• 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.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2002년도 창립10주년기념 및 국립독성연구원 의약품동등성평가부서 신설기념 국재학술대회:생물학적 동등성과 의약품 개발 전략을 위한 국제심포지움
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• pp.205-205
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• 2002

Two formulations of tiropramide {(${\pm}$)${\alpha}$-(benzoylamino)-4-[2-(diethylamino)-ethoxy]-N,N-dipropyl-benzenepropanamide hydrochloride}, an antispasmodic agent, were orally administered to 16 healthy Korean male volunteers by Latin crossover design with the purpose of evaluating bioeqivalence and phamacokinetics of tiropramide. Tiropramide in human plasma was determined by a gas chromatography/nitrogen phosphorus detector. Detection limit of tiropramide was 5 ng/ml. C$\_$max/ values in test and reference formulations were 93.9 ${\pm}$ 54.3 and 96.4 ${\pm}$ 51.6 ng/ml, respectively. AUC$\_$0\longrightarrowlast/ and AUC$\_$0\longrightarrowinf/ were, respectively, 330.7 ${\pm}$ 193.9 and 349.5 ${\pm}$ 205.3 ng.hr/ml for test formulation, 348.9 ${\pm}$ 207.7 and 380.8 ${\pm}$ 239.0 ng.hr/ml for reference formulation. Terminal half-life was 2.3-2.6 hr. Bioavailability differences for C/aub max/ and AUC$\_$0\longrightarrowlast/ were 2.48% and 5.22%, respectively. Minimum detection differences were less than 20% in both C$\_$max/ AUC. Based on this results, two formulations of tiropramide were considered to be bioequivalent

• 약학회지
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• 제44권2호
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• pp.189-192
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• 2000

We examined a dermal pharmacokinetics for a new SYC-fentanyl patch in rabbits. Determination of fentanyl in the plasma was performed using a gas chromatography with nitrogen-phosphorus detection system and a capillary column. One patch per animal (fentanyl 2.5 mg) was applied to clipped back skin for 72 hours. The plasma fentanyl concentration profile of SYC-patch was similar to that of a conventional patch (Durogesi $c^{R}$, Janssen Co.). No significant difference was observed in the pharmacokinetic parameters, the area under the concentration-time curve (AU $C^{0-}$72hrs/) and the total area under the first moment-time curve (AUM $C^{0-}$7hrs/), between the two patch types. The AU $C^{0-}$7hrs/ and AUM $C^{0-}$72hrs/ of durogesi $c^{R}$ were 183.3$\pm$46.28 ng＊hr/ml and 6,450$\pm$1,939ng＊h$r^2$/ml, and those of SYC-fentany patch were 217.2$\pm$50.51$\pm$ng＊hr/ml and 8,022$\pm$2,245ng＊h$r^2$/ml, respectively (n=3). This result indicates that the new SYC patch has a similar bioavailability compared to durogesi $c^{R}$ patch. Therefore, the SYC-patch may be considered as a bioequivalent fentanyl patch.patch.tch.

• Asian-Australasian Journal of Animal Sciences
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• 제12권2호
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• pp.203-208
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• 1999

An 8 week feeding trial was conducted with 864 ISA Brown laying hens, 48 weeks old, to determine if microbial phytase $(Natuphos^{(R)})$ supplementation can reduce non-phytate phosphorus (NPP) level in laying diets. The experiment consisted of four dietary treatments: T1, control diet with 0.26% NPP (0.55% total P) wand no supplementary phytase; T2, 0.21% NPP (0.50% total P) diet with 250 U of phytase/kg of diet; T3, 0.16% NPP (0.45% total P) diet with 250 U of phytase/kg of diet; and T4, 0.11% NPP (0.40% total P) diet with 250 U of phytase/kg of diet. T3 showed the highest egg production and egg weight and the lowest feed conversion while T4 gave the lowest egg production and the highest feed conversion and mortality. Daily feed consumption ranged from 130.4 g (T4) to 132.7 g (T2). T1 and T2 were not significantly different in the production parameters. Eggshell strength, egg specific gravity, and eggshell thickness were not significantly different among treatments. However, broken egg ratio was significantly lower in T2 and T4 than in T1. Retentions of Ca, P, Mg, and Cu were greater in phytase supplemented treatments (T2, T3, and T4) than the control (T1), and those in T3 and T4 were greater than in T2. Excretions of P in phytase supplemented treatments (T2, T3, and T4) were significantly (p<0.05) smaller than in T1 but excretions of N were not significantly different among the treatments. Contents of ash in tibiae were not significantly affected by treatments, but contents of Ca, P, Mg, and Zn was increased and that of Cu decreased by phytase supplementation. It is concluded that the NPP concentration in the diet of Brown layers consuming about 130 g/d of feed can be safely lowered from 0.26% (0.55% total P) to 0.16% (0.45% total P). The excretion of P was reduced by the inclusion of 250 U phytase/kg of diet.

• 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.

• 한국축산식품학회지
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• 제33권3호
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• pp.369-376
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• 2013

Milk is well known to be rich in some nutrients such as protein, calcium, phosphorus, and vitamins. In particular, absorption and bioavailability of calcium receive lots of attention because calcium is very little absorbed until it is changed to the ionized form in the intestine. In this study, concentration of the soluble calcium was determined in the commercial bovine milk products, which were processed by different heat-treatment methods for pasteurization. As for general constituents, lactose, fat, protein, and mineral were almost same in the liquid milk products by different processors. Ultrafiltration of the skimmed milk caused little change in the permeate as for lactose content but both fat and protein decreased. pH values ranges from 6.57-6.62 at room temperature and slightly increase after centrifugation, 10,000 g, 10 min. Rennet-coagulation activity was the lowest in the ultra high temperature (UHT-)milk compared to the low temperature long time (LTLT-) and high temperature short time (HTST-)milk products. Each bovine milk products contains 1056.5-1111.3 mg/kg of Ca. The content of sulfhydryl group was the lowest in raw milk compared to the commercial products tested. For the skimmed milks after ultrafiltration with a membrane (Mw cut-off, 3 Kd), soluble Ca in the raw milk was highest at 450.2 mg/kg, followed by LTLT-milk 336.4-345.1 mg/kg, HTST-milk 305.5-313.3 mg/kg, UHT-milk 370.3-380.2 mg/kg in the decreasing order. After secondary ultrafiltration with a membrane (Mw cut-off, 1 kD), total calcium in raw milk had a highest of 444.2 mg/kg, and those in the market milk products. As follow: UHT-milk, 371.3 to 378.2 mg/kg; LTLT-milk, 333.3 to 342.2 mg/kg; HTST-milk 301.9 to 311.2 mg/kg in a decreasing order.

• 자원환경지질
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• 제42권5호
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• pp.495-500
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• 2009

액티나이드(actinide)의 지질물질로의 수착은 핵에너지와 핵무기 개발로 인하여 인위적으로 자연환경에 노출된 핵종 원소의 이동성과 생물이용가능성을 낮추어 줄 수 있을 것으로 기대된다. 3가의 액티나이드 이온은 3가의 란타나이드(lanthanide) 이온과 유사한 화학적 성질을 띠므로, 3가 액티나이드의 산화광물 표면수착량은, 중성 또는 약한 산성의 pH 상태에서 3가의 란타나이드처럼, 인산염 이온(${PO_4}^{3-}$) 이 수착된 상태에서 크게 증가될 것으로 사료된다. 본 연구에서는 3가액티나이드 이온의 화학적 동족체인 3가 유로피움 이온($Eu^{3+}$)이 인산염이 수착된 상태의 뵈마이트 (${\gamma}$-AlOOH; boehmite) 표면에 수착되는 삼성분 수착계를 X선 흡수분석(EXAFS)을 통하여 관찰하였다. Eu X-선 흡수분석은 Eu-$PO_4$-뵈마이트 삼성분 수착계에서 뵈마이트 표면에 $EuPO_4$ 표면침전물이 형성되는 것을 지시하여 준다. 인산염이 뵈마이트 표면에서 $EuPO_4$ 표면침전물을 형성할 뿐 아니라 두자리 단핵 표면착물을 형성한 것을 P X-선 흡수분석을 통하여 확인하였다.