• Title/Summary/Keyword: Membrane chromatography

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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
    • Journal of Animal Reproduction and Biotechnology
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    • v.37 no.3
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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.

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

  • Lee, Sang-Hwan;Seo, Jeong-Su;Kim, Na-Yeong;Eom, Hye-Gyeong;Wi, Hyo-Jin;Park, Seong-Il;Jeong, Jun-Gi
    • Journal of fish pathology
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    • v.17 no.2
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    • pp.131-137
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    • 2004
  • In the present study, phosphatidylcholine (PC) hydrolyzing enzyme had been isolated from membrane of flounder liver. PC hydrolyzing enzyme solubilized in 1% Triton X -100 from membrane was partially purified by sequential chromatography on Heparin Sepharose CL-6B and Heparin-5PW columns. The products by membrane-bound hydrolyzing enzyme were identified as phosphatidic acid and choline, but in the presence of primary alcohol, phosphatidylethanol was produced at the expense of phosphatidic acid. These data suggest that membrane-bound enzyme may be a PC-phosphoipase D (PLD) type. The enzyme had pH optimum at below 6.0 and temperature optimum at $37^\circ{C}$. The activity of PC-PLD was dose-dependently increased by $Ca^{2+}$ but not $Mg^{2+}$. The activity of PC-PLD was stimulate by PC, PIP2 and PE.

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

  • Kang, Na-Ra;Hong, Jong-Rak;Choung, Pill-Hoon
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.30 no.3
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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.

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

  • Koo, Jin-Sun;Kwak, Noh-Seok;Hwang, Taek-Sung
    • Polymer(Korea)
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    • v.36 no.5
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    • pp.564-572
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    • 2012
  • A terpolymer of vinylbenzyl chloride-co-ethyl methacrylate-co-styrene (VBC-EMA-St) was prepared for membrane capacitive deionization (MCDI) by radical polymerization and amination reaction of various amination times. Nonfluoro aminated VBC-EMA-St anion-exchange membranes were characterized by Fourier transform infrared (FTIR) spectrometry. Molecular weight, polydispersity and thermal stability were obtained by gel permeation chromatography (GPC) and thermogravimetric analysis (TGA). The basic properties such as water uptake, ion exchange capacity, electrical resistance and CDI charge-discharge current were measured. The optimal values of ion exchange capacity, water uptake, electrical resistance and molecular weight of synthesized anion-exchange membrane were 1.69 meq/g, 23.7%, 1.61 ${\Omega}{\cdot}cm$ and $3.4{\times}10^4$ g/mol, respectively. As compared with conventional membrane, the pattern of cyclic charge-discharge current of synthesized anion-exchange membrane indicated efficient electrosorption and desorption.

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

  • 정건용;정동진;김천호;신현수;민병렬;김래현
    • Membrane Journal
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    • v.13 no.2
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    • pp.81-87
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    • 2003
  • The permeation experiments f3r the aqueous dextran solutions of which the molecular weights were distributed between several thousands and million daltons were carried out using the dead-end cell system equipped with sheet membrane to determine the molecular weight cut-off (MWCO) of ultrafiltration membranes. The dextran rejections with respect to molecular weight were obtained by GPC analysis of feed and permeate solutions, then the MWCO was decided as the dextran molecular weight corresponding to the 90% rejection. The MWCO of PBTK membrane manufactured by Millipore was varied between 63,000 and 68,000 daltons as the pressure increased from 0.5 to 2.0 bar. However, the MWCO of PBQK(Millipore) and UE 1812(Saehan) under the above operating pressure increased to 3.5 and 4.3 times, respectively. Also, the MWCO of PBTK increased to 25% as the membrane permeation increased from 10 to 40% of the feed solution.

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

  • KIM, HYUN JIN;YOO, DONG JIN
    • Transactions of the Korean hydrogen and new energy society
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    • v.33 no.3
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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.

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

  • 전홍성;차영주
    • KSBB Journal
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    • v.10 no.1
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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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    • v.14 no.3
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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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    • v.5 no.1
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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
    • Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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    • 2003.10a
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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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