Recombinant L-Asparaginase from Zymomonas mobilis: A Potential New Antileukemic Agent Produced in Escherichia coli

• Published in: PloS one Vol. 11; no. 6; p. e0156692
• Main Authors: Einsfeldt, Karen, Baptista, Isis Cavalcante, Pereira, Juliana Christina Castanheira Vicente, Costa-Amaral, Isabele Campos, Costa, Elaine Sobral da, Ribeiro, Maria Cecília Menks, Land, Marcelo Gerardin Poirot, Alves, Tito Lívio Moitinho, Larentis, Ariane Leites, Almeida, Rodrigo Volcan
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 02.06.2016; Public Library of Science (PLoS)
• Subjects: Acute lymphoblastic leukemia; Amino acids; Analysis; Anticancer properties; Antineoplastic Agents - pharmacology; Antineoplastic Agents - therapeutic use; Apoptosis; Apoptosis - drug effects; Asparaginase; Asparaginase - genetics; Asparaginase - pharmacology; Asparaginase - therapeutic use; Bacteria; Base Sequence; Biology and Life Sciences; Bioreactors; Cancer; Cell culture; Cell cycle; Cell Cycle - drug effects; Cell Line, Tumor; Cell Nucleus Shape - drug effects; Cell Proliferation - drug effects; Cell Survival - drug effects; Child; Child, Preschool; Cloning; Cloning, Molecular; Codons; Computer Simulation; Cytotoxicity; Drug resistance; Drug therapy; E coli; Enzymes; Escherichia coli; Escherichia coli - metabolism; Female; Genetic aspects; Genetic engineering; Helicobacter pylori; Histidine; Humans; Infant; Kinases; L-asparaginase; Leukemia; Leukemia - drug therapy; Leukemia - pathology; Lymphatic leukemia; Male; Pediatrics; Phylogeny; Physiological aspects; Polyethylene glycol; Proteins; Recombinant Proteins - genetics; Recombinant Proteins - pharmacology; Recombinant Proteins - therapeutic use; Research and Analysis Methods; Toxicity; Zymomonas; Zymomonas - enzymology; Zymomonas mobilis; Brazil; United States > US; Rio de Janeiro Brazil
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0156692

L-asparaginase is an enzyme used as a chemotherapeutic agent, mainly for treating acute lymphoblastic leukemia. In this study, the gene of L-asparaginase from Zymomonas mobilis was cloned in pET vectors, fused to a histidine tag, and had its codons optimized. The L-asparaginase was expressed extracellularly and intracellularly (cytoplasmically) in Escherichia coli in far larger quantities than obtained from the microorganism of origin, and sufficient for initial cytotoxicity tests on leukemic cells. The in silico analysis of the protein from Z. mobilis indicated the presence of a signal peptide in the sequence, as well as high identity to other sequences of L-asparaginases with antileukemic activity. The protein was expressed in a bioreactor with a complex culture medium, yielding 0.13 IU/mL extracellular L-asparaginase and 3.6 IU/mL intracellular L-asparaginase after 4 h of induction with IPTG. The cytotoxicity results suggest that recombinant L-asparaginase from Z. mobilis expressed extracellularly in E.coli has a cytotoxic and cytostatic effect on leukemic cells.