scholarly journals Glucosylation of phosphorylpolyisoprenol and sterol at the plasma membrane of soya-bean (Glycine max) protoplasts

1980 ◽  
Vol 186 (2) ◽  
pp. 411-421 ◽  
Author(s):  
C M Chadwick ◽  
D H Northcote
Keyword(s):  
Plasma Membrane ◽  
Glycine Max ◽  
Neutral Lipids ◽  
Deae Cellulose ◽  
Soya Bean ◽  
Radioactive Material ◽  
Optimum Conditions ◽  
Radioactive Label ◽  
Acidic Fraction ◽  
Chloroform Methanol

Protoplasts were prepared from cells of soya-bean (Glycine max) suspension cultures and the plasma membrane was labelled with diazotized [G-3H]sulphanilic acid. Homogenates were fractionated by differential and isopycnic centrifugation, and membrane fractions in a density gradient were characterized by enzymic markers and the radioactive label. When fractions containing a large amount of protein were incubated with UDP-[U-14C]glucose, radioactive material soluble in chloroform/methanol was formed and this separated into acidic and neutral fractions on ion-exchange chromatograms of DEAE-cellulose. The acidic fraction was shown to consist of dolichol phosphate glucose, and the neutral fraction sterol glucosides and acylsterol glucosides. Optimum conditions for glucosylation of dolichol phosphate were established as 5 mM-MgCl2, pH 6.0, and the enzyme had a Michaelis constant of 1.5 × 10(-5) m-UDP-glucose. Optimum conditions for glucosylation of sterol were 5 mM-MgCl2, pH 8.0 GDP-[U-14C]glucose was a poor substrate for the synthesis of both acidic and neutral lipids. Although the synthesis of dolichol phosphate glucose and sterol glucosides occurred throughout the sucrose gradient, the specific activities of both glucosyltransferases were greatest in a fraction coincident with the radioactively labelled plasma membrane. Results are discussed in relation to the likely role fo these transglucosylase activities.

scholarly journals Dolichyl monophosphate and its sugar derivatives in plants

1977 ◽  
Vol 161 (1) ◽  
pp. 93-101 ◽  
Author(s):  
C T Brett ◽  
L F Leloir
Keyword(s):  
Enzyme Preparation ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Sodium Deoxycholate ◽  
Soya Bean ◽  
Pea Seedlings ◽  
Lipid Moiety ◽  
Sugar Derivatives ◽  
Chloroform Methanol ◽  
Cellulose Column

A glucose acceptor was isolated from soya beans by extraction with chloroform/methanol (2:1, v/v), followed by DEAE-cellulose column chromatography of the extract. This acceptor could not be distinguished from liver dolichyl monophosphate by t.l.c. It could replace dolichyl monophosphate as a mannose acceptor with a liver enzyme and its glucosylated derivative could replace dolichyl monophosphate glucose as a glucose donor in the same system. These results, together with those already reported [Pont Lezica, Brett, Romero Martinez & Dankert (1975) Biochem, Biophys. Res. Commun. 66, 980-987], indicate that the acceptor from soya bean is a dolichyl monophosphate. Gel filtration of its glucosylated derivative on Sephadex G-75 in the presence of sodium deoxycholate indicated that the acceptor contained 17 or 18 isoprene units. An enzyme preparation from pea seedlings was shown to use endogenous acceptors to form lipid phosphate sugars containing mannose and N-acetylglucosamine from GDP-mannose and UDP-N-acetylglucosamine. Chromatographic and degradative techniques indicated that the compounds formed were lipid monophosphate mannose, lipid pyrophosphate N-acetylglucosamine, lipid pyrophosphate chitobiose and a series of lipid pyrophosphate oligosaccharides containing both mannose and N-acetylglucosamine. None of these compounds was degraded by catalytic hydrogenation, and so the lipid moiety in each case was probably an alpha-saturated polyprenol. The endogenous acceptors for mannose and N-acetylglucosamine in peas may therefore be dolichyl monophosphate, as has been found in mammalian systems.

scholarly journals Androgen-sensitive spermine-binding protein of rat ventral prostate. Purification of the protein and characterization of the hormonal effect

1979 ◽  
Vol 184 (2) ◽  
pp. 431-440 ◽  
Author(s):  
G Mezzetti ◽  
R Loor ◽  
S Liao
Keyword(s):  
Binding Protein ◽  
Sodium Dodecyl ◽  
Deae Cellulose ◽  
Binding Activity ◽  
Ventral Prostate ◽  
Hormonal Effect ◽  
Rat Ventral Prostate ◽  
Acidic Fraction ◽  
Androgen Effect

The rat ventral prostate contains a cytosol protein that can non-covalently bind spermine much more tightly than spermidine or other natural diamines. The protein has been purified to homogeneity, as judged by electrophoresis in urea- and sodium dodecyl sulphate-containing polyacrylamide gels. The protein, with or without spermine bound to it, sediments at 3 S in a sucrose gradient with or without 0.4 M-KCl. The molecular weight of the protein is about 30 000. Each molecule of the binding protein can bind one molecule of spermine. In the prostate of rats injected with cycloheximide, the protein appears to have a half-life of about 3.5 h. The spermine-binding activity of an acidic fraction obtained by DEAE-cellulose chromatography of the prostate cytosol proteins is reduced by about 40–60% within 20–40 h after castration. This effect is reversed very rapidly within 15–30 min by intraperitoneal injection of 5 alpha-dihydrotestosterone. The hormonal effect is androgen-specific and is not mimicked by dexamethasone or oestradiol-17 beta. The androgen effect was reduced significantly when rats were injected with cycloheximide or actinomycin D, suggesting that the acidic protein may be one of the earliest proteins induced by androgen in the rat ventral prostate.

scholarly journals Chemical synthesis of all-trans-β-retinoyl phosphate

1976 ◽  
Vol 159 (3) ◽  
pp. 799-801 ◽  
Author(s):  
J P Frot-Coutaz ◽  
L M de Luca
Keyword(s):  
Retinoic Acid ◽  
Chemical Synthesis ◽  
Cellulose Acetate ◽  
Silica Gel ◽  
Absorption Maximum ◽  
Ammonium Acetate ◽  
Deae Cellulose ◽  
Thin Layers ◽  
Molar Ratio ◽  
Chloroform Methanol

all-trans-β-Retinoic acid is phosphorylated to retinoyl phosphate by bis(triethylamine) phosphate with yields of 10-15%. The product is soluble in methanol and is eluted from DEAE-cellulose acetate at a concentration of 0.1M-ammonium acetate in 99% (v/v) methanol. Its phosphate/retinoic acid molar ratio is 1. Retinoyl phosphate has an absorption maximum at 360nm in methanol, whereas retinoic acid has a maximum at 350 nm. The compound is hydrolysed at pH2 and pH13 for 20 min at 37 degrees C, but is relatively stable under the same conditions at pH4, 6, 8 and 10. Retinoyl phosphate (RF 0.1) can be separated from retinyl phosphate (RF 0.2) by chromatography on thin layers of silica gel in chloroform/methanol/water (60:25:4, by vol.).

scholarly journals Extraction and purification of L-Asparaginase II from local isolate of Proteus vulgaris

2011 ◽  
Vol 8 (1) ◽  
pp. 509-518
Author(s):  
Baghdad Science Journal
Keyword(s):  
Enzyme Production ◽  
Quantitative Methods ◽  
Specific Activity ◽  
Deae Cellulose ◽  
Ion Exchanger ◽  
Optimum Conditions ◽  
Proteus Vulgaris ◽  
Clinical Specimens ◽  
Extraction And Purification ◽  
High Level

Forty one isolates of genus Proteus were collected from 140 clinical specimens such as urine, stool, wound, burn, and ear swabs from patients of both sex. These isolates were identified to three Proteus spp. P. mirabilis, P. vulgaris and P. penneri .The ability of these bacteria to produce L-asparaginase II by using semi quantitative and quantitative methods was determined. P. vulgaris Pv.U.92 was distinguished for high level of L-asparaginase II production with specific activity 1.97 U/mg. Optimum conditions for enzyme production were determined; D medium with 0.3% of L-asparagine at pH 7.5 with temperature degree 35°C for incubation. Ultrasonication was used to destroy the P. vulgaris Pv.U.92 cells then ASNase II was extracted and purified throughout several purification steps including precipitation with (NH4)2SO4(60-80%), DEAE-cellulose ion exchanger chromatography followed by Sephacryl S-300 filtration. The specific activity was 155.6 U/ mg and the purification fold was 27.3 with 10.4% yield.

scholarly journals Supercritical CO2 Extraction of Nannochloropsis sp.: A Lipidomic Study on the Influence of Pretreatment on Yield and Composition

Molecules ◽  
2018 ◽  
Vol 23 (8) ◽  
pp. 1854 ◽  
Author(s):  
Kathy Elst ◽  
Miranda Maesen ◽  
Griet Jacobs ◽  
Leen Bastiaens ◽  
Stefan Voorspoels ◽  
...  
Keyword(s):  
Supercritical Co2 ◽  
Neutral Lipids ◽  
Lipid Extraction ◽  
Fatty Acid Distribution ◽  
Extraction Yield ◽  
Partial Hydrolysis ◽  
Complex Nature ◽  
Supercritical Co2 Extraction ◽  
Optimized Conditions ◽  
Chloroform Methanol

Algal lipids have gained wide interest in various applications ranging from biofuels to nutraceuticals. Given their complex nature composed of different lipid classes, a deep knowledge between extraction conditions and lipid characteristics is essential. In this paper, we investigated the influence of different pretreatments on lipid extraction with supercritical CO2 by a lipidomic approach. Pretreatment was found to double the total extraction yield, thereby reaching 23.1 wt.% comparable to the 26.9 wt.% obtained with chloroform/methanol. An increase in acylglycerides was concurrently observed, together with a nearly doubling of free fatty acids indicative of partial hydrolysis. Moreover, an alteration in the distribution of glyco- and phospholipids was noted, especially promoting digalactosyldiglycerides and phosphatidylcholine as compared to monogalactosyldiglycerides and phosphatidylglycerol. At optimized conditions, supercritical CO2 extraction provided a lipid extract richer in neutral lipids and poorer in phospholipids as compared to chloroform/methanol, though with a very similar fatty acid distribution within each lipid class.

Effect of Panchagavya, Humic Acid and Micro herbal Fertilizer on the Yield and Post Harvest Soil of Soya Bean (Glycine max L.)

2012 ◽  
Vol 11 (2) ◽  
pp. 83-86 ◽  
Author(s):  
B. Vijayakuma ◽  
R. Hiranmai Yadav ◽  
P. Gowri ◽  
L.S. Kandari
Keyword(s):  
Glycine Max ◽  
Humic Acid ◽  
Soya Bean ◽  
Post Harvest ◽  
Glycine Max L

Soya Bean (Glycine max L. Merr.) Genotype Response to Early-season Flooding: I. Root and Nodule Development

2007 ◽  
Vol 193 (3) ◽  
pp. 177-188 ◽  
Author(s):  
T. L. Henshaw ◽  
R. A. Gilbert ◽  
J. M. S. Scholberg ◽  
T. R. Sinclair
Keyword(s):  
Glycine Max ◽  
Soya Bean ◽  
Nodule Development ◽  
Early Season ◽  
Glycine Max L ◽  
Genotype Response

Elicitor-binding proteins and signal transduction in the activation of a phytoalexin defense response

1995 ◽  
Vol 73 (S1) ◽  
pp. 506-510 ◽  
Author(s):  
Jürgen Ebel ◽  
Markus Feger ◽  
Ulrich Kissel ◽  
Axel Mithöfer ◽  
Tom Waldmüller ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Glycine Max ◽  
Defense Response ◽  
Binding Sites ◽  
Binding Proteins ◽  
Cultured Cells ◽  
Phytophthora Sojae ◽  
Phytophthora Megasperma ◽  
Early Stages ◽  
Signalling Process

Inducible plant defenses against potential pathogens are thought to be activated by signal compounds released during early stages of the infection process. In the incompatible interaction between soybean (Glycine max L.) and the oomycete Phytophthora megasperma f.sp. glycinea (= Phytophthora sojae) a rapid, localized phytoalexin response is activated at the level of transcription. The phytoalexin response is also stimulated in various soybean tissues, including cultured cells, following treatment with an elicitor derived from the cell walls of the fungus. The best characterized elicitors of P. megasperma for soybean are the branched (1→3)- and (1→6)-linked β-glucans, structural polysaccharides of the hyphal walls. The glucans are naturally released during the early stages of germination of the fungal cysts in a host-independent manner. Cyclic β-glucans of Bradyrhizobium japonicum USDA 110, a symbiont of soybean, arc not active in inducing phytoalexin production in soybean. When tested in combination, B. japonicum β-glucans inhibited stimulation of phytoalexin accumulation by the fungal glucans. Surface-localized glucan-binding proteins exist in soybean cells that display high affinity and specificity for the fungal β-glucans, including an elicitor-active hepta-β-glucoside fragment derived from the polysaccharide, suggesting that elicitor action involves a transmembrane signalling process. The main component of the soybean β-glucan binding sites appears to be a 70-kDa protein. Hepta-β-glucoside binding sites exist in several other legumes, such as bean (Phaseolus vulgaris L.), pea (Pisum sativum L.), and lupine (Lupinus albus L.). The signalling process initiated by the β-glucan elicitor, which leads to the activation of the phytoalexin defense response in soybean, involves changes in the permeability of the plasma membrane to Ca2+ and H+. Chloride channel antagonists are more efficient than calcium channel antagonists in inhibiting both the phytoalexin response and the inducible ion fluxes. The results present evidence that the observed permeability changes of the plasma membrane are primary events in the transduction of the elicitor signal(s) by the challenged soybean cells. Key words: soybean (Glycine max), Phytophthora megasperma f.sp. glycinea, β-glucan elicitor, elicitor-binding proteins, phytoalexins, Ca2+.

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