• Title/Summary/Keyword: Acid phytase

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Effect of Polar Amino Acid Residue Substitution by Site-Directed Mutagenesis in the N-terminal Domain of Pseudomonas sp. Phytase on Enzyme Activity

  • Lee, Ga Hye;Jang, Won Je;Kim, Soyeong;Kim, Yoonha;Kong, In-Soo
    • Journal of Microbiology and Biotechnology
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    • v.30 no.7
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    • pp.1104-1107
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    • 2020
  • The N-terminal domain of the Pseudomonas sp. FB15 phytase increases low-temperature activity and catalytic efficiency. In this study, the 3D structure of the N-terminal domain was predicted and substitutions for the amino acid residues of the region assumed to be the active site were made. The activity of mutants, in which alanine (A) was substituted for the original residue, was investigated at various temperatures and pH values. Significant differences in enzymatic activity were observed only in mutant E263A, suggesting that the amino acid residue at position 263 of the N-terminal domain is important in enzyme activity.

Effects of Phytase and Carbohydrases Supplementation to Diet with a Partial Replacement of Soybean Meal with Rapeseed Meal and Cottonseed Meal on Growth Performance and Nutrient Digestibility of Growing Pigs

  • Shim, Y.H.;Chae, B.J.;Lee, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.9
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    • pp.1339-1347
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    • 2003
  • An experiment was conducted to investigate the effects of microbial phytase ($Natuphos^{(R)}$) supplementation in combination with carbohydrases (composed of enzymes targeted to soybean meal (SBM) dietary components such as $\alpha$-galactosides and galactomannans; $Endo-Power^{(R)}$) to corn-soybean meal based diet (CSD) and complex diet (CD) with a partial replacement of SBM with rape seed meal (RSM) and cotton seed meal (CSM) on growth performance and nutrient digestibility of growing pigs. A total of 168 growing pigs averaging $13.18{\pm}1.77kg$ of initial body weight was arranged as a $2{\times}2$ factorial design with main effects of diet types (corn-SBM based diet (CSD) and complex diets (CD; 5% of SBM was replaced with 2.5% of RSM and 2.5% of CSM in diet for phase I (0 to 3 weeks) and 6% of SBM was replaced with 3% of RSM and 3% of CSM in diet for phase II (4 to 7 weeks))) and enzyme supplementation (none and 0.1% of phytase (500 FTU/kg diet) and 0.1% of carbohydrases). The diet with enzyme application were formulated to have a 0.18% unit lower aP than diets without enzyme application. Each treatment had three replicates with 14 pigs per replicate. To determine supplementation effect of phytase and carbohydrases on ileal amino acid digestibility of SBM, RSM and CSM, a total of 18 T-cannulated pigs (initial body weight; $13.52{\pm}1.24kg$) were assigned to six dietary treatments in the present study. Dietary treatments in metabolic trial included 1) SBM diet, 2) SBM diet+with enzymes (phytase (500 FTU/kg) and carbohydrases at 0.1%, respectively), 3) CSM diet, 4) CSM diet+enzymes, 5) RSM diet and 6) RSM diet+enzymes. During whole experimental period (0 to 7 wks), there was no difference in growth performance between diets (CSD and CD). However, dietary phytase and carbohydrases supplementation significantly improved gain/feed ratio (G:F) of growing pigs. During the phase II (4-7 weeks), dietary phytase and carbohydrases supplementation significantly improved all fecal nutrient digestibilities (Dry matter (DM), gross energy (GE), crude protein (CP), crude fat (CF), calcium (Ca) and phosphorus (P)). Dietary phytase and carbohydrases supplementation improved significantly overall ileal amino acid digestibilities of SBM, RSM and CSM based diets (p<0.05). The simultaneous inclusion of phytase and carbohydrases in both of CSD and CD reduced feed cost per kg body weight gain (FCG). Also, results suggest that 2.5 to 3% of RSM and CSM, respectively, might be used as a protein source in growing pig diets without having an adverse effect on the growth performance and nutrient digestibility and simultaneous phytase and carbohydrases addition improves nutritional value of SBM, RSM and CSM by improving ileal amino acid digestibilities.

Effects of Phytase and Enzyme Complex Supplementation to Diets with Different Nutrient Levels on Growth Performance and Ileal Nutrient Digestibility of Weaned Pigs

  • Shim, Y.H.;Chae, B.J.;Lee, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.4
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    • pp.523-532
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    • 2004
  • An experiment was conducted to investigate the effect of microbial phytase ($Natuphos^{R}$) supplementation in combination with enzyme complex (composed of enzymes targeted to SBM dietary components such as $\alpha$-galactosides and galactomannans; $Endo-Power^{R}$) to diet with low nutrient levels on growth performance and ileal nutrient digestibility of weaned pigs. A total of 210 crossbred weaned pigs (Landrace$\times$Yorkshire$\times$Duroc), 6.68$\pm$0.98 kg of initial body weight, were randomly allotted to five dietary treatments, based on weight and age, according to a randomized complete block design. There were three pens per treatment and 14 pigs per pen. The dietary treatments were 1) CON (Control diet with no phytase and enzyme complex (EC)), 2) LP+EC 100 (Control diet with 0.15% unit lower available phosphorus (aP) level+0.1% phytase (500 FTU/kg diet) and 0.1% enzyme complex), 3) LP+EC 80 (Control diet with 0.15% unit lower aP level+0.08% phytase (400 FTU/kg diet) and 0.08% enzyme complex, 4) LPEA+EC 100 (Control diet with 0.15% unit lower aP and 3% lower ME and amino acid levels (lysine, methionine, threonine and typtophan)+0.1% phytase (500 FTU/kg diet) and 0.1% enzyme complex), 5) LPEA+EC 80 (Control diet with 0.15% unit lower aP and 3% lower ME and amino acid levels+0.08% phytase (400 FTU/ kg diet) and 0.08% enzyme complex). For the determination of ileal nutrients digestibility, a total of 15 T-cannulated pigs (initial body weight; 7.52$\pm$1.24 kg; 3 replicates per treatment) were used in the present study. Piglets were weighted and allotted into same dietary treatments as one in growth trial and phase I experimental diets were provided for ileal digestibility study. There was no significant difference (p>0.05) in average daily gain (ADG) and average daily feed intake (ADFI) among dietary treatments during the whole experimental period (0 to 5 weeks). However, piglets in LP+EC 100 group had a significantly higher gain/feed ratio (G:F) than piglets had in control (p<0.05). Crude protein, energy and phosphorus digestibilities were significantly improved when both of phytase and enzyme complex were supplemented at the revel of 0.1%, respectively to diets with low nutrient level (aP or (and) ME and amino acids) (p<0.05). Piglets in LP+EC 100 and LPEA+EC 100 groups showed significantly higher phosphorus content (%) in bone than that of piglets in control group (p<0.05). Supplementation of both of phytase and enzyme complex at 0.1%, respectively, to diet with low nutrient levels (aP or (and) ME and amino acids) significantly improved total ileal essential amino acid and nonessential amino acid digestibilities compared to control group (p<0.05). In conclusion, the results from the present study suggest that the simultaneous inclusion of phytase and enzyme complex to diets at recommended level is advantageous with respect to improving growth performance and nutrient digestibility of weaned pigs and may contribute to increased economic return when added to corn-soy based weaned pig diets.

A Study on the Variation of Phytase Activity in the River Environment (하천환경에서의 phytsae 활성변화에 관한 연구)

  • 김영진;오남순;정문호
    • Journal of Environmental Health Sciences
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    • v.27 no.4
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    • pp.1-8
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    • 2001
  • From August 2000 to August 2001, 9 variables of physicochemical factors and phytase activity were investigated at 4 sites in the River Yungpyung and the influences of Physicochemical factors to Phytase activity were analyzed. Phytase activities of Site 1, Site 2, Site 3, and Site 4 varied between N.D ∼566 nmol/ ι /hr, N.D \" 434 nmol/ ι /hr, N.D ∼557 nmol/ ι /hr, and N.D ∼723 nmol/ ι /hr, respectively. The activities of summer season were higher than those of other season. But the activities were not detected on the winter season. The phytase activity and temperature showed high correlation. The correlation coefficients of Site 1, Site 2, Site 3, and Site 4 were 0.82(p<0.01).0.92(p<0.01),0.87(p<0.01), and 0.88(p<0.01), respectively. The phytase activity and NOI₃/sup -/ ion showed negative relation(r=-0.59, p<0.05) at Site 1. And the phytase activity had relation with Zn/sup 2+/at Site 2(r=().57, p<0.05) and Site 3(r=0.7E, p<7.07).

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Evaluation of the impact of phytase supplementation on growth performance, nutrient digestibility, and fecal score of growing pigs

  • Shanshui Gao;Md Mortuza Hossain;In Ho Kim
    • Korean Journal of Agricultural Science
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    • v.50 no.4
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    • pp.653-661
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    • 2023
  • Phytase enhances phosphorus availability in pig diets by breaking down phytic acid, which is challenging for animals to digest. This study aimed to assess the impact of dietary phytase supplementation on the growth performance, nutrient digestibility, and fecal score of growing pigs. Sixty-four growing pigs ([Yorkshire × Landrace] × Duroc, average body weight 21.20 ± 0.18 kg) were randomly assigned to one of two treatment groups. The dietary treatments were CON, basal diet, and PHY, basal diet + 0.0025% phytase. Results indicated a tendency for increased average daily gain in the phytase-supplemented diet group (p < 0.10) compared to the control diet group. The average daily feed intake and feed conversion ratio were not affected by the addition of 0.0025% phytase. Furthermore, dry matter digestibility, nitrogen content, and digestible energy were not influenced (p > 0.05) by phytase supplementation. Fecal score remained similar (p > 0.05) in growing pigs fed phytase-supplemented and control diets. These findings suggest that supplementing the diet of growing pigs with 0.0025% phytase may lead to improved average daily gain without adverse effects on nutrient digestibility and fecal score. This implies the potential to enhance growth performance without compromising overall health or digestion in pigs. Further investigations into the optimal levels of phytase supplementation for growing pigs are recommended.

Isolation, Characterization, and Molecular Cloning of the cDNA Encoding a Novel Phytase from Aspergillus niger 113 and High Expression in Pichia pastoris

  • Xiong, Ai Sheng;Yao, Quan-Hong;Peng, Ri-He;Li, Xian;Fan, Hui-Qin;Guo, Mei-Jin;Zhang, Si-Liang
    • BMB Reports
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    • v.37 no.3
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    • pp.282-291
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    • 2004
  • Phytases catalyze the release of phosphate from phytic acid. Phytase-producing microorganisms were selected by culturing the soil extracts on agar plates containing phytic acid. Two hundred colonies that exhibited potential phytase activity were selected for further study. The colony showing the highest phytase activity was identified as Aspergillus niger and designated strain 113. The phytase gene from A. niger 113 (phyI1) was isolated, cloned, and characterized. The nucleotide and deduced amino acid sequence identity between phyI1 and phyA from NRRL3135 were 90% and 98%, respectively. The identity between phyI1 and phyA from SK-57 was 89% and 96%. A synthetic phytase gene, phyI1s, was synthesized by successive PCR and transformed into the yeast expression vector carrying a signal peptide that was designed and synthesized using P. pastoris biased codon. For the phytase expression and secretion, the construct was integrated into the genome of P. pastoris by homologous recombination. Over-expressing strains were selected and fermented. It was discovered that ~4.2 g phytase could be purified from one liter of culture fluid. The activity of the resulting phytase was 9.5 U/mg. Due to the heavy glycosylation, the expressed phytase varied in size (120, 95, 85, and 64 kDa), but could be deglycosylated to a homogeneous 64 kDa species. An enzymatic kinetics analysis showed that the phytase had two pH optima (pH 2.0 and pH 5.0) and an optimum temperature of $60^{\circ}C$.

Enzyme Kinetic Study for Degradation of Phytic acid by Phytase (Phytase에 의한 phytic acid의 분해 반응속도 연구)

  • Kim, Jae-Seok;Lee, Jae-Heung
    • KSBB Journal
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    • v.26 no.4
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    • pp.300-304
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    • 2011
  • Phytases are hydrolytic enzymes that catalyze the sequential hydrolysis of phytic acid (myo-inositol-1,2,3,4,5,6-hexakisphosphate) to myo-inositols with lower numbers of phosphate groups. Two types of phytases have been identified which initiate hydrolysis of the phytic acid at either the 3- or 6- position of the inositol ring. In the present investigation, a mathematical model was proposed and computed to estimate maximum enzyme reaction rate constants which fit the experimental data obtained by other authors. Although the data points were scattered to some extent, good agreement was found between the model and the experiment data. It appears that the maximum rate constants of removal of the first, second, and third phosphate groups were not equal. Also there was neither a steady trend upward or downward in the rate constants with the stepwise hydrolysis reactions.

Identification of Novel Phytase Genes from an Agricultural Soil-Derived Metagenome

  • Tan, Hao;Mooij, Marlies J.;Barret, Matthieu;Hegarty, Pardraig M.;Harrington, Catriona;Dobson, Alan D.W.;O'Gara, Fergal
    • Journal of Microbiology and Biotechnology
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    • v.24 no.1
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    • pp.113-118
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    • 2014
  • Environmental microorganisms are emerging as an important source of new enzymes for wide-scale industrial application. In this study, novel phytase genes were identified from a soil microbial community. For this, a function-based screening approach was utilized for the identification of phytase activity in a metagenomic library derived from an agricultural soil. Two novel phytases were identified. Interestingly, one of these phytases is an unusual histidine acid phosphatase family phytase, as the conserved motif of the active site of PhyX possesses an additional amino acid residue. The second phytase belongs to a new type, which is encoded by multiple open reading frames (ORFs) and is different to all phytases known to date, which are encoded by a single ORF.

Thermal Stabilization of Aspergillus phytase by L-Arginine

  • Sunghoon Ryu;Park, Tae-Gwan
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.3 no.1
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    • pp.32-34
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    • 1998
  • Phytase from Aspergillus species is a very heat unstable enzyme which inactivates to a great extent during the thermal processing of animal feed formulation. Various protein stabilization additives were tested to improve its heat stability. Among them, a basic amino acid, L-arginine remarkably increased the thermal stability of phytase in an aqueous solution state.

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Response of broiler chickens to diets containing different levels of sodium with or without microbial phytase supplementation

  • Akter, Marjina;Graham, Hadden;Iji, Paul Ade
    • Journal of Animal Science and Technology
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    • v.61 no.2
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    • pp.87-97
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    • 2019
  • Phytate induced excessive mineral excretion through poultry litter leads to poor performance and environmental pollution. Exogenous microbial phytase supplementation to poultry diets reduce the environmental excretion of nutrient and improve bird's performance. However, excessive dietary sodium (Na) level may hinder the phytase-mediated phytate hydrolysis and negate the beneficial effects of phytase. Therefore, this experiment was conducted to investigate the effects of different concentration dietary Na on phytase activity and subsequent impact on broiler performance, bone mineralisation and nutrient utilisation. In this study, six experimental diets, consisting of three different levels of Na (1.5, 2.5, or 3.5 g/kg) and two levels of microbial phytase (0 or 500 U/kg) were formulated by using $3{\times}2$ factorial design. The six experimental diets were offered to 360 day-old Ross 306 male chicks for 35 days, where, each experimental diet consisted of 6 replicates groups with 10 birds. Along with growth performance, nutrient utilization, intestinal enzyme activity, dry matter (DM) content of litter and mineral status in bone were analysed. Dietary Na and phytase had no effect on bode weight gain and feed intake. Birds on the low Na diet showed higher (p < 0.05) feed conversion ratio (FCR) than the mid-Na diets. High dietary Na adversely affected (p < 0.001) excreta DM content. Phytase supplementation to the high-Na diet increased (p < 0.01) the litter ammonia content. High dietary Na with phytase supplementation improved ($Na{\times}phytase$, p < 0.05) the AME value and ileal digestibility of Ca and Mg. The total tract retention of Ca, P, and Mg was reduced with high Na diet, which was counteracted by phytase supplementation ($Na{\times}phytase$, p < 0.001). The diets containing mid-level of Na improved (p < 0.001) the function of Na-K-ATPase and Mg-ATPase in the jejunum. The overall results indicate that high dietary Na did not affect phytase activity but influenced the nutrient utilization of birds, which was not reflected in bird overall performance.