• 제목/요약/키워드: membrane chromatography

검색결과 262건 처리시간 0.028초

Effect of bicarbonate and progesterone on plasma membrane integrity, acrosome reaction and proportion of fatty acids in boar sperm

  • Park, Choon-Keun;Lee, Sang-Hee
    • 한국동물생명공학회지
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    • 제37권3호
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    • pp.202-208
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    • 2022
  • This study investigated the influence of sodium bicarbonate (NaHCO3) and progesterone on acrosome reaction and proportion of polyunsaturated fatty acid (PUFA) composition boar sperm. The sperm were diluted with semen extender and incubated with NaHCO3 and progesterone at 38℃, 5% CO2 for 6 h. Plasma membrane integrity and acrosome reaction were analyzed using SYBR14/propidium iodide (PI) and FITC-PNA/PI doubling staining method, and proportion of PUFA was analyzed using gas chromatography. In results, Plasma membrane integrity was significantly decreased in 50 mM NaHCO3 group and acrosome reaction was significantly increased by over the 100 mM NaHCO3 group compared to control group (p < 0.05). In addition, progesterone significantly increased decreased plasma membrane integrity at 100 mM progesterone and acrosome reaction at over the 5.0 µM progesterone (p < 0.05), but there was no difference among the 5.0 to 100 µM groups. PUFAs were significantly decreased in 100 mM NaHCO3 and 50 µM progesterone treatments compared to control group. In summary NaHCO3 and progesterone induce acrosome reaction and reduce PUFA composition in boar sperm, therefore, the results maybe help to understand basically knowledge for the acrosome reaction and PUFA composition in boar sperm.

넙치 간에 있어 가수분해 효소의 부분정제 및 특성규명 (Partial purification and characterization of phosphatidylcholine hydrolyzing enzyme from liver membrane of flounder , Paralichtys olivervaceus)

  • 이상환;서정수;김나영;엄혜경;위효진;박성일;정준기
    • 한국어병학회지
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    • 제17권2호
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    • pp.131-137
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    • 2004
  • 본 실험은 넙치 (Paralichthys olivaceus) 간으로부터 membrane부분에 존재하는 PC 가수분해 효소의 특성에 대해 조사하였다. 먼저 간 조직을 초고속 원심 분리기와 nonion-detergent인 1% triton X - 100을 이용하여 membrane 부분을 분리하였으며, Heprain-Sepharose CL-6B 칼럼과 Heparin-5PW 칼럼을 이용하여 분리정제 하였다. 얻어진 PC 가수분해효소에 대한 반응 ․ 생성물을 확인하기 위해 autoradiography를 실시하였다. 지용성 부분의 결과에서 transphosphatidylation 반응의 결과물인 PEtOH을 형성하는 것으로 보아 PC-PLD임을 알 수 있었다. 얻어진 PC 가수분해효소에 대한 생화학적 특성을 조사한 결과 적정 pH가 6.5이하인 산성 조건 및 $37^\circ{C}$의 배양온도에서 최고 활성을 나타내었으며, 이가 이온들에 대한 영향의 경우 칼슘은 1.67mM 농도에서 최고 활성을 나타냈으나, 마그네슘은 활성에 영향을 미치지 않았다. 각종 세포막 기질에 대한 영향을 조사한 결과 PC는 $0.75\mu{M}$, PIP2는 $2.35\mu{M}$, PE는 $26.8\mu{M}$ 농도에서 최고 활성을 나타내었다. 이상의 결과부터 넙치 간조직의 막층부분에 존재하는 PC를 가수분해효소는 PC-PLD가 존재함을 알 수 있었다.

사람치아 단백질을 분리 흡착한 PVDF막의 생체반응에 관한 연구 (BIOASSAY OF HUMNA TOOTH PROTEIN BLOTTED POLYVINYLIDENE DIFLUORIDE(PVDF)MEMBRANE)

  • 강나라;홍종락;정필훈
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제30권3호
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    • pp.186-192
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    • 2004
  • Purpose: Human tooth proteins are highly heterogeneous, comprising diverse proteins derived from a number of genes. The attempts to identify protein for activity of tooth matrix proteins have been defied by several factors. First, the amount of proteins within teeth is very small relative to many extracellular matrix proteins of other tissues. Second, the bioassay system is tedious and needed for long time. Therefore we tried to find easy techniques, which increase the product rate, and an assay of small proteins, with which amino acid sequence is possible without additional procedures. Materials and Methods: Total protein were extracted from 300 g enamel removed teeth and 600 g teeth with 4 mol/L guanidine HCl and purified by gel chromatography. Aliquot of proteins was implanted into muscle pouches in Sprague-Dawley rats for bioassay. By SDS-PAGE and membrane blotting, molecular weight of each protein was estimated and a partial amino acid sequence was obtained. Each fraction blotted on the membrane was cut out and inserted in rat ectopic model. Results: In dissociative method, total tooth proteins were obtained 1mg/ml from enamel removed teeth and 3.5 mg/ml from teeth. In SDS-PAGE, four clear bands at the sites corresponding to 66, 40, 20 and 18 kD. Especially The 66 kD band was clearly exhibited. Amino acid sequencing from tooth could be possible using PVDF membrane blotting technique. In amino acid sequencing, 66 kD protein was identified as albumin. Conclusion: Compared with conventional method for extraction of teeth protein and bioassay of proteins, the methods in this study were easy, time-saving and more productive technique. The matured tooth proteins omitting additional procedure of mechanical removal of enamel were simply analyzed using blotted PVDF membrane. This method seems to make a contribution as a technique for bioassay and amino acid sequencing of protein.

막 축전식 탈염용 비불소계 아민화 Poly(vinylbenzyl chloride-co-ethyl methacrylate-co-styrene) 음이온교환막의 합성 및 특성 (Synthesis and Properties of Nonfluoro Aminated Poly(vinylbenzyl chloride-co-ethyl methacrylate-co-styrene) Anion Exchange Membranes for MCDI Process)

  • 구진선;곽노석;황택성
    • 폴리머
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    • 제36권5호
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    • pp.564-572
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    • 2012
  • 본 연구에서는 막축전식 탈염(membrane capacitive deionization, MCDI) 공정용 음이온교환막의 제조를 위하여 vinylbenzyl chloride-co-ethyl methacrylate-co-styrene(VBC-EMA-St) 공중합체를 합성하였으며, 아민화 반응과 열처리를 통하여 음이온교환막을 제조하였다. 구조확인을 위하여 FTIR 분석을 하였고, GPC와 TGA를 통하여 합성한 고분자의 분자량과 분자분포, 열안정성을 분석하였으며, 함수율 및 이온교환용량을 측정하였다. 또한 LCR meter로 전기저항을 측정하고, MCDI 공정에 적용하기 위하여 제조한 음이온교환막을 충방전 시험 측정하였다. 이온교환용량, 함수율, 전기저항, 분자량은 각각 1.69 meq/g, 23.7%, 1.61 ${\Omega}{\cdot}cm$, $3.4{\times}10^4$ g/mol이었으며, CDI 충방전 시험 결과 상용화막인 AMX보다 우수한 성능을 나타내었다.

GPC를 이용한 한외여과막의 평가 (Evaluation of Ultrafiltration Membrane Using Gel Permeation Chromatography)

  • 정건용;정동진;김천호;신현수;민병렬;김래현
    • 멤브레인
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    • 제13권2호
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    • pp.81-87
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    • 2003
  • 한외여과막의 분획분자량을 결정하기 위하여 dead-end형 셀내에 평막을 설치하고 분자량 분포가 수천 내지 수백만의 혼합 dextran 수용액으로 투과 실험하였다. 원료 용액과 투과액을 GPC로 분석하여 각 분자량에 대한 배제율을 구하고 90% 배제율에 해당하는 dextran분자량을 분획분자량를 결정하였다. 투과압력을 0.5에서 2.0 bar까지 증가시킬 경우, Millipore사의 PBTK막은 63,000 내지 68,000 daltons로 10% 이내에서 변화하였지만 Millipore사의 PBQK 막 또는 (주)새한의 UE1812막의 분획분자량은 각각 3.5 및 4.3 배 증가하였다. 또한 투과액을 원료용액의 10내지 40%까지 분리막을 증가시키면서 배제율을 측정한 결과, PBTK의 분획분자량은 25% 증가하였다.

수지상 폴리(알릴렌 이써 설폰)에 도입된 지방족 알킬사슬 연결자길이에 따른 음이온교환막의 특성 연구 (A Study on the Characteristics of Anion Exchange Membrane According to Aliphatic Alkyl Chain Spacer Length Introduced into Branched Poly (Arylene Ether Sulfone))

  • 김현진;유동진
    • 한국수소및신에너지학회논문집
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    • 제33권3호
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    • pp.209-218
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    • 2022
  • Recently, research on the development of anion exchange membranes (AEMs) has received considerable attention from the scientific community around the world. Here, we fabricated a series of AEMs with branched structures with different alkyl spacers and conducted comparative evaluations. The introduction of these branched structures is an attempt to overcome the low ionic conductivity and stability problems that AEMs are currently facing. To this end, branched polymers with different spacer lengths were synthesized and properties of each membrane prepared according to the branched structure were compared. The chemical structure of the polymer was investigated by proton nuclear magnetic resonance, Fourier transform infrared, and gel permeation chromatography, and the thermal properties were investigated using thermogravimetric analysis. The branched anion exchange membrane with (CH2)3 and (CH2)6 spacers exhibited ionic conductivities of 8.9 mS cm-1 and 22 mS cm-1 at 90℃, respectively. This means that the length of the spacer affects the ionic conductivity. Therefore, this study showing the effect of the spacer length on the ionic conductivity of the membrane in the polymer structure constituting the ion exchange membrane is judged to be very useful for future application studies of AEM fuel cells.

Acetobater sp.KM Alcohol Dehydrogenase의 분리 및 특성 (Purification and Characterization of Alcohol Dehydrogenase from Acetobacter sp. KM)

  • 전홍성;차영주
    • KSBB Journal
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    • 제10권1호
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    • pp.30-37
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    • 1995
  • Membrane-bound alcohol dehydrogenase(ADH) was purified to homogeneity from the acetic acid producing bacteria, Acetobacter sp. KM. The enzyme was solubilized and extracted with Triton X-100 and purified using the Mono-Q ion exchange chromatography and Superose 12 gel filtration chromatography. The enzyme was purified to 12-fold with a yield of 30%. The molecular weight of the purified enzyme was to be 335 KDa. SDS-PAGE of the enzyme showed two subunits with molecular weights of 79 KDa and 49 KDa. It indicated that the enzyme consisted of three subunits of the 79 KDa and two subunits of the 49 KDa. The purified .ADH preferentially oxidized straight chain aliphatic alcohol except methanol. Formaldehyde, acetaldehyde and glutaraldehyde were also oxidized. The apparent Km for ethanol was 1.04 mM and the optimum pH and temperature were 5.0∼6.0 and 32$^{\circ}C$, respectively. V2O5 and divalent cation such as ZnCl2 and NiCl2 inhibited enzymatic activity.

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Solubilization of an Angiotensin II Binding Site from Rat Liver

  • Chung, Sung-Hyun;Ravi Iyengar
    • Archives of Pharmacal Research
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    • 제14권3호
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    • pp.231-236
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    • 1991
  • The high affinity binding sites for angiotensin II were solubilized from rat liver membranes by treatment with CHAPS. The binding protein was also partially purified by angiotensin III inhibitor-coupled Affi-gel affinity chromatography. Binding to the intact membrances as well as to the solubilized preparation was specific and saturable. According to the Scatchard plot, the membrane preparations exhibited a single class of high affinity binding sites with a Kd OF 0.71 nM. The solubilized preparation also showed the presence of a single class of bindings sites with less affinity (Kd of 14 nM). Meanwhile the competition studies using angiotensin II analogues represented two separate binding sites for angiotensin II and single binding site for antagonist. These latter findings were correlated to the results provided by Garrison's research group. More works are needed to clarify this discrepancy.

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Phospholipid Analysis by Nanoflow Liquid Chromatography-Tandem Mass Spectrometry

  • Moon, Myeong Hee
    • Mass Spectrometry Letters
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    • 제5권1호
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    • pp.1-11
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    • 2014
  • Lipids play important roles in biological systems; they store energy, play a structural role in the cell membrane, and are involved in cell growth, signal transduction, and apoptosis. Phospholipids (PLs) in particular have received attention in the medical and lipidomics research fields because of their involvement in human diseases such as diabetes, obesity, atherosclerosis, and many cancers associated with lipid metabolic disorders. Here I review experimental strategies for PL analysis based on nanoflow liquid chromatography-electrospray ionization-tandem mass spectrometry (nLC-ESI-MSn). In particular, discussed are lipid extraction methods, nanoflow LC separation of PLs, effect of ionization modifiers on the ESI of PLs, influence of chain lengths and unsaturation degree of acyl chains of PLs on MS intensity, structural determination of the molecular structure of PLs and their oxidized products, and quantitative profiling of PLs from biological samples such as tissue, urine, and plasma in relation to cancer and coronary artery disease.

Purification and Characterization of an Angiotensin Converting Enzyme Inhibitor from Squid Ink

  • Kim, So-youn;Kim, Sun-hye;Song, Kyung-Bin
    • 한국식품저장유통학회:학술대회논문집
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    • 한국식품저장유통학회 2003년도 제23차 추계총회 및 국제학술심포지움
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    • pp.135.2-135
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    • 2003
  • Angiotensin converting enzyme (ACE) converts angiotensin I into angiotensin II by cleaving C-terminal dipeptide of angiotensin I and inactivates bradykinin. ACE inhibitors have been screened from various food sources since the inhibitors decrease blood pressure. Therefore, in this study, an ACE inhibitor was isolated and purified from squid ink using membrane filtration, gel permeation chromatography, normal phase HPLC, and fast protein liquid chromatography. The purified inhibitor was identified to be a molecular mass of 294 by mass spectrometry, and to have IC$\sub$50/ value of 4.9 $\mu\textrm{g}$/mL.

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