• Journal of Microbiology and Biotechnology
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• v.9 no.2
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• pp.223-226
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• 1999

A bacterial strain producing high level of an extracellular phytase was isolated from cooked rice and identified as a strain of Bacillus sp. and designated as Bacillus sp. KHU-10. Optimum culture conditions were investigated for the maximum productivity of phytase by Bacillus sp. KHU-10. 1.0% Maltose and 1.0% peptone with 0.5% beef extract were the best carbon source and nitrogen source, respectively. The addition of $CaCl_2$, stimulated the enzyme productivity with concentration between 0.01% and 0.2%, in the medium. Although sodium phosphate increased the cell mass, the enzyme activity decreased. Calcium phytate and wheat bran containing phytate did not enhance the enzyme production. Under the optimum medium, the production of the phytase reached the highest level of 0.2 unit/ml after 4 days of incubation.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.10
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• pp.1479-1487
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• 2015

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 ${\geq}750FTU$ resulted in AID of total AA commensurate to that of PC fed birds and at ${\geq}1,000FTU$ 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.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.6
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• pp.576-581
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• 2005

Phytase improves the bioavailability of phytate phosphorus in plant foods to humans and animals, and reduces the phosphorus pollution of animal waste. We have engineered the cell surface of the yeast. Saccharomyces cerevisiae, by anchoring active fungal phytase on its cell wall, in order to apply it as a dietary supplement containing bioconversional functions in animal foods and a whole cell bio-catalyst for the treatment of waste. The phytase gene (phyA) of Aspergillus niger with a signal peptide of rice amylase 1A (Ramy1A) was fused with the gene encoding the C-terminal half (320 amino acid residues from the C-terminus) of yeast ${\alpha}-agglutinin$, a protein which is involved in mating and is covalently anchored to the cell wall. The resulting fusion construct was introduced into S. cerevisiae and expressed under the control of the constitutive glyceraldehydes-3-phosphate dehydrogenase (GPD) promoter. Phytase plate assay revealed that the surface-engineered cell exhibited a catalytically active opaque zone which was restricted to the margin of the colony. Additionally, the phytase activity was detected in the cell fraction, but was not detected in the culture medium when it was grown in liquid. These results indicate that the phytase was successfully anchored to the cell surface of yeast and was displayed as its active form. The amount of recombinant phytase on the surface of yeast cells was estimated to be 16,000 molecules per cell.

• Korean Journal of Poultry Science
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• v.36 no.4
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• pp.351-356
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• 2009

This work was carried out to investigate the effects of CDDGS (corn distillers dried grains with solubles) colors and phytase levels on the amino acid digestibility of broilers for 7 days. One hundred eight Ross broilers were used in 9 treatment groups with a $3\;{\times}\;3$ factorial design as three colors of DDGSs (DDGS1, DDGS2, and DDGS3) in combination with phytase (0, 500, and 1,000 FTU/kg). The $L^*$ values of DDGS1, DDGS2, and DDGS3 were 55.2, 39.8, and 28.3; the $a^*$ values, 8.8, 7.5, and 6.1; and the $b^*$ values, 41.3, 27.1, and 15.4, respectively. The ileal digestibility of amino acids was decreased as DDGS color was dark, and increased as additive phytase level was increased in the diet. Average digestibilities of essential amino acids were 67.6~80.0%, and those of nonessential amino acids 67.4~77.3. Amino acid digestibility was low in the dark DDGS with no phytase in the diet.

• Journal of Microbiology and Biotechnology
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• v.20 no.11
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• pp.1491-1499
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• 2010

Rhizosphere microorganisms possessing phytase activity are considered important for rendering phytate-phosphorus (P) available to plants. In the present study, the Citrobacter braakii phytase gene (appA) was overexpressed in rhizobacteria possessing plant growth promoting (PGP) traits, for increasing their potential as bioinoculants. AppA was cloned under the lac promoter in the broadhost-range expression vector pBBR1MCS-2. Transformation of the recombinant construct pCBappA resulted in high constitutive phytase activity in all of the eight rhizobacterial strains belonging to genera Pantoea, Citrobacter, Enterobacter, Pseudomonas (two strains), Rhizobium (two strains), and Ensifer that were studied. Transgenic rhizobacterial strains were found to display varying levels of phytase activity, ranging from 10-folds to 538-folds higher than the corresponding control strains. The transgenic derivative of Pseudomonas fluorescens CHA0, a well-characterized plant growth promoting rhizobacterium, showed the highest expression of phytase (~8 U/mg) activity in crude extracts. Although all transformants showed high phytase activity, rhizobacteria having the ability to secrete organic acid showed significantly higher release of P from Ca-phytate in buffered minimal media. AppA overexpressing rhizobacteria showed increased P content, and dry weight (shoot) or shoot/ root ratio of mung bean (Vigna radiata) plants, to different extents, when grown in semisolid agar (SSA) medium containing Na-phytate or Ca-phytate as the P sources. This is the first report of the overexpression of phytase in rhizobacterial strains and its exploitation for plant growth enhancement.

• Korean Journal of Microbiology
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• v.40 no.2
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• pp.81-86
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• 2004

Among the annotated ORFs of Streptomyces coelicolor, SCO7697 was supposed to encode for phytase (myo-inositol hexakisphosphate phosphohydrolase). The DNA fragment containing SCO7697 was cloned by the PCR from the chromosomal DNA of S.coelicolor A3(2)M. The cloned fragment was introduced into E. coli expres-sion vector, pET28a(+), to yield two recombinant plasmids, pET28-SP and pET28-LP, which were designed to encode different length of proteins. When the pET28-SP and pET28-LP were introduced into E. coli BL21, the transformants successfully overexpressed recombinant proteins, but the molecular weights of the expressed pro-teins were appeared bigger than those of expected in SDS-polyacrylamide gel electrophoresis. The shift of cul-tural temperature from 37 to $30^{\circ}C$ made most of expressed protein be solubilized. The expressed protein, however, did not show any phytase activity. When the DNA fragment with its own promoter placed on the E. coli-Streptomyces vector, pWHM3, and introduced into S. lividans, the phytase activity was not detected either. These results suggest that even though the SCO7697 was annotated as a probable phytase with high probability (E value is $6e^{-89}$), the real product doest not have phytase activity.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.5
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• pp.691-700
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• 2019

Objective: This study was conducted to assess increasing doses of phytase added to broiler diets formulated with different levels of available phosphorus (avP), calcium (Ca), and sodium (Na), and the respective effects on performance parameters, quantitative carcass characteristics, ash and phosphorus deposition in tibia and weight of organs. Methods: Three different matrices were assumed for phytase with the following nutritional values: matrix A (MT A): 0.165% Ca, 0.150% avP, and 0.035% Na; matrix B (MT B): 0.215% Ca, 0.195% avP, and 0.045% Na; matrix C (MT C): 0.245% Ca, 0.225% avP, and 0.053% Na. There were six different diets: No phytase (formulated to meet the nutritional requirements); phytase 500 FTU/kg+MT A; phytase 1,000 FTU/kg+MT A; phytase 1,500 FTU/kg+MT A; phytase 1,000 FTU/kg+MT B and phytase 1,500 FTU/kg+MT C. Results: There was no significant phytase influence on performance, quantitative carcass characteristics, ash and phosphorus deposition in tibia and weight of the organ throughout the study period, however, it was possible to observe a tendency of improvement in body weight corrected feed conversion for broilers fed the phytase 1,500+MT C diet, where potentially these birds were more efficient on utilize phytic phosphorus and other nutrients bounded to phytate molecule, translating into improvement in performance, and there was also a non significant numerical improvement in body weight corrected feed conversion of broilers fed this diet. Conclusion: Broilers fed with diets formulated with different levels of avP, Ca, and Na and increasing doses of phytase have shown no change on performance, quantitative carcass characteristics, ash and phosphorus deposition in tibia and weight of organs.

• Animal Bioscience
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• v.36 no.12
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• pp.1869-1879
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• 2023

Objective: This study was to evaluate the effects of individual or combinational use of phytase, protease, and xylanase on total tract digestibility of corn, soybean meal, and distillers dried grains with soluble (DDGS) fed to pigs. Methods: Each experiment had four 4×4 Latin squares using 16 barrows. Each period had 5-d adaptation and 3-d collection. All experiments had: CON (no enzyme); Phy (CON+phytase); Xyl (CON+xylanase); Pro (CON+protease); Phy+Xyl; Phy+Pro, Xyl+Pro, Phy+Xyl+Pro. Each Latin square had 'CON, Phy, Xyl, and Phy+Xyl'; 'CON, Phy, Pro, and Phy+Pro'; 'CON, Pro, Xyl, and Xyl+Pro'; and 'Phy+Xyl, Phy+Pro, Xyl+Pro, Phy+Xyl+Pro'. Results: The digestible energy (DE), metabolizable energy (ME), and nitrogen retention (NR) of corn were not affected by enzymes but the apparent total tract digestibility (ATTD) of phosphorus (P) was improved (p<0.01) by Phy. The DE and ATTD dry matter (DM) in soybean meal were increased (p<0.05) by Phy+Pro and the ATTD P was improved (p<0.01) by Phy, Phy+Pro, and Phy+Xyl. The DE, ME, and ATTD DM in DDGS were improved (p<0.05) by Phy+Xyl and the ATTD P was improved (p<0.01) by Phy, Phy+Pro, and Phy+Xyl. Conclusion: Phytase individually or in combination with xylanase and protease improved the Ca and P digestibility of corn, soybean meal, and DDGS, from the hydrolysis of phytic acid. The supplementation of protease was more effective when combined with phytase and xylanase in the soybean meal and DDGS possibly due to a higher protein content in these feedstuffs. Xylanase was more effective in DDGS diets due to the elevated levels of non-starch polysaccharides in these feedstuffs. However, when xylanase was combined with phytase, it demonstrated a higher efficacy improving the nutrient digestibility of pigs. Overall, combinational uses of feed enzymes can be more efficient for nutrient utilization in soybean meal and DDGS than single enzymes.

• Microbiology and Biotechnology Letters
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• v.10 no.2
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• pp.133-144
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• 1982

The distribution of acid phosphatase activity was investigated with 141 microorganisms from the type culture collection of Chong Kun Dang laboratory and the 41 strains isolated from natural sources. The phytase activity was detected mainly with fungal strains. A fungus isolated from soil and identified as Aspergillus niger had shown the highest phytase activity. The environmental conditions for the enzyme formation by the isolate and some properties of the enzyme were also studied. The results obtained were as follows: (1) The highest phytase production was observed when the fungus was cultivated at 28$^{\circ}C$ for 5 days in the corn starch based medium using the cells incubated at 34$^{\circ}C$ for 3 days as a seed. (2) The optimal initial pH of the culture medium was found to around 2 for the formation of phytase. (3) Sucrose was proved to be one of the most effective carbon sources tested for the enzyme production. (4) As an inorganic nitrogen source, potassium nitrate was found to give a good result in the production of phytase. (5) Synthesis of phytase was significantly increased by the supplement with 0.2 % corn steep liquor to the basal medium as an organic nitrogen source. (6) At the concentration of 40-80 mg inorganic phosphate per liter of the culture medium, the enzyme formation revealed the highest level. But as the phosphate was increased above this optimum concentration the phytase activity was drastically decreased although the cell density showed to be still increasing

• Asian-Australasian Journal of Animal Sciences
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• v.12 no.2
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• pp.305-327
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• 1999

In the paper insight is given in the legislation policy to restrain environmental pollution by pig husbandry, focused on The Netherlands (Mineral Accounting System). Besides, nutritional measures are presented to reduce environmental pollution by lowering excretion of N and P, emphasizing (multi) phase feeding, the use of low protein, synthetic amino acids supplemented diets, phytase and its effect on phosphorus and calcium digestibility, its interaction with phytic acid and proteins, and the environmental impact of the use of phytase in pig diets. Also, nutritional means are indicated to reduce ammonia volatilization from pig operations. It is concluded that nutrition management can substantially contribute to reduction of N and P excretion by pigs, mainly by lowering dietary protein levels, (multi) phase feeding and the use of microbial phytase, and that the use of phytase on a large scale in The Netherlands has a tremendous environmental impact. In 20 years the excretion of P in growing-finishing pigs has more than halved. Ammonia emission from manure of pigs can be reduced substantially by lowering dietary protein content, but also by including additional non-starch polysaccharides in the diet. A very promising method to reduce ammonia emission is to manipulate dietary cation-anion difference, e.g. by adding acidifying salts to the diet, which will lower pH of urine substantially. Further research is desirable. This also applies to determining dietary factors influencing the odour release from manure. Finally, some speculation on the future of pig farming from an environmental viewpoint is presented.