An Experimental Design of L-Asparaginase Excreted by Pseudomonas aeruginosa Isolated from Poincianella pyramidalis Rhizosphere

L-Asparaginase enzyme (E.C. 3.5.1.1) present in bacteria, fungi, plants and animals, absent in
humans, is essential for normal cell cycle progress. The L-asparaginase is important for the treatment
of leukemic diseases and Hodgkin's disease. Among the bacteria producing this enzyme is the genus
Pseudomonas which have the ability to produce L-asparaginase type I and / or II. The objective of this
study was to understand better the parameters for optimization and partial characterization of Lasparaginase,
secreted by Pseudomonas aeruginosa isolated from the rhizosphere of Poincianella
pyramidalis, representative of the Caatinga biome, unique to Brazil. In the present study to optimize
the production of L-asparaginase complex it was performed a central experimental design compound
(23) with four central points and variables pH values (4, 7 and 10), temperature (30°, 40° and 50°C)
and concentration of L-asparagine (0.2%, 0.5% and 0.8%). Later the statistical analysis of the
variables and the variance (ANOVA) of the results were evaluated by the Statistica software, version
7.0 (StatSoft Co., USA), enzymatic characterization at different pH values, temperatures, ions,
substrates and finally bacterial identification at morphological and molecular levels. The results in the
optimization of L-asparaginase activity assays were in concentration of L-asparagine 0.8%, pH 4 and
30°C, with activity values 0.5604 U/mL to 0.0632 mg protein/mL; the optimum pH of the enzyme was
7, with the best activity at 30°C. L-Asparagine showed a better effect on various substrates such as Lglutamine,
sodium nitrate (NaNO3), mercury chloride (HgCl2), magnesium chloride (MgCl2) and urea.
Regarding the bacterial identification met rod-shaped bacteria having flagella and species level was
designated as being Pseudomonas aeruginosa. Therefore, the results of characterized Lasparaginase,
isolated from P. aeruginosa showed important data for the biotechnology industry, thus
presenting the viability and reliability of the study performed.