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...

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.

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.

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.

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 ...

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.

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

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 ...

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.

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

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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.

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.

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 ...

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

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.

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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.

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

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.

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

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.

Effect of phytase supplementation on growing pigs performance

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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

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

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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.

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

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

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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 ...

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.

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.

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

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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.

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.

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

Directory of Open Access Journals (Sweden)

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.

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

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...

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....

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

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

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).

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...

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.

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

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

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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.

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...

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...

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.

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

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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

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

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

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%.

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...

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....

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.

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.

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.

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...

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.

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

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 ...

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

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.

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.

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

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.

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....

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 ...

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.

Phytase for Food Application

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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.

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.

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...

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....

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....

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

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

Science.gov (United States)

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.

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.

Biotechnological Aspects of Microbial Extracellular Electron Transfer

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Kato, Souichiro

2015-01-01

Extracellular electron transfer (EET) is a type of microbial respiration that enables electron transfer between microbial cells and extracellular solid materials, including naturally-occurring metal compounds and artificial electrodes. Microorganisms harboring EET abilities have received considerable attention for their various biotechnological applications, in addition to their contribution to global energy and material cycles. In this review, current knowledge on microbial EET and its application to diverse biotechnologies, including the bioremediation of toxic metals, recovery of useful metals, biocorrosion, and microbial electrochemical systems (microbial fuel cells and microbial electrosynthesis), were introduced. Two potential biotechnologies based on microbial EET, namely the electrochemical control of microbial metabolism and electrochemical stimulation of microbial symbiotic reactions (electric syntrophy), were also discussed. PMID:26004795

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....

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...

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 ...

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.

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

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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

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.

Phytase in broiler diets with reduced nonphytate phosphorus levels

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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.

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 ...

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

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

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.

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.

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 ...

Effect of nonphytate phosphorus and phytase levels on broiler femur.

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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.

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

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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.

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...

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...

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.

Phosphorus and phytase levels for layer hens

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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.

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

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...

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

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

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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.

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 ...

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 ...

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

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...

Exocellular electron transfer in anaerobic microbial communities.

Science.gov (United States)

Stams, Alfons J M; de Bok, Frank A M; Plugge, Caroline M; van Eekert, Miriam H A; Dolfing, Jan; Schraa, Gosse

2006-03-01

Exocellular electron transfer plays an important role in anaerobic microbial communities that degrade organic matter. Interspecies hydrogen transfer between microorganisms is the driving force for complete biodegradation in methanogenic environments. Many organic compounds are degraded by obligatory syntrophic consortia of proton-reducing acetogenic bacteria and hydrogen-consuming methanogenic archaea. Anaerobic microorganisms that use insoluble electron acceptors for growth, such as iron- and manganese-oxide as well as inert graphite electrodes in microbial fuel cells, also transfer electrons exocellularly. Soluble compounds, like humic substances, quinones, phenazines and riboflavin, can function as exocellular electron mediators enhancing this type of anaerobic respiration. However, direct electron transfer by cell-cell contact is important as well. This review addresses the mechanisms of exocellular electron transfer in anaerobic microbial communities. There are fundamental differences but also similarities between electron transfer to another microorganism or to an insoluble electron acceptor. The physical separation of the electron donor and electron acceptor metabolism allows energy conservation in compounds as methane and hydrogen or as electricity. Furthermore, this separation is essential in the donation or acceptance of electrons in some environmental technological processes, e.g. soil remediation, wastewater purification and corrosion.

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....

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.

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.

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.

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.

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,...

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.

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.

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.

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.

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.

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

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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.

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.

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.

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.

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

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...

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

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 ...

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

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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.

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.

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.

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.

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.

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.

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...

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

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

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

Directory of Open Access Journals (Sweden)

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

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.

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.

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

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

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.

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

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

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.

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.

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.

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.

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.

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

Directory of Open Access Journals (Sweden)

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.

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...

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.

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.

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

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,.

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.

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.

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)

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.

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

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

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.

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.

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...

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.

Microbially-reduced graphene scaffolds to facilitate extracellular electron transfer in microbial fuel cells.

Science.gov (United States)

Yuan, Yong; Zhou, Shungui; Zhao, Bo; Zhuang, Li; Wang, Yueqiang

2012-07-01

A one-pot method is exploited by adding graphene oxide (GO) and acetate into an microbial fuel cell (MFC) in which GO is microbially reduced, leading to in situ construction of a bacteria/graphene network in the anode. The obtained microbially reduced graphene (MRG) exhibits comparable conductivity and physical characteristics to the chemically reduced graphene. Electrochemical measurements reveal that the number of exoelectrogens involved in extracellular electron transfer (EET) to the solid electrode, increases due to the presence of graphene scaffolds, and the EET is facilitated in terms of electron transfer kinetics. As a result, the maximum power density of the MFC is enhanced by 32% (from 1440 to 1905 mW m(-2)) and the coulombic efficiency is improved by 80% (from 30 to 54%). The results demonstrate that the construction of the bacteria/graphene network is an effective alternative to improve the MFC performance. Copyright © 2012 Elsevier Ltd. All rights reserved.

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.

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

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

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.

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.

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.

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.

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.

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

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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.

Phytase-Producing Bacteria from Extreme Regions in Indonesia

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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.

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.

An overview of electron acceptors in microbial fuel cells

DEFF Research Database (Denmark)

Ucar, Deniz; Zhang, Yifeng; Angelidaki, Irini

2017-01-01

Microbial fuel cells (MFC) have recently received increasing attention due to their promising potential in sustainable wastewater treatment and contaminant removal. In general, contaminants can be removed either as an electron donor via microbial catalyzed oxidization at the anode or removed at t...... acceptors (e.g., nitrate, iron, copper, perchlorate) and mediators....

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.

Exocellular electron transfer in anaerobic microbial communities

NARCIS (Netherlands)

Stams, A.J.M.; Bok, de F.A.M.; Plugge, C.M.; Eekert, van M.H.A.; Dolfing, J.; Schraa, G.

2006-01-01

Exocellular electron transfer plays an important role in anaerobic microbial communities that degrade organic matter. Interspecies hydrogen transfer between microorganisms is the driving force for complete biodegradation in methanogenic environments. Many organic compounds are degraded by obligatory

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.

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.

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.

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 ...

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.

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.

Localized electron transfer rates and microelectrode-based enrichment of microbial communities within a phototrophic microbial mat

Directory of Open Access Journals (Sweden)

Jerome eBabauta

2014-01-01

Full Text Available Phototrophic microbial mats frequently exhibit sharp, light-dependent redox gradients that regulate microbial respiration on specific electron acceptors as a function of depth. In this work, a benthic phototrophic microbial mat from Hot Lake, a hypersaline, epsomitic lake located near Oroville in north-central Washington, was used to develop a microscale electrochemical method to study local electron transfer processes within the mat. To characterize the physicochemical variables influencing electron transfer, we initially quantified redox potential, pH and dissolved oxygen gradients by depth in the mat under photic and aphotic conditions. We further demonstrated that power output of a mat fuel cell was light-dependent. To study local electron transfer processes, we deployed a microscale electrode (microelectrode with tip size ~20 µm. To enrich a subset of microorganisms capable of interacting with the microelectrode, we anodically polarized the microelectrode in the mat. Subsequently, to characterize the microelectrode-associated community and compare it to the neighboring mat community, we performed amplicon sequencing of the V1-V3 region of the 16S gene. Differences in Bray-Curtis beta diversity, illustrated by large changes in relative abundance at the phylum level, suggested successful enrichment of specific mat community members on the microelectrode surface. The microelectrode-associated community exhibited substantially reduced alpha diversity and elevated relative abundances of Prosthecochloris, Loktanella, Catellibacterium, other unclassified members of Rhodobacteraceae, Thiomicrospira, and Limnobacter, compared with the community at an equivalent depth in the mat. Our results suggest that local electron transfer to an anodically polarized microelectrode selected for a specific microbial population, with substantially more abundance and diversity of sulfur-oxidizing phylotypes compared with the neighboring mat community.

Localized electron transfer rates and microelectrode-based enrichment of microbial communities within a phototrophic microbial mat.

Science.gov (United States)

Babauta, Jerome T; Atci, Erhan; Ha, Phuc T; Lindemann, Stephen R; Ewing, Timothy; Call, Douglas R; Fredrickson, James K; Beyenal, Haluk

2014-01-01

Phototrophic microbial mats frequently exhibit sharp, light-dependent redox gradients that regulate microbial respiration on specific electron acceptors as a function of depth. In this work, a benthic phototrophic microbial mat from Hot Lake, a hypersaline, epsomitic lake located near Oroville in north-central Washington, was used to develop a microscale electrochemical method to study local electron transfer processes within the mat. To characterize the physicochemical variables influencing electron transfer, we initially quantified redox potential, pH, and dissolved oxygen gradients by depth in the mat under photic and aphotic conditions. We further demonstrated that power output of a mat fuel cell was light-dependent. To study local electron transfer processes, we deployed a microscale electrode (microelectrode) with tip size ~20 μm. To enrich a subset of microorganisms capable of interacting with the microelectrode, we anodically polarized the microelectrode at depth in the mat. Subsequently, to characterize the microelectrode-associated community and compare it to the neighboring mat community, we performed amplicon sequencing of the V1-V3 region of the 16S gene. Differences in Bray-Curtis beta diversity, illustrated by large changes in relative abundance at the phylum level, suggested successful enrichment of specific mat community members on the microelectrode surface. The microelectrode-associated community exhibited substantially reduced alpha diversity and elevated relative abundances of Prosthecochloris, Loktanella, Catellibacterium, other unclassified members of Rhodobacteraceae, Thiomicrospira, and Limnobacter, compared with the community at an equivalent depth in the mat. Our results suggest that local electron transfer to an anodically polarized microelectrode selected for a specific microbial population, with substantially more abundance and diversity of sulfur-oxidizing phylotypes compared with the neighboring mat community.

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.

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.

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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

Quantifying electron fluxes in methanogenic microbial communities

NARCIS (Netherlands)

Junicke, H.

2015-01-01

Anaerobic digestion is a widely applied process in which close interactions between different microbial groups result in the formation of renewable energy in the form of biogas. Nevertheless, the regulatory mechanisms of the electron transfer between acetogenic bacteria and methanogenic archaea in

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.

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...

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

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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.

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.

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...

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.

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.

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.

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

Electron microscopy study of microbial mat in the North Fiji basin hydrothermal vent

Science.gov (United States)

Park, H.; Kim, J. W.; Lee, J. W.

2017-12-01

Hydrothermal vent systems consisting of hydrothermal vent, hydrothermal sediment and microbial mat are widely spread around the ocean, particularly spreading axis, continental margin and back-arc basin. Scientists have perceived that the hydrothermal systems, which reflect the primeval earth environment, are one of the best places to reveal the origin of life and extensive biogeochemical process of microbe-mineral interaction. In the present study multiline of analytical methods (X-Ray Diffraction (XRD), Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM)) were utilized to investigate the mineralogy/chemistry of microbe-mineral interaction in hydrothermal microbial mat. Microbial mat samples were recovered by Canadian scientific submersible ROPOS on South Pacific North Fiji basin KIOST hydrothermal vent expedition 1602. XRD analysis showed that red-colored microbial mat contains Fe-oxides and Fe-oxyhydroxides. Various morphologies of minerals in the red-colored microbial mat observed by SEM are mainly showed sheath shaped, resembled with Leptothrix microbial structure, stalks shaped, similar with Marioprofundus microbial structure and globule shaped microbial structures. They are also detected with DNA analysis. The cross sectional observation of microbial structures encrusted with Fe-oxide and Fe-oxyhydroxide at a nano scale by Transmission Electron Microscopy (TEM) and Focused Ion Beam (FIB) technique was developed to verify the structural/biogeochemical properties in the microbe-mineral interaction. Systematic nano-scale measurements on the biomineralization in the microbial mat leads the understandings of biogeochemical environments around the hydrothermal vent.

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

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.

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.

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

Directory of Open Access Journals (Sweden)

Cecep Hidayat

2014-10-01

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

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.

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.

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 ...

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.

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

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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

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.

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.

Microbial electron transport and energy conservation – the foundation for optimizing bioelectrochemical systems

Science.gov (United States)

Kracke, Frauke; Vassilev, Igor; Krömer, Jens O.

2015-01-01

Microbial electrochemical techniques describe a variety of emerging technologies that use electrode–bacteria interactions for biotechnology applications including the production of electricity, waste and wastewater treatment, bioremediation and the production of valuable products. Central in each application is the ability of the microbial catalyst to interact with external electron acceptors and/or donors and its metabolic properties that enable the combination of electron transport and carbon metabolism. And here also lies the key challenge. A wide range of microbes has been discovered to be able to exchange electrons with solid surfaces or mediators but only a few have been studied in depth. Especially electron transfer mechanisms from cathodes towards the microbial organism are poorly understood but are essential for many applications such as microbial electrosynthesis. We analyze the different electron transport chains that nature offers for organisms such as metal respiring bacteria and acetogens, but also standard biotechnological organisms currently used in bio-production. Special focus lies on the essential connection of redox and energy metabolism, which is often ignored when studying bioelectrochemical systems. The possibility of extracellular electron exchange at different points in each organism is discussed regarding required redox potentials and effect on cellular redox and energy levels. Key compounds such as electron carriers (e.g., cytochromes, ferredoxin, quinones, flavins) are identified and analyzed regarding their possible role in electrode–microbe interactions. This work summarizes our current knowledge on electron transport processes and uses a theoretical approach to predict the impact of different modes of transfer on the energy metabolism. As such it adds an important piece of fundamental understanding of microbial electron transport possibilities to the research community and will help to optimize and advance bioelectrochemical

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.

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

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Leny Yuanita

2012-01-01

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

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

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...

Microbial growth and sensory quality of dried potato slices irradiated by electrons

International Nuclear Information System (INIS)

Kim, Hyun-Jin; Song, Hyeon-Jeong; Song, Kyung-Bin

2011-01-01

Electron beam irradiation was applied to secure the microbial safety of dried purple sweet potato. After purple sweet potato slices had been dehydrated with 20% (w/w) maltodextrin solution, the samples were irradiated at doses 2, 4, 6, 8, and 10 kGy and then stored at 20 o C for 60 days. Microbiological data indicated that the populations of total aerobic bacteria and of yeast and molds significantly decreased with increase in irradiation dosage. Specifically, microbial load was reduced by about three log cycles at 6 kGy compared to those of the control. Based on the color measurement of the potato slices, electron beam irradiation treatment did not affect the color quality. Sensory evaluation results also showed that electron beam irradiation did not affect overall sensory scores during storage. These results suggest that electron beam irradiation could be useful for improving microbial safety without impairing the quality of the potato slices during storage.

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 ...

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...

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.

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

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Xian-Ren Jiang

2015-04-01

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

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

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.

Electron-beam and microwave treatment of some microbial strains

International Nuclear Information System (INIS)

Martin, D.; Ferdes, O.S.; Minea, R.; Tirlea, A.; Badea, M.; Plamadeala, S.; Ferdes, M.

1998-01-01

The experimental results concerning the combined effects of microwaves and accelerated electron beams on various microbial strains such as E. coli, Salmonella sp. and Monascus purpureus are presented. A special designed microwave applicator with a 2.45 GHz frequency CW magnetron of 850 maximum output power and with associate electronics that allow to control the microwave power, the current intensity, and the exposure time was used. The electron-beam irradiation was performed at different irradiation doses and at a dose rate of 1.5 - 2.0 kGy/min by using a linac at a mean electron energy about 6 MeV, mean bean current of 10 μA, pulse period of 3.5 μs and repetition frequency 100 Hz. The experiments were carried out in 5 variants: microwave treatment; electron-beam irradiation; microwaves followed by electron beam; electrons followed by microwaves; and simultaneous application of microwaves and electron beam. The microbiocidal effect was found to be enhanced by additional use of microwave energy to electron beam irradiation. Enhancement of inactivation rate is only remarkable for the microwave treatment or simultaneous electron beam and microwave irradiation at a temperature above the critical value at which microorganisms begin to perish by heat. Simultaneous irradiation with electron beam and microwaves results in a reduction of temperature and time as well as in the decrease of the upper limit of required electron beam absorbed dose for an assumed microbiological quality parameter. The results obtained indicate the occurrence of a synergistic effect of the two physical fields on a non-thermal basis. Hence, combined microwave-electron beam treatment may be applied as an effective method to reduce microbial load

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.

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

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

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

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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.

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

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

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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.

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

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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.

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

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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.

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

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

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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.

Imaging hydrated microbial extracellular polymers: Comparative analysis by electron microscopy

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Dohnalkova, A.C.; Marshall, M. J.; Arey, B. W.; Williams, K. H.; Buck, E. C.; Fredrickson, J. K.

2011-01-01

Microbe-mineral and -metal interactions represent a major intersection between the biosphere and geosphere but require high-resolution imaging and analytical tools for investigating microscale associations. Electron microscopy has been used extensively for geomicrobial investigations and although used bona fide, the traditional methods of sample preparation do not preserve the native morphology of microbiological components, especially extracellular polymers. Herein, we present a direct comparative analysis of microbial interactions using conventional electron microscopy approaches of imaging at room temperature and a suite of cryogenic electron microscopy methods providing imaging in the close-to-natural hydrated state. In situ, we observed an irreversible transformation of the hydrated bacterial extracellular polymers during the traditional dehydration-based sample preparation that resulted in their collapse into filamentous structures. Dehydration-induced polymer collapse can lead to inaccurate spatial relationships and hence could subsequently affect conclusions regarding nature of interactions between microbial extracellular polymers and their environment.

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

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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

Set anode potentials affect the electron fluxes and microbial community structure in propionate-fed microbial electrolysis cells

KAUST Repository

Rao, Hari Ananda

2016-12-09

Anode potential has been shown to be a critical factor in the rate of acetate removal in microbial electrolysis cells (MECs), but studies with fermentable substrates and set potentials are lacking. Here, we examined the impact of three different set anode potentials (SAPs; −0.25, 0, and 0.25 V vs. standard hydrogen electrode) on the electrochemical performance, electron flux to various sinks, and anodic microbial community structure in two-chambered MECs fed with propionate. Electrical current (49–71%) and CH4 (22.9–41%) were the largest electron sinks regardless of the potentials tested. Among the three SAPs tested, 0 V showed the highest electron flux to electrical current (71 ± 5%) and the lowest flux to CH4 (22.9 ± 1.2%). In contrast, the SAP of −0.25 V had the lowest electron flux to current (49 ± 6%) and the highest flux to CH4 (41.1 ± 2%). The most dominant genera detected on the anode of all three SAPs based on 16S rRNA gene sequencing were Geobacter, Smithella and Syntrophobacter, but their relative abundance varied among the tested SAPs. Microbial community analysis implies that complete degradation of propionate in all the tested SAPs was facilitated by syntrophic interactions between fermenters and Geobacter at the anode and ferementers and hydrogenotrophic methanogens in suspension.

Ethanol tolerance in Aspergillus niger and Escherichia coli phytase

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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...

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.

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

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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.

Characterization of phytase enzymes as feed additive for poultry and feed

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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.

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.

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

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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.

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

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

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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.

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

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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.

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

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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.

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.

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.

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...

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.

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.

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, ...

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

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

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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.

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.

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 ...

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

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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.

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

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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.

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

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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%.

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.

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.

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

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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.

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

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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.

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

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Taheri HR

2015-06-01

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

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

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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.

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.

Bar-coded pyrosequencing reveals the responses of PBDE-degrading microbial communities to electron donor amendments.

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Meiying Xu

Full Text Available Polybrominated diphenyl ethers (PBDEs can be reductively degraded by microorganisms under anaerobic conditions. However, little is known about the effect of electron donors on microbial communities involved in PBDEs degradation. Here we employed 454 Titanium pyrosequencing to examine the phylogenetic diversity, composition, structure and dynamics of microbial communities from microcosms under the conditions of different electron donor amendments. The community structures in each of the five alternate electron donor enrichments were significantly shifted in comparison with those of the control microcosm. Commonly existing OTUs between the treatment and control consortia increased from 5 to 17 and more than 50% of OTUs increased around 13.7 to 186 times at least in one of the microcosms after 90-days enrichment. Although the microbial communities at different taxonomic levels were significantly changed by different environmental variable groups in redundancy analysis, significant correlations were observed between the microbial communities and PBDE congener profiles. The lesser-brominated PBDE congeners, tri-BDE congener (BDE-32 and hexa-BDE, were identified as the key factors shaping the microbial community structures at OTU level. Some rare populations, including the known dechlorinating bacterium, Dehalobacter, showed significant positive-correlation with the amounts of PBDE congeners in the consortia. The same results were also observed on some unclassified bacteria. These results suggest that PBDEs-degrading microbial communities can be successfully enriched, and their structures and compositions can be manipulated through adjusting the environmental parameters.

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.

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

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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.

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

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.

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

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Ralf Greiner

2009-12-01

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

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.

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

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

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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.

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

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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.

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%.

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).

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.

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...

[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.

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.

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

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

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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.

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

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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

High dietary zinc supply reduces the digestibility of phosphorus in pig diets1

DEFF Research Database (Denmark)

Poulsen, Hanne Damgaard; Blaabjerg, Karoline; Sørensen, Kristina Ulrich

2016-01-01

. However, divalent metal ions may bind phytate, leaving phytate less accessible to phytase-induced hydrolysis. The objective was to examine the effects of increased Zn supply on the apparent total tract digestibility (ATTD) of P in pig diets supplemented with microbial phytase. A wheat, barley, and soy–based...... diet (4.3 g P/kg DM, 3.1 g phytate P/kg DM, and no feed phosphate) were heat treated at 81°C (plant phytase diet was divided into 3 batches that were supplemented with 0, 100 or 2,500 mg...... a significant (P diet decreased the ATTD of plant P by about 16% in microbial phytase–supplemented diets....

Outward electron transfer by Saccharomyces cerevisiae monitored with a bi-cathodic microbial fuel cell-type activity sensor.

Science.gov (United States)

Ducommun, Raphaël; Favre, Marie-France; Carrard, Delphine; Fischer, Fabian

2010-03-01

A Janus head-like bi-cathodic microbial fuel cell was constructed to monitor the electron transfer from Saccharomyces cerevisiae to a woven carbon anode. The experiments were conducted during an ethanol cultivation of 170 g/l glucose in the presence and absence of yeast-peptone medium. First, using a basic fuel-cell type activity sensor, it was shown that yeast-peptone medium contains electroactive compounds. For this purpose, 1% solutions of soy peptone and yeast extract were subjected to oxidative conditions, using a microbial fuel cell set-up corresponding to a typical galvanic cell, consisting of culture medium in the anodic half-cell and 0.5 M K(3)Fe(CN)(6) in the cathodic half-cell. Second, using a bi-cathodic microbial fuel cell, it was shown that electrons were transferred from yeast cells to the carbon anode. The participation of electroactive compounds in the electron transport was separated as background current. This result was verified by applying medium-free conditions, where only glucose was fed, confirming that electrons are transferred from yeast cells to the woven carbon anode. Knowledge about the electron transfer through the cell membrane is of importance in amperometric online monitoring of yeast fermentations and for electricity production with microbial fuel cells. Copyright (c) 2009 John Wiley & Sons, Ltd.

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

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.

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.

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...

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

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.

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.

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...

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.

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.

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...

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.

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.

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.

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

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.

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.

Microbial community structure elucidates performance of Glyceria maxima plant microbial fuel cell

OpenAIRE

Timmers, R.A.; Rothballer, M.; Strik, D.P.B.T.B.; Engel, M.; Schulz, M.; Hartmann, A.; Hamelers, H.V.M.; Buisman, C.J.N.

2012-01-01

The plant microbial fuel cell (PMFC) is a technology in which living plant roots provide electron donor, via rhizodeposition, to a mixed microbial community to generate electricity in a microbial fuel cell. Analysis and localisation of the microbial community is necessary for gaining insight into the competition for electron donor in a PMFC. This paper characterises the anode-rhizosphere bacterial community of a Glyceria maxima (reed mannagrass) PMFC. Electrochemically active bacteria (EAB) w...

Microbial community structure elucidates performance of Glyceria maxima plant microbial fuel cell

OpenAIRE

Timmers, Ruud A.; Rothballer, Michael; Strik, David P. B. T. B.; Engel, Marion; Schulz, Stephan; Schloter, Michael; Hartmann, Anton; Hamelers, Bert; Buisman, Cees

2012-01-01

The plant microbial fuel cell (PMFC) is a technology in which living plant roots provide electron donor, via rhizodeposition, to a mixed microbial community to generate electricity in a microbial fuel cell. Analysis and localisation of the microbial community is necessary for gaining insight into the competition for electron donor in a PMFC. This paper characterises the anode–rhizosphere bacterial community of a Glyceria maxima (reed mannagrass) PMFC. Electrochemically active bacteria (EAB) w...

The decrease of microbial charge in some spices by electron beam irradiation

International Nuclear Information System (INIS)

Ferdes, Mariana; Ferdes, Ovidiu

2008-01-01

The radiations of primary interest in food preservation are ionizing radiations; the use of electron accelerators offers certain advantages like high efficiency and flexibility in the choice of surface and depth treatments for a variety of food items. The paper presents the influence of ionising radiation on the microbial content in different spices: nutmeg, paprika, savory, ginger and oregano. The electron beam irradiation at doses within the range of 1-10 kGy reduced the values of CFU/g of total counts of aerobic germs and also of yeasts and moulds in these food products. The D 10 and D L for each sample were determined from the inactivation curves. For each food item the D L values for total aerobic microflora are higher than the corresponding D L values for fungi. In conclusion, values doses of 10 kGy are sufficient to reduce the microbial load of these spices under the values permitted by law. (authors)

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.

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

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.

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.

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.

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

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.

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.

Electron transfer mechanisms, new applications, and performance of biocathode microbial fuel cells

KAUST Repository

Huang, Liping; Regan, John M.; Quan, Xie

2011-01-01

Broad application of microbial fuel cells (MFCs) requires low cost and high operational sustainability. Microbial-cathode MFCs, or cathodes using only bacterial catalysts (biocathodes), can satisfy these demands and have gained considerable attention in recent years. Achievements with biocathodes over the past 3-4. years have been particularly impressive not only with respect to the biological aspects but also the system-wide considerations related to electrode materials and solution chemistry. The versatility of biocathodes enables us to use not only oxygen but also contaminants as possible electron acceptors, allowing nutrient removal and bioremediation in conjunction with electricity generation. Moreover, biocathodes create opportunities to convert electrical current into microbially generated reduced products. While many new experimental results with biocathodes have been reported, we are still in the infancy of their engineering development. This review highlights the opportunities, limits, and challenges of biocathodes. © 2010 Elsevier Ltd.

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...

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

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.

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.

Dose-effect curves for electron-beam irradiation of some collection microbial strains

International Nuclear Information System (INIS)

Ferdes, O.; Dumitru, E.; Catargiu, L.; Ferdes, M.; Minea, R.; Oproiu, C.; Niculescu, A.

1994-01-01

There were electron-beam irradiated some microbial strains of B.subtilis ICA I-60 both in germination and in sporulated forms. The irradiation were performed at the IPTRD's electron accelerator at 6 MeV, and in the dose range between 0.1-5.0 kGy, at different dose-rate varying from 50 Gy/minute to 100 Gy/minute. The dosimetry was carried out by a PTW medical dosemeter. There were established the dose-effect relationships and curves, the inactivation dose (factor) and the optimum domain for electron-beam mutagenesis. There were obtained some mutant strains with 2-3.5 higher biosynthesis potential, which are in the IFC's collection. (Author)

Microbial community structure elucidates performance of Glyceria maxima plant microbial fuel cell.

Science.gov (United States)

Timmers, Ruud A; Rothballer, Michael; Strik, David P B T B; Engel, Marion; Schulz, Stephan; Schloter, Michael; Hartmann, Anton; Hamelers, Bert; Buisman, Cees

2012-04-01

The plant microbial fuel cell (PMFC) is a technology in which living plant roots provide electron donor, via rhizodeposition, to a mixed microbial community to generate electricity in a microbial fuel cell. Analysis and localisation of the microbial community is necessary for gaining insight into the competition for electron donor in a PMFC. This paper characterises the anode-rhizosphere bacterial community of a Glyceria maxima (reed mannagrass) PMFC. Electrochemically active bacteria (EAB) were located on the root surfaces, but they were more abundant colonising the graphite granular electrode. Anaerobic cellulolytic bacteria dominated the area where most of the EAB were found, indicating that the current was probably generated via the hydrolysis of cellulose. Due to the presence of oxygen and nitrate, short-chain fatty acid-utilising denitrifiers were the major competitors for the electron donor. Acetate-utilising methanogens played a minor role in the competition for electron donor, probably due to the availability of graphite granules as electron acceptors.

Microbial community structure elucidates performance of Glyceria maxima plant microbial fuel cell

Energy Technology Data Exchange (ETDEWEB)

Timmers, Ruud A.; Strik, David P.B.T.B.; Hamelers, Bert; Buisman, Cees [Wageningen Univ. (Netherlands). Sub-dept. of Environmental Technology; Rothballer, Michael; Hartmann, Anton [Helmholtz Zentrum Muenchen, German Research Center for Environmental Health, Neuherberg (Germany). Dept. Microbe-Plant Interactions; Engel, Marion; Schulz, Stephan; Schloter, Michael [Helmholtz Zentrum Muenchen, German Research Center for Environmental Health, Neuherberg (Germany). Dept. Terrestrial Ecogenetics

2012-04-15

The plant microbial fuel cell (PMFC) is a technology in which living plant roots provide electron donor, via rhizodeposition, to a mixed microbial community to generate electricity in a microbial fuel cell. Analysis and localisation of the microbial community is necessary for gaining insight into the competition for electron donor in a PMFC. This paper characterises the anode-rhizosphere bacterial community of a Glyceria maxima (reed mannagrass) PMFC. Electrochemically active bacteria (EAB) were located on the root surfaces, but they were more abundant colonising the graphite granular electrode. Anaerobic cellulolytic bacteria dominated the area where most of the EAB were found, indicating that the current was probably generated via the hydrolysis of cellulose. Due to the presence of oxygen and nitrate, short-chain fatty acid-utilising denitrifiers were the major competitors for the electron donor. Acetate-utilising methanogens played a minor role in the competition for electron donor, probably due to the availability of graphite granules as electron acceptors. (orig.)

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

Microbial community structure elucidates performance of Glyceria maxima plant microbial fuel cell

NARCIS (Netherlands)

Timmers, R.A.; Rothballer, M.; Strik, D.P.B.T.B.; Engel, M.; Schulz, M.; Hartmann, A.; Hamelers, H.V.M.; Buisman, C.J.N.

2012-01-01

The plant microbial fuel cell (PMFC) is a technology in which living plant roots provide electron donor, via rhizodeposition, to a mixed microbial community to generate electricity in a microbial fuel cell. Analysis and localisation of the microbial community is necessary for gaining insight into

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

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.

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

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.

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.

Stay connected: Electrical conductivity of microbial aggregates.

Science.gov (United States)

Li, Cheng; Lesnik, Keaton Larson; Liu, Hong

2017-11-01

The discovery of direct extracellular electron transfer offers an alternative to the traditional understanding of diffusional electron exchange via small molecules. The establishment of electronic connections between electron donors and acceptors in microbial communities is critical to electron transfer via electrical currents. These connections are facilitated through conductivity associated with various microbial aggregates. However, examination of conductivity in microbial samples is still in its relative infancy and conceptual models in terms of conductive mechanisms are still being developed and debated. The present review summarizes the fundamental understanding of electrical conductivity in microbial aggregates (e.g. biofilms, granules, consortia, and multicellular filaments) highlighting recent findings and key discoveries. A greater understanding of electrical conductivity in microbial aggregates could facilitate the survey for additional microbial communities that rely on direct extracellular electron transfer for survival, inform rational design towards the aggregates-based production of bioenergy/bioproducts, and inspire the construction of new synthetic conductive polymers. Copyright © 2017 Elsevier Inc. All rights reserved.

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.

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 .

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...

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.

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)

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

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

Directory of Open Access Journals (Sweden)

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.

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.

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 ...

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...

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.

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

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)

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)

New method for characterizing electron mediators in microbial systems using a thin-layer twin-working electrode cell.

Science.gov (United States)

Hassan, Md Mahamudul; Cheng, Ka Yu; Ho, Goen; Cord-Ruwisch, Ralf

2017-01-15

Microbial biofilms are significant ecosystems where the existence of redox gradients drive electron transfer often via soluble electron mediators. This study describes the use of two interfacing working electrodes (WEs) to simulate redox gradients within close proximity (250µm) for the detection and quantification of electron mediators. By using a common counter and reference electrode, the potentials of the two WEs were independently controlled to maintain a suitable "voltage window", which enabled simultaneous oxidation and reduction of electron mediators as evidenced by the concurrent anodic and cathodic currents, respectively. To validate the method, the electrochemical properties of different mediators (hexacyanoferrate, HCF, riboflavin, RF) were characterized by stepwise shifting the "voltage window" (ranging between 25 and 200mV) within a range of potentials after steady equilibrium current of both WEs was established. The resulting differences in electrical currents between the two WEs were recorded across a defined potential spectrum (between -1V and +0.5V vs. Ag/AgCl). Results indicated that the technique enabled identification (by the distinct peak locations at the potential scale) and quantification (by the peak of current) of the mediators for individual species as well as in an aqueous mixture. It enabled a precise determination of mid-potentials of the externally added mediators (HCF, RF) and mediators produced by pyocyanin-producing Pseudomonas aeruginosa (WACC 91) culture. The twin working electrode described is particularly suitable for studying mediator-dependent microbial electron transfer processes or simulating redox gradients as they exist in microbial biofilms. Copyright © 2016 Elsevier B.V. All rights reserved.

Electron acceptors for anaerobic oxidation of methane drive microbial community structure and diversity in mud volcanoes.

Science.gov (United States)

Ren, Ge; Ma, Anzhou; Zhang, Yanfen; Deng, Ye; Zheng, Guodong; Zhuang, Xuliang; Zhuang, Guoqiang; Fortin, Danielle

2018-04-06

Mud volcanoes (MVs) emit globally significant quantities of methane into the atmosphere, however, methane cycling in such environments is not yet fully understood, as the roles of microbes and their associated biogeochemical processes have been largely overlooked. Here, we used data from high-throughput sequencing of microbial 16S rRNA gene amplicons from six MVs in the Junggar Basin in northwest China to quantify patterns of diversity and characterize the community structure of archaea and bacteria. We found anaerobic methanotrophs and diverse sulfate- and iron-reducing microbes in all of the samples, and the diversity of both archaeal and bacterial communities was strongly linked to the concentrations of sulfate, iron and nitrate, which could act as electron acceptors in anaerobic oxidation of methane (AOM). The impacts of sulfate/iron/nitrate on AOM in the MVs were verified by microcosm experiments. Further, two representative MVs were selected to explore the microbial interactions based on phylogenetic molecular ecological networks. The sites showed distinct network structures, key species and microbial interactions, with more complex and numerous linkages between methane-cycling microbes and their partners being observed in the iron/sulfate-rich MV. These findings suggest that electron acceptors are important factors driving the structure of microbial communities in these methane-rich environments. © 2018 The Authors. Environmental Microbiology published by Society for Applied Microbiology and John Wiley & Sons Ltd.

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.

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.

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,

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

Influence of Bicarbonate, Sulfate, and Electron Donors on Biological reduction of Uranium and Microbial Community Composition

Energy Technology Data Exchange (ETDEWEB)

Luo, Wensui [ORNL; Zhou, Jizhong [ORNL; Wu, Weimin [ORNL; Yan, Tingfen [ORNL; Criddle, Craig [ORNL; Jardine, Philip M [ORNL; Gu, Baohua [ORNL

2007-01-01

A microcosm study was performed to investigate the effect of ethanol and acetate on uranium(VI) biological reduction and microbial community changes under various geochemical conditions. Each microcosm contained an uranium-contaminated sediment (up to 2.8 g U/kg) suspended in buffer with bicarbonate at concentrations of either 1 mM or 40 mM and sulfate at either 1.1 or 3.2 mM. Ethanol or acetate was used as an electron donor. Results indicate that ethanol yielded in significantly higher U(VI) reduction rates than acetate. A low bicarbonate concentration (1 mM) was favored for U(VI) bioreduction to occur in sediments, but high concentrations of bicarbonate (40 mM) and sulfate (3.2 mM) decreased the reduction rates of U(VI). Microbial communities were dominated by species from the Geothrix genus and Proteobacteria phylum in all microcosms. However, species in the Geobacteraceae family capable of reducing U(VI) were significantly enriched by ethanol and acetate in low bicarbonate buffer. Ethanol increased the population of unclassified Desulfuromonales, while acetate increased the population of Desulfovibrio. Additionally, species in the Geobacteraceae family were not enriched in high bicarbonate buffer, but the Geothrix and the unclassified Betaproteobacteria species were enriched. This study concludes that ethanol could be a better electron donor than acetate for reducing U(VI) under given experimental conditions, and electron donor and geoundwater geochemistry alter microbial communities responsible for U(VI) reduction.

Influence of bicarbonate, sulfate, and electron donors on biological reduction of uranium and microbial community composition

Energy Technology Data Exchange (ETDEWEB)

Luo Wensui [Oak Ridge Inst. for Science and Education, TN (United States); Oak Ridge National Lab., TN (United States). Environmental Sciences Div.; Wu Wei-Min; Criddle, C.S. [Stanford Univ., CA (United States). Dept. of Civil and Environmental Engineering; Yan Tingfen [Oak Ridge Inst. for Science and Education, TN (United States); Jardine, P.M.; Gu Baohua [Oak Ridge National Lab., TN (United States). Environmental Sciences Div.; Zhou Jizhong [Oklahoma Univ., Norman, OK (United States). Dept. of Botany and Microbiology

2007-12-15

A microcosm study was performed to investigate the effect of ethanol and acetate on uranium(VI) biological reduction and microbial community changes under various geochemical conditions. Each microcosm contained an uranium-contaminated sediment (up to 2.8 g U/kg) suspended in buffer with bicarbonate at concentrations of either 1 or 40 mM and sulfate at either 1.1 or 3.2 mM. Ethanol or acetate was used as an electron donor. Results indicate that ethanol yielded in significantly higher U(VI) reduction rates than acetate. A low bicarbonate concentration (1 mM) was favored for U(VI) bioreduction to occur in sediments, but high concentrations of bicarbonate (40 mM) and sulfate (3.2 mM) decreased the reduction rates of U(VI). Microbial communities were dominated by species from the Geothrix genus and Proteobacteria phylum in all microcosms. However, species in the Geobacteraceae family capable of reducing U(VI) were significantly enriched by ethanol and acetate in low-bicarbonate buffer. Ethanol increased the population of unclassified Desulfuromonales, while acetate increased the population of Desulfovibrio. Additionally, species in the Geobacteraceae family were not enriched in high-bicarbonate buffer, but the Geothrix and the unclassified Betaproteobacteria species were enriched. This study concludes that ethanol could be a better electron donor than acetate for reducing U(VI) under given experimental conditions, and electron donor and groundwater geochemistry alter microbial communities responsible for U(VI) reduction. (orig.)

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.

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.

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.

The effect of flavin electron shuttles in microbial fuel cells current production

Energy Technology Data Exchange (ETDEWEB)

Velasquez-Orta, Sharon B. [Newcastle Univ., Newcastle upon Tyne (United Kingdom). School of Civil Engineering and Geosciences; Newcastle Univ., Newcastle upon Tyne (United Kingdom). School of Chemical Engineering and Advanced Materials; Head, Ian M.; Curtis, Thomas P. [Newcastle Univ., Newcastle upon Tyne (United Kingdom). School of Civil Engineering and Geosciences; Scott, Keith [Newcastle Univ., Newcastle upon Tyne (United Kingdom). School of Chemical Engineering and Advanced Materials; Lloyd, Jonathan R.; Canstein, Harald von [Manchester Univ. (United Kingdom). School of Earth, Atmospheric and Environmental Sciences

2010-02-15

The effect of electron shuttles on electron transfer to microbial fuel cell (MFC) anodes was studied in systems where direct contact with the anode was precluded. MFCs were inoculated with Shewanella cells, and flavins used as the electron shuttling compound. In MFCs with no added electron shuttles, flavin concentrations monitored in the MFCs' bulk liquid increased continuously with FMN as the predominant flavin. The maximum concentrations were 0.6 {mu}M for flavin mononucleotide and 0.2 {mu}M for riboflavin. In MFCs with added flavins, micro-molar concentrations were shown to increase current and power output. The peak current was at least four times higher in MFCs with high concentrations of flavins (4.5-5.5 {mu}M) than in MFCs with low concentrations (0.2-0.6 {mu}M). Although high power outputs (around 150 mW/m{sup 2}) were achieved in MFCs with high concentrations of flavins, a Clostridium-like bacterium along with other reactor limitations affected overall coulombic efficiencies (CE) obtained, achieving a maximum CE of 13%. Electron shuttle compounds (flavins) permitted bacteria to utilise a remote electron acceptor (anode) that was not accessible to the cells allowing current production until the electron donor (lactate) was consumed. (orig.)

Evaluation of phytate-degrading Lactobacillus culture administration to broiler chickens.

Science.gov (United States)

Askelson, Tyler E; Campasino, Ashley; Lee, Jason T; Duong, Tri

2014-02-01

Probiotics have been demonstrated to promote growth, stimulate immune responses, and improve food safety of poultry. While widely used, their effectiveness is mixed, and the mechanisms through which they contribute to poultry production are not well understood. Microbial phytases are increasingly supplemented in feed to improve digestibility and reduce antinutritive effects of phytate. The microbial origin of these exogenous enzymes suggests a potentially important mechanism of probiotic functionality. We investigated phytate degradation as a novel probiotic mechanism using recombinant Lactobacillus cultures expressing Bacillus subtilis phytase. B. subtilis phyA was codon optimized for expression in Lactobacillus and cloned into the expression vector pTRK882. The resulting plasmid, pTD003, was transformed into Lactobacillus acidophilus, Lactobacillus gallinarum, and Lactobacillus gasseri. SDS-PAGE revealed a protein in the culture supernatants of Lactobacillus pTD003 transformants with a molecular weight similar to that of the B. subtilis phytase. Expression of B. subtilis phytase increased phytate degradation of L. acidophilus, L. gasseri, and L. gallinarum approximately 4-, 10-, and 18-fold over the background activity of empty-vector transformants, respectively. Phytase-expressing L. gallinarum and L. gasseri were administered to broiler chicks fed a phosphorus-deficient diet. Phytase-expressing L. gasseri improved weight gain of broiler chickens to a level comparable to that for chickens fed a control diet adequate in phosphorus, demonstrating proof of principle that administration of phytate-degrading probiotic cultures can improve performance of livestock animals. This will inform future studies investigating whether probiotic cultures are able to provide both the performance benefits of feed enzymes and the animal health and food safety benefits traditionally associated with probiotics.

Reduced Nutrient Excretion and Environmental Microbial Load with the Addition of a Combination of Enzymes and Direct-Fed Microbials to the Diet of Broiler Chickens

Directory of Open Access Journals (Sweden)

MFFM Praes

2016-03-01

Full Text Available Abstract This study evaluated the effects of the dietary inclusion of an enzyme blend and a direct-fed microbials in broiler diets on litter production and quality. In total, 900 Cobb 500(r broiler chicks were distributed according to a completely randomized design into 4 treatments and 9 replicates of 25 birds each. Broilers were reared from 1 to 42 days of age. The treatments consisted of the following diets: NC: negative control; DFM: NC + 500 ppm of direct-fed microbials product (DFM, containing Bacillus subtilis and Bacillus licheniformis; ENZ: diet formulated with an enzyme blend (20 ppm phytase, 200 ppm protease and 200 ppm of xylanase; DFM+E: ENZ + DFM. Birds and litter were weighed at the start and end of the rearing period, for litter production and waste ratio (Rw determination. Litter samples were analyzed for dry matter (DM content, total and thermotolerant coliform counts, nutrient composition (nitrogen (N, phosphorous (P and potassium (K, and fiber fraction (neutral detergent fiber (NDF, acid detergent fiber (ADF and lignin. The dietary inclusion of the evaluated additivesdid not influence litter production or Rw; however, ADF (%, NDF (kg and kg/kg DM litter, and total and thermotolerant coliform counts were reduced, and N content increased in the litter. The diets containing enzymes (ENZ and DFM+E reduced litter P content. The addition of exogenous enzymes and their combination with a DFM based on Bacillus spp .Did not affect waste production, and reduced litter microbial load, and the contents of P and insoluble fiber in the litter.

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.

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.

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.

Microbial Anaerobic Ammonium Oxidation Under Iron Reducing Conditions, Alternative Electron Acceptors

Science.gov (United States)

Ruiz-Urigüen, M.; Jaffe, P. R.

2015-12-01

Autotrophic Acidimicrobiaceae-bacterium named A6 (A6), part of the Actinobacteria phylum have been linked to anaerobic ammonium (NH4+) oxidation under iron reducing conditions. These organisms obtain their energy by oxidizing NH4+ and transferring the electrons to a terminal electron acceptor (TEA). Under environmental conditions, the TEAs are iron oxides [Fe(III)], which are reduced to Fe(II), this process is known as Feammox. Our studies indicate that alternative forms of TEAs can be used by A6, e.g. iron rich clays (i.e. nontronite) and electrodes in bioelectrochemical systems such as Microbial Electrolysis Cells (MECs), which can sustain NH4+removal and A6 biomass production. Our results show that nontronite can support Feammox and promote bacterial cell production. A6 biomass increased from 4.7 x 104 to 3.9 x 105 cells/ml in 10 days. Incubations of A6 in nontronite resulted in up to 10 times more NH4+ removal and 3 times more biomass production than when ferrihydrite is used as the Fe(III) source. Additionally, Fe in nontronite can be reoxidized by aeration and A6 can reutilize it; however, Fe is still finite in the clay. In contrast, in MECs, A6 harvest electrons from NH4+ and use an anode as an unlimited TEA, as a result current is produced. We operated multiple MECs in parallel using a single external power source, as described by Call & Logan (2011). MECs were run with an applied voltage of 0.7V and different growing mediums always containing initial 5mM NH4+. Results show that current production is favored when anthraquinone-2,6-disulfonate (AQDS), an electron shuttled, is present in the medium as it facilitates the transfer of electrons from the bacterial cell to the anode. Additionally, A6 biomass increased from 1 x 104 to 9.77 x 105cells/ml in 14 days of operation. Due to Acidimicrobiaceae-bacterium A6's ability to use various TEAs, MECs represent an alternative, iron-free form, for optimized biomass production of A6 and its application in NH4

Electron shuttles in biotechnology.

Science.gov (United States)

Watanabe, Kazuya; Manefield, Mike; Lee, Matthew; Kouzuma, Atsushi

2009-12-01

Electron-shuttling compounds (electron shuttles [ESs], or redox mediators) are essential components in intracellular electron transfer, while microbes also utilize self-produced and naturally present ESs for extracellular electron transfer. These compounds assist in microbial energy metabolism by facilitating electron transfer between microbes, from electron-donating substances to microbes, and/or from microbes to electron-accepting substances. Artificially supplemented ESs can create new routes of electron flow in the microbial energy metabolism, thereby opening up new possibilities for the application of microbes to biotechnology processes. Typical examples of such processes include halogenated-organics bioremediation, azo-dye decolorization, and microbial fuel cells. Herein we suggest that ESs can be applied widely to create new microbial biotechnology processes.

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

Effects of gamma ray and electron beam irradiation on reduction of microbial load and antioxidant properties of Chum-Hed-Thet (Cassia alata (L.) Roxb.)

Science.gov (United States)

Prakhongsil, P.; Pewlong, W.; Sajjabut, S.; Chookaew, S.

2017-06-01

Considering the growing demands of herbal medicines, Cassia alata (L.) Roxb. has been reported to have various phytochemical activities. It has also been called in Thai as Chum-Hed-Thet. In this study, C. alata (L.) Roxb. powder were exposed to gamma and electron beam irradiation at doses of 0, 5, 10, 15 and 20 kGy. At the dose of 10 kGy, both of gamma and electron beam irradiation were sufficient in reducing microbial load of irradiated samples as specified in Thai pharmacopoeia (2005). These include the total aerobic microbial count of bacteria of 0.05). Therefore, both of radiation by gamma ray or electron beam at 10 kGy was sufficient in elimination of microbial flora and did not significantly affected the total phenolic content and antioxidant activities of C. alata (L.) Roxb.

The presence of inositol phosphates in gastric pig digesta is affected by time after feeding a nonfermented or fermented liquid wheat- barley-based diet

DEFF Research Database (Denmark)

Blaabjerg, Karoline; Jørgensen, H.; Tauson, Anne-Helene

2011-01-01

The objective was to quantify the retention of digesta and evaluate the degradation of phytate or inositol hexakisphosphate (InsP(6)) and lower inositol phosphates (InsP(5), InsP(4), InsP(3), and InsP(2)) in the stomach at different times after feeding pigs a fermented liquid diet with microbial...... collection of gastric digesta. For each pig, the digesta was collected once daily at 1, 2, 3, or 5 h after feeding the morning meal. A basal wheat- and barley-based diet was steam-pelleted at 90°C. The dietary treatments were a nonfermented basal diet (NF-BD), the NF-BD with microbial phytase (750 phytase...... over time (P

Electronic Nose Technology to Measure Soil Microbial Activity and Classify Soil Metabolic Status

OpenAIRE

Fabrizio De Cesare; Elena Di Mattia; Simone Pantalei; Emiliano Zampetti; Vittorio Vinciguerra; Antonella Macagnano

2011-01-01

The electronic nose (E-nose) is a sensing technology that has been widely used to monitor environments in the last decade. In the present study, the capability of an E-nose, in combination with biochemical and microbiological techniques, of both detecting the microbial activity and estimating the metabolic status of soil ecosystems, was tested by measuring on one side respiration, enzyme activities and growth of bacteria in natural but simplified soil ecosystems over 23 days of incubation thr...

Humin as an electron donor for enhancement of multiple microbial reduction reactions with different redox potentials in a consortium.

Science.gov (United States)

Zhang, Dongdong; Zhang, Chunfang; Xiao, Zhixing; Suzuki, Daisuke; Katayama, Arata

2015-02-01

A solid-phase humin, acting as an electron donor, was able to enhance multiple reductive biotransformations, including dechlorination of pentachlorophenol (PCP), dissimilatory reduction of amorphous Fe (III) oxide (FeOOH), and reduction of nitrate, in a consortium. Humin that was chemically reduced by NaBH4 served as an electron donor for these microbial reducing reactions, with electron donating capacities of 0.013 mmol e(-)/g for PCP dechlorination, 0.15 mmol e(-)/g for iron reduction, and 0.30 mmol e(-)/g for nitrate reduction. Two pairs of oxidation and reduction peaks within the humin were detected by cyclic voltammetry analysis. 16S rRNA gene sequencing-based microbial community analysis of the consortium incubated with different terminal electron acceptors, suggested that Dehalobacter sp., Bacteroides sp., and Sulfurospirillum sp. were involved in the PCP dechlorination, dissimilatory iron reduction, and nitrate reduction, respectively. These findings suggested that humin functioned as a versatile redox mediator, donating electrons for multiple respiration reactions with different redox potentials. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

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

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.

The impact of phosphorus on the immune system and the intestinal microbiota with special focus on the pig.

Science.gov (United States)

Heyer, Charlotte M E; Weiss, Eva; Schmucker, Sonja; Rodehutscord, Markus; Hoelzle, Ludwig E; Mosenthin, Rainer; Stefanski, Volker

2015-06-01

There is increasing interest in dietary ingredients that are appropriate to support digestive and immune functions, but also maintain a stable microbial ecosystem in the gastrointestinal tract (GIT), particularly in weaned pigs. P is an essential nutrient for both microbes and their host, as it is involved, for example, in bone formation, energy metabolism, cellular signalling and stabilisation of cell membranes. Non-ruminant animals have limited access to phytate, the main storage form of P in plant seeds. The release of P bound to phytate requires phytase activity of plant or microbial origin, resulting in the formation of variable phosphorylated inositol phosphates (InsPs). The present review focuses on interactions between variations in dietary P supply, the immune system of the host, and the intestinal microbial ecosystem. Although results on the interaction between P and the immune system are inconsistent, several studies in different species have shown a positive impact of dietary P and phytase addition on the adaptive immune response. Recent studies with pigs suggest that P supply may influence intestinal microbial composition and activity. Individual InsPs or phosphate may also affect properties of pathogenic micro-organisms, such as metabolism or virulence. In conclusion, P may be considered as part of an integrated approach to support immune functions and maintain a stable microbial ecosystem in the GIT, thereby providing a barrier against potential pathogens. Within this regard, differences in phytate-P content and intrinsic phytase activity of plant feedstuffs, as well as the formation of individual InsPs, have to be taken into account.

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.

Influence of Gamma and Electron Beam Irradiation on Microbial Load of Pueraria mirifica

International Nuclear Information System (INIS)

Eamsiri, J.; Pewlong, W.; Sajjabut, S.; Chookaew, S.

2014-01-01

The purpose of this study is to investigate the effect of gamma ray and electron beam on the microbial load of Pueraria mirifica at selected storage period post exposure to irradiation. The samples were irradiated at 0, 5, 10, 15 and 20 kGy and then analyzed for the total bacteria, total yeast and mold, and the presence of Coliform bacteria, Escherichia coli, Salmonella sp., Bacillus cereus and Clostridium perfringens after 0, 3, 6, 9 and 12 months of storage. Results demonstrated that both irradiation techniques significantly reduced microbial contamination. As the reduction in bacteria count decreased linearly with the absorbed dose, the dose of 5 kGy was found to be sufficient in eliminating pathogens with the total bacteria count decreased to the value accepted by the Thai Industrial standard 1441/2552. In addition, we found that total bacteria, total yeast and mold and pathogens did not change significantly after storage up to 12 months post irradiation.

Multiple paths of electron flow to current in microbial electrolysis cells fed with low and high concentrations of propionate

KAUST Repository

Rao, Hari Ananda; Katuri, Krishna; Gorron, Eduardo; Logan, Bruce E.; Saikaly, Pascal

2016-01-01

Microbial electrolysis cells (MECs) provide a viable approach for bioenergy generation from fermentable substrates such as propionate. However, the paths of electron flow during propionate oxidation in the anode of MECs are unknown. Here, the paths

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.

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.

Multiple paths of electron flow to current in microbial electrolysis cells fed with low and high concentrations of propionate

KAUST Repository

Rao, Hari Ananda

2016-03-03

Microbial electrolysis cells (MECs) provide a viable approach for bioenergy generation from fermentable substrates such as propionate. However, the paths of electron flow during propionate oxidation in the anode of MECs are unknown. Here, the paths of electron flow involved in propionate oxidation in the anode of two-chambered MECs were examined at low (4.5 mM) and high (36 mM) propionate concentrations. Electron mass balances and microbial community analysis revealed that multiple paths of electron flow (via acetate/H2 or acetate/formate) to current could occur simultaneously during propionate oxidation regardless of the concentration tested. Current (57–96 %) was the largest electron sink and methane (0–2.3 %) production was relatively unimportant at both concentrations based on electron balances. At a low propionate concentration, reactors supplemented with 2-bromoethanesulfonate had slightly higher coulombic efficiencies than reactors lacking this methanogenesis inhibitor. However, an opposite trend was observed at high propionate concentration, where reactors supplemented with 2-bromoethanesulfonate had a lower coulombic efficiency and there was a greater percentage of electron loss (23.5 %) to undefined sinks compared to reactors without 2-bromoethanesulfonate (11.2 %). Propionate removal efficiencies were 98 % (low propionate concentration) and 78 % (high propionate concentration). Analysis of 16S rRNA gene pyrosequencing revealed the dominance of sequences most similar to Geobacter sulfurreducens PCA and G. sulfurreducens subsp. ethanolicus. Collectively, these results provide new insights on the paths of electron flow during propionate oxidation in the anode of MECs fed with low and high propionate concentrations.

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.

- Invited Review - Calcium Digestibility and Metabolism in Pigs*

Science.gov (United States)

González-Vega, J. C.; Stein, H. H.

2014-01-01

Calcium (Ca) and phosphorus (P) are minerals that have important physiological functions in the body. For formulation of diets for pigs, it is necessary to consider an appropriate Ca:P ratio for an adequate absorption and utilization of both minerals. Although both minerals are important, much more research has been conducted on P digestibility than on Ca digestibility. Therefore, this review focuses on aspects that are important for the digestibility of Ca. Only values for apparent total tract digestibility (ATTD) of Ca have been reported in pigs, whereas values for both ATTD and standardized total tract digestibility (STTD) of P in feed ingredients have been reported. To be able to determine STTD values for Ca it is necessary to determine basal endogenous losses of Ca. Although most Ca is absorbed in the small intestine, there are indications that Ca may also be absorbed in the colon under some circumstances, but more research to verify the extent of Ca absorption in different parts of the intestinal tract is needed. Most P in plant ingredients is usually bound to phytate. Therefore, plant ingredients have low digestibility of P due to a lack of phytase secretion by pigs. During the last 2 decades, inclusion of microbial phytase in swine diets has improved P digestibility. However, it has been reported that a high inclusion of Ca reduces the efficacy of microbial phytase. It is possible that formation of insoluble calcium-phytate complexes, or Ca-P complexes, not only may affect the efficacy of phytase, but also the digestibility of P and Ca. Therefore, Ca, P, phytate, and phytase interactions are aspects that need to be considered in Ca digestibility studies. PMID:25049919

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...

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.

Microbial fuel cell operation using monoazo and diazo dyes as terminal electron acceptor for simultaneous decolourisation and bioelectricity generation.

Science.gov (United States)

Oon, Yoong-Sin; Ong, Soon-An; Ho, Li-Ngee; Wong, Yee-Shian; Oon, Yoong-Ling; Lehl, Harvinder Kaur; Thung, Wei-Eng; Nordin, Noradiba

2017-03-05

Monoazo and diazo dyes [New coccine (NC), Acid orange 7 (AO7), Reactive red 120 (RR120) and Reactive green 19 (RG19)] were employed as electron acceptors in the abiotic cathode of microbial fuel cell. The electrons and protons generated from microbial organic oxidation at the anode which were utilized for electrochemical azo dye reduction at the cathodic chamber was successfully demonstrated. When NC was employed as the electron acceptor, the chemical oxygen demand (COD) removal and dye decolourisation efficiencies obtained at the anodic and cathodic chamber were 73±3% and 95.1±1.1%, respectively. This study demonstrated that the decolourisation rates of monoazo dyes were ∼50% higher than diazo dyes. The maximum power density in relation to NC decolourisation was 20.64mW/m 2 , corresponding to current density of 120.24mA/m 2 . The decolourisation rate and power output of different azo dyes were in the order of NC>AO7>RR120>RG19. The findings revealed that the structure of dye influenced the decolourisation and power performance of MFC. Azo dye with electron-withdrawing group at para substituent to azo bond would draw electrons from azo bond; hence the azo dye became more electrophilic and more favourable for dye reduction. Copyright © 2016 Elsevier B.V. All rights reserved.

Are Microbial Nanowires Responsible for Geoelectrical Changes at Hydrocarbon Contaminated Sites?

Science.gov (United States)

Hager, C.; Atekwana, E. A.; Gorby, Y. A.; Duris, J. W.; Allen, J. P.; Atekwana, E. A.; Ownby, C.; Rossbach, S.

2007-05-01

Significant advances in near-surface geophysics and biogeophysics in particular, have clearly established a link between geoelectrical response and the growth and enzymatic activities of microbes in geologic media. Recent studies from hydrocarbon contaminated sites suggest that the activities of distinct microbial populations, specifically syntrophic, sulfate reducing, and dissimilatory iron reducing microbial populations are a contributing factor to elevated sediment conductivity. However, a fundamental mechanistic understanding of the processes and sources resulting in the measured electrical response remains uncertain. The recent discovery of bacterial nanowires and their electron transport capabilities suggest that if bacterial nanowires permeate the subsurface, they may in part be responsible for the anomalous conductivity response. In this study we investigated the microbial population structure, the presence of nanowires, and microbial-induced alterations of a hydrocarbon contaminated environment and relate them to the sediments' geoelectrical response. Our results show that microbial communities varied substantially along the vertical gradient and at depths where hydrocarbons saturated the sediments, ribosomal intergenic spacer analysis (RISA) revealed signatures of microbial communities adapted to hydrocarbon impact. In contrast, RISA profiles from a background location showed little community variations with depth. While all sites showed evidence of microbial activity, a scanning electron microscope (SEM) study of sediment from the contaminated location showed pervasive development of "nanowire-like structures" with morphologies consistent with nanowires from laboratory experiments. SEM analysis suggests extensive alteration of the sediments by microbial Activity. We conclude that, excess organic carbon (electron donor) but limited electron acceptors in these environments cause microorganisms to produce nanowires to shuttle the electrons as they seek for

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

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

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.

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,...

Microbial fuel cell operation using monoazo and diazo dyes as terminal electron acceptor for simultaneous decolourisation and bioelectricity generation

International Nuclear Information System (INIS)

Oon, Yoong-Sin; Ong, Soon-An; Ho, Li-Ngee; Wong, Yee-Shian; Oon, Yoong-Ling; Lehl, Harvinder Kaur; Thung, Wei-Eng; Nordin, Noradiba

2017-01-01

Highlights: • Monoazo and diazo dyes were used as electron acceptor in the abiotic cathode of MFC. • Simultaneous decolourisation and bioelectricity generation were achieved. • Azo dye structures influenced the decolourisation performance. • Positive relation between decolourisation rate and power performance. - Abstract: Monoazo and diazo dyes [New coccine (NC), Acid orange 7 (AO7), Reactive red 120 (RR120) and Reactive green 19 (RG19)] were employed as electron acceptors in the abiotic cathode of microbial fuel cell. The electrons and protons generated from microbial organic oxidation at the anode which were utilized for electrochemical azo dye reduction at the cathodic chamber was successfully demonstrated. When NC was employed as the electron acceptor, the chemical oxygen demand (COD) removal and dye decolourisation efficiencies obtained at the anodic and cathodic chamber were 73 ± 3% and 95.1 ± 1.1%, respectively. This study demonstrated that the decolourisation rates of monoazo dyes were ∼50% higher than diazo dyes. The maximum power density in relation to NC decolourisation was 20.64 mW/m"2, corresponding to current density of 120.24 mA/m"2. The decolourisation rate and power output of different azo dyes were in the order of NC > AO7 > RR120 > RG19. The findings revealed that the structure of dye influenced the decolourisation and power performance of MFC. Azo dye with electron-withdrawing group at para substituent to azo bond would draw electrons from azo bond; hence the azo dye became more electrophilic and more favourable for dye reduction.

Microbial fuel cell operation using monoazo and diazo dyes as terminal electron acceptor for simultaneous decolourisation and bioelectricity generation

Energy Technology Data Exchange (ETDEWEB)

Oon, Yoong-Sin [Water Research Group (WAREG), School of Environmental Engineering, Universiti Malaysia Perlis, 02600, Arau, Perlis (Malaysia); Ong, Soon-An, E-mail: ongsoonan@yahoo.com [Water Research Group (WAREG), School of Environmental Engineering, Universiti Malaysia Perlis, 02600, Arau, Perlis (Malaysia); Ho, Li-Ngee [School of Materials Engineering, Universiti Malaysia Perlis, 02600, Arau, Perlis (Malaysia); Wong, Yee-Shian; Oon, Yoong-Ling; Lehl, Harvinder Kaur; Thung, Wei-Eng [Water Research Group (WAREG), School of Environmental Engineering, Universiti Malaysia Perlis, 02600, Arau, Perlis (Malaysia); Nordin, Noradiba [School of Materials Engineering, Universiti Malaysia Perlis, 02600, Arau, Perlis (Malaysia)

2017-03-05

Highlights: • Monoazo and diazo dyes were used as electron acceptor in the abiotic cathode of MFC. • Simultaneous decolourisation and bioelectricity generation were achieved. • Azo dye structures influenced the decolourisation performance. • Positive relation between decolourisation rate and power performance. - Abstract: Monoazo and diazo dyes [New coccine (NC), Acid orange 7 (AO7), Reactive red 120 (RR120) and Reactive green 19 (RG19)] were employed as electron acceptors in the abiotic cathode of microbial fuel cell. The electrons and protons generated from microbial organic oxidation at the anode which were utilized for electrochemical azo dye reduction at the cathodic chamber was successfully demonstrated. When NC was employed as the electron acceptor, the chemical oxygen demand (COD) removal and dye decolourisation efficiencies obtained at the anodic and cathodic chamber were 73 ± 3% and 95.1 ± 1.1%, respectively. This study demonstrated that the decolourisation rates of monoazo dyes were ∼50% higher than diazo dyes. The maximum power density in relation to NC decolourisation was 20.64 mW/m{sup 2}, corresponding to current density of 120.24 mA/m{sup 2}. The decolourisation rate and power output of different azo dyes were in the order of NC > AO7 > RR120 > RG19. The findings revealed that the structure of dye influenced the decolourisation and power performance of MFC. Azo dye with electron-withdrawing group at para substituent to azo bond would draw electrons from azo bond; hence the azo dye became more electrophilic and more favourable for dye reduction.

Fundamental Insights into Propionate Oxidation in Microbial Electrolysis Cells Using a Combination of Electrochemical, Molecular biology and Electron Balance Approaches

KAUST Repository

Rao, Hari Ananda

2016-11-01

Increasing demand for freshwater and energy is pushing towards the development of alternative technologies that are sustainable. One of the realistic solutions to address this is utilization of the renewable resources like wastewater. Conventional wastewater treatment processes can be highly energy demanding and can fails to recover the full potential of useful resources such as energy in the wastewater. As a consequence, there is an urgent necessity for sustainable wastewater treatment technologies that could harness such resources present in wastewaters. Advanced treatment process based on microbial electrochemical technologies (METs) such as microbial fuel cells (MFCs) and microbial electrolysis cells (MECs) have a great potential for the resources recovery through a sustainable wastewater treatment process. METs rely on the abilities of microorganisms that are capable of transferring electrons extracellularly by oxidizing the organic matter in the wastewater and producing electrical current for electricity generation (MFC) or H2 and CH4 production (MEC). Propionate is an important volatile fatty acid (VFA) (24-70%) in some wastewaters and accumulation of this VFA can cause a process failure in a conventional anaerobic digestion (AD) system. To address this issue, MECs were explored as a novel, alternative wastewater treatment technology, with a focus on a better understanding of propionate oxidation in the anode of MECs. Having such knowledge could help in the development of more robust and efficient wastewater treatment systems to recover energy and produce high quality effluents. Several studies were conducted to: 1) determine the paths of electron flow in the anode of propionate fed MECs low (4.5 mM) and high (36 mM) propionate concentrations; 2) examine the effect of different set anode potentials on the electrochemical performance, propionate degradation, electron fluxes, and microbial community structure in MECs fed propionate; and 3) examine the temporal

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

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

Quorum sensing alters the microbial community of electrode-respiring bacteria and hydrogen scavengers toward improving hydrogen yield in microbial electrolysis cells

International Nuclear Information System (INIS)

Cai, Weiwei; Zhang, Zhaojing; Ren, Ge; Shen, Qiuxuan; Hou, Yanan; Ma, Anzhou; Deng, Ye; Wang, Aijie; Liu, Wenzong

2016-01-01

Highlights: • Enhanced hydrogen yield has been achieved with addition of AHL. • AHL regulated exoelectrogens resulting in electrochemical activity enhancement. • Microbial community shift in cathodic biofilm inhibited hydrogen loss. - Abstract: Quorum sensing has been widely applied to enhance the energy recovery of bioelectrochemical system as a sustainable pathway to enhance communication between cells and electrodes. However, how signalling molecules (acyl-homoserine lactones, AHLs) regulate the microbial community to improve hydrogen generation in microbial electrolysis cells (MECs) is not well understood, especially the subsequent influence on interspecies relationships among not only electrode-respiring bacteria but also hydrogen scavengers. Understanding AHL regulation in a complicated and actual biofilm system will be valuable for future applications of microbial electrochemical technology. Herein, we added short-chain AHLs (3OC6) to regulate the biofilm community on bio-electrodes in MECs. As a result, hydrogen yields were enhanced with AHL addition, increasing by 5.57%, 38.68%, and 81.82% with varied external voltages (0.8 V, 0.6 V, and 0.4 V, respectively). Accordingly, overall reactor performance was enhanced, including coulombic efficiency, electron recovery efficiency, and energy efficiency. Based on an electrochemical impedance spectra analysis, the structured biofilm under simple nutrient conditions (acetate) showed a lower internal resistance with AHL addition, indicating that the microbial communities were altered to enhance electron transfer between the biofilm and electrode. The change in the cathodic microbial structure with more electrochemically active bacteria and fewer hydrogen scavengers could contribute to a higher electron recovery and hydrogen yield with AHL addition. The regulation of the microbial community structure by AHLs represents a potential strategy to enhance electron transfer and hydrogen generation in

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

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.

Free-living spirochetes from Cape Cod microbial mats detected by electron microscopy

Science.gov (United States)

Teal, T. H.; Chapman, M.; Guillemette, T.; Margulis, L.

1996-01-01

Spirochetes from microbial mats and anaerobic mud samples collected in salt marshes were studied by light microscopy, whole mount and thin section transmission electron microscopy. Enriched in cellobiose-rifampin medium, selective for Spirochaeta bajacaliforniensis, seven distinguishable spirochete morphotypes were observed. Their diameters ranged from 0.17 micron to > 0.45 micron. Six of these morphotypes came from southwest Cape Cod, Massachusetts: five from Microcoleus-dominated mat samples collected at Sippewissett salt marsh and one from anoxic mud collected at School Street salt marsh (on the east side of Eel Pond). The seventh morphotype was enriched from anoxic mud sampled from the north central Cape Cod, at the Sandy Neck salt marsh. Five of these morphotypes are similar or identical to previously described spirochetes (Leptospira, Spirochaeta halophila, Spirochaeta bajacaliforniensis, Spirosymplokos deltaeiberi and Treponema), whereas the other two have unique features that suggest they have not been previously described. One of the morphotypes resembles Spirosymplokos deltaeiberi (the largest free-living spirochete described), in its large variable diameter (0.4-3.0 microns), cytoplasmic granules, and spherical (round) bodies with composite structure. This resemblance permits its tentative identification as a Sippewissett strain of Spirosymplokos deltaeiberi. Microbial mats samples collected in sterile Petri dishes and stored dry for more than four years yielded many organisms upon rewetting, including small unidentified spirochetes in at least 4 out of 100 enrichments.

Effects of gamma and electron beam irradiation on the microbial quality of steamed tofu rolls

International Nuclear Information System (INIS)

Jia, Qian; Gao, Meixu; Li, Shurong; Wang, Zhidong

2013-01-01

The effectiveness of two kinds of radiation processing, gamma and electron beam (ebeam) irradiation, for the inactivation of Staphylococcus aureus, Salmonella enteritidis and Listeria innocua which were inoculated in pre-sterilised steamed tofu rolls was studied. The corresponding effects of both irradiation types on total bacterial counts (TBCs) in commercial steamed tofu rolls available in the market were also examined. The microbiological results demonstrated that gamma irradiation yielded D 10 values of 0.20, 0.24 and 0.22 kGy for S. aureus, S. enteritidis and L. innocua, respectively. The respective D 10 values for ebeam irradiation were 0.31, 0.35 and 0.27 kGy. Gamma and ebeam irradiation yielded D 10 values of 0.48 and 0.43 kGy for total bacterial counts in commercial steamed tofu rolls, respectively. The results suggest that ebeam irradiation has similar effect on decreasing TBCs in steamed tofu rolls, and gamma irradiation is slightly more effective than ebeam irradiation in reducing the populations of pathogenic bacteria. The observed differences in D 10 -values between them might be due to the significant differences in dose rate applied, and radiation processing of soybean products to improve their microbial quality could be available for other sources of protein. - Highlights: ► Our research material is steamed tofu rolls, a kind of soybean products. ► We compared the effects of gamma ray and electron beam irradiation. ► Total bacterial and three strains of pathogens are studied in our research. ► We reported electron beam has similar decontamination effect as gamma ray. ► Radiation processing of soybean products to improve their microbial quality could be available for other sources of protein.

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.

Establishment and metabolic analysis of a model microbial community for understanding trophic and electron accepting interactions of subsurface anaerobic environments

Directory of Open Access Journals (Sweden)

Yang Zamin K

2010-05-01

Full Text Available Abstract Background Communities of microorganisms control the rates of key biogeochemical cycles, and are important for biotechnology, bioremediation, and industrial microbiological processes. For this reason, we constructed a model microbial community comprised of three species dependent on trophic interactions. The three species microbial community was comprised of Clostridium cellulolyticum, Desulfovibrio vulgaris Hildenborough, and Geobacter sulfurreducens and was grown under continuous culture conditions. Cellobiose served as the carbon and energy source for C. cellulolyticum, whereas D. vulgaris and G. sulfurreducens derived carbon and energy from the metabolic products of cellobiose fermentation and were provided with sulfate and fumarate respectively as electron acceptors. Results qPCR monitoring of the culture revealed C. cellulolyticum to be dominant as expected and confirmed the presence of D. vulgaris and G. sulfurreducens. Proposed metabolic modeling of carbon and electron flow of the three-species community indicated that the growth of C. cellulolyticum and D. vulgaris were electron donor limited whereas G. sulfurreducens was electron acceptor limited. Conclusions The results demonstrate that C. cellulolyticum, D. vulgaris, and G. sulfurreducens can be grown in coculture in a continuous culture system in which D. vulgaris and G. sulfurreducens are dependent upon the metabolic byproducts of C. cellulolyticum for nutrients. This represents a step towards developing a tractable model ecosystem comprised of members representing the functional groups of a trophic network.

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 ...

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.

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.

MbT-Tool: An open-access tool based on Thermodynamic Electron Equivalents Model to obtain microbial-metabolic reactions to be used in biotechnological process.

Science.gov (United States)

Araujo, Pablo Granda; Gras, Anna; Ginovart, Marta

2016-01-01

Modelling cellular metabolism is a strategic factor in investigating microbial behaviour and interactions, especially for bio-technological processes. A key factor for modelling microbial activity is the calculation of nutrient amounts and products generated as a result of the microbial metabolism. Representing metabolic pathways through balanced reactions is a complex and time-consuming task for biologists, ecologists, modellers and engineers. A new computational tool to represent microbial pathways through microbial metabolic reactions (MMRs) using the approach of the Thermodynamic Electron Equivalents Model has been designed and implemented in the open-access framework NetLogo. This computational tool, called MbT-Tool (Metabolism based on Thermodynamics) can write MMRs for different microbial functional groups, such as aerobic heterotrophs, nitrifiers, denitrifiers, methanogens, sulphate reducers, sulphide oxidizers and fermenters. The MbT-Tool's code contains eighteen organic and twenty inorganic reduction-half-reactions, four N-sources (NH4 (+), NO3 (-), NO2 (-), N2) to biomass synthesis and twenty-four microbial empirical formulas, one of which can be determined by the user (CnHaObNc). MbT-Tool is an open-source program capable of writing MMRs based on thermodynamic concepts, which are applicable in a wide range of academic research interested in designing, optimizing and modelling microbial activity without any extensive chemical, microbiological and programing experience.

Extracellular enzymes facilitate electron uptake in biocorrosion and bioelectrosynthesis.

Science.gov (United States)

Deutzmann, Jörg S; Sahin, Merve; Spormann, Alfred M

2015-04-21

Direct, mediator-free transfer of electrons between a microbial cell and a solid phase in its surrounding environment has been suggested to be a widespread and ecologically significant process. The high rates of microbial electron uptake observed during microbially influenced corrosion of iron [Fe(0)] and during microbial electrosynthesis have been considered support for a direct electron uptake in these microbial processes. However, the underlying molecular mechanisms of direct electron uptake are unknown. We investigated the electron uptake characteristics of the Fe(0)-corroding and electromethanogenic archaeon Methanococcus maripaludis and discovered that free, surface-associated redox enzymes, such as hydrogenases and presumably formate dehydrogenases, are sufficient to mediate an apparent direct electron uptake. In genetic and biochemical experiments, we showed that these enzymes, which are released from cells during routine culturing, catalyze the formation of H2 or formate when sorbed to an appropriate redox-active surface. These low-molecular-weight products are rapidly consumed by M. maripaludis cells when present, thereby preventing their accumulation to any appreciable or even detectable level. Rates of H2 and formate formation by cell-free spent culture medium were sufficient to explain the observed rates of methane formation from Fe(0) and cathode-derived electrons by wild-type M. maripaludis as well as by a mutant strain carrying deletions in all catabolic hydrogenases. Our data collectively show that cell-derived free enzymes can mimic direct extracellular electron transfer during Fe(0) corrosion and microbial electrosynthesis and may represent an ecologically important but so far overlooked mechanism in biological electron transfer. The intriguing trait of some microbial organisms to engage in direct electron transfer is thought to be widespread in nature. Consequently, direct uptake of electrons into microbial cells from solid surfaces is assumed

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.

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

Directory of Open Access Journals (Sweden)

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

Effects of combination of ethylenediaminetetraacetic acid and ...

African Journals Online (AJOL)

Administrator

2010-12-27

Dec 27, 2010 ... Key words: Ethylene diamine tetraacetic acid, microbial phytase, zinc, copper, manganese. .... At day 44, chrome oxide (Cr2O3) was added to all diets at 0.1% level as a .... The hydrolysis of phytate phosphorus by chicks and.

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

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.

Effect of anode polarization on biofilm formation and electron transfer in Shewanella oneidensis/graphite felt microbial fuel cells.

Science.gov (United States)

Pinto, David; Coradin, Thibaud; Laberty-Robert, Christel

2018-04-01

In microbial fuel cells, electricity generation is assumed by bacterial degradation of low-grade organics generating electrons that are transferred to an electrode. The nature and efficiency of the electron transfer from the bacteria to the electrodes are determined by several chemical, physical and biological parameters. Specifically, the application of a specific potential at the bioanode has been shown to stimulate the formation of an electro-active biofilm, but the underlying mechanisms remain poorly understood. In this study, we have investigated the effect of an applied potential on the formation and electroactivity of biofilms established by Shewanella oneidensis bacteria on graphite felt electrodes in single- and double-chamber reactor configurations in oxic conditions. Using amperometry, cyclic voltammetry, and OCP/Power/Polarization curves techniques, we showed that a potential ranging between -0.3V and +0.5V (vs. Ag/AgCl/KCl sat.) and its converse application to a couple of electrodes leads to different electrochemical behaviors, anodic currents and biofilm architectures. For example, when the bacteria were confined in the anodic compartment of a double-chamber cell, a negative applied potential (-0.3V) at the bioanode favors a mediated electron transfer correlated with the progressive formation of a biofilm that fills the felt porosity and bridges the graphite fibers. In contrast, a positive applied potential (+0.3V) at the bioanode stimulates a direct electron transfer resulting in the fast-bacterial colonization of the fibers only. These results provide significant insight for the understanding of the complex bacteria-electrode interactions in microbial fuel cells. Copyright © 2017 Elsevier B.V. All rights reserved.

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

Directory of Open Access Journals (Sweden)

Valéria Vânia Rodrigues

2011-02-01

Full Text Available Two experiments were conducted to evaluate the performance, the energy and nutrient balance and economic viability of production of growing pigs fed diets with reduced levels and phytase. In experiment 1, 60 barrows and gilts (initial weight 25 ± 1.72 kg were housed in a randomized block design with three diets, ten replicates and two animals per experimental unit (one male and one female. One of the diets was formulated with a reduced level with phytase and evaluated in comparison to a positive-control diet that was formulated according to the nutritional recommendations for animals of this breed, and a negative-control diet without phytase. The different diets did not affect feed intake, but the diet with phytase promoted better final weight, daily weight gain and feed conversion, although it did not differ from the positive control diet. In experiment 2, 27 barrows (initial weight 49 ± 3.67 kg were housed in random blocks with three diets and nine replicates with experimental plot represented by one animal. The diets did not affect the balance of energy and nutrients but had a significant effect on the efficiency of utilization of calcium and phosphorus, which was the lowest in animals that received the positive-control diet. Economic viability improved when nutrients were reduced and phytase was added in the ration. The addition of phytase (500 ftu/kg in rations formulated with nutritional levels reduced for growing swines improves rearing productivity and it is economical viable.Dois experimentos foram conduzidos para avaliar o desempenho, o balanço de energia e nutrientes e a viabilidade econômica da produção de suínos em crescimento alimentados com rações com níveis reduzidos e fitase. No experimento 1, 60 suínos machos castrados e fêmeas (peso inicial de 25 ± 1,72 kg foram alojados em delineamento de blocos casualizados, com três dietas, dez repetições e dois animais por unidade experimental (macho e fêmea. Uma das

MbT-Tool: An open-access tool based on Thermodynamic Electron Equivalents Model to obtain microbial-metabolic reactions to be used in biotechnological process

Directory of Open Access Journals (Sweden)

Pablo Araujo Granda

2016-01-01

Full Text Available Modelling cellular metabolism is a strategic factor in investigating microbial behaviour and interactions, especially for bio-technological processes. A key factor for modelling microbial activity is the calculation of nutrient amounts and products generated as a result of the microbial metabolism. Representing metabolic pathways through balanced reactions is a complex and time-consuming task for biologists, ecologists, modellers and engineers. A new computational tool to represent microbial pathways through microbial metabolic reactions (MMRs using the approach of the Thermodynamic Electron Equivalents Model has been designed and implemented in the open-access framework NetLogo. This computational tool, called MbT-Tool (Metabolism based on Thermodynamics can write MMRs for different microbial functional groups, such as aerobic heterotrophs, nitrifiers, denitrifiers, methanogens, sulphate reducers, sulphide oxidizers and fermenters. The MbT-Tool's code contains eighteen organic and twenty inorganic reduction-half-reactions, four N-sources (NH4+, NO3−, NO2−, N2 to biomass synthesis and twenty-four microbial empirical formulas, one of which can be determined by the user (CnHaObNc. MbT-Tool is an open-source program capable of writing MMRs based on thermodynamic concepts, which are applicable in a wide range of academic research interested in designing, optimizing and modelling microbial activity without any extensive chemical, microbiological and programing experience.

Batteryless, wireless sensor powered by a sediment microbial fuel cell.

Science.gov (United States)

Donovan, Conrad; Dewan, Alim; Heo, Deukhyoun; Beyenal, Haluk

2008-11-15

Sediment microbial fuel cells (SMFCs) are considered to be an alternative renewable power source for remote monitoring. There are two main challenges to using SMFCs as power sources: 1) a SMFC produces a low potential at which most sensor electronics do not operate, and 2) a SMFC cannot provide continuous power, so energy from the SMFC must be stored and then used to repower sensor electronics intermittently. In this study, we developed a SMFC and a power management system (PMS) to power a batteryless, wireless sensor. A SMFC operating with a microbial anode and cathode, located in the Palouse River, Pullman, Washington, U.S.A., was used to demonstrate the utility of the developed system. The designed PMS stored microbial energy and then started powering the wireless sensor when the SMFC potential reached 320 mV. It continued powering until the SMFC potential dropped below 52 mV. The system was repowered when the SMFC potential increased to 320 mV, and this repowering continued as long as microbial reactions continued. We demonstrated that a microbial fuel cell with a microbial anode and cathode can be used as an effective renewable power source for remote monitoring using custom-designed electronics.

Extracellular Electron Uptake: Among Autotrophs and Mediated by Surfaces

DEFF Research Database (Denmark)

Tremblay, Pier-Luc; Angenent, Largus T.; Zhang, Tian

2017-01-01

Autotrophic microbes can acquire electrons from solid donors such as steel, other microbial cells, or electrodes. Based on this feature, bioprocesses are being developed for the microbial electrosynthesis (MES) of useful products from the greenhouse gas CO2. Extracellular electron-transfer mechan......Autotrophic microbes can acquire electrons from solid donors such as steel, other microbial cells, or electrodes. Based on this feature, bioprocesses are being developed for the microbial electrosynthesis (MES) of useful products from the greenhouse gas CO2. Extracellular electron......; or (iii) mediator-generating enzymes detached from cells. This review explores the interactions of autotrophs with solid electron donors and their importance in nature and for biosustainable technologies....

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

Competitive microbial reduction of perchlorate and nitrate with a cathode directly serving as the electron donor

International Nuclear Information System (INIS)

Xie, Daohai; Yu, Hui; Li, Chenchen; Ren, Yuan; Wei, Chaohai; Feng, Chunhua

2014-01-01

Microbial reduction of perchlorate with an electrode as the electron donor represents an emerging technology for remediation of perchlorate contamination; it is important to know how perchlorate reduction behaves when nitrate, a co-contaminant of perchlorate is present. We reported that electrons derived from the electrode can be directly transferred to the bacteria with perchlorate or nitrate as the sole electron acceptor. The presence of nitrate, even at the 0.07 mM level, can slow reduction of perchlorate (0.70 mM) as a poised potential of -0.50 V (vs. SCE) was applied to the inoculated cathode. Increasing the concentration of nitrate resulted in a noticeable inhibitory effect on perchlorate reduction. When the nitrate concentration was 2.10 mM, reduction of 0.70 mM perchlorate was totally inhibited. Bacterial community analyses based on 16S rDNA gene analysis with denaturing gradient gel electrophoresis (DGGE) revealed that most of the bacteria newly enriched on the nitrate and/or perchlorate biocathodes were the known electrochemically active denitrifiers, which possibly prefer to reduce nitrate over perchlorate. These results show that nitrate is a more favorable electron acceptor than perchlorate in the bioelectrochemical system where the cathode directly serves as the electron donor

Microbial quality evaluation and effective decontamination of nutraceutically valued lotus seeds by electron beams and gamma irradiation

International Nuclear Information System (INIS)

Bhat, Rajeev; Sridhar, K.R.; Karim, A.A.

2010-01-01

Lotus seeds are nutraceutically valued natural plant produce, which succumbs to microbial contamination, predominantly to toxigenic moulds. Results of the present study revealed seed coat portion to harbor higher proportion of microbial load, particularly fungi than cotyledon portion. Among the mycotoxins analyzed, aflatoxins (B 1 , B 2 , G 1 and G 2 ) were below detectable limits, while the seeds were devoid of Ochratoxin-A (OTA). Application of different doses of electron beam and gamma irradiation (0, 2.5, 5, 7.5, 10, 15 and 30 kGy) for decontamination purpose revealed significant dose-dependent decrease in the fungal contaminants (P<0.05). However, the contaminant yeasts could survive up to 10 kGy dose, which could be completely eliminated at 15 kGy. From the results obtained, a dose range between 10 and 15 kGy is recommended for complete decontamination, as these doses have also been shown earlier to have minimal effects on nutritional and functional properties of lotus seeds.

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...

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

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.

Studies about behavior of microbial degradation of organic compounds

International Nuclear Information System (INIS)

Ohtsuka, Makiko

2003-02-01

Some of TRU waste include organic compounds, thus these organic compounds might be nutrients for microbial growth at disposal site. This disposal system might be exposed to high alkali condition by cement compounds as engineering barrier material. In the former experimental studies, it has been supposed that microbial exist under pH = 12 and the microbial activity acclimated to high alkali condition are able to degrade asphalt under anaerobic condition. Microbes are called extremophile that exist in cruel habitat as high alkali or reductive condition. We know less information about the activity of extremophile, though any recent studies reveal them. In this study, the first investigation is metabolic pathway as microbial activity, the second is microbial degradation of aromatic compounds in anaerobic condition, and the third is microbial activity under high alkali. Microbial metabolic pathway consist of two systems that fulfill their function each other. One system is to generate energy for microbial activities and the other is to convert substances for syntheses of organisms' structure materials. As these systems are based on redox reaction between substances, it is made chart of the microbial activity region using pH, Eh, and depth as parameter, There is much report that microbe is able to degrade aromatic compounds under aerobic or molecular O 2 utilizing condition. For degradation of aromatic compounds in anaerobic condition, supplying electron acceptor is required. Co-metabolism and microbial consortia has important role, too. Alcalophile has individual transporting system depending Na + and acidic compounds contained in cell wall. Generating energy is key for survival and growth under high alkali condition. Co-metabolism and microbial consortia are effective for microbial degradation of aromatic compounds under high alkali and reductive condition, and utilizable electron acceptor and degradable organic compounds are required for keeping microbial activity and

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...

Effects of combination of ethylenediaminetetraacetic acid and ...

African Journals Online (AJOL)

This experiment was conducted to evaluate the combined effects of ethylene diamine tetraacetic acid (EDTA) and microbial phytase (MP) on the serum concentration and digestibility of some minerals in broiler chicks. This experiment was conducted using 360 Ross-308 male broiler chicks in a completely randomized ...

Utilization of subsurface microbial electrochemical systems to elucidate the mechanisms of competition between methanogenesis and microbial iron(III)/humic acid reduction in Arctic peat soils

Science.gov (United States)

Friedman, E. S.; Miller, K.; Lipson, D.; Angenent, L. T.

2012-12-01

High-latitude peat soils are a major carbon reservoir, and there is growing concern that previously dormant carbon from this reservoir could be released to the atmosphere as a result of continued climate change. Microbial processes, such as methanogenesis and carbon dioxide production via iron(III) or humic acid reduction, are at the heart of the carbon cycle in Arctic peat soils [1]. A deeper understanding of the factors governing microbial dominance in these soils is crucial for predicting the effects of continued climate change. In previous years, we have demonstrated the viability of a potentiostatically-controlled subsurface microbial electrochemical system-based biosensor that measures microbial respiration via exocellular electron transfer [2]. This system utilizes a graphite working electrode poised at 0.1 V NHE to mimic ferric iron and humic acid compounds. Microbes that would normally utilize these compounds as electron acceptors donate electrons to the electrode instead. The resulting current is a measure of microbial respiration with the electrode and is recorded with respect to time. Here, we examine the mechanistic relationship between methanogenesis and iron(III)- or humic acid-reduction by using these same microbial-three electrode systems to provide an inexhaustible source of alternate electron acceptor to microbes in these soils. Chamber-based carbon dioxide and methane fluxes were measured from soil collars with and without microbial three-electrode systems over a period of four weeks. In addition, in some collars we simulated increased fermentation by applying acetate treatments to understand possible effects of continued climate change on microbial processes in these carbon-rich soils. The results from this work aim to increase our fundamental understanding of competition between electron acceptors, and will provide valuable data for climate modeling scenarios. 1. Lipson, D.A., et al., Reduction of iron (III) and humic substances plays a major

Diversity and function of the microbial community on anodes of sediment microbial fuel cells fueled by root exudates

Energy Technology Data Exchange (ETDEWEB)

Cabezas da Rosa, Angela

2010-11-26

Anode microbial communities are essential for current production in microbial fuel cells. Anode reducing bacteria are capable of using the anode as final electron acceptor in their respiratory chain. The electrons delivered to the anode travel through a circuit to the cathode where they reduce oxygen to water generating an electric current. A novel type of sediment microbial fuel cell (SMFC) harvest energy from photosynthetically derived compounds released through the roots. Nothing is known about anode microbial communities of this type of microbial fuel cell. This work consists of three parts. The first part focuses on the study of bacterial and archaeal community compositions on anodes of SMFCs fueled by rice root exudates. By using terminal restriction fragment length polymorphism (T-RFLP), a profiling technique, and cloning / sequencing of 16S rRNA, we determined that the support type used for the plant (vermiculite, potting soil or rice field soil) is an important factor determining the composition of the microbial community. Finally, by comparing microbial communities of current producing anodes and non-current producing controls we determined that Desulfobulbus- and Geobacter-related populations were probably most important for current production in potting soil and rice field soil SMFCs, respectively. However, {delta}-proteobacterial Anaeromyxobacter spp., unclassified {delta}-proteobacteria and Anaerolineae were also part of the anode biofilm in rice field soil SMFCs and these populations might also play a role in current production. Moreover, distinct clusters of Geobacter and Anaeromyxobacter populations were stimulated by rice root exudates. Regarding Archaea, uncultured Euryarchaea were abundant on anodes of potting soil SMFCs indicating a potential role in current production. In both, rice field soil and potting soil SMFCs, a decrease of Methanosaeta, an acetotrophic methanogen, was detected on current producing anodes. In the second part we focused

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

Directory of Open Access Journals (Sweden)

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.

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

Microbial ecology to manage processes in environmental biotechnology.

Science.gov (United States)

Rittmann, Bruce E

2006-06-01

Microbial ecology and environmental biotechnology are inherently tied to each other. The concepts and tools of microbial ecology are the basis for managing processes in environmental biotechnology; and these processes provide interesting ecosystems to advance the concepts and tools of microbial ecology. Revolutionary advancements in molecular tools to understand the structure and function of microbial communities are bolstering the power of microbial ecology. A push from advances in modern materials along with a pull from a societal need to become more sustainable is enabling environmental biotechnology to create novel processes. How do these two fields work together? Five principles illuminate the way: (i) aim for big benefits; (ii) develop and apply more powerful tools to understand microbial communities; (iii) follow the electrons; (iv) retain slow-growing biomass; and (v) integrate, integrate, integrate.

Impact of the electron donor on in situ microbial nitrate reduction in Opalinus Clay: results from the Mont Terri rock laboratory (Switzerland)

Energy Technology Data Exchange (ETDEWEB)

Bleyen, N.; Smets, S. [Belgian Nuclear Research Centre SCK-CEN, Mol (Belgium); Small, J. [National Nuclear Laboratory NLL, Warrington (United Kingdom); and others

2017-04-15

At the Mont Terri rock laboratory (Switzerland), an in situ experiment is being carried out to examine the fate of nitrate leaching from nitrate-containing bituminized radioactive waste, in a clay host rock for geological disposal. Such a release of nitrate may cause a geochemical perturbation of the clay, possibly affecting some of the favorable characteristics of the host rock. In this in situ experiment, combined transport and reactivity of nitrate is studied inside anoxic and water-saturated chambers in a borehole in the Opalinus Clay. Continuous circulation of the solution from the borehole to the surface equipment allows a regular sampling and online monitoring of its chemical composition. In this paper, in situ microbial nitrate reduction in the Opalinus Clay is discussed, in the presence or absence of additional electron donors relevant for the disposal concept and likely to be released from nitrate-containing bituminized radioactive waste: acetate (simulating bitumen degradation products) and H{sub 2} (originating from radiolysis and corrosion in the repository). The results of these tests indicate that - in case microorganisms would be active in the repository or the surrounding clay - microbial nitrate reduction can occur using electron donors naturally present in the clay (e.g. pyrite, dissolved organic matter). Nevertheless, non-reactive transport of nitrate in the clay is expected to be the main process. In contrast, when easily oxidizable electron donors would be available (e.g. acetate and H{sub 2}), the microbial activity will be strongly stimulated. Both in the presence of H{sub 2} and acetate, nitrite and nitrogenous gases are predominantly produced, although some ammonium can also be formed when H{sub 2} is present. The reduction of nitrate in the clay could have an impact on the redox conditions in the pore-water and might also lead to a gas-related perturbation of the host rock, depending on the electron donor used during denitrification

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

Directory of Open Access Journals (Sweden)

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.

Microbial Energy Conversion

Energy Technology Data Exchange (ETDEWEB)

Buckley, Merry [American Society for Microbiology (ASM), Washington, DC (United States); Wall, Judy D. [Univ. of Missouri, Columbia, MO (United States)

2006-10-01

natural gas from the subsurface. The participants discussed--key microbial conversion paths; overarching research issues; current funding models and microbial energy research; education, training, interdisciplinary cooperation and communication. Their recommendations are--Cellulose and lignocellulose are the preferred substrates for producing liquid transportation fuels, of which ethanol is the most commonly considered example. Generating fuels from these materials is still difficult and costly. A number of challenges need to be met in order to make the conversion of cellulose and lignocellulose to transportation fuels more cost-competitive. The design of hydrogen-producing bioreactors must be improved in order to more effectively manage hydrogen removal, oxygen exclusion, and, in the case of photobioreactors, to capture light energy more efficiently. Methane production may be optimized by fine-tuning methanogenic microbial communities. The ability to transfer electrons to an anode in a microbial fuel cell is probably very broadly distributed in the bacterial world. The scientific community needs a larger inventory of cultivated microorganisms from which to draw for energy conversion development. New and unusual organisms for manufacturing fuels and for use in fuel cells can be discovered using bioprospecting techniques. Particular emphasis should be placed on finding microbes, microbial communities, and enzymes that can enhance the conversion of lignocellulosic biomass to usable sugars. Many of the microbial processes critical to energy conversion are carried out by complex communities of organisms, and there is a need to better understand the community interactions that make these transformations possible. Better understanding of microbial community structure, robustness, networks, homeostasis, and cell-to-cell signaling is also needed. A better understanding of the basic enzymology of microorganisms is needed in order to move forward more quickly with microbial energy

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.

Interconnection of Key Microbial Functional Genes for Enhanced Benzo[a]pyrene Biodegradation in Sediments by Microbial Electrochemistry.

Science.gov (United States)

Yan, Zaisheng; He, Yuhong; Cai, Haiyuan; Van Nostrand, Joy D; He, Zhili; Zhou, Jizhong; Krumholz, Lee R; Jiang, He-Long

2017-08-01