Wyniki wyszukiwania - AGRO

1

Purification and general properties of pectin methyl esterase from Curvularia inaequalis NRRL 13884 in solid state culture using orange peels as an inducer

100%

Afifi A. F., Fawzi E. M., Foaad M. A.

Acta Microbiologica Polonica

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2002

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tom 51

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nr 3

2

The application of FPLC chromatography [Mono Q] for the purification of wheat RNA polymerases II and III

100%

Glanc P., Galbas M., Dullin P., Szalata M., Slomski R.

Cellular and Molecular Biology Letters

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2003

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tom 08

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nr 3

Transcription is the main step in the regulation of gene expression. To study this process in vitro, it is necessary to obtain highly purified RNA polymerases. Here, we describe a method of RNA polymerase purification using a Mono Q FPLC column. Using Mono Q column chromatography accelerates the purification process and separates RNA polymerase II from RNA polymerase III with good yield.

3

Purification and some properties of exo-1,4-beta-glucanase from Chaetomium olivaceum

100%

El-Gindy A. A., Saad R. R., Fawzi E.

Acta Microbiologica Polonica

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2003

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tom 52

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nr 1

4

Purification of arginase from Aspergillus nidulans

100%

Dzikowska A., Le Caer J. P., Jonczyk P., Weglenski P.

Acta Biochimica Polonica

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1994

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tom 41

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nr 4

Arginase (EC 3.5.3.1) of Aspergillus nidulans, the enzyme which enables the fungus to use arginine as the sole nitrogen source was purified to homogeneity. Molecular mass of the purified arginase subunit is 40 kDa and is similar to that reported for the Neurospora crussa (38.3 kDa) and Saccharomyces cerevisiae (39 kDa) enzymes. The native molecular mass of arginase is 125 kDa. The subunit/nati ve molecular mass ratio suggests a trimeric form of the protein. The arginase protein was cleaved and partially sequenced. Two out of the six polypeptides sequenced show a high degree of homology to conserved domains in arginases from other species.

5

Purification and characterization of xylanase from a thermophilic Streptomyces sp.K37

100%

Mansour F. A., Shereif A. A., Nour El-Dein M. M., Abou-Dobara M. I., Ball A. S.

Acta Microbiologica Polonica

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2003

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tom 52

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nr 2

6

Transforming growth factors (TGFs) produced by R-XC cells

88%

Klein A., Branny J.

Folia Histochemica et Cytobiologica

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1985

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tom 23

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nr 4

7

Characterization and mass spectrometry analysis of aminopeptidase N from Pseudomonas putida Lup

88%

Jankiewicz U., Swiontek-Brzezinska M., Gorska E. B., Kowalczyk P.

Polish Journal of Microbiology

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2013

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tom 62

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nr 4

An intracellular aminopeptidase N synthesized by Pseudomonas putida Lup was purified and characterized. The approx. 150-fold purified enzyme showed highest activity against A-β-naphthylamide at pH 7.5 and at temperature 40°C and was 100% thermostable for 240 min at 40°C. P. putida lup aminopeptidase N is a monomer with molecular mass approx. 99 kDa determined by SDS-PAGE and gel permeation chromatography. The enzyme has broad substrate specificity, but is the most active against protein substrates with N-terminal alanine and arginine. The activity of P. putida Lup aminopeptidase N is strongly inhibited in the presence of specific metallopeptidase inhibitors and is partly recovered in the presence of Zn²⁺ and Co²⁺ ions. Co²⁺, Mg²⁺ and Ca²⁺ ions increased the activity of the enzyme. Moreover, the enzyme was inhibited by inhibitors of cysteine enzymes. Analysis of fragments of the amino acid sequence of the purified enzyme demonstrated high similarity to PepN of Pseudomonas putida GB-1.

8

One-step purification of vitronectin from human plasma by affinity chromatography on phage-displayed peptides

88%

Skurzynski S.

Acta Biochimica Polonica

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2010

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tom 57

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nr 1

89-93

A novel affinity purification method for rapid isolation of vitronectin (VN) from human plasma is described. Recently we have used phage display technology to obtain clones expressing peptides with high binding activity for VN. The isolated "strong VN binders" were covalently coupled to CNBr-activated Sepharose. Human plasma was applied to the column and bound VN was eluted using 0.5 M acetic acid, giving purity exceeding 90 %. The developed method is a convenient alternative to conventional antibody-antigen affinity chromatography techniques for purification of VN, as it offers low ligand cost, is rapid and ensures good protein recovery from human plasma.

9

Production of bacteriocin E50-52 by small ubiquitin-related modifier fusion in Escherichia coli

88%

Wang Q., Fu W., Ma Q., Yu Z., Zhang R.

Polish Journal of Microbiology

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2013

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tom 62

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nr 4

Bacteriocin E50-52, a class IIa bacteriocin with a wide antibacterial spectrum, and has a huge potential to be a substitute for conventional antibiotics. In this research, the bacteriocin E50-52 gene was cloned into the expression vector pET SUMO (small ubiquitin-related modifier) and introduced into Escherichia coli BL21 (DE3). The recombinant fusion protein SUMO-bacteriocin E50-52 expressed in a soluble form was purified to a purity of more than 90% by Ni-NTA sepharose column and 117 mg fusion protein was obtained per liter of fermentation culture. The fusion protein was cleaved with SUMO protease and re-applied to a Ni-NTA Sepharose column. Finally, about 16 mg recombinant bacteriocin E50-52 (rbE50-52) was obtained from a 1-liter fermentation culture with no less than 95% purity. The rbE50-52 had similar antimicrobial properties and molecular weight as the native bacteriocin E50-52 and showed very low hemolytic activity.

10

Purification and kinetics of extra and intracellular phospholipase-A of Salmonella enteritidis

88%

Kapoor S., Sharma D. K., Sharma S., Kalra M. S.

Bulletin of the Veterinary Institute in Puławy

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2005

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tom 49

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nr 4

11

Purification and characterization of trypsin inhibitor from seeds of faba bean [Vicia faba L.]

88%

Gupta P., Dhawan K., Malhotra S. P., Singh R.

Acta Physiologiae Plantarum

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2000

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tom 22

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nr 4

12

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Attempt to use cellulose nitrate membrane for the purification of diffusion juice

88%

Regiec P.

Acta Agrophysica

|

2003

|

tom 02

|

nr 2[96]

13

Preparation of endotoxin-free bacteriophages

88%

Boratynski J., Syper D., Weber-Dabrowska B., Lusiak-Szelachowska M., Pozniak G., Gorski A.

Cellular and Molecular Biology Letters

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2004

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tom 09

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nr 2

Bacteriophages (phages) are bacterial viruses that interact with bacterial walls and invade bacterial cells. Moreover, they disturb bacterial metabolism and lead to bacteria lysis. In the case of Gram-negative bacteria crude phage cultures, apart from the phages themselves, the bacterial debris, bacterial proteins and nucleic acids contain endotoxins. These endotoxins (lipopolysaccharides) posses a high degree of toxicity in vitro and in vivo, and their removal is essential for safety in antibacterial bacteriophage therapy. An effective, scaleable purification of bacteriophages from endotoxins was accomplished by sequential ultrafiltration through polysulfone membrane (30 nm) followed by chromatography on sepharose 4B and Matrex Cellulofine Sulfate. The phage fraction after gel filtration chromatography routinely contained endotoxins in the 150-2500 EU/mL range. The procedure yielded bacteriophages contaminated with as little as 0.4-7 EU/ml (Limulus assay). This value lies within the permitted level for intravenous applications (5 EU/kg/h by European Pharmacopoeia, 1997)

14

Purification and characterization of a glutathione S-transferase from Mucor mucedo

88%

Hamed R. R., Abu-Shady M. R., El-Beih F. M., Abdalla A. M. A., Afifi O. M.

Polish Journal of Microbiology

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2005

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tom 54

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nr 2

An intracellular glutathione transferase was purified to homogenity from the fungus, Mucor mucedo, using DEAE-cellulose ion-exchange and glutathione affinity chromatography. Gel filtration chromatography and SDS-PAGE revealed that the purified GST is a homodimer with approximate native and subunit molecular mass of 53 kDa and 23.4 kDa, respectively. The enzyme has a pI value of 4.8, a pH optimum at pH 8.0 and apparent activation energy (Ea) of 1.42 kcal mol⁻¹. The purified GST acts readily on CDNB with almost negligible peroxidase activity and the activity was inhibited by Cibacron Blue (IC₅₀ 0.252 μM) and hematin (IC₅₀ 3.55 μM). M. mucedo GST displayed a non-Michaelian behavior. At Iow (0.1-0.3 mM) and high (0.3-2 mM) substrate concentration, Km(GSH) was calculated to be 0.179 and 0.65 mM, whereas Km(CDNB) was 0.531 and 11 mM and kcat was 39.8 and 552 s⁻¹, respectively. The enzyme showed apparent pKa values of 6-6.5 and 8.0.

15

Purification of autocrine growth factor from conditioned medium of rat sarcoma [XC] cells

88%

Checiowna D., Klein A.

Folia Histochemica et Cytobiologica

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1996

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tom 34

|

nr 3-4

16

The detection and purification of a protein that recognizes a line 1 DNA repetitive sequence

75%

Lukyanov D., Rogolinski J., Podgornaya O., Rzeszowska-Wolny J.

Cellular and Molecular Biology Letters

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2000

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tom 05

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nr 4

551-521

A protein which binds specifically to MspI8 (a 454 bp long repeated sequence highly homologous to the 5’ untranslated region (5'UTR) of the LINE 1 sequence) was found and identified in nuclear extracts of rat liver cells. This protein was detected using the electrophoretic mobility shift assay (EMSA) and was purified by Q-Sepharose and DNA affinity chromatography. Its molecular mass was estimated by SDS electrophoresis as 29 kDa. The possibility that this protein (p29) is the rat analogue of human L1PBP-A, specific for the human LINE sequence LPE1, is discussed.

17

Kinetics properties of polyphenol oxidase in hawthorn (Crataegus spp.)

75%

Saeidian S.

International Letters of Natural Sciences

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2014

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tom 20

Polyphenol oxidase (PPO) from hawthorn was extracted and partially purified through (NH4)2SO4 precipitation, dialysis and ion exchange chromatography. The activity of polyphenol oxidase was investigated in Crataegus spp. Spectrophotometric method was used to assay the enzyme activity and the kinetic constants - maximum enzyme velocity (Vmax) and Michealis - Menten constant (Km). Of the substrates tested, catechol was the best substrate for PPO with a Km value of 2.2 mM. The optimum pH for PPO activity was found to be 7. The enzyme showed high activity over a broad pH range of 4 - 8. The optimal pH and temperature for enzyme activity were found to be 7 and 40-45 °C, respectively. km value for hawthorn PPO is calculated 22 mM for catechol and 6.7 mM for pyrogallol and 9.7 mM for L-dopa. As can be seen, affinity of PPOs for various substrates varies widely. The enzyme showed a broad activity over a broad pH and temperature range. The thermal inactivation studies showed that the enzyme is heat resistant. The enzyme showed the highest activity toward pyrogallol and no activity toward tyrosine. Of the inhibitors tested, the most potent inhibitors were kojic acid, cysteine and glycine , respectively.

18

Production and applications of xylanases – an overview

75%

Malhotra G., Chapadgaonkar S. S.

BioTechnologia. Journal of Biotechnology Computational Biology and Bionanotechnology

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2018

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tom 99

|

nr 1

19

Purification and characterization of an antibacterial protein from dried fruiting bodies of the wild mushroom Clitocybe synopica

75%

Zheng S., Liu Q., Zhang G., Wang H., Ng T. B.

Acta Biochimica Polonica

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2010

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tom 57

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nr 1

43-48

A novel antibacterial protein with a molecular mass of 44 kDa has been isolated from dried fruiting bodies of the wild mushroom Clitocybe sinopica. Sodium dodecyl sulfate/polyacrylamide gel electrophoresis showed that the protein was composed of two subunits each with a molecular mass of 22 kDa. Its N-terminal amino-acid sequence, SVQATVNGDKML, has not been reported for other antimicrobial proteins. The purification protocol included ion exchange chromatography on DEAE-cellulose, CM-cellulose and Q-Sepharose, and gel filtration by fast protein liquid chromatography on Superdex 75. The antibacterial protein was adsorbed on all three ion exchangers. The antimicrobial activity profile of the protein against tested bacterial and fungal strains disclosed that it possessed potent antibacterial activity against Agrobacterium rhizogenes, A. tumefaciens, A. vitis, Xanthomonas oryzae and X. malvacearum with a minimum inhibitory concentration mostly below 0.6 μM. However, it had no antibacterial activity against Pseudomonas batatae, Erwinia herbicola, Escherichia coli, and Staphylococcus aureus, and no antifungal activity against Setosphaeria turcica, Fusarium oxysporum, Verticillium dahliae, Bipolaris maydis, and B. sativum. The antibacterial antivity against A. tumefaciens was stable after exposure to 20-60°C for 30 min and to pH 4-9 for 1 h.

20

Polyphenol oxidase from wheat bran is a serpin

75%

Yamasaki Y., Konno H., Noda K.

Acta Biochimica Polonica

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2008

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tom 55

|

nr 2

325-328

Polyphenol oxidase (PPO; EC 1.10.3.2) was isolated from wheat bran by a procedure that included ammonium sulfate fractionation, batch adsorption by DEAE-cellulofine, CM-cellulofine column chromatography, DEAE-cellulofine column chromatography, preparative isoelectric focusing, adsorption on the membrane of a Vivapure Q Maxi H spin column, and heat treatment. These procedures led to 150-fold purification with 4.2% recovery. The PPO was homogeneous by SDS/PAGE. The relative molecular weight of the PPO was estimated to be 37000 based on its mobility in SDS/PAGE. The isoelectric point of the PPO was 4.4. The Kmvalues of the PPO for caffeic acid, chlorogenic acid, pyrocatechol, 4-methyl catechol and l-DOPA as substrates were 0.077, 0.198, 1.176, 1.667 and 4.545 mM. The PPO was strongly inhibited by tropolone. The Kivalue for tropolone is 2.2 × 10–7M. The sequence of the 15 N-terminal amino-acid residues was determined to be ATDVRLSIAHQTRFA, which was identical to those of serpin from Triticum aestivum and protein Z from Hordeum vulgare. The PPO strongly inhibited the activity of trypsin, which is an enzyme of serine proteases; 50% inhibition was observed with 1.5 × 10–7M PPO. The Kivalue for PPO is 2.3 × 10–8M. The wheat bran PPO should be a very important protein for protecting wheat against disease, virus, insect and herbivore damages by both the activities of PPO and protease inhibitor.

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