• Asian-Australasian Journal of Animal Sciences
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• 제16권1호
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• pp.124-135
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• 2003

In order to prevent pollution from animal waste, the excretion of nutrients should be reduced through proper nutritional management. Among the many nutrients of concern, such as N, P, Cu, Zn and K, P is one of the most concerned nutrients to be managed. Seven feeding trials, three with layers and four with broilers, were conducted to determine if microbial phytase supplementation can reduce non-phytate phosphorus (NPP) level in diets and results in concomitant reductions of P excretion. The results showed that microbial phytase can be successfully used to achieve these purposes. Activity of natural phytase in certain plant feedstuffs is high enough to be considered in feed formulation. Three experiments have been conducted to study the characteristics of plant phytase and its application to feeding of broilers. Selected brands of wheat bran could be successfully used as a source of phytase in broiler feeding.

• Journal of Animal Science and Technology
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• 제44권4호
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• pp.407-418
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• 2002

Phytase 공급원으로 밀과 밀기울의 첨가가 육계 생산성 및 P이용률에 미치는 영향을 측정하기 위해 5주간의 사양 실험을 실시하였다. 갓 부화한 1,000수의 병아리(Ross$^{(R)}$)를 20pen에 50수씩(암수 각 25수) 공시하여 5처리 4반복으로 완전임의 배치하였다. T1은 대조구로 정상수준의 nonphytate-P(NPP)를 함유하였고, T2는 T1 －0.1% NPP의 低 NPP구, T3는 T2 + 사료 kg당 600IU microbial phytase(NOVO$^{(R)}$)구, T4는 T2 + 밀과 밀기울로 사료 kg당 600IU plant phytase 공급구, T5는 T2 + 밀과 수침 후 건조 처리한 밀기울로 사료 kg당 600IU plant phytase 공급구 였다. NPP 수준을 0.1% 감소시킨(T2) 처리에서 생산성과 사료효율이 감소되었고 식물성 phytase 처리시(T4와 T5)에서는 생산성이 회복되었다. 식물성 phytase 처리구들(T4와 T5)이 microbial phytase 처리구(T3)보다 더 나은 결과를 보였다. 정상 밀기울구(T4)와 수침 후 건조 처리한 밀기울구(T5)와는 차이를 보이지 않았다. 폐사율은 低  NPP(T2)에서 가장 높았다. 육성 사료에서 조지방과 조회분 이용률에 일반 밀기울 처리구(T4)가 가장 높았다. Ca과 P의 이용률 또한 T4가 가장 높고 T3와 T5가 다음으로 높았다. Phytase 처리구 (T3, T4와 T5) 들은 Mg, Fe와 Zn의 이용률을 크게 증가시켰다. Ca, P, Mg, Fe와 Zn의 배설량은 microbial phytase가 가장 적었다. 혈청내 Ca과 Mg의 함량은 低 NPP에서 가장 높았고, 경골내 조회분 함량은 T2와 T3가 T1, T4와 T5보다 낮았다. 그러나 경골내 Ca 함량은 T1과 T2가 다른 처리구들보다 높았다. 경골내 P과 Mg의 함량은 T1구가 가장 높았다. 결론적으로 밀기울을 통한 식물성 phytase의 공급은 P 이용률 향상에 효과적이었다. 수침 후 건조처리하는 것은 이용률 향상에 도움이 되지 않았다.

• Asian-Australasian Journal of Animal Sciences
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• 제10권6호
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• pp.551-566
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• 1997

Phosphorus has been known as an essential component of animal body. However, the requirement has not been determined precisely because of the variable bioavailabilities of feedstuffs from plant origin. The bioavailability of P in various feedstuffs of plant origin varies from 10 to 60%. Digestibility and availability of the P differed considerably depending on the feed. The lowest values were found for maize (under 20%), the highest for wheat and triticale (over 50%). This is due to the proportion of phytate and the presence of intrinsic phytase. And the digestive tract of monogastric animals does not contain sufficient amounts of phytase, an enzyme that hydrolyses the unavailable phytate complexes to available, inorganic orthophosphates. Microbial phytase supplementation improves the P availability, and both intrinsic plant and microbial phytase improves the availability of P in feedstuffs of plant origin. In a mixture of feeds with low and high activity of intrinsic phytase and/or supplemented by commercial phytase, the P availability is additive. However, in the light of current results it seems that exceeding the P availability equal to 60-70% is unrealizable even at large microbial phytase doses.

• Journal of Microbiology and Biotechnology
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• 제20권11호
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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.

• Journal of Animal Science and Technology
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• 제44권1호
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• pp.55-60
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• 2002

본 연구는 식물성 phytase 처리수준이 산란계의 생산성에 미치는 영향 및 인 이용률에 미치는 영향을 검토하기 위하여 실시하였다. NRC 사양표준의 유효 인 요구량을 충족시킨 대조구와 무기태인 사용량을 50%로 제한한 처리구로 나누고, 각각의 처리구에 식물성 phytase를 각각 0.05%, 0.1% 및 0.2% 첨가하였다. 58주령의 갈색 산란계(ISA-Brown) 240수를 공시하여 총 6주간 사양실험을 실시하였고 산란종계를 이용하여 인 및 질소의 이용성을 측정하였다. 그 결과를 요약하면 다음과 같다. 1. 50% 무기태인 수준에서 식물성 phytase의 첨가에 따른 일당증체량, 산란율의 저하는 나타나지 않았다. 2. 50% 무기태인 수준에서 식물성 phytase 0.05% 첨가수준에서 일시적으로 난각질 저하 현상이 나타났으나 0.1% 이상 첨가수준에서는 난각질의 변화가 관찰되지 않았다. 3. 효소의 첨가수준이 증가함에 따라 P 배설량의 감소와 P 이용률이 개선되었다(P<0.05). 4. 식물성 phytase 첨가는 산란계의 성적 저하없이 사료내 무기태인의 사용량을 줄일 수 있음이 시사되었다.

• Asian-Australasian Journal of Animal Sciences
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• 제27권1호
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• pp.1-9
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• 2014

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.

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권6호
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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.

• Journal of Microbiology and Biotechnology
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• 제11권6호
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• pp.915-921
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• 2001

Phytase improves the bioavailability of phytate phosphorus in plant foods to humans and animals, and reduces the phosphorus pollution of animal waste. In order to express a high level of fungal phytase in Saccharomyces cerevisiae, various expression vectors were constructed with different combinations of promoters, translation enhancers, signal peptides, and terminator. Three different promoters fused to the phytase gene (phyA) from Aspergillus niger were tested: a galactokinase (GAL1) promoter, glyceraldehyde-3-phosphate dehydrogenase (GPD) promoter, and yeast hybrid ADH2-GPD promoter consisting of alcohol dehydrogenase II (ADH2) and a GPD promoter. The signal peptides of phytase, glucose oxidase (GO), and rice amylase 1A(RAmy1A) were included. Plus, the translation enhancers of the ${\Omega}$ sequence and UTR70 from the tobacco mosaic virus (TMV) and spinach, respectively, were also tested. Among the recombinant vectors, pGphyA06 containing the GPD promoter, the ${\Omega}$ sequence, RAmy1A, and GAL7 terminator expressed the highest phytase activity in a culture filtrate, which was estimated at 20 IU/ml. An intracellular localization of the expressed phytase activity in a culture filtrate, which was estimated at 20 IU/ml. An intracellular localization of the expressed phytase was also performed by inserting an endoplasmic reticulum (ER) retention signal, KDEL sequence, into the C-terminus of the phytase within the vector pHphyA-6. It appeared that the KDEL sequence directed most of the early expression of phytase into the intracellular compartment yet more than $60\%$ of the total phytase activity was still retained within the cell even after the prolonged (>3 days) incubation of the transformant. However, the intracellular enzyme activity of the transformant without a KDEL sequence was as high as that of the extracellular one, thereby strongly suggesting that the secretion of phytase in S. cerevisiae appeared to be the rate-limiting step for the expression of a large amount of extracellular recombinant phytase, when compared with other yeasts.

• The Plant Pathology Journal
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• 제17권6호
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• pp.378.1-378
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• 2001

• Asian-Australasian Journal of Animal Sciences
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• 제27권1호
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• pp.77-82
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• 2014

The study aimed to evaluate the potential effects of feeding with phytase transgenic corn (PTC) on organ weight, serum biochemical parameters and nutrient digestibility, and to determine the fate of the transgenic DNA in laying hens. A total of 144 50-week-old laying hens were grouped randomly into 2 treatments, with 8 replicates per treatment and 9 hens per replicate. Each treatment group of hens was fed with diets containing 62.4% non-transgenic conventional corn (CC) or PTC for 16 weeks. The phytase activity for CC was 37 FTU/kg of DM, whereas the phytase activity for PTC was 8,980 FTU/kg of DM. We observed that feeding PTC to laying hens had no adverse effect on organ weight or serum biochemical parameters (p>0.05). A fragment of a poultry-specific ovalbumin gene (ov) was amplified from all tissues of hens showing that the DNA preparations were amenable to PCR amplification. Neither the corn-specific invertase gene (ivr) nor the transgenic phyA2 gene was detected in the breast muscle, leg muscle, ovary, oviduct and eggs. The digestibility data revealed no significant differences between the hens that received the CC- and PTC-based diets in the digestibility of DM, energy, nitrogen and calcium (p>0.05). Phosphorus digestibility of hens fed the PTC-based diet was greater than that of hens fed the CC-based diet (58.03% vs 47.42%, p<0.01). Based on these results, it was concluded that the PTC had no deleterious effects on the organ weight or serum biochemical parameters of the laying hens. No recombinant phyA2 gene was detected in muscle tissues and reproductive organs of laying hens. The novel plant phytase was efficacious in improving the phosphorus digestibility of laying hens.