Studies on pH and thermal stability of novel purified L-asparaginase from Pectobacterium carotojorum MTCC 1428.
ABSTRACT Glutaminase free L-asparaginase is known to be an excellent anticancer agent. In the present study, the combined effect of pH and temperature on the performance of purified novel L-asparaginase from Pectobacterium carotovorum MTCC 1428 was studied under assay conditions using response surface methodology (RSM). Deactivation studies and thermodynamic parameters of this therapeutically important enzyme were also investigated. The optimum pH and temperature of the purified L-asparaginase were found to be 8.49 and 39.3 degrees C, respectively. The minimum deactivation rate constant (k(d)) and maximum half life (t1/2) were found to be 0.041 min(-1) and 16.9 h, respectively at pH of 8.6 and 40 degreesC. Thermodynamic parameters (deltaG, deltaH, deltaS, and activation energies) were also evaluated for purified L-asparaginase. The probable mechanism of deactivation of purified L-asparaginase was explained to an extent on the basis of deactivation studies and thermodynamic parameters.
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ABSTRACT: L-Asparaginase is known to catalyze the hydrolysis of L-asparagine to L-aspartic and ammonia, but little is known about its action on peptides. When we incubated L-asparaginases purified either from Escherichia coli or Erwinia chrysanthemi - commonly used as chemotherapeutic agents because of their antitumour activity - with eight small beta-aspartylpeptides such as beta-aspartylserineamide, beta-aspartylalanineamide, beta-aspartylglycineamide and beta-aspartylglycine, we found that both L-asparaginases could catalyze the hydrolysis of five of them yielding L-aspartic acid and amino acids or peptides. Our data show that L-asparaginases can hydrolyze beta-aspartylpeptides and suggest that L-asparaginase therapy may affect the metabolism of beta-aspartylpeptides present in human body.FEBS Letters 10/2002; 528(1-3):130-2. DOI:10.1016/S0014-5793(02)03273-8 · 3.34 Impact Factor
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ABSTRACT: Acrylamide formation in French fries was investigated in relation to blanching and asparaginase soaking treatments before final frying. Par-fried potatoes of Bintje variety were prepared by cutting strips (0.8 × 0.8 × 5 cm) which were blanched at 75 °C for 10 min. Unblanched strips were used as the control. Control or blanched strips were then dried at 85 °C for 10 min and immediately partially fried at 175 °C for 1 min. Finally, frozen par-fried potatoes were fried at 175 °C for 3 min to obtain French fries. Pre-drying of raw or blanched potato strips did not generate acrylamide formation as expected. Partial frying of pre-dried control potato strips generated 370 μg/kg of acrylamide and the final frying determined French fries with 2075 μg/kg of acrylamide. When control potato strips were treated with a 10000 ASNU/l asparaginase solution at 40 °C for 20 min, the acrylamide formation in French fries was reduced by 30%. When blanched potato strips were treated in the same way, the produced French fries have 60% less acrylamide content than blanched strips without the enzyme treatment. Soaking of blanched potato strips (75 °C, 10 min) in an 10000 ASNU/l asparaginase solution at 40 °C for 20 min is an effective way to reduce acrylamide formation after frying by reducing the amount of one of its important precursors such as asparagine.Food Chemistry 07/2008; DOI:10.1016/j.foodchem.2007.12.057 · 3.26 Impact Factor
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ABSTRACT: The association of magnetic nanoparticles, which could be controlled by a magnetic field and have dimensions which facilitate their penetration in cells/tissues, with hydrogel type biopolymeric shells confer them compatibility and the capacity to retain and deliver bioactive substances. The main objective of this work is the development of a new system based on a biocompatible polymer with organic-inorganic structure capable of vectoring support for biologic active agents (L: -asparaginase, e.g.). Characterization of size and morphology of the hydrogel-magnetic nanoparticles with entrapped L: -asparaginase was made using Dynamic Light Scattering method, Transmission Electron Microscopy and Confocal Microscopy. The structure of magnetic nanoparticles coated with hydrogel was characterized by Fourier Transformed Infrared Spectroscopy. The cytotoxicity of nanoparticles was evaluated and also the interactions with microorganisms. We obtained hydrogel-magnetic nanoparticles with L: -asparaginase entrapped, with sizes below 30 nm in dried stage, capable to penetrate the cells and tissues.Journal of Materials Science Materials in Medicine 02/2009; 20(6):1307-14. DOI:10.1007/s10856-008-3684-y · 2.38 Impact Factor