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.
[Show abstract][Hide abstract] 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.
[Show abstract][Hide abstract] 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 · 2.59 Impact Factor
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