An Experimental Design of L-Asparaginase Excreted by Pseudomonas aeruginosa Isolated from Poincianella pyramidalis Rhizosphere | Chapter 2 | Current Strategies in Biotechnology and Bioresource Technology Vol. 3

 The L-Asparaginase enzyme (E.C. 3.5.1.1) present in humans, bacteria, fungi , plants and animals, is essential for the normal progress of the cell cycle. L-asparaginase is important for the treatment of Hodgkin's and leukaemia diseases. The genus Pseudomonas, which has the capacity to produce L-asparaginase type I and/or II, is among the bacteria that produce this enzyme. The aim of this study was to better understand the optimization and partial characterization parameters of lasparaginase secreted by Pseudomonas aeruginosa isolated from the Poincianella pyramidalis rhizosphere, a representative of the Brazilian Caatinga biome. A central experimental design compound (23) with four central points and variables pH values (4, 7 and 10), temperature (30 °, 40 ° and 10 °) was performed in the current study to optimise the production of L-asparaginase complex  (50 ° C) and L-asparagine concentrations (0.2 percent, 0.5 percent and 0.8 percent). 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 were evaluated later in the statistical analysis of the variables and variance (ANOVA) of the results. The results of the optimization of L-asparaginase activity assays were 0.8 percent, pH 4 and 30 ° C in the concentration of L-asparagine, with activity values of 0.5604 U / mL to 0.0632 mg protein / mL; the optimum pH of the enzyme was 7, 7, and 7, respectively. With the best 30 ° C activity. In different substrates such as Lglutamine, sodium nitrate (NaNO3), mercury chloride (HgCl2), magnesium chloride (MgCl2) and urea, L-Asparagine has shown a better effect. With regard to the identification of bacteria, rod-shaped bacteria with flagella and species level were designated as Pseudomonas aeruginosa. The results of the characterised lasparaginase isolated from P. aeruginosa therefore revealed important data for the biotechnology industry, thus showing the feasibility and reliability of the study carried out.

Author(s) Details

Iasmim Lucas da Silva
Departamento de Antibióticos, Centro de Biociências (CB), Universidade Federal de Pernambuco, UFPE, Avenida Arthur de Sá, S/N, Cidade Universitária, Recife, 50740-520, PE, Brazil.

Wellma de Oliveira da Silva
Departamento de Antibióticos, Centro de Biociências (CB), Universidade Federal de Pernambuco, UFPE, Avenida Arthur de Sá, S/N, Cidade Universitária, Recife, 50740-520, PE, Brazil.

Leonor Alves de Oliveira da Silva
Departamento de Antibióticos, Centro de Biociências (CB), Universidade Federal de Pernambuco, UFPE, Avenida Arthur de Sá, S/N, Cidade Universitária, Recife, 50740-520, PE, Brazil and Departamento de Biologia Molecular, Centro de Ciências Exatas e da Natureza, Universidade Federal da Paraíba, Campus I, João Pessoa, 58051-900, PB, Brazil.

Luana Cassandra Breitenbach Barroso Coelho
Departamento de Bioquímica, CB, UFPE, Avenida Moraes Rego, S/N, Cidade Universitária, Recife, 50670-420, PE, Brazil.

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