• 제목/요약/키워드: phosphate buffer

검색결과 683건 처리시간 0.045초

New Retention Mechanism of Mononucleotides with Buffer Concentrations in Ion-suppressing RP-HPLC

  • Lee, Ju-Weon;Row, Kyung-Ho
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제6권1호
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    • pp.37-41
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    • 2001
  • HPLC separation of ionic samples tends to be more complicated and difficult to understand than that of non-ionic compounds. On the other hand, band spacing is much more easily manipulated for ionic than for neutral samples. Ion-suppression RP-HPLC method was used with organic modifier and aqueous buffer solution. In this work, five mononucleotides of cytidine-5-monophosphate (5-CMP) disodium salt, uridine-5-monophosphate disodium salt (5-UMP), guanosine-5-monophosphate disodium salt (5-GMP), inosine-5-monophosphate disodium salt (5-IMP), and adenosine-5-monophosphate disodium salt (5-AMP) were examined. Acetic acid and sodium phosphate were used as buffers, and methanol as an organic modifier. A new relationship between the retention factor and the buffer concentration at a fixed modifier content (5% of methanol) could be expressed by following: K = (k(sub)-1 + k(sub)0 (k(sub)B/k(sub)S)/(1 + (k(sub)B/k(sub)S) C(sub)B(sup)a), where C(sub)B was the concentration of buffer. Using this relationship, the calculated values closely matched the experimental data. The derived relationship showed that as the buffer concentration increased, the retention factor approached a certain value, and this was buffer dependent.

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Seasonal Phosphorus Dynamics in a Forest Stream Water Following Different Harvests

  • Park, Byung Bae
    • 한국산림과학회지
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    • 제97권2호
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    • pp.181-186
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    • 2008
  • Even small changes in phosphorus concentrations in stream water could cause eutrophication because of very low level of phosphorus concentrations in natural waters. I investigated the impact of strip cut and clear cut on phosphorus concentrations in stream water at the Hubbard Brook Experimental Forest and investigated stream water phosphorus concentrations as a function of flow rate and season (as well as cutting history). Mean phosphate concentrations in the control (undisturbed forest) increased $1.9{\mu}g\;L^{-1}\;to\;2.6{\mu}g\;L^{-1}$, while strip cut treatment increased phosphate concentrations in stream water $2.2{\mu}g\;L^{-1}\;to\;3.7{\mu}g\;L^{-1}$ during the same period. There was no significant effect of clear cut treatment on phosphate concentrations in stream water. No relationships were found between discharge rate and phosphate concentrations, but the magnitude of fluctuation were increased during two decades in undisturbed forest: $1-5{\mu}g\;L^{-1}$ from 1963 to 1975 and $1-12{\mu}g\;L^{-1}$ from 1983 to 1995. Based on this study, forest harvests with buffer zone will not make a problem by imported phosphate to cause eutrophication in natural water.

도열병균의 원형질체 나출 및 세포벽 재생 (Purification and Cell Wall Regeneration of Protoplasts from Pyricularia oryzae Cav.)

  • 한성숙;이영희;유재당;이은종
    • 한국식물병리학회지
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    • 제3권2호
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    • pp.124-130
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    • 1987
  • 수도 도열병균(Pyricularia oryzae)의 균사체로부터 다량의 원형질체의 분리 및 세포벽 재생을 하기 위하여 필요한 몇가지 조건을 선발하였다. 나출을 위하여 기본용액으로 0.02M potassium phosphate buffer와 pH 5.2, 0.6M KCl로 삼투압을 조절하였고 분해효소는 ml당 각각 20mg Cellulase R-10, 5mg Macerozyme $-10, 10mg Driselase를 사용하였는데 각각의 단독처리구보다 3가지 효소의 복합처리구에서 원형질체 나출 정도가 우수하였다. 또한 선발된 복합효소액에 2일간 액체배양된 어린 균사체를 3시간, $30^{\circ}C$ 항온기에서 진탕했을 때 가장 많은 원형질체가 분리되었다. 원형질체로부터 세포벽의 재생은 순수 정제한 원형질체를 0.2M potassium phosphate와 0.6M KCl을 삼투압을 조절한 감자한철배지에 접종시켜 가장 높은 재생율을 얻을 수 있었다.

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Capillary electrophoresis 및 근적외선분광분석기를 이용한 황기의 원산지 판별 (Discrimination of Geographical Origin for Astragalus Root (Astragalus membranaceus) by Capillary Electrophoresis and Near-Infrared Spectroscopy)

  • 김은영;김정현;이남윤;김수정;류미라
    • 한국식품과학회지
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    • 제35권5호
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    • pp.818-824
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    • 2003
  • 외국농산물의 국내 유입증가와 이에 따른 신속한 원산지 판별법 확립이 요구되는 가운데 최근 수입이 급증한 품목 중 하나인 황기를 선택, CE 및 NIRS를 이용하여 분석조건을 확립하고 원산지판별에의 적용 가능성을 검토하였다. CE를 이용하여 분석 시 추출은 methanol: 0.1M phosphate buffer(pH 2.5)(3:7)를 사용하였으며 uncoated fused silica capillary$(50\;{\mu}m\;I.D.{\times}27cm)$를 이용하여 $45^{\circ}C$, 14 kV로 분석, 200 nm에서 검출하였다. 분석 buffer는 0.1 M phosphate buffer(pH 2.5)에 20% methoxy ethanol과 40 mM HSA를 첨가하여 사용하였으며, 8초간 pressure injection 하였다. Peak의 재현성을 증대시키기 위하여 시료 주입 전 분석 buffer를 1분간 분석 시와 같은 방향으로(F) 흘려주고 0.1 M phosphoric acid와 1 M sodium hydroxide는 각각 4분, 5분간 반대방향으로(R) 홀려주었다. 증류수를 다시 1분간 흘려주고(R) 분석 buffer로 2분간 평형화(F) 시킨 후 시료를 주입하였다. 이상의 조건으로 국내산(97점)과 수입(113점) 황기를 분석한 결과 전체 peak의 양상은 유사하였으나 약 $11{\sim}13$분에 용출되는 2개의 peak(peak am-1, am-2)의 면적 비율에서 차이가 나타나 국산은 peak am-2가 peak am-1의 약 4배인 반면, 수입 산은 10배로 나타나 원산지 판별이 가능하였으며, 약 80%의 편별율을 나타내었다. NIRS는 국산 및 수입산 황기 raw 스펙트럼의 2차 미분 스펙트럼 R값이 0.915로 비교적 안정된 값을 얻을 수 있었고, SEP는 약 14.3%로 나타났다. 이를 국산과 수입산 황기에 적용 시 전체 판별율이 약 97%로 비교적 높은 판별율을 보였다. 또한 NIRS로 판별이 불가능한 시료가 CE로는 판별이 가능하여 이 두 기기를 함께 사용 시 상호보완하여 신속 정확한 원산지 판별법의 개발 가능성이 시사되었다.

Adsorption of Globular Proteins to Vaccine Adjuvants

  • Jang, Mi-Jin;Cho, Il-Young;Callahan, Patricia
    • BMB Reports
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    • 제30권5호
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    • pp.346-351
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    • 1997
  • The maximum adsorption/desorption conditions and the adsorption mechanism of globular proteins to vaccine adjuvants were determined. The maximum adsorption ratio of protein to the $Al^{3+}$ content of aluminum oxyhydroxide and the optimal adsorption pH are 2:1 (${\mu}g:{\mu}g$) for bovine serum albumin (BSA) at pH 6.0 and 2.5:1 (${\mu}g:{\mu}g$) for immunoglobulin G (IgG) at pH 7.0, respectively. The maximum adsorption ratio onto aluminum phosphate gel was 1.5:1 (${\mu}g$ Protein:${\mu}g$ $Al^{3+}$) at pH 5.0 for both BSA and IgG. Adsorption of the native globular proteins, BSA and IgG, to aluminum oxyhydroxide and aluminum phosphate gel was reversible as a function of pH. Complete desorption of these proteins from aluminum phosphate gel was observed at alkaline pH, whereas only 80~90% removal from aluminum oxyhydroxide was achieved with alkaline pH and 50 mM phosphate buffer. We conclude that electrostatic and hydrogen bonding interactions between the native proteins and adjuvants are important binding mechanisms for adsorption, and that the surface charge of the protein and the colloid components control the maximum adsorption conditions.

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Ultrasonic Studies of Proton-Exchange Reaction Between Hydrogen Phosphate Ions and Imidazole

  • Choi, Chang-Ha;Chung, Myung-Kiu
    • The Journal of the Acoustical Society of Korea
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    • 제16권1E호
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    • pp.24-28
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    • 1997
  • Ultrasonic relaxation measurements for imidazole and its derivative in phosphate buffer exhibit a high peak of absorption at neutral pH. Near neutral pH, protolysis and hydrosis may be neglected and the essential reaction only consists of a direct proton-exchange. The kinetics constants and the volume changes for the proton transfer reaction with the protonated imidazole and 2-methylimidazole have been determined at 25℃. The kinetics constants are 7.2×108s-1M-1for imidazole and 1.7×108s-1M-1 for 2-methylimidazole. The kinetics constants are used to estimate the spectrum of relaxation times and acoustic relaxation amplitude associated with intermolecular and intramolecular proton-exchange reactions in bilogical media. It is concluded that the magnitude of the acoustic absorption reasonalbly attributable to the perturbation of proton-transfer equilibria between imidazole and inorganic phosphate is comparable in magnitude with the acoustic absorption observed in some intact tissues.

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Ascorbic Acid와 Pyrophosphate로부터 Ascorbic Acid-2-Phosphate의 효소적 생산

  • 최현일;이상협;방원기
    • 한국미생물·생명공학회지
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    • 제24권5호
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    • pp.613-618
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    • 1996
  • Microorganisms capable of producing ascorbic acid-2-phosphate (AsA2P) from ascorbic acid (AsA) and pyrophosphate (PPi) were screened from the culture collection of this laboratory. Among them, Cellulomonas sp. AP-7 showed the highest productivity of AsA2P. The optimal conditions for the production of AsA2P from AsA and PPi with cell-free extract as an enzyme source were investigated. The reaction mixture for the maximal production of AsA2P consisted of 21 g protein of cell-free extract per liter as the enzyme source, 250 mM AsA, 200 mM sodium pyrophosphate, 150 mM sodium acetate buffer (pH 4.5). By using this reaction mixture, 31.9 mM of AsA2P, which corresponded to a 12.76% yield based on AsA, was produced after incubation of 48 hr at 33$\circ$C.

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Optimization of Protein Extraction for Lichen Thalli

  • Kondratiuk, Anna S.;Savchuk, Oleksiy M.;Hur, Jae-Seoun
    • Mycobiology
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    • 제43권2호
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    • pp.157-162
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    • 2015
  • Lichen-forming fungal proteins have been seldom searched due to many difficulties in their extraction. Phenols, quinones, proteases, and other components released during cell disruption have been known to be the greatest challenges related to protein extraction from lichens. To overcome these problems and maintain good electrophoretic resolution and high protein concentration, an extraction buffer containing polyvinylpolypyrrolidone, ascorbic acid, Triton X-100, polyethylene glycol, proteinase, and oxidase inhibitors in sodium phosphate buffer was developed. This extraction buffer showed high efficiency for all lichen species tested in the study.

Refolding of Fusion Ferritin by Gel Filtration Chromatography(GFC)

  • Kim, Hyung-Won;Kim, In-Ho
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제10권6호
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    • pp.500-504
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    • 2005
  • Fusion ferritin (heavy chain ferritin, $F_H+$ light chain ferritin, $F_L$), an iron-binding protein, was primarily purified from recombinant Escherichia coli by two-step sonications with urea [1]. Unfolded ferritin was refolded by gel filtration chromatography (GFC) with refolding enhancer, where 50 mM Na-phosphate (pH 7.4) buffer containing additives such as Tween 20, PEG, and L-arginine was used. Ferritin is a multimeric protein that contains approximately 20 monomeric units for full activity. Fusion ferritin was expressed in the form of inclusion bodies (IBs). The IBs were initially solubilized in 4 M urea denaturant. The refolding process was then performed by decreasing the urea concentration on the GFC column to form protein multimers. The combination of the buffer-exchange effect of GFC and the refolding enhancers in refolding buffer resulted in an efficient route for producing properly folded fusion ferritin.

고온기 육계의 수송 스트레스에 대한 버퍼제 급여에 따른 닭고기 품질 및 PSE 발생에 미치는 영향 (Effects of Dietary Buffer Material for Chicken Meat Grades and PSE Incidence in Broilers under Transport Heat Stress)

  • 채현석;최희철;나재천;장애라;김민지;방한태;강환구;김동욱;서옥석;박성복;함준상
    • 한국가금학회지
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    • 제37권2호
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    • pp.131-137
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    • 2010
  • 고온기 때 육계 수송 스트레스를 줄이기 위한 대비책으로 농가에서 출하 2일전에 버퍼제(sodium phosphate, sodium bicarbonate, magnesium sulfate)를 급여함으로 육계의 품질에 미치는 영향을 구명하기 위해 수행하였다 닭고기의 품질 $1^+$ 등급 평가에서 버퍼제를 급여하지 않는 대조구에서 4%를 차지하였으나 sodium phosphate 급여구가 8%, sodium bicarbonate 급여구 14%, magnesium sulfate 급여구가 28%로 대조구에 비하여 버퍼제를 급여한 처리구에서 $1^+$등급 출현율이 증가하였다. PSE 발생율은 대조구에서 경증 및 중증 모두 합하여 92%를 차지하였으나, sodium phosphate 급여구가 88%, sodium bicarbonate 급여구 28%, magnesium sulfate 급여구가 56%로 대조구에 비하여 버퍼제를 급여한 처리구에서 PSE 발생율이 현저히 저하되었다. 닭고기 외관의 멍은 대조구에서는 1~6 cm의 멍이 32%를 차지하였으나, sodium phosphate 급여구는 22%, sodium bicarbonate 급여구 24%, magnesium sulfate 급여구가 44%로 magnesium sulfate 급여구를 제외하고는 대조구에 비해 멍 발생율의 8~10% 정도 저하하였다. 닭고기의 명도($L^*$) 값의 변화는 가슴 근육 부위에서 대조구가 67.88을 나타내었고, sodium phosphate 급여구가 67.05, sodium bicarbonate 급여구 66.27, magnesium sulfate 급여구가 65.89로 magnesium sulfate 급여구에 명도가 가장 낮은 경향을 나타내었고, 전체적으로 대조구에 비해 버퍼제를 급여한 구에서 명도 값이 저하되었다. 상기와 같이 고온기 때 육계 수송 스트레스를 줄이기 위하여 버퍼제(sodium phosphate, sodium bicarbonate, magnesium sulfate)를 급여함으로 사계 및 닭고기의 이상 도체를 줄일 수 있을 것으로 사료된다.