scholarly journals Preparation, purification and analyses of thirteen alkali-stable dinucleotides from ribonucleic acid

1970 ◽  
Vol 116 (4) ◽  
pp. 589-598 ◽  
Author(s):  
A. R. Trim ◽  
Janet E. Parker
Keyword(s):  
Nucleotide Sequence ◽  
Ion Exchange ◽  
Ribonucleic Acid ◽  
Deae Cellulose ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Spectroscopic Constants ◽  
Commercial Yeast ◽  
High Degree ◽  
Hydrolysis Of

Of the 16 alkali-stable dinucleotides known to be obtained by hydrolysis of commercial yeast RNA with alkali, 13 were prepared in quantities of the order of 10mg or more. The samples, with only one exception, contain at least 90% of dinucleotide, and spectroscopic constants and nucleotide-sequence determinations, although not conclusive, indicate a high degree of purity of these products. The small dinucleotide fraction in 150g of RNA hydrolysed with alkali (1–2% of the total nucleotides) was separated from the mononucleotides by stepwise ion-exchange chromatography on DEAE-cellulose columns and resolved into seven fractions containing from one to four different dinucleotides by electrophoresis on paper at pH3.0. These fractions were resolved into their constituent dinucleotides by chromatography in ammonium sulphate. Contamination of the products by impurities from the paper was minimized by washing it before using it for chromatography or electrophoresis and, by using a thick grade of paper (Whatman no. 17), it was possible to handle and purify relatively large quantities of nucleotides.

THE SEPARATION AND PURIFICATION OF HUMAN LUTEINIZING AND THYROTROPHIC HORMONES

1964 ◽  
Vol 29 (1) ◽  
pp. 61-69 ◽  
Author(s):  
ANNE STOCKELL HARTREE ◽  
W. R. BUTT ◽  
K. E. KIRKHAM
Keyword(s):  
Luteinizing Hormone ◽  
Ion Exchange ◽  
Deae Cellulose ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Separation And Purification ◽  
Hormone Activity ◽  
Human Pituitary ◽  
Thyrotrophic Hormone

SUMMARY Ion-exchange chromatography was used to further purify a human pituitary fraction rich in thyrotrophic and luteinizing hormone activities. Approximately twofold concentration of both activities was obtained by chromatography on IRC-50 at pH 7·5, but the hormones were not separated. Subsequent chromatography on DEAE-cellulose at pH 9·5 led to a tenfold concentration of the luteinizing hormone in a fraction practically free of thyrotrophic activity and to a fourfold concentration of the thyrotrophic hormone in a fraction still exhibiting substantial luteinizing hormone activity.

Kristallstruktur von Cs2[B6H5(SCN)]/ Crystal Structure of Cs2[B6H5(SCN)]

1998 ◽  
Vol 53 (8) ◽  
pp. 816-818 ◽  
Author(s):  
W. Preetz ◽  
S. Zander ◽  
C. Bruhn
Keyword(s):  
Crystal Structure ◽  
Ion Exchange ◽  
Structure Determination ◽  
Deae Cellulose ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Orthorhombic Space Group ◽  
Space Group ◽  
X Ray

Abstract By reaction of [B6H6]2-with (SCN)2 in dichloromethane at -80 C° the thiocyanatohexaborate anion is formed and can be isolated by ion exchange chromatography on diethylaminoethyl (DEAE) cellulose. The X-ray structure determination of Cs2[B6H5(SCN)] (orthorhombic, space group Pbca with a = 9.506(5), b = 10.644(5), c = 21.857(5) Å, Z = 8) reveals that the SCN substituent is bonded via the S atom with the B-S distance of 1.885(9) Å and the B-S-C angle of 99.8(5)°. The SCN group is nearly linear (179.9(9)°).

scholarly journals Isolation of albumin from whole human plasma and fractionation of albumin-depleted plasma

1976 ◽  
Vol 157 (2) ◽  
pp. 301-306 ◽  
Author(s):  
J Travis ◽  
J Bowen ◽  
D Tewksbury ◽  
D Johnson ◽  
R Pannell
Keyword(s):  
Ion Exchange ◽  
Human Plasma ◽  
Plasma Proteins ◽  
Deae Cellulose ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Affinity Column ◽  
Plasma Albumin ◽  
Cibacron Blue ◽  
Blue Sepharose

The dye Cibacron Blue F-3-GA was conjugated to Sepharose to provide an affinity column for serum albumin. Passage of whole human plasma through a column of Cibacron Blue-Sepharose results in the removal of approx. 98% of the albumin. The latter can be quantitatively recovered by desorption with NaSCN. Albumin-depleted plasma can be readily resolved into discrete fractions by a combination of conventional biochemical techniques. In particular, the resolution of plasma proteins with properties similar to those of native human plasma albumin can readily be accomplished by ion-exchange chromatography of the Sepharose-dye-treated plasma on DEAE-cellulose.

The diguanosine nucleotides: do they exist in aquatic fungi?

1977 ◽  
Vol 55 (8) ◽  
pp. 841-846 ◽  
Author(s):  
A. H. Warner ◽  
D. des S. Thomas ◽  
V. Shridhar ◽  
H. D. McCurdy
Keyword(s):  
Ion Exchange ◽  
Deae Cellulose ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Aquatic Fungi ◽  
Enzyme Hydrolysis ◽  
Achlya Ambisexualis ◽  
Cast Doubt ◽  
Blastocladiella Emersonii ◽  
Metabolic Regulators

The aquatic fungi Achlya ambisexualis and Blastocladiella emersonii were grown in the presence of 32Pi and the 32P-labeled acid-soluble nucleotide fractions were analyzed by ion-exchange chromatography on DEAE-cellulose. Selected column fractions containing diguanosine tri- and tetra-phosphates (Gp3G and Gp4G) added as chromatographic markers were analyzed further for 32P by chromatography and (or) enzyme hydrolysis. The results of these experiments clearly indicate that neither Gp4G nor Gp3G is synthesized during vegetative growth of these organisms and cast doubt on the hypothesis that diguanosine nucleotides are important metabolic regulators in fungi.

scholarly journals PEMISAHAN ASAM AMINO DARI LIMBAH CAIR PABRIK KELAPA SAWIT DENGAN KROMATOGRAFI PENUKAR ION

2018 ◽  
Vol 6 (2) ◽  
pp. 69
Author(s):  
Indah Rahmi Sari ◽  
Ade Ayu Oksari ◽  
Irma Kresnawaty
Keyword(s):  
Amino Acid ◽  
Ion Exchange ◽  
Liquid Waste ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Enzyme Hydrolysis ◽  
Protease Enzyme ◽  
Absorbance Reading ◽  
Amino Acid Levels ◽  
Hydrolysis Of

Separation of Amino acid from Liquid waste of Oil palm Factory with Ion Exchange Chromatography Research on Separation of Amino Acid Liquid Waste mills with Ion Exchange Chromatography was carried out from October to November 2015. The results of  hydrolysis of 6 N HCl results showed  that the highest absorbance reading was obtained at a concentration of eluent of 0,2 and 0,6 M NaCl, while the results of the protease enzyme hydrolysis the highest absorbance reading at NaCl eluent 0,2 and 1 M. There was no significant difference in the results of separation by ion exchange chromatography, showed that the concentration of NaCl eluent is not very influential, so for subsequent analysis used only one concentration of the eluent. Results of linear regression obtained was equal to 0,9946, these results indicate that the series standard amino acid lysine has a value that is linear as it approaches 1. The amino acid levels obtained on the sample results LCPKS hydrolysis with 6 N HCl which was about 0 to 8.82 ppm and samples of the protease enzyme hydrolysis of about 0 to 4.31 ppm. Amino acid levels obtained were still far from the expected.Keywords: Amino Acid, Oil Palm, Liquid Waste, Ion Exchange Chromatography ABSTRAKPenelitian mengenai Pemisahan Asam Amino dari Limbah Cair Pabrik Kelapa Sawit dengan Kromatografi Penukar Ion telah dilaksanakan dari bulan Oktober sampai November 2015. Hasil hidrolisis HCl 6 N menunjukkan bahwa pembacaan absorbansi tertinggi diperoleh pada konsentrasi eluen NaCl 0,2 dan 0,6 M, sedangkan hasil hidrolisis enzim protease pembacaan absorbansi tertinggi pada eluen NaCl 0,2 dan 1 M. Tidak ada perbedaan yang signifikan pada hasil pemisahan dengan kromatografi penukar ion ini, menunjukkan bahwa konsentrasi eluen NaCl tidak terlalu berpengaruh, sehingga untuk analisis selanjutnya digunakan hanya satu konsentrasi eluen. Hasil regresi linear yang diperoleh yaitu sebesar 0,9946, hasil tersebut menunjukkan bahwa deret standar asam amino lisin mempunyai nilai yang linear karena mendekati 1. Kadar asam amino yang diperoleh pada sampel hasil hidrolisis LCPKS dengan HCl 6N yaitu sekitar 0 – 8,82 ppm dan sampel hasil hidrolisis enzim protease sekitar 0 – 4,31 ppm. Kadar asam amino yang diperoleh masih jauh dari yang diharapkan.Kata Kunci: Asam Amino, Minyak Kelapa Sawit, Limbah Cair, Kromatografi Penukar Ion

Catabolism of (R)-Amygdalin and (R)-Vicianin by Partially Purified ß-Glycosidases from Prunus serotina Ehrh. and Davallia trichomanoides

1984 ◽  
Vol 39 (3-4) ◽  
pp. 232-239 ◽  
Author(s):  
Gary Kuroki ◽  
Pauline A. Lizotte ◽  
Jonathan E. Poulton
Keyword(s):  
Ion Exchange ◽  
Deae Cellulose ◽  
Hydrolytic Activity ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Prunus Serotina ◽  
Artificial Substrates ◽  
Black Cherry ◽  
Significant Activity ◽  
Optimum Ph

Mature black cherry (Prunus serotina Ehrh.) seeds accum ulate high levels of the cyanogenic disaccharide (R)-amygdalin. Extracts from these seeds contain two β-glycosidases which have been identified and completely resolved by DEAE-cellulose ion-exchange chromatography. Amygdalin hydrolase hydrolyzed (R)-am ygdalin at an optimum pH of 5.5, releasing (R)-prunasin and D-glucose. This enzyme showed highest activity towards (R)-am ygdalin and failed to hydrolyze (R)-prunasin. linamarin, β-gentiobiose and cellobiose. A distinct β-glycosidase, prunasin hydrolase, displayed a pronounced preference for (R)-prunasin, hydrolyzing this cyanogenic monosaccharide at an optimum pH of 6.5 to mandelonitrile and D-glucose. Prunasin hydrolase was inactive towards (R)-am ygdalin, linamarin, and β-gentiobiose. Both enzymes showed significant activity towards the artificial substrates β-ONPGlu and β-PNPGlu but did not hydrolyze α-PNPGlu. In view of the pronounced specificity of these enzymes towards endogenous cyanogens, it is concluded that upon disruption of black cherry seeds (R)- amygdalin is catabolized to mandelonitrile in a stepwise manner (the sequential mechanism) by amygdalin hydrolase and prunasin hydrolase with (R)-prunasin serving as intermediate. Young fronds of Davallia trichomanoides are rich sources of (R)-vicianin (the β-vicianoside of (R)-mandelonitrile). A β-glycosidase, vicianin hydrolase, has been partially purified from frond extracts by ion-exchange chromatography. At the optimum pH of 6.0, this enzyme showed highest hydrolytic activity with (R)-vicianin, although both (R)-am ygdalin and (R)-prunasin could be hydrolyzed at approximately 15% of the rate observed with (R)-vicianin. It failed to hydrolyze β-gentiobiose, cellobiose, linamarin and α-PNPGlu. Closer exam ination revealed that (R)-vicianin and (R)-amygdalin were hydrolyzed at the aglycone-disaccharide bond (the simultaneous mechanism) yielding mandelonitrile and the respective disaccharides vicianose and β-gentiobiose

Purification of glysojanin, an antifungal protein, from the black soybean Glycine soja

2003 ◽  
Vol 81 (6) ◽  
pp. 387-394 ◽  
Author(s):  
Patrick H.K Ngai ◽  
T B Ng
Keyword(s):  
Liquid Chromatography ◽  
Ion Exchange ◽  
Glycine Soja ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Fast Protein Liquid Chromatography ◽  
Antifungal Protein ◽  
Black Soybean

A monomeric protein, with a molecular mass of 25 kDa and an N-terminal sequence resembling a segment of chitin synthase, was isolated from the seeds of the black soybean Glycine soja. The protein, designated glysojanin, demonstrated potent antifungal activity against the fungi Fusarium oxysporum and Mycosphaerella arachidicola. It inhibited HIV-1 reverse transcriptase with an IC50 of 47 µmol/L, [methyl-3H]thymidine incorporation by mouse spleen cells with an IC50 of 175 µmol/L, and translation in the rabbit reticulocyte lysate with an IC50 of 20 µmol/L. Glysojanin was purified using a procedure that involved ion-exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel, ion-exchange chromatography by fast protein liquid chromatography on Mono S, and gel filtration by fast protein liquid chromatography on Superdex 75.Key words: antifungal protein, seeds, soybean, purification.

scholarly journals The N-acetylhexosaminidase components of the ram testis and epididymis

1973 ◽  
Vol 133 (3) ◽  
pp. 593-599 ◽  
Author(s):  
Sarah Bullock ◽  
Bryan Winchester
Keyword(s):  
Molecular Weight ◽  
Ion Exchange ◽  
Isoelectric Focusing ◽  
Catalytic Properties ◽  
Deae Cellulose ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Multiple Forms ◽  
The Individual

Three and four N-acetylhexosaminidase components, from ram testis and epididymis respectively, have been separated by ion-exchange chromatography on DEAE-cellulose. Although they all have the same molecular weight (approx. 140000) and very similar catalytic properties towards the synthetic substrates, 4-methylumbelliferyl N-acetyl-β-glucosaminide and N-acetyl-β-galactosaminide, isoelectric focusing of the individual components showed that each had a distinct pI value. Isoelectric focusing has also been used to demonstrate the occurrence of multiple forms in ejaculated ram semen.

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