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Biomedical subjects

T B Ng

Publications and source records attributed to T B Ng.

At least 37 records · Page 2Linked to original sources

Isolation of a ribonuclease from sanchi ginseng (Panax pseudoginseng) flowers distinct from other ginseng ribonucleases.

A single-chained ribonuclease was isolated from the aqueous extract of sanchi ginseng (Panax pseudoginseng) flowers. It exhibited a molecular mass of 23 kDa, an N-terminal sequence with some similarity to other enzymes involved in RNA metabolism but different from known ribonucleases, and considerably higher activity toward poly U than poly C and only slight activity toward poly A and poly G. The purification protocol entailed ion exchange chromatography on diethylaminoethyl (DEAE)-cellulose, affinity chromatography on Affi-gel blue gel, ion exchange chromatography on carboxymethyl (CM)-cellulose, and gel filtration on Superdex 75. The ribonuclease was unadsorbed on DEAE-cellulose and adsorbed on Affi-gel blue gel and CM-cellulose. Maximal activity of the ribonuclease was attained at pH 7. On either side of this pH the enzyme activity underwent a drastic decline. The enzyme activity was at its highest at 50 degrees C and dropped to about 20% of the maximal activity when the temperature was decreased to 20 degrees C or elevated to 80 degrees C. The characteristics of sanchi ginseng flower ribonuclease were different from those of the ribonucleases previously purified from sanchi ginseng and Chinese ginseng roots including ribonuclease from Chinese ginseng flowers which are morphologically very similar to sanchi ginseng flowers.

Amino Acid Sequence↗

Smilaxin, a novel protein with immunostimulatory, antiproliferative, and HIV-1-reverse transcriptase inhibitory activities from fresh Smilax glabra rhizomes.

A protein, with a novel N-terminal amino acid sequence and a molecular mass of 30 kDa, was purified from fresh Smilax glabra rhizomes by adsorption on DEAE-cellulose, CM-cellulose, Con A-Sepharose, and Mono S, and by fast protein liquid chromatography-gel filtration on Superdex 75. The protein, designated as smilaxin, stimulated uptake of [methyl-3H]thymidine by murine splenocytes, peritoneal macrophages, and bone marrow cells, and production of nitric oxide by peritoneal macrophages. It inhibited uptake of [methyl-3H]thymidine by MBL2 and PU5 tumor cells but not uptake by S180 and L1210 cells. Smilaxin augmented glucose uptake into rat adipose tissue. It attenuated the activity of HIV-1-reverse transcriptase with an IC50 of 5.6 microM. However, it did not display hemagglutinating, antifungal or translation-inhibitory activities, indicating that it is not a lectin, an antifungal protein, or a ribosome-inactivating protein.

Animals↗

Concurrent isolation of a Kunitz-type trypsin inhibitor with antifungal activity and a novel lectin from Pseudostellaria heterophylla roots.

A simple purification protocol, involving ion exchange chromatography on DEAE-cellulose and CM-cellulose and fast protein liquid chromatography-gel filtration on Superdex 75, was employed to isolate a Kunitz-type trypsin inhibitor with antifungal activity and a novel lectin from Pseudostellaria heterophylla roots. Both the trypsin inhibitor and the lectin were unadsorbed on DEAE-cellulose and adsorbed on CM-cellulose. They could be separated from one another by gel filtration on Superdex 75 in which the 36-kDa lectin appeared as the first peak and the 20.5-kDa trypsin inhibitor as the second peak. P. heterophylla trypsin inhibitor exhibited a trypsin inhibitory potency similar to that of soybean trypsin inhibitor. It also demonstrated antifungal activity toward Fusarium oxysporum like aprotinin and Kunitz-type trypsin inhibitors from soybeans and lima beans. P. heterophylla lectin was devoid of antifungal activity and exhibited low thermostability and also lability in the presence of acid and alkali. The novel aspects of the present report include demonstration of antifungal activity in Kunitz-type trypsin inhibitors and isolation of a novel lectin as well as a trypsin inhibitor from roots.

Amino Acid Sequence↗

Limenin, a defensin-like peptide with multiple exploitable activities from shelf beans.

From the seeds of the shelf bean, an antifungal peptide with a molecular mass of 6.5 kDa was isolated. The isolation procedure comprised affinity chromatography on Affi-gel blue gel, ion exchange chromatography on Mono S, and gel filtration on Superdex 75. The peptide was adsorbed on Affi-gel blue gel and Mono S. It potently suppressed mycelial growth in Botrytis cinerea, Fusarium oxysporum, and Mycosphaerella arachidicola with an IC(50) of 2.9, 2.1, and 0.34 microM, respectively. It exerted antibacterial activity toward several bacterial species with an IC(50) approximating 100 microM. [Methyl-(3)H]-thymidine incorporation into isolated mouse splenocytes was stimulated. [Methyl-(3)H]-thymidine incorporation into M1 (myeloma) and L1210 (leukemia) cells was inhibited. The peptide reduced the activity of HIV-1 reverse transcriptase and also inhibited translation in a cell-free rabbit reticulocyte lysate system.

Amino Acid Sequence↗

A novel ribonuclease from fresh fruiting bodies of the portabella mushroom Agaricus bisporus.

A 14 kDa ribonuclease with a novel N-terminal sequence was isolated from fresh fruiting bodies of the portabella mushroom. It was adsorbed on DEAE-cellulose and carboxymethyl-cellulose, and demonstrated the highest ribonucleolytic potency toward poly (A), 60% as much activity toward poly (C), 40% as much activity toward poly (U), and the least activity (7% as much) toward poly (G). It exhibited a pH optimum at pH 4.5 and a temperature optimum at 60 degrees C. Its activity at 100 degrees C was higher than that at 20 degrees C.

Agaricus↗

A polysaccharopeptide complex and a condensed tannin with antioxidant activity from dried rose (Rosa rugosa) flowers.

In this study, the fraction (P) from an aqueous extract of dried rose (Rosa rugosa) flowers was obtained by ethanol precipitation. P was chromatographed on DEAE-cellulose. The components retained on DEAE-cellulose were eluted with a linear gradient of 0-2 M NaCl solution. Two fractions, eluted at concentrations of 0.5 M NaCl and 1 M NaCl, respectively, were obtained. These two components were designated as P1 and P2, respectively. P1 was further purified using gel filtration on Sephadex G-200. P(1) yielded two peaks, and the two components were designated as P(1-a) and P(1-b), respectively. P(1-a) was a polysaccharide-peptide complex, and P(1-b) exhibited chemical properties of a condensed tannin as revealed by FTIR and NMR assay of carbohydrate and protein contents and HPLC-ESI-MS. The molecular masses of P(1-a) and P(1-b) were 150 kDa and 8 kDa, respectively. Both P(1-a) and P(1-b) possessed antioxidant activity, with the activity of P(1-b) higher than that of P(1-a). This study demonstrated that different components from rose flowers exhibited antioxidant activity.

Animals↗

Pharmacological activity of sanchi ginseng (Panax notoginseng).

The pharmacological activity and constituents of the sanchi ginseng Panax notoginseng have been reviewed. The bulk of pharmacological findings have been based on the saponins or steryl glycosides, although polysaccharides with immunopotentiating activity, proteins with antifungal, ribonuclease and xylanase activity, and a triacylglycerol (trilinolein) with antioxidant activity have been reported. Protective actions against cerebral ischaemia, beneficial effects on the cardiovascular system, and haemostatic, antioxidant, hypolipidaemic, hepatoprotective, renoprotective and estrogen-like activities have been described. Various methods for authentication of P. notoginseng are available.

Animals↗

Compounds from rose (Rosa rugosa) flowers with human immunodeficiency virus type 1 reverse transcriptase inhibitory activity.

The aqueous extracts and ethanol precipitates of aqueous extracts of 18 medicinal herbs traditionally used in China were screened for their ability to inhibit human immunodeficiency virus type-1 reverse transcriptase (HIV-1 RT) in-vitro. Among the samples screened at a concentration of 500 microg mL-1, dried rose (Rosa rugosa) flowers showed the strongest inhibition. The ethanol precipitate of the aqueous extract of R. rugosa was processed and two components (P1 and P2) were obtained after ion exchange chromatography on DEAE-cellulose. Then, P1-a (Mr 150 kDa) and P1-b (Mr 8 kDa) were isolated from P1 by gel filtration on Sephadex G-200. They inhibited the activity of HIV-1 RT with an IC50 of 158 nM and 148.16 microg mL-1 (18.5 microM), respectively. Further structural analyses revealed that P1-a was a polysaccharide-peptide complex, and P1-b was a polymer consisting of acteoside and acteoside derivatives identified by Fourier transform infrared spectroscopy, nuclear magnetic resonance, assays of carbohydrate and protein contents and high-performance liquid chromatography electrospray ionization mass spectrometry.

Chromatography, Gel↗

A homotetrameric agglutinin with antiproliferative and mitogenic activities from haricot beans.

A homotetrameric agglutinin with a molecular mass of 130 kDa was isolated from seeds of the haricot bean. The agglutinin was isolated using a procedure that involved ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel and gel filtration by fast protein liquid chromatography on Superdex 200. Haricot bean agglutinin was adsorbed on DEAE-cellulose and Affi-gel blue gel. The hemagglutinating activity of the agglutinin was stable up to 40 degrees C. It underwent a 40% decline when the temperature was raised to 50 degrees C and a complete loss when the temperature was further increased to 80 degrees C. The hemagglutinating activity exhibited a time-dependent loss in activity when the agglutinin was incubated at 100 degrees C for different durations. No activity was discernible when the agglutinin was left at 100 degrees C for 1 min. The activity also underwent a decline in the presence of 500 mM FeCl(3) and CaCl(2). Haricot bean agglutinin manifested a weaker mitogenic activity than concanavalin A toward mouse splenocytes. It exhibited antiproliferative activity toward the tumor cell lines M1 [leukemia], HepG2 [hepatoma] and L1210 [leukemia] cells.

Animals↗

A novel homodimeric lactose-binding lectin from the edible split gill medicinal mushroom Schizophyllum commune.

A homodimeric lactose-binding lectin with a molecular mass of 64kDa was isolated from fresh fruiting bodies of the split gill mushroom Schizophyllum commune. The N-terminal sequence of the lectin is similar to a part of the sequence of the cell division protein from Gleobacter violaceus. The lectin was isolated by using a procedure which involved ion exchange chromatography on DEAE-cellulose, CM-cellulose, and Q-Sepharose, and gel filtration by fast protein liquid chromatography on Superdex 75. The hemagglutinating activity of the lectin was stable at temperatures up to 40 degrees C, and in concentrations of NaOH and HCl solution up to 125 and 25mM, respectively. The lectin exhibited potent mitogenic activity toward mouse splenocytes, antiproliferative activity toward tumor cell lines, and inhibitory activity toward HIV-1 reverse transcriptase. It was devoid of antifungal activity.

Amino Acid Sequence↗

Isolation and characterization of a glucose/mannose-specific lectin with stimulatory effect on nitric oxide production by macrophages from the emperor banana.

Emperor banana (Musa basjoo cv. 'Emperor Banana') is a banana cultivar that has not been studied previously. In this study, a glucose/mannose-specific lectin has been purified from the emperor banana by affinity chromatography on Affi-gel blue gel, ion exchange chromatography on Mono S and gel filtration by fast protein liquid chromatography on Superdex 75. This lectin was composed of two identical 15-kDa subunits with N-terminal amino acid sequence similarity to other lectins from other Musa species. Emperor banana lectin stimulated [3H-methyl]-thymidine uptake by mouse splenocytes and nitric oxide production by mouse macrophages. In contrast to Con A, the mitogenic activity of emperor banana lectin toward mouse splenocytes but not its stimulatory effect on nitric oxide production by mouse macrophages could be abrogated by 200 mM glucose. Emperor banana lectin also inhibited proliferation of leukemia cell (L1210) and the activity of HIV-1 reverse transcriptase. In summary, this is the first report of the macrophage-stimulating, antiproliferative and HIV-1 reverse transcriptase inhibiting activities of a banana lectin.

Amino Acid Sequence↗

Purification of a laccase from fruiting bodies of the mushroom Pleurotus eryngii.

A purification scheme involving ion exchange chromatography on diethylaminoethyl (DEAE) cellulose, carboxymethyl (CM) cellulose, and Q-Sepharose, followed by fast protein liquid chromatography-gel filtration on Superdex 75, was utilized to isolate a laccase from the fruiting bodies of the mushroom Pleurotus eryngii. The laccase was adsorbed on DEAE-cellulose and Q-Sepharose and unadsorbed on CM-cellulose. The molecular mass of the laccase in sodium dodecyl sulfate-polyacrylamide gel electrophoresis was 34 kDa. The laccase activity was maximal at 70 degrees C and remained high at 80 degrees C. High activity of the laccase was maintained at ph 3 to 5. The activity underwent an abrupt decline at pH 6 and 7 and was indiscernible at ph 8 and 9. The laccase exhibited an inhibitory activity toward HIV-1 reverse transcriptase with an IC50 of 2.2 microM. Its N-terminal sequence was dissimilar to those of laccase isoenzymes previously isolated from cultured mycelia of the same species.

Amino Acid Sequence↗

Ganodermin, an antifungal protein from fruiting bodies of the medicinal mushroom Ganoderma lucidum.

A 15-kDa antifungal protein, designated ganodermin, was isolated from the medical mushroom Ganoderma lucidum. The isolation procedure utilized chromatography on DEAE-cellulose, Affi-gel blue gel, CM-Sepharose and Superdex 75. Ganodermin was unadsorbed on DEAE-cellulose and adsorbed on Affi-gel blue gel and CM-Sepharose. Ganodermin inhibited the mycelial growth of Botrytis cinerea, Fusarium oxysporum and Physalospora piricola with an IC50 value of 15.2 microM, 12.4 microM and 18.1 microM, respectively. It was devoid of hemagglutinating, deoxyribonuclease, ribonuclease and protease inhibitory activities.

Amino Acid Sequence↗

An antifungal protein from flageolet beans.

A protein with antifungal and hemagglutinating activities was isolated from dried flageolet beans (Phaseolus vulgaris cv. 'Flageolet Bean'). The protein was unadsorbed on DEAE-cellulose but adsorbed on Affi-gel blue gel and CM-cellulose. The protein demonstrated antifungal activity against Mycophaerella arachidicola with an IC50 of 9.8 microM, but was inactive toward Fusarium oxysporum and Botrytis cinerea. Its hemagglutinating activity could not be inhibited by a variety of the sugars tested. The activity was stable up to 60 degrees C. At 70 degrees C, 75% of the hemagglutinating activity remained while no activity was discernible at and above 100 degrees C. The hemagglutinating activity was stable in the presence of a variety of monovalent, divalent and trivalent chlorides, and also when the ambient pH changed from 3 to 12. It did not exert any mitogenic activity on mouse splenocytes in vitro. Neither did it inhibit HIV-1 reverse transcriptase. It inhibited [3H-methyl]-thymidine incorporation into leukemia L1210 cells with an IC50 of about 4 microM.

Animals↗

Isarfelin, a peptide with antifungal and insecticidal activities from Isaria felina.

Isarfelin, a peptide with inhibitory activity on mycelial growth in Rhizoctonia solani and Sclerotinia sclerotiorum and insecticidal activity toward Leucania separata, was isolated from the mycelia of Isaria felina. The IC50 value of its antifungal activity against R. solani was 3.1 microg mL(-1). However, it was devoid of activity toward several bacterial species including Bacillus subtilis, E. coli and Staphylococcus aureus. The isolation procedure involved ethanol extraction, adsorption on YPR II macropore adsorption resin, ethyl acetate extraction, petroleum ether precipitation and recrystallization from ethyl acetate.

Animals↗

Isolation and characterization of a glucose/mannose/rhamnose-specific lectin from the knife bean Canavalia gladiata.

A lectin with specificity toward mannose, glucose, and rhamnose has been isolated from the legumes of the knife bean Canavalia gladiata. The lectin is composed of two identical 30-kDa subunits with substantial N-terminal sequence similarity to Concanavalin A (Con A). It was purified by affinity chromatography on Affi-gel blue gel, ion exchange chromatography on Mono S, and gel filtration by fast protein liquid chromatography on Superdex 200. Compared with Con A, knife bean lectin started to stimulate [methyl-(3)H]thymidine uptake by mouse splenocytes at a lower concentration, and more potently inhibited proliferation of L1210 leukemia cells. In contrast to Con A, the mitogenic activity of knife bean lectin toward mouse splenocytes, but not its antiproliferative activity toward tumor cells, could be abrogated by 250 mM glucose. Both mitogenic and antiproliferative activities of Con A were abolished by glucose. The lectin inhibited HIV-1 reverse transcriptase with an IC(50) of 35 microM and cell-free translation in a rabbit reticulocyte lysate system with an IC(50) of 2.08 microM. The lectin did not exhibit antifungal activity.

Animals↗

Pleurostrin, an antifungal peptide from the oyster mushroom.

A 7kDa peptide, with inhibitory activity on mycelial growth in the fungi Fusaerium oxysporum, Mycosphaerella arachidicola and Physalospora piricola, was isolated from fresh fruiting bodies of the oyster mushroom. The isolation procedure entailed extraction with an aqueous buffer, ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel and gel filtration by fast protein liquid chromatography on Superdex 75. The protein was unadsorbed on DEAE-cellulose and adsorbed on Affi-gel blue gel. It demonstrated an N-terminal sequence different from known antifungal proteins and peptides.

Antifungal Agents↗

Antioxidant activity of gallic acid from rose flowers in senescence accelerated mice.

The activities of the antioxidant enzymes catalase (CAT) and glutathione peroxidase (GPx) in the blood and liver of the aging model induced by injection of different doses of D-gal into normal mice, and in senescence accelerated mice (SAM) of different ages, were determined. The results showed that the enzyme activities in the D-gal treated mice did not alter appreciably but the enzyme activities in blood of the SAM declined significantly with the increase in age, especially in the 9-month-old SAM. When gallic acid purified from rose flowers was used to treat the 9-month-old male SAM, it not only reinstated the activities of CAT and GPx but also significantly reduced the amount of malondialdehyde (MDA) in the liver, brain and kidney. The results demonstrated that 9-month-old male SAM represent an appropriate animal model to evaluate the antioxidant activities of natural products.

Aging↗