scholarly journals Acid phosphatases and ribonucleases from Dactylis glomerata seeds. I. Chromatographic and electrophoretic heterogeneity of the enzymes

2015 ◽  
Vol 47 (4) ◽  
pp. 441-453 ◽  
Author(s):  
Eleonora Wieczorek ◽  
Janina Wiśniewska ◽  
Bronisława Morawiecka
Keyword(s):  
Acid Phosphatase ◽  
Sodium Acetate ◽  
Specific Activity ◽  
Deae Cellulose ◽  
Dactylis Glomerata ◽  
Molecular Forms ◽  
Acid Phosphatases ◽  
Sodium Acetate Buffer ◽  
Optimal Activity ◽  
Optimum Ph

Acid phosphatase and ribonuclease extracted with 0.1 M sodium acetate buffer, pH 5.1 from Dactylis glomerata seeds, and partially purified by means of 70% ethanol precipitation showed electrophoretic and Chromatographic heterogeneity. After chromatography on DEAE-cellulose acid phosphatase and ribonuclease were separated into four peaks. Nonadsorbing acid phosphatase on DEAE-cellulose (peak I) was separated into four peaks on CM-cellulose. The highest activity (11 units/mg) was found in fraction b (acid phosphatase Ib). The enzyme was activated by Mg<sup>2+</sup>, Ca<sup>2+</sup>, Li<sup>+</sup>, Cs<sup>+</sup>, K<sup>+</sup> ions and inhibited by Cu<sup>2+</sup>, Zu<sup>2+</sup>, F<sup>-</sup> and Mo<sup>-6</sup> at optimum pH 5.0. Strong absorbing ribonuclease on DEAE-cellulose (peak IV) was further separated on G-200 Sephadex into two molecular forms: RN-asa1 and RN-ase2. Ribonuclease l, a thermolabile enzyme with specific activity 807 units/mg, showed an optimal activity at pH 4.8-5.1.

scholarly journals Two acid RNases from Dactylis glomerata seeds. Purification, properties and effect of polyamines and lectins on their activity

2014 ◽  
Vol 54 (3) ◽  
pp. 241-253 ◽  
Author(s):  
Janina Wiśniowska ◽  
Bronisława Morawiecka
Keyword(s):  
Specific Activity ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Dactylis Glomerata ◽  
Hydrolytic Activity ◽  
Relative Activity ◽  
Exchange Chromatography ◽  
Lentil Lectin ◽  
Optimum Ph ◽  
Rnase Ii

Two glycoproteidic acid RNases (RNase I and RNase II) were obtained and purified from the seeds of <em>Dactylis glomerata</em> by extraction with acetate buffer, fractionation with ammonium sulfate, ion-exchange chromatography on DEAE-cellulose, DEAE-Sphadex, affinity chromatography on Con A-Sepharose and gel filtration on Bio-Gel P60. RNase I with a specific activity of 2582 U•mg<sup>-1</sup> protein and an optimum pH of 4.9 and RNase II with a specific activity of 1928 U• mg<sup>-1</sup> protein and optimum pH of 4.6, were isolated. They lacked nuclease, phosphodi- and monoesterase activities. Both forms of the enzyme hydrolyzed pyrimidine homopolymers with a preference for poly U and exhibited a low specificity for purine homopolymers (poly G and poly A). RNase I acted with a 3-fold higher hydrolytic activity on poly C homopolymer than RNase IL The hydrolytic activity of both enzymes was inhibited by Zn<sup>+2</sup>, Fe<sup>+2</sup>, Cu<sup>+2</sup> ions when yeast RNA was the substrate. The amines spermine, spermidine and tyramine at a concentration of 0.1 mM increased the enzymatic activity of both RNases by 20 to 60% of the relative activity. The hydrolytic activity of RNases I and II was stimulated by the presence of lentil lectin (LL), soybean lectin (SBA) and potato lectin (STA), and inhibited by the presence of concanavalin A. The 20-200% stimulation and 40-60% inhibition depended on the proportion, on a weight basis, of enzyme to lectin and were reversible in the presence of receptor sugars.

scholarly journals Phosphatase activity of Poa pratensis seeds. I. Preliminary studies on acid phosphatase II

2015 ◽  
Vol 42 (3) ◽  
pp. 369-377
Author(s):  
I. Lorenc-Kubis ◽  
B. Morawiecka
Keyword(s):  
Acid Phosphatase ◽  
High Activity ◽  
Phosphatase Activity ◽  
Sodium Acetate ◽  
Poa Pratensis ◽  
Deae Cellulose ◽  
Acetate Buffer ◽  
Sodium Acetate Buffer ◽  
Nitrophenyl Phosphate ◽  
Low Activity

Acid phosphatase (EC 3.1.3.2) was extracted with 0.1 M sodium acetate buffer pH 5.1 from <i>Poa pratensis</i> seeds, and separated into three fractions by chromatography on DEAE cellulose. The highest activity was found in fraction Il-b (acid phosphatase II). The activity of the enzyme was optimal at pH 4.9. It hydrolyzed p-nitrophenyl phosphate most readily among the various phosphomonoesters examined. Acid phosphatase II showed also a high activity toward β-naphtyl phosphate and phenyl phosphate, very low activity towards β-glycero phosphate, 5'-GMP and no activity with glucose-1 phosphate. The enzyme was inhibited by Ca<sup>2+</sup> and fluoride, but activated by Mg<sup>2+</sup>. EDTA had no influence on the activity of the enzyme.

scholarly journals [Phosphotyrosine]protein phosphatase in rat brain. A major [phosphotyrosine]protein phosphatase is a 23 kDa protein distinct from acid phosphatase

1986 ◽  
Vol 239 (1) ◽  
pp. 155-162 ◽  
Author(s):  
M Okada ◽  
K Owada ◽  
H Nakagawa
Keyword(s):  
Acid Phosphatase ◽  
Molecular Mass ◽  
Rat Brain ◽  
Column Chromatography ◽  
Protein Phosphatase ◽  
Specific Activity ◽  
Deae Cellulose ◽  
Sodium Vanadate ◽  
Nitrophenyl Phosphate ◽  
Phosphotyrosine Protein Phosphatase

A [phosphotyrosine]protein phosphatase (PTPPase) was purified almost to homogeneity from rat brain, with [32P]p130gag-fps, an oncogene product of Fujinami sarcoma virus, as substrate. The characteristics of the purified preparation of PTPPase were as follows: the enzyme was a monomer with a molecular mass of 23 kDa; its optimum pH was 5.0-5.5; its activity was not dependent on bivalent cations; its activity was strongly inhibited by sodium vanadate, but was not inhibited by ZnCl2, L(+)-tartrate or NaF; it catalysed the dephosphorylation of [32P]p130gag-fps, [[32P]Tyr]casein, p-nitrophenyl phosphate and L-phosphotyrosine, but did not hydrolyse [[32P]Ser]tubulin, L-phosphoserine, DL-phosphothreonine, 5′-AMP, 2′-AMP or beta-glycerophosphate significantly. During the purification, most of the PTPPase activity was recovered in distinct fractions from those of conventional low-molecular-mass acid phosphatase (APase), which was reported to be a major PTPPase [Chernoff & Li (1985) Arch. Biochem. Biophys. 240, 135-145], from DE-52 DEAE-cellulose column chromatography, and those two enzymes could be completely separated by Sephadex G-75 column chromatography. APase also showed PTPPase activity with [32P]p130gag-fps, but the specific activity was lower than that of PTPPase with molecular mass of 23 kDa, and it was not sensitive to sodium vanadate. These findings suggested that PTPPase (23 kDa) was the major and specific PTPPase in the cell.

A fundamental thermostable cyanide-sensitive phosphoenolpyruvate acid carboxylase in photosynthetic and other organisms

1971 ◽  
Vol 49 (4) ◽  
pp. 631-643 ◽  
Author(s):  
David Pan ◽  
E. Roy Waygood
Keyword(s):  
Heat Treatment ◽  
Zea Mays ◽  
Acid Phosphatase ◽  
Plant Species ◽  
Phosphoenolpyruvate Carboxylase ◽  
Deae Cellulose ◽  
Bundle Sheath ◽  
Evolutionary Significance ◽  
Optimum Temperature ◽  
Optimum Ph

A thermostable 'phosphoenolpyruvate carboxylase' has been isolated from leaves of Zea mays different from phosphoenolpyruvate carboxylase (EC. 4.1.1.31) in that its optimum pH is 5.4, it does not liberate orthophosphate during the reaction, and it is inhibited by cyanide. The enzymic reaction has an optimum temperature of 70–75C and has been purified through steps including acidification to pH 4.6, heat treatment to 50C, and DEAE-cellulose and Sephadex G-200 column chromatography. Three fractions were active in the Sephadex eluate, but only fraction III was free from a thermostable acid phosphatase which catalyzes the liberation of orthophosphate from the substrate and the end product which is suggested to be a C4 phosphocarbonyl compound, although phosphohydroxypyruvate appears by either spontaneous or enzymic decarboxylation. The enzyme is assayed by the formation of a phenyl-hydrazone at 325 nm. The enzyme is localized and tightly bound in both the parenchyma bundle sheath and mesophyll chloroplasts, which are free from the thermostable acid phosphatase. Similar concentrations of the enzyme have been found in all plant species tested including C3 plants, ferns, bryophytes, algae, fungi, and even in calf liver. The enzyme must have considerable evolutionary significance.

Comparison of the Thermostability Properties of Three Acid Phosphatases from Molds: Aspergillus fumigatusPhytase, A. niger Phytase, and A. nigerpH 2.5 Acid Phosphatase

1998 ◽  
Vol 64 (11) ◽  
pp. 4446-4451 ◽  
Author(s):  
Markus Wyss ◽  
Luis Pasamontes ◽  
Roland Rémy ◽  
Josiane Kohler ◽  
Eric Kusznir ◽  
...  
Keyword(s):  
Acid Phosphatase ◽  
Aspergillus Niger ◽  
Enzymatic Activity ◽  
Conformational Changes ◽  
Animal Feed ◽  
Heat Denaturation ◽  
Acid Phosphatases ◽  
Active Conformation ◽  
Feed Supplements ◽  
Optimum Ph

ABSTRACT Enzymes that are used as animal feed supplements should be able to withstand temperatures of 60 to 90°C, which may be reached during the feed pelleting process. The thermostability properties of three histidine acid phosphatases, Aspergillus fumigatus phytase,Aspergillus niger phytase, and A. niger optimum pH 2.5 acid phosphatase, were investigated by measuring circular dichroism, fluorescence, and enzymatic activity. The phytases ofA. fumigatus and A. niger were both denatured at temperatures between 50 and 70°C. After heat denaturation at temperatures up to 90°C, A. fumigatus phytase refolded completely into a nativelike, fully active conformation, while in the case of A. niger phytase exposure to 55 to 90°C was associated with an irreversible conformational change and with losses in enzymatic activity of 70 to 80%. In contrast to these two phytases,A. niger pH 2.5 acid phosphatase displayed considerably higher thermostability; denaturation, conformational changes, and irreversible inactivation were observed only at temperatures of ≥80°C. In feed pelleting experiments performed at 75°C, the recoveries of the enzymatic activities of the three acid phosphatases were similar (63 to 73%). At 85°C, however, the recovery of enzymatic activity was considerably higher for A. fumigatusphytase (51%) than for A. niger phytase (31%) or pH 2.5 acid phosphatase (14%). These findings confirm that A. niger pH 2.5 acid phosphatase is irreversibly inactivated at temperatures above 80°C and that the capacity of A. fumigatus phytase to refold properly after heat denaturation may favorably affect its pelleting stability.

Biochemical and histochemical studies of alkaline and acid phosphatases in a digenetic trematode,Pegosomum egretti

1986 ◽  
Vol 60 (4) ◽  
pp. 293-298 ◽  
Author(s):  
Indra Rajvanshi ◽  
K. L. Mali
Keyword(s):  
Alkaline Phosphatase ◽  
Enzyme Activity ◽  
Acid Phosphatase ◽  
Raw Materials ◽  
Digenetic Trematode ◽  
Acid Phosphatases ◽  
Alkaline Phosphatases ◽  
Optimum Ph ◽  
Standard Techniques ◽  
Alkaline And Acid Phosphatases

ABSTRACTThe biochemistry and histochemistry ofPegosomum egrettihave been studied using standard techniques. Phosphatases were analysed colorimetrically; the optimum pH for acid phosphatase activity was 5·0 and for alkaline phosphatase was 10·0. The results were compared with those of other trematodes. Histochemical localization of acid and alkaline phosphatases revealed differences in enzyme activity in various tissues. These differences in the site and pattern of distribution of the two enzymes have been discussed in relation to transport of raw materials and the metabolism of the cell concerned.

scholarly journals Presence of multiple acid phosphatases activity in seedlings of cucumber, radish and rocket salad

2008 ◽  
Vol 38 (3) ◽  
pp. 650-657 ◽  
Author(s):  
Luciane Almeri Tabaldi ◽  
Raquel Ruppenthal ◽  
Luciane Belmonte Pereira ◽  
Denise Cargnelutti ◽  
Jamile Fabbrin Gonçalves ◽  
...  
Keyword(s):  
Acid Phosphatase ◽  
Phosphate Esters ◽  
Acid Phosphatases ◽  
Simple Preparation ◽  
Toxicological Studies ◽  
Plant Enzymes ◽  
Optimum Ph ◽  
Ph Range ◽  
Hydrolysis Of ◽  
Assay Conditions

Acid phosphatases (3.1.3.2) are a group of enzymes widely distributed in nature, which catalyze the hydrolysis of a variety of phosphate esters in the pH range of 4-6. We confirmed the presence of acid phosphatases in seedlings of cucumber (Cucumis sativus), radish (Raphanus sativus) and rocket salad (Eruca vesicaria) under different assay conditions using a rapid and simple preparation. The results showed that the optimum pH and temperature used for all species were close to 5.5 and 35°C, respectively. The enzyme was inhibited by molybdate, fluoride, azide, levamisole, orthovanadate, Zn2+ and Cu2+. Suramin had no effect on enzyme activity. The acid phosphatase from cucumber, radish and rocket salad hydrolyzed a wide variety of phosphate esters and the highest activity was observed with PPi, ATP and GTP. These results demonstrate that the enzyme investigated in this study is different from well known ester phosphate cleaving plant enzymes (apyrase and inorganic pyrophosphatases) and this preparation could be a useful tool to future toxicological studies and to study initially all isoforms of acid phosphatase.

Cellulase-gold labelling of cellulose in forages

1989 ◽  
Vol 47 ◽  
pp. 1012-1013
Author(s):  
Janet H. Woodward
Keyword(s):  
Sodium Acetate ◽  
Cynodon Dactylon ◽  
Dactylis Glomerata ◽  
Sodium Acetate Buffer ◽  
Total Dietary Fiber ◽  
Coastal Bermudagrass ◽  
Cell Wall Constituent ◽  
Structural Differences ◽  
Gold Labelling ◽  
Insight Into

Current fiber analyses provide information on the chemical and structural composition of the fiber as a whole. However, these techniques do not offer insight into the compositional and structural differences between specific fiber tissues which may affect digestibility. The purpose of this study was to: (1) ultrastructurally localize one cell wall constituent, cellulose, in four forages; and (2) apply this technique as a tool for the evaluation of biodegradation in forages.Insoluble fibers were prepared using a modified AOAC Total Dietary Fiber Technique. Two millimeter sections cut from leaflets of alfalfa (Medicago sativa L.) and 1-76 1espedeza [Sericea cuneata (Dumont)] and leaves of coastal bermudagrass (Cynodon dactylon L. Pers.) and orchardgrass (Dactylis glomerata L.) were incubated stepwise with: (1) α-amylase, pH 6.0 for ½ h at 100°C; (2) protease, pH 7.5 for ½ h at 60°C; (3) amyloglucosidase, pH 4.5 for ½ h at 60°C, and (4) 1% (w/v) cellulase in 0.05 M sodium acetate buffer, pH 5.0 for 24 h at 37°C.

scholarly journals Phosphatase activity of Poa pratensis seeds. II. Purification and characterization of acid phosphatase Ia2 and Ia3

2015 ◽  
Vol 44 (2) ◽  
pp. 255-263 ◽  
Author(s):  
I. Lorenc-Kubis ◽  
B. Morawiecka ◽  
M. Niezgódka ◽  
A. Hebrowska
Keyword(s):  
Acid Phosphatase ◽  
Mixed Type ◽  
Poa Pratensis ◽  
Maximal Activity ◽  
Molecular Forms ◽  
Acid Phosphatases ◽  
Purification And Characterization ◽  
Inorganic Pyrophosphate ◽  
Nitrophenyl Phosphate

Two acid phosphatases (Ia2, Ia3) have been isolated from <i>Poa pratensis</i> seeds and partially purified. Both enzymes showed maximal activity at pH 4,9. They exhibited high activity towards p-nitrophenyl phosphate, inorganic pyrophosphate and phenyl phosphate, much less activity towards glucose-6 phosphate, and mononucleotides. Phosphatases a<sub>2</sub> and a<sub>3</sub> differed in their activity towards ADP. Orthophosphate, fluoride and Zn<sup>2+</sup> were effective inhibitors. EDTA, β-mercaptoethanol and Mg<sup>2+</sup> activated phophatase a<sub>2</sub> but had no effect on phosphatase a<sub>3</sub>. Zn<sup>2+</sup> inhibited the activity of phosphatase a<sub>2</sub> noncompetitively, whereas phosphatase a<sub>3</sub> showed inhibition of mixed type. Trypsin, chymotrypsin and pronase had no effect on the enzyme activities of both molecular forms.

The Demonstration of Human Prostatic Acid Phosphatase (Pap) With Phosphorylcholine

1976 ◽  
Vol 34 ◽  
pp. 88-89
Author(s):  
José A. Serrano ◽  
Hannah L. Wasserkrug ◽  
Anna A. Serrano ◽  
Arnold M. Seligman
Keyword(s):  
Acid Phosphatase ◽  
Prostate Gland ◽  
Prostatic Acid Phosphatase ◽  
Human Prostate ◽  
Rat Tissues ◽  
Specific Substrate ◽  
Acid Phosphatases ◽  
Lysosomal Acid ◽  
Cacodylate Buffer

As previously reported (1, 2) phosphorylcholine (PC) is a specific substrate for prostatatic acid phosphatase (PAP) as opposed to other acid phosphatases, e.g., lysosomal acid phosphatase. The specificity of PC for PAP is due to the pentavalent nitrogen in PC, a feature that renders PC resistant to hydrolysis by all other acid phosphatases. Detailed comparative cytochemical results in rat tissues are in press. This report deals with ultracytochemical results applying the method to normal and pathological human prostate gland.Fresh human prostate was obtained from 7 patients having transurethral resections or radical prostatectomies. The tissue was fixed in 3% glutaraldehyde- 0.1 M cacodylate buffer (pH 7.4) for 15 min, sectioned at 50 μm on a Sorvall TC-2 tissue sectioner, refixed for a total of 2 hr, and rinsed overnight in 0.1 M cacodylate buffer (pH 7.4)-7.5% sucrose.

Sign in / Sign up

Export Citation Format

Share Document