scholarly journals Novel galactonic acid-binding hexameric lectin from Hibiscus mutabilis seeds with antiproliferative and potent HIV-1 reverse transcriptase inhibitory activities.

2009 ◽  
Vol 56 (4) ◽  
Author(s):  
Sze Kwan Lam ◽  
Tzi Bun Ng
Keyword(s):  
Reverse Transcriptase ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Inhibitory Activities ◽  
Binding Lectin ◽  
Hiv 1 ◽  
Hepatoma Hepg2 ◽  
Mcf 7

A hexameric 150-kDa lectin was isolated from dried Hibiscus mutabilis seeds using a chromatographic protocol that involved ion exchange chromatography on SP-Sepharose, and gel filtration on Superdex 75 and Superdex 200. The lectin was not adsorbed on SP-Sepharose and was eluted from the Superdex 75 column in the void volume. It was eluted in the first peak from Superdex 200. It was strongly adsorbed on DEAE-cellulose and Q-Sepharose and could not be easily desorbed. The hemagglutinating activity of the lectin, which was stable at pH 4-7 and up to 50 degrees C, could be inhibited by 25 mM galactonic acid. This is the first report of a galactonic acid-binding lectin. It potently inhibited HIV-1 reverse transcriptase with an IC(50) of 0.2 microM. It exhibited weak antiproliferative activity towards both hepatoma HepG2 cells (40% inhibition) and breast cancer MCF-7 cells (50% inhibition) at 100 microM concentration of the lectin. It did not inhibit mycelial growth of a number of fungi tested.

scholarly journals A Laccase with Antiproliferative and HIV-I Reverse Transcriptase Inhibitory Activities from the Mycorrhizal FungusAgaricus placomyces

2012 ◽  
Vol 2012 ◽  
pp. 1-8 ◽  
Author(s):  
Jian Sun ◽  
Qing-Jun Chen ◽  
Qing-Qin Cao ◽  
Ying-Ying Wu ◽  
Li-Jing Xu ◽  
...  
Keyword(s):  
Reverse Transcriptase ◽  
Mycorrhizal Fungus ◽  
Gel Filtration ◽  
Exchange Chromatography ◽  
Hep G2 ◽  
Immunodeficiency Virus ◽  
Terminal Amino ◽  
Hiv 1 ◽  
Mcf 7

A novel 68 kDa laccase was purified from the mycorrhizal fungusAgaricus placomycesby utilizing a procedure that comprised three successive steps of ion exchange chromatography and gel filtration as the final step. The monomeric enzyme exhibited the N-terminal amino acid sequence of DVIGPQAQVTLANQD, which showed only a low extent of homology to sequences of other fungal laccases. The optimal temperature forA. placomyceslaccase was 30°C, and optimal pH values for laccase activity towards the substrates 2,7′-azinobis[3-ethylbenzothiazolone-6-sulfonic acid] diammonium salt (ABTS) and hydroquinone were 5.2 and 6.8, respectively. The laccase displayed, at 30°C and pH 5.2,Kmvalues of 0.392 mM towards hydroquinone and 0.775 mM towards ABTS. It potently suppressed proliferation of MCF 7 human breast cancer cells and Hep G2 hepatoma cells and inhibited human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) activity with an IC50of 1.8 μM, 1.7 μM, and 1.25 μM, respectively, signifying that it is an antipathogenic protein.

scholarly journals A Laccase with HIV-1 Reverse Transcriptase Inhibitory Activity from the Broth of Mycelial Culture of the MushroomLentinus tigrinus

2012 ◽  
Vol 2012 ◽  
pp. 1-7 ◽  
Author(s):  
LiJing Xu ◽  
HeXiang Wang ◽  
TziBun Ng
Keyword(s):  
Reverse Transcriptase ◽  
Inhibitory Activity ◽  
Laccase Activity ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Exchange Chromatography ◽  
Mycelial Culture ◽  
Lentinus Tigrinus ◽  
Terminal Amino ◽  
Hiv 1

A 59 kDa laccase with inhibitory activity against HIV-1 reverse transcriptase (IC50=2.4 μM) was isolated from the broth of mycelial culture of the mushroomLentinus tigrinus. The isolation procedure involved ion exchange chromatography on DEAE-cellulose and CM-cellulose, and gel filtration by fast protein liquid chromatography on Superdex 75. The laccase was adsorbed on both types of ion exchangers. About 95-fold purification was achieved with a 25.9% yield of the enzyme. The procedure resulted in a specific enzyme activity of 76.6 U/mg. Its N-terminal amino acid sequence was GIPDLHDLTV, which showed little similarity to other mushroom laccase and otherLentinus tigrinusstrain laccase. Its characteristics were different from previously reported laccase of otherLentinus tigrinusstrain. Maximal laccase activity was observed at a pH of 4 and at a temperature of 60°C, respectively. This study yielded the information about the potentially exploitable activities ofLentinus tigrinuslaccase.

scholarly journals A Novel Laccase with Potent Antiproliferative and HIV-1 Reverse Transcriptase Inhibitory Activities from Mycelia of MushroomCoprinus comatus

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
Shuang Zhao ◽  
Cheng-Bo Rong ◽  
Chang Kong ◽  
Yu Liu ◽  
Feng Xu ◽  
...  
Keyword(s):  
Reverse Transcriptase ◽  
Ph Value ◽  
Sequence Similarity ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Exchange Chromatography ◽  
High Sequence Similarity ◽  
Immunodeficiency Virus ◽  
Filtration Step ◽  
Hiv 1

A novel laccase was isolated and purified from fermentation mycelia of mushroomCoprinus comatuswith an isolation procedure including three ion-exchange chromatography steps on DEAE-cellulose, CM-cellulose, and Q-Sepharose and one gel-filtration step by fast protein liquid chromatography on Superdex 75. The purified enzyme was a monomeric protein with a molecular weight of 64 kDa. It possessed a unique N-terminal amino acid sequence of AIGPVADLKV, which has considerably high sequence similarity with that of other fungal laccases, but is different from that ofC. comatuslaccases reported. The enzyme manifested an optimal pH value of 2.0 and an optimal temperature of 60°C using 2,2′-azinobis(3-ethylbenzothiazolone-6-sulfonic acid) diammonium salt (ABTS) as the substrate. The laccase displayed, at pH 2.0 and 37°C,Kmvalues of 1.59 mM towards ABTS. It potently suppressed proliferation of tumor cell lines HepG2 and MCF7, and inhibited human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) with an IC50value of 3.46 μM, 4.95 μM, and 5.85 μM, respectively, signifying that it is an antipathogenic protein.

scholarly journals The isocitrate dehydrogenases of Acinetobacter lwoffi. Separation and properties of two nicotinamide–adenine dinucleotide phosphate-linked isoenzymes

1972 ◽  
Vol 130 (1) ◽  
pp. 211-219 ◽  
Author(s):  
Colin H. Self ◽  
P. David J. Weitzman
Keyword(s):  
Molecular Weight ◽  
Ph Dependence ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Nicotinamide Adenine Dinucleotide Phosphate ◽  
Molecular Size ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Low Concentrations ◽  
Stimulation Of

Two isoenzymes of NADP-linked isocitrate dehydrogenase have been identified in Acinetobacter lwoffi and have been termed isoenzyme-I and isoenzyme-II. The isoenzymes may be separated by ion-exchange chromatography on DEAE-cellulose, by gel filtration on Sephadex G-200, or by zonal ultracentrifugation in a sucrose gradient. Low concentrations of glyoxylate or pyruvate effect considerable stimulation of the activity of isoenzyme-II. The isoenzymes also differ in pH-dependence of activity, kinetic parameters, stability to heat or urea and molecular size. Whereas isoenzyme-I resembles the NADP-linked isocitrate dehydrogenases from other organisms in having a molecular weight under 100000, isoenzyme-II is a much larger enzyme (molecular weight around 300000) resembling the NAD-linked isocitrate dehydrogenases of higher organisms.

THYROID-STIMULATING AND LIPOLYTIC ACTIVITIES OF PURIFIED PREPARATIONS OF HUMAN THYROID-STIMULATING HORMONE

1972 ◽  
Vol 53 (1) ◽  
pp. 95-100 ◽  
Author(s):  
ANNE STOCKELL HARTREE ◽  
MARJORIE THOMAS ◽  
BRIDGET E. FURNIVAL ◽  
T. W. BURNS ◽  
P. LANGLEY
Keyword(s):  
Lipolytic Activity ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Protein Molecule ◽  
Thyroid Stimulating Hormone ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Human Thyroid ◽  
Rat Adipose Tissue ◽  
Stimulating Hormone

SUMMARY A partially purified fraction of human thyroid-stimulating hormone (DEAE-II) was further purified by ion-exchange chromatography on IRC-50, gel-filtration on Sephadex G-100 and finally chromatography on DEAE-cellulose. Two fractions were obtained which were high in thyroid-stimulating activity (8·3 and 7·3 units human Research Standard A/mg) and were comparable in potency to other preparations of the human hormone reported in the literature. They were also electrophoretically heterogeneous as were the preparations of other workers. Lipolytic activity toward cells obtained from human or rat adipose tissue was demonstrated for all fractions containing thyroid-stimulating activity, the two activities being roughly parallel. It is concluded that both thyroid-stimulating and lipolytic activities are probably present in the same protein molecule, but it is unlikely that the latter activity is of physiological significance.

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.

Comparative studies of manganese(II)-, nickel(II)-, zinc(II)-, copper(II)-, cadmium(II)-, and iron(III)-binding components in human cord and adult sera

1984 ◽  
Vol 62 (6) ◽  
pp. 449-455 ◽  
Author(s):  
Show-Jy Lau ◽  
Bibudhendra Sarkar
Keyword(s):  
Molecular Weight ◽  
Trace Metals ◽  
Ion Exchange ◽  
Comparative Studies ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Low Molecular Weight ◽  
High Molecular Weight Proteins

The binding of six trace metals, Mn(II), Ni(II), Zn(II), Cu(II), Cd(II) and Fe(III), to human cord serum has been studied by Sephadex G-100 gel filtration at physiological pH, using radioisotopes as tracers. The results are compared with those obtained from adult serum. In both cord and adult sera, extensive amounts of the metals are bound to high molecular weight proteins. Among them, Fe(III) is mostly bound to transferrin; Ni(II), Zn(II), Cu(II), and Cd(II) are bound to albumin and other macro-molecules. The binding of Mn(II) either to transferrin or albumin is not resolved. Small fractions of Zn(II), Cu(II), and Cd(II) and large fractions of Mn(II) and Ni(II) are found to be associated with low molecular weight components of both sera. The distribution varies from metal to metal. However, the low molecular weight component of the size 1500 – 10 000 is present in all the metals studied. Further purification of this component was attempted by DEAE-cellulose ion-exchange chromatography. The possible identity as well as the biological role played by this particular component of serum in the transport of metals in blood and across membranes is discussed.

Isolation of deoxyribonuclease II from bovine intestinal mucosa

1980 ◽  
Vol 58 (9) ◽  
pp. 749-753 ◽  
Author(s):  
D. Stephen Keys ◽  
S. H. Zbarsky
Keyword(s):  
Gel Filtration ◽  
Deae Cellulose ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Degraded Dna ◽  
Major Peak ◽  
Dnase Ii ◽  
Minor Peak ◽  
Supernatant Solution ◽  
Bovine Small Intestine

Mucosa from bovine small intestine was homogenized in Krebs–Ringer phosphate buffer, pH 7.8, the homogenate centrifuged at 16 300 × g, and the supernatant solution filtered through cheesecloth to remove lipid material. The filtrate was centrifuged at 105 000 × g and the supernatant solution chromatographed on DEAE-cellulose. The major peak of DNase II activity, eluted with 20 mM phosphate – 10 mM EDTA buffer, pH 7.8, was purified further by ion-exchange chromatography on CM-cellulose and gel filtration on Sephadex G-100. The enzyme was purified 78-fold in 13% yield. Evidence was adduced to indicate that the second minor peak of DNase II activity, eluted from the DEAE-cellulose by a potassium chloride gradient in the 20 mM phosphate – 10 mM EDTA buffer, was an artifact arising from the presence of significant amounts of DNA in the 105 000 × g supernatant. The enzyme degraded DNA endonucleolytically to 3′-PO4, 5′-OH oligonucleotides and is similar in its properties to DNase II from other tissues.

scholarly journals Antitumor and HIV-1 Reverse Transcriptase Inhibitory Activities of a Hemagglutinin and a Protease Inhibitor from Mini-Black Soybean

2011 ◽  
Vol 2011 ◽  
pp. 1-12 ◽  
Author(s):  
Xiu Juan Ye ◽  
Tzi Bun Ng
Keyword(s):  
Breast Cancer ◽  
Reverse Transcriptase ◽  
Inhibitory Activity ◽  
Breast Cancer Cells ◽  
Deae Cellulose ◽  
Defense Proteins ◽  
Black Soybean ◽  
Trypsin Inhibitory Activity ◽  
Inhibitory Activities ◽  
Hiv 1

Protease inhibitors (PIs) and hemagglutinins are defense proteins produced by many organisms. From Chinese mini-black soybeans, a 17.5-kDa PI was isolated using chromatography on Q-Sepharose, SP-Sepharose, and DEAE-cellulose. A 25-kDa hemagglutinin was purified similarly, but using Superdex 75 instead of DEAE-cellulose in the final step. The PI inhibited trypsin and chymotrypsin (IC50= 7.2 and 8.8 μM). Its trypsin inhibitory activity was stable from pH 2 to pH 13 and from 0∘C to 70∘C. The hemagglutinin activity of the hemagglutinin was stable from pH 2 to pH 13 and from 0∘C to 75∘C. The results indicated that both PI and hemagglutinin were relatively thermostable and pH-stable. The trypsin inhibitory activity was inhibited by dithiothreitol, signifying the importance of the disulfide bond to the activity. The hemagglutinating activity was inhibited most potently by D (+)-raffinose and N-acetyl-D-galactosamine, suggesting that the hemagglutinin was specific for these two sugars. Both PI and hemagglutinin inhibited HIV-1 reverse transcriptase (IC50= 3.2 and 5.5 μM), proliferation of breast cancer cells (IC50= 9.7 and 3.5 μM), and hepatoma cells (IC50= 35 and 6.2 μM), with relatively high potencies.

Isolation and characterization of a mannose-binding lectin from leaves of the Chinese daffodil Narcissus tazetta

1998 ◽  
Vol 76 (4) ◽  
pp. 601-608 ◽  
Author(s):  
Linda SM Ooi ◽  
Hexiang Wang ◽  
T B Ng ◽  
Vincent EC Ooi
Keyword(s):  
Gel Filtration ◽  
Deae Cellulose ◽  
Exchange Chromatography ◽  
Mannose Binding Lectin ◽  
Narcissus Tazetta ◽  
Isolation And Characterization ◽  
Mannose Binding ◽  
Narcissus Pseudonarcissus ◽  
Considerable Homology ◽  
Binding Lectin

A mannose-binding lectin was isolated from leaves of the Chinese daffodil Narcissus tazetta (family Amaryllidaceae) using a procedure that comprised extraction with aqueous buffer, ammonium sulfate precipitation, ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel Blue gel and mannose-agarose, and FPLC-gel filtration on Superose 12. The lectin was adsorbed on mannose-agarose and unadsorbed on DEAE-cellulose and Affi-gel Blue gel. It was an unglycosylated homodimer with a molecular mass of 26 kDa. Analysis of the N-terminal sequence of the N. tazetta lectin revealed considerable homology to lectins from the daffodil Narcissus pseudonarcissus, the snowdrop Galanthus nivalis (family Amaryllidaceae), the tulip Tulipa, and Kidachi aloe Aloe arborescens (family Liliaceae), and the orchid lectins (family Orchidaceae). The most striking likeness exists among the Amaryllidaceae lectins. The N. tazetta lectin exhibits hemagglutinating activity toward rabbit erythrocytes.Key words: daffodil, mannose-binding lectin, agglutinin.

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