect of different inclusion levels of rice bran in diets supplemented with phytase for broiler chickens

    Directory of Open Access Journals (Sweden)

    Antônio Robério Vieira

    2007-07-01

    Full Text Available O experimento foi realizado com o objetivo de avaliar o desempenhozootécnico, rendimento de carcaça e a deposição de fósforo e cinzas na tíbia de frangos de corte submetidos a dietas contendo quantidades crescentes de farelo de arroz integral (FAI, suplementados com fitase. Foram utilizados 600 frangos de corte, machos, da linhagemRoss, alojados em boxes de acordo com um delineamento inteiramente casualizado, nos seguintes tratamentos: T1-ração referência (milho e farelo de soja; T2-ração referência com 750 FTU g-1; T3-ração com 3,5% de FAI com 750 FTU g-1; T4-ração com 7,5% de FAI com 750 FTU g-1; T5-ração com 10,5% FAI com 750 FTU g-1 e T6-ração com 14% FAIcom 750 FTU g-1. A inclusão de fitase, com a utilização de FAI até o nível de 14% e redução do fósforo inorgânico na formulação da ração, não afetou o desempenho zootécnico e características de carcaça dos frangos de corte, porém reduziu a deposição de fósforo e cinzas na tíbia dos ossos.The experiment was carried with the objective of evaluating the productive performance, carcass yield, phosphorus availability and tibia ash in broilers subjected to diets containing increasing amounts of whole rice bran (WRB, supplemented with phytase. Six hundred broilers (Ross, all males, were allocated in boxesaccording to a completely randomized design, being fed the following regimens: T1 standard feed (corn and soybean meal; T2 standard feed with 750 FTU g-1; T3 feed with 3.5% of WRB with 750 FTU g-1; T4 feed with 7.5% of WRB with 750 FTU g-1 T5 feed with 10.5% WRB with 750 FTU g-1 and T6 feed with 14% WRB with 750 FTU g-1. The inclusion of phytase with the use of WRB to the level of 14% and the reduction of the inorganic P in the formulation of the feed did not affect the performance and carcass characteristics of the broiler chickens; however, it did decrease the presence of phosphorus and ash in the tibia.

  15. Metatranscriptomics reveals the diversity of genes expressed by eukaryotes in forest soils.

    Directory of Open Access Journals (Sweden)

    Coralie Damon

    Full Text Available Eukaryotic organisms play essential roles in the biology and fertility of soils. For example the micro and mesofauna contribute to the fragmentation and homogenization of plant organic matter, while its hydrolysis is primarily performed by the fungi. To get a global picture of the activities carried out by soil eukaryotes we sequenced 2×10,000 cDNAs synthesized from polyadenylated mRNA directly extracted from soils sampled in beech (Fagus sylvatica and spruce (Picea abies forests. Taxonomic affiliation of both cDNAs and 18S rRNA sequences showed a dominance of sequences from fungi (up to 60% and metazoans while protists represented less than 12% of the 18S rRNA sequences. Sixty percent of cDNA sequences from beech forest soil and 52% from spruce forest soil had no homologs in the GenBank/EMBL/DDJB protein database. A Gene Ontology term was attributed to 39% and 31.5% of the spruce and beech soil sequences respectively. Altogether 2076 sequences were putative homologs to different enzyme classes participating to 129 KEGG pathways among which several were implicated in the utilisation of soil nutrients such as nitrogen (ammonium, amino acids, oligopeptides, sugars, phosphates and sulfate. Specific annotation of plant cell wall degrading enzymes identified enzymes active on major polymers (cellulose, hemicelluloses, pectin, lignin and glycoside hydrolases represented 0.5% (beech soil-0.8% (spruce soil of the cDNAs. Other sequences coding enzymes active on organic matter (extracellular proteases, lipases, a phytase, P450 monooxygenases were identified, thus underlining the biotechnological potential of eukaryotic metatranscriptomes. The phylogenetic affiliation of 12 full-length carbohydrate active enzymes showed that most of them were distantly related to sequences from known fungi. For example, a putative GH45 endocellulase was closely associated to molluscan sequences, while a GH7 cellobiohydrolase was closest to crustacean sequences, thus

  16. Planos nutricionais com a utilização de aminoácidos e fitase para frangos de corte mantendo o conceito de proteína ideal nas dietas Nutritional plans with use of amino acids and phytase for broiler chickens maintaining the ideal protein profile in the diets

    Directory of Open Access Journals (Sweden)

    Elisangela Minati Gomide

    2007-12-01

    Full Text Available Objetivou-se neste estudo avaliar o desempenho e as características de carcaça de frangos de corte, recebendo rações com níveis reduzidos de PB, P disponível (Pd e Ca, suplementadas com fitase e aminoácidos. As fases de crescimento avaliadas foram: fase 1, 1 - 21dias de idade e fase 2, 22 - 42 dias de idade. O delineamento experimental foi inteiramente casualizado, sendo os tratamentos constituídos de cincos planos nutricionais (PN, com seis repetições de 25 aves cada. Os planos nutricionais são: PN1 - ração basal (21,4% PB - fase 1 + ração basal (19,3% PB - fase 2; PN2 - ração a basal (fase 1 + ração com 18% PB (fase 2; PN3 - ração basal (fase 1 + ração com 16% PB (fase 2; PN4 - ração com 19% PB (fase 1 + ração com 16% PB (fase 2; e PN5 = ração com 17 % PB (fase 1 + ração com 16% PB (fase 2. Exceto para a ração basal, reduziu-se o nível de Pd para 0,34 e 0,30% e o de Ca para 0,80 e 0,70% nas fases 1 e 2, respectivamente; nestas rações foram adicionados 500 FTU de fitase/kg de ração. As aves alimentadas com PN2 e PN3 apresentaram desempenho semelhante ao daquelas que receberam PN1, enquanto o das aves que receberam os PN4 e PN5 permaneceu inalterado. Os rendimentos de carcaça e peito não foram influenciados pelos planos nutricionais. As aves que consumiram a ração basal nas duas fases de criação (PN1 apresentaram maior rendimento de coxa+sobre-coxa. Houve aumento da gordura abdominal quando as aves receberam os PN3, 4 e 5. É possível reduzir a PB da ração até 16% na fase 2, suplementada com fitase e aminoácidos, desde que a ração da primeira fase seja formulada com níveis nutricionais recomendados.This study was carried out to evaluate the performance and characteristics of carcass of broiler chickens fed diets with reduced levels of CP, available P (aP and Ca, supplemented with phytase and amino acids. The growth phases evaluated were: Phase1, 1 - 21 days old, phase 2, 22 - 42 days old. A

  17. Níveis de fósforo disponível e cálcio em rações suplementadas com fitase para frangos de corte nas fases de crescimento e final Calcium and available phosphorus levels in diets supplemented with phytase for broilers in the growing and finishing phases

    Directory of Open Access Journals (Sweden)

    Luziane Moreira dos Santos

    2011-11-01

    Full Text Available Avaliaram-se os níveis de cálcio e fósforo disponível (Pdisp em rações com fitase para frangos de corte da linhagem Cobb nas fases de crescimento (22 a 35 dias e final (36 a 42 dias. Utilizou-se um esquema fatorial 3 × 4 + 1, composto de três níveis de Pdisp (0,36; 0,31; 0,26% na fase de crescimento e 0,33; 0,28; 0,23% na final, quatro níveis de cálcio (0,82; 0,72; 0,62; 0,52% para a fase de crescimento e 0,76; 0,66; 0,56; 0,46% para a final e um nível de suplementação da fitase (500 ftu/kg. As rações controle não foram suplementadas com fitase e continham, respectivamente, os seguintes níveis de Pdisp e cálcio: 0,41 e 0,82% (fase de crescimento e 0,38 e 0,76% (final. Nos ensaios de desempenho, aos 35 e 42 dias de idade, avaliaram-se o consumo de ração, o ganho de peso, a conversão alimentar e o teor de cinzas na tíbia. Nos ensaios de metabolismo, foram determinados os valores energéticos das rações (EMAn e os coeficientes de metabolizabilidade da matéria seca (CMMS. Na fase de crescimento, a redução do cálcio e Pdisp para 0,52 e 0,26%, respectivamente, não comprometeu o desempenho, a mineralização óssea, a EMAn e o CMMS. Na fase final, a redução do cálcio e fósforo disponível para 0,56 e 0,28%, respectivamente, não comprometeu o desempenho, a EMAn e o CMMS e melhorou a mineralização óssea. Os níveis de cálcio e fósforo disponível em rações para frangos de corte podem ser reduzidos para 0,52 e 0,26% na fase de crescimento e 0,56 e 0,28% na fase final desde que essa redução seja combinada com a suplementação de fitase no nível de 500 ftu/kg de ração.Calcium and available phosphorus (aP levels in diets with phytase for Cobb broilers in the growing (22 to 35 days and finishing phases (36 to 42 days of age were evaluated. A 3 × 4 + 1 factorial scheme, with three aP levels (0.36, 0.31, 0.26% in the growing phase and 0.33, 0.28, 0.23% in the finishing phase, four calcium levels (0.82, 0.72, 0

  18. Níveis de cálcio e fósforo disponível em rações com fitase para frangos de corte nas fases pré-inicial e inicial Calcium and available phosphorus levels in diets with phytase for broilers in the pre-starter and starter phases

    Directory of Open Access Journals (Sweden)

    Luziane Moreira dos Santos

    2011-11-01

    carried out with broilers to evaluate levels of calcium and available phosphorus (aP in diets supplemented with 500 units of phytase activity/kg of diet. Each experiment consisted of a performance and a metabolic assay, and was conducted in a factorial schedule (3 × 4 + 1, with three aP levels (0.42, 0.37, 0.32% in the pre-starter phase and 0.39, 0.34, 0.29% in the starter phase, four calcium levels (0.94, 0.84, 0.74, 0.64% in the pre-starter phase and 0.88; 0.78, 0.68, 0.58% in the starter phase plus a control diet. The control corresponded to the only diet without phytase and contained 0.47% of aP and 0.94% of calcium (pre-starter phase or 0.44% of aP and 0.88% of calcium (starter phase. In the performance assays, at 7 and 21 days of age the feed intake, weight gain, feed conversion and the ash content in the tibia were evaluated. In the metabolic assays the apparent metabolizable energy corrected for nitrogen balance (AMEn of the feed, and dry matter digestibility coefficients (DMDC were determined. Calcium levels influenced the feed intake, AMEn and DMDC in the pre-starter phase, and in the starter phase they influenced bone ash, AMEn and DMDC. The aP levels influenced the content of tibia ash, AMEn and DMDC in the pre-starter phase, and bone ash and DMDC in the starter phase. By adding 500 units of phytase activity/kg of diet, it is possible to reduce, respectively, the calcium and aP levels to 0.64% and 0.37% in the pre-starter phase, and to 0.58% and 0.29% in the starter phase, since this reduction does not cause a negative effect on performance or bone mineralization of broilers in the evaluated periods.

  19. Levels of available phosphorus and calcium for broilers from 8 to 35 days of age fed rations containing phytase Níveis de fósforo disponível e de cálcio para frangos de corte de 8 a 35 dias de idade alimentados com rações contendo fitase

    Directory of Open Access Journals (Sweden)

    Adimar Cardoso Júnior

    2010-06-01

    Full Text Available Two experiments using Cobb broiler chicks from 8 to 35 days of age were carried out to evaluate the effect of reducing levels of calcium and available phosphorus in diets supplemented with 500 ftu phytase/kg on the performance of the birds, the content of ashes in the tibia, the metabolizable energy of the diet, and the metabolizability of the dry matter and crude protein. It was used 1,404 broiler chicks in the experiment 1 to evaluate feed intake, weight gain, food conversion and percentage of the ashes in the tibia. In the experiment 2, simultaneously carried out with experiment 1, a total of 390 birds were transferred to a metabolism room to determine the metabolizable energy and metabolizability of the dry matter and crude protein. It was used in the two experiments, a complete random design in a 3 × 4 + 1 factorial scheme with three levels of available phosphorus (0.375; 0.325; and 0.275% and four levels of calcium (0.85; 0.75; 0.65 and 0.55% in the diets, supplemented with phytase. It was also used a control diet without phytase, based on corn and soybean bran, formulated with 0.425% of available phosphorus and 0.85% of calcium. In the experiment 1, the reduction of levels of calcium and phosphorus did not cause a significant difference on the performance and percentage of bone ashes. The levels of calcium and available phosphorus used in the experiment 2 can be reduced down to 0.65 and 0.325%, respectively, because the effects are similar to those obtained with the currently suggested levels.Dois experimentos com pintos de corte da linhagem Cobb no período de 8 a 35 dias de idade foram realizados para avaliar o efeito da redução dos níveis de cálcio e fósforo disponível (Pdisp em rações suplementadas com 500 ftu de fitase/kg sobre o desempenho das aves, o teor de cinzas na tíbia, a energia metabolizável (EMAn da dieta e a metabolizabilidade da matéria seca (MS e da proteína bruta (PB. No experimento 1, foram utilizados 1

  20. Chromatin loops, gene positioning, and gene expression

    NARCIS (Netherlands)

    Holwerda, S.; de Laat, W.

    2012-01-01

    Technological developments and intense research over the last years have led to a better understanding of the 3D structure of the genome and its influence on genome function inside the cell nucleus. We will summarize topological studies performed on four model gene loci: the alpha- and beta-globin

  1. Your Genes, Your Choices

    Science.gov (United States)

    Table of Contents Your Genes, Your Choices describes the Human Genome Project, the science behind it, and the ethical, legal, and social issues that are ... Nothing could be further from the truth. Your Genes, Your Choices points out how the progress of ...