title: Recombinant L-Asparaginase from Zymomonas mobilis: A Potential New Antileukemic Agent Produced in Escherichia coli.
abstract: 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.
authors.affiliation: Programa de Engenharia Química, COPPE, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil.|Programa de Pós-Graduação em Bioquímica, Instituto de Química, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil.
authors.forename: Karen
authors.initials: K
authors.lastname: Einsfeldt
authors.affiliation: Programa de Engenharia Química, COPPE, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil.|Programa de Pós-Graduação em Clínica Médica, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil.
authors.forename: Isis Cavalcante
authors.initials: IC
authors.lastname: Baptista
authors.affiliation: Programa de Engenharia Química, COPPE, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil.|Programa de Pós-Graduação em Bioquímica, Instituto de Química, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil.
authors.forename: Juliana Christina Castanheira Vicente
authors.initials: JC
authors.lastname: Pereira
authors.affiliation: Programa de Saúde Pública e Meio Ambiente, Escola Nacional de Saúde Pública Sergio Arouca (ENSP), Centro de Estudos da Saúde do Trabalhador e Ecologia Humana (CESTEH), Fundação Oswaldo Cruz (FIOCRUZ), Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Isabele Campos
authors.initials: IC
authors.lastname: Costa-Amaral
authors.affiliation: Programa de Pós-Graduação em Clínica Médica, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil.
authors.forename: Elaine Sobral da
authors.initials: ES
authors.lastname: Costa
authors.affiliation: Programa de Pós-Graduação em Clínica Médica, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil.
authors.forename: Maria Cecília Menks
authors.initials: MC
authors.lastname: Ribeiro
authors.affiliation: Programa de Pós-Graduação em Clínica Médica, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil.
authors.forename: Marcelo Gerardin Poirot
authors.initials: MG
authors.lastname: Land
authors.affiliation: Programa de Engenharia Química, COPPE, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil.
authors.forename: Tito Lívio Moitinho
authors.initials: TL
authors.lastname: Alves
authors.affiliation: Programa de Saúde Pública e Meio Ambiente, Escola Nacional de Saúde Pública Sergio Arouca (ENSP), Centro de Estudos da Saúde do Trabalhador e Ecologia Humana (CESTEH), Fundação Oswaldo Cruz (FIOCRUZ), Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Ariane Leites
authors.initials: AL
authors.lastname: Larentis
authors.affiliation: Programa de Pós-Graduação em Bioquímica, Instituto de Química, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil.
authors.forename: Rodrigo Volcan
authors.initials: RV
authors.lastname: Almeida
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology / Q000627:therapeutic use*; D017209:Apoptosis / Q000187:drug effects; D001215:Asparaginase / Q000235:genetics / Q000494:pharmacology / Q000627:therapeutic use*; D001483:Base Sequence; D019149:Bioreactors; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D053144:Cell Nucleus Shape / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002648:Child; D002675:Child, Preschool; D003001:Cloning, Molecular; D003198:Computer Simulation; D004926:Escherichia coli / Q000378:metabolism*; D005260:Female; D006801:Humans; D007223:Infant; D007938:Leukemia / Q000188:drug therapy* / Q000473:pathology; D008297:Male; D010802:Phylogeny; D011994:Recombinant Proteins / Q000235:genetics / Q000494:pharmacology / Q000627:therapeutic use*; D017355:Zymomonas / Q000201:enzymology*
chemical_list: D000970:Antineoplastic Agents; D011994:Recombinant Proteins; D001215:Asparaginase