Partial purification and properties of apoplastic β-1,3 glucanases of groundnut leaves treated with glucan isolated from a biocontrol agent, Acremonium obclavatum

2000 ◽  
Vol 78 (2) ◽  
pp. 168-174 ◽  
Author(s):  
M Sathiyabama ◽  
R Balasubramanian
Keyword(s):  
Arachis Hypogaea ◽  
Partial Purification ◽  
Ph Optimum ◽  
Native Page ◽  
Puccinia Arachidis ◽  
Ammonium Sulphate Precipitation ◽  
Arachis Hypogaea L ◽  
Purification And Properties ◽  
Molecular Masses ◽  
Electrophoretic Techniques

Apoplastic β-1,3 glucanases (G1, G2) of Arachis hypogaea L. (peanut) leaves treated with glucan have been partially purified by ammonium sulphate precipitation, sephadex G-100, CM-Sephadex, DEAE-Sephadex chromatography, and preparative native PAGE electrophoretic techniques. The pI values of purified enzymes G1 and G2 were near 8 and 4, respectively. The apparent molecular masses of purified glucanases G1 and G2 from glucan-treated peanut leaves were 36 and 34 kDa, respectively. Both isoforms (G1 and G2) showed their pH optimum of 5.0 and temperature optimum of 40°C. The partially purified enzymes hydrolysed laminarin better than other substrates and inhibited uredospore germination of Puccinia arachidis. Both isoforms (G1 and G2) inhibited spore germination of some biocontrol agents such as Acremonium obclavatum W. Gams, Myrothecium verrucaria (Alb. Schw.) Ditmer, Fusarium solani (Mart.) Sacc., and Trichoderma harzianum Rifai.Key words: Acremonium obclavatum, Arachis hypogaea, β-1,3 glucanase, glucan, inhibition, Puccinia arachidis.

scholarly journals Extraction, Characterization and Partial Purification of L-Asparaginase from the Leaves of Arachis Hypogaea L.

2019 ◽  
Vol 16 (3) ◽  
pp. 681-691
Author(s):  
Niranjana. J Niranjana ◽  
Kandasamy Arun Gandhi ◽  
D. Sunmathi ◽  
P. Nanthavanan
Keyword(s):  
Metal Ions ◽  
Arachis Hypogaea ◽  
Specific Activity ◽  
Lymphoblastic Leukemia ◽  
Gel Filtration ◽  
Size Exclusion ◽  
Partial Purification ◽  
Crude Enzyme Extract ◽  
Ammonium Sulphate Precipitation ◽  
Arachis Hypogaea L

L-asparaginase has been a promising therapeutic agent in the treatment of acute lymphoblastic leukemia and lymphoma. In recent times, due to the side effects of commercially available bacterial L-asparaginase and its unavoidable importance, plants are being explored as the source of L-asparaginase. The enzyme L-asparaginase was partially purified from Arachis hypogaea L. The crude enzyme extract was subjected to different purification steps including ammonium sulphate precipitation, dialysis followed by separation on Sephadex G-100 gel filtration (size exclusion chromatography) to obtain partially pure form of L - asparaginase. The enzyme was partially purified to 118 folds and contained specific activity of 4686.86 U/mg with 9.85% yield. SDS-PAGE electrophoresis of the partially purified enzyme revealed that it was a single protein with molecular weight of 70 kDa. The study on physiochemical properties showed that L - asparaginase from Arachis hypogaea L. was potassium-dependent in nature, where its optimum pH of enzyme activity was found to be 8.0 and temperature as 40°/50°C with reaction time of 15 - 20 minutes. Also it was observed that the L-asparaginase activity increased with the presence of metal ions such as Na+, Mg++, making it an enzyme dependent on metal ions for its reaction. In addition to this, it was revealed that the enzyme was partially inhibited in presence of certain chelators. The specificity of L-asparaginase obtained from Arachis hypogaea L. with lack of urease activity and minimal glutaminase activity along with less cytotoxicity on human blood indicated it as an efficient chemotherapeutic agent that could be investigated further in future studies.

scholarly journals UDP-glucose:solasodine glucosyltransferase from eggplant (Solanum melongena L.) leaves: partial purification and characterization.

1997 ◽  
Vol 44 (1) ◽  
pp. 43-53 ◽  
Author(s):  
C Paczkowski ◽  
M Kalinowska ◽  
Z A Wojciechowski
Keyword(s):  
Molecular Mass ◽  
Solanum Melongena ◽  
Divalent Metal ◽  
Partial Purification ◽  
Divalent Metal Ions ◽  
Purification And Characterization ◽  
Ph Optimum ◽  
Cytosol Fraction ◽  
Ammonium Sulphate Precipitation ◽  
Unripe Fruits

Uridine 5'-diphosphoglucose-dependent glucosyltransferase which catalyzes the glycosylation of solasodine i.e. UDP-glucose:solasodine glucosyltransferase, is present in leaves, roots, unripe fruits and unripe seeds of eggplant (Solanum melongena L.). The glucosylation product is chromatographically identical with authentic solasodine 3 beta-D-monoglucoside, a putative intermediate in the biosynthesis of solasodine-based glycoalkaloids characteristic of the eggplant. The enzyme was purified about 50-fold from crude cytosol fraction of eggplant leaves by ammonium sulphate precipitation and column chromatography on Q-Sepharose and Sephadex G-100. The native enzyme has a molecular mass of approx. 55 kDa and pH optimum of 8.5. Divalent metal ions are not required for its activity but the presence of free-SH groups is essential. Besides solasodine (Km = 0.04 microM), the enzyme effectively glucosylates tomatidine, another steroidal alkaloid of the spirosolane type, but it is virtually inactive towards the solanidane-type steroidal alkaloids such as solanidine or demissidine. The enzyme is specific for UDP-glucose (Km = 2.1 microM) since unlabelled ADP-, GDP-, CDP- or TDP-glucose could not effectively compete with UDP-[14C]glucose used as the sugar donor for solasodine glucosylation. Moreover, no synthesis of labelled solasodine galactoside was observed when UDP-[14C]glucose was replaced with UDP-[14C]galactose.

Partial purification and properties of an L-arabinose dehydrogenase from Azospirillum brasiliense

1983 ◽  
Vol 29 (2) ◽  
pp. 242-246 ◽  
Author(s):  
Norman J. Novick ◽  
Max E. Tyler
Keyword(s):  
Molecular Weight ◽  
Divalent Cation ◽  
Gel Filtration ◽  
Reducing Agent ◽  
Partial Purification ◽  
Ph Optimum ◽  
Glycine Buffer ◽  
Purification And Properties ◽  
Filtration Technique

An L-arabino-aldose dehydrogenase responsible for the oxidation of L-arabinose to L-arabino-γ-lactone has been purified 59-fold from L-arabinose grown cells of Azospirillum brasiliense. The dehydrogenase was found to be specific for substrates with the L-arabino-configuration at carbons 2, 3, and 4. Km values for L-arabinose of 75 and 140 μM were found with NADP and NAD as coenzymes, respectively. The enzyme had a pH optimum of 9.5 in glycine buffer and was stable when heated to 55 °C for 5 min. No enhancement of activity in the presence of any divalent cation or reducing agent tested was found. L-Arabinose dehydrogenase had a molecular weight of 175 000 as measured by the gel filtration technique.

scholarly journals Leucine Aminopeptidase from Arachis hypogaea L. Seeds Partial Purification and Characterization

2017 ◽  
Vol 33 (5) ◽  
pp. 2676-2680 ◽  
Author(s):  
Taghreed U. Mohammd ◽  
Layla O. Farhan ◽  
Ashgan S. Dawood ◽  
Bushra F. Hasan
Keyword(s):  
Arachis Hypogaea ◽  
Leucine Aminopeptidase ◽  
Partial Purification ◽  
Purification And Characterization ◽  
Arachis Hypogaea L

scholarly journals Evaluation of Improved Valencia Peanut Varieties for Production in Haiti

2020 ◽  
Vol 47 (1) ◽  
pp. 1-8
Author(s):  
A.M. Fulmer ◽  
T.B. Brenneman ◽  
R.C. Kemerait ◽  
R. Macajoux ◽  
D.A. Carroll ◽  
...  
Keyword(s):  
Arachis Hypogaea ◽  
Leaf Spot ◽  
Yield Potential ◽  
Average Increase ◽  
Cercosporidium Personatum ◽  
Central Plateau ◽  
Late Leaf Spot ◽  
Puccinia Arachidis ◽  
Arachis Hypogaea L ◽  
Plot Design

ABSTRACT Late leaf spot (Cercosporidium personatum) and peanut rust (Puccinia arachidis) are the most important diseases of peanut (Arachis hypogaea L.) in Haiti. Traditional Haitian peanut varieties are not only susceptible to these diseases but are also typically grown without benefit of a fungicide program. Five trials were conducted from 2015 to 2017 to evaluate the performance of six Valencia varieties in Quartier-Morin, Haiti (with an additional trial in 2017 at the Central Plateau) with respect to yield, resistance to rust and leaf spot diseases, and response to a fungicide program. A split-plot design with four or six replications was used in these studies. In each, “variety” was the whole plot and presence or absence of a fungicide program was the subplot. Valencia market types 309 Red, 309 Tan, M2, M3, SGV0801 and a local landrace were compared with and without Muscle ADV (tebuconazole + chlorothalonil, Sipcam) (2.3 L/ha) applied at 45, 60 and 75 days after planting (DAP). Final disease ratings (late leaf spot and peanut rust) were assessed approximately 94 DAP and plots were harvested the day following. In all trials, 309 Tan variety had the least amount of leaf spot and rust, but resulted in the lowest yield in four out of five trials, averaging 1727 kg/ha across fungicide treatments. M3, M2 and 309 Red were generally the numerically highest-yielding varieties, averaging 2906, 2864 and 2541 kg/ha across fungicide treatments, respectively, but were not statistically higher than the local Haitian Valencia, averaging 2374 kg/ha. Three fungicide applications during the season significantly increased yields in most trials for all varieties except 309 Tan. The highest and lowest average increase in yield from fungicide was for 309 Red (1126 kg/ha) and 309 Tan (103 kg/ha), respectively. The results from this study conducted over 2 years and 4 seasons document that while resistance to late leaf spot and rust is available in Valencia varieties, yield potential is not directly associated with that resistance. Also, use of fungicide improves yield potential in more susceptible varieties.

scholarly journals Solubilization, partial purification and properties of N-methylglutamate dehydrogenase from Pseudomonas aminovorans

1977 ◽  
Vol 161 (2) ◽  
pp. 357-370 ◽  
Author(s):  
C W Bamforth ◽  
P J Large
Keyword(s):  
Cytochrome C ◽  
Electron Acceptor ◽  
Specific Activity ◽  
Partial Purification ◽  
Ph Optimum ◽  
Transport Chain ◽  
Purification And Properties ◽  
Solubilized Enzyme ◽  
Triton X ◽  
Sepharose 4B

1. Extracts of amine-grown Pseudomonas aminovorans contained a particle-bound N-methylglutamate dehydrogenase (EC 1.5.99.5). The enzyme was not present in succinate-grown cells, and activity appeared before growth began in succinate-grown cells which had been transferred to methylamine growth medium. 2. Membrane-containing preparations from methylamine-grown cells catalysed an N-methylglutamate-dependent uptake of O2 or reduction of cytochrome c, which was sensitive to inhibitors of the electron-transport chain. 3. N-Methylglutamate dehydrogenase activity with phenazine methosulphate or 2,6-dichlorophenol-indophenol as electron acceptor could be solubilized with 1% (w/v) Triton X-100. The solubilized enzyme was much less active with cytochrome c as electron acceptor and did not sediment in 1 h at 150000g. Solubilization was accompanied by a change in the pH optimum for activity. 4. The solubilized enzyme was partially purified by Sepharose 4B and hydroxyapatite chromatograpy to yield a preparation 22-fold increased in specific activity over the crude extract. 5. The partially-purified enzyme was active with sarcosine, N-methylalanine and N-methylaspartate as well as with N-methylglutamate. Evidence suggesting activity with N-methyl D-amino acids as well as with the L-forms was obtained. 6. The enzyme was inhibited by p-chloromercuribenzoate, iodoacetamide and by both ionic and non-ionic detergents. 2-Oxoglutarate and formaldehyde were also inhibitors. 7. Kinetic analysis confirmed previous workers' observations of a group transfer (Ping Pong) mechanism. 8. Spectral observations suggested that the partially purified preparation contained flavoprotein and a b-type cytochrome. 9. The role of the enzyme in the oxidation of methylamine is discussed.

scholarly journals Partial purification and properties of an AMP-specific soluble 5′-nucleotidase from pigeon heart

1990 ◽  
Vol 268 (1) ◽  
pp. 117-122 ◽  
Author(s):  
A C Skladanowski ◽  
A C Newby
Keyword(s):  
Ionic Strength ◽  
Molecular Mass ◽  
Formation Rate ◽  
Linear Increase ◽  
High Ionic Strength ◽  
Partial Purification ◽  
Ph Optimum ◽  
Purification And Properties ◽  
Wide Range ◽  
Alpha Beta

A soluble 5′-nucleotidase was purified 200-fold from pigeon heart. The enzyme (1) had an apparent molecular mass close to 150 kDa, (2) had a neutral pH optimum and hydrolysed a wide range of nucleoside 5′-monophosphates with a 15-fold preference for AMP over IMP, (3) at near-physiological concentrations of AMP was activated by ADP but not by ATP, (4) was inhibited by high Mg2+ concentration and high ionic strength, (5) was weakly inhibited by p-nitrophenol phosphate and Pi, and (6) was non-competitively inhibited more potently by 5′-deoxy-5′-isobutylthioinosine than by 5′-deoxy-5′-isobutylthioadenosine, but not by [alpha, beta-methylene]ADP. The data show that the enzyme is distinct from the ecto-5′-nucleotidase and from the previously purified IMP-specific 5′-nucleotidase. They also predict that the enzyme is activated during ATP catabolism and hence will generate a more-than-linear increase in the adenosine-formation rate in response to an increase in cytosolic AMP concentration.

scholarly journals Effect of Temperature on Urediniospore Production and Germinability in Puccinia arachidis1

1994 ◽  
Vol 21 (2) ◽  
pp. 79-81
Author(s):  
P. V. Subba Rao ◽  
P. Subrahmanyam ◽  
D. McDonald
Keyword(s):  
Arachis Hypogaea ◽  
Effect Of Temperature ◽  
Optimal Temperature ◽  
Puccinia Arachidis ◽  
Arachis Hypogaea L ◽  
Different Temperatures ◽  
Percent Germination ◽  
Detached Leaves

Abstract Effect of temperature on urediniospore production in Puccinia arachidis was investigated under monocyclic infection using detached leaves of the susceptible peanut (Arachis hypogaea L.) cultivar TMV 2. Urediniospores produced at different temperatures were also examined for their germinability in vitro. The optimal temperature for urediniospore production was at about 20 and 25 C. Temperatures below 20 C or above 30 C were highly detrimental to urediniospore production. There were also marked differences in the percent germination of urediniospores produced at different temperatures. Urediniospores produced at 20 and 25 C showed the highest germination percentages. The interaction of temperature with urediniospore production and germinability is important in understanding the development of peanut rust epidemics.

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