title: Therapeutic l-asparaginase: upstream, downstream and beyond.
abstract: l-asparaginase (l-asparagine amino hydrolase, E.C.3.5.1.1) is an enzyme clinically accepted as an antitumor agent to treat acute lymphoblastic leukemia and lymphosarcoma. It catalyzes l-asparagine (Asn) hydrolysis to l-aspartate and ammonia, and Asn effective depletion results in cytotoxicity to leukemic cells. Microbial l-asparaginase (ASNase) production has attracted considerable attention owing to its cost effectiveness and eco-friendliness. The focus of this review is to provide a thorough review on microbial ASNase production, with special emphasis to microbial producers, conditions of enzyme production, protein engineering, downstream processes, biochemical characteristics, enzyme stability, bioavailability, toxicity and allergy potential. Some issues are also highlighted that will have to be addressed to achieve better therapeutic results and less side effects of ASNase use in cancer treatment: (a) search for new sources of this enzyme to increase its availability as a drug; (b) production of new ASNases with improved pharmacodynamics, pharmacokinetics and toxicological profiles, and (c) improvement of ASNase production by recombinant microorganisms. In this regard, rational protein engineering, directed mutagenesis, metabolic flux analysis and optimization of purification protocols are expected to play a paramount role in the near future.
authors.affiliation: a Department of Biochemical and Pharmaceutical Technology , School of Pharmaceutical Sciences, University of São Paulo , São Paulo , Brazil.
authors.forename: André Moreni
authors.initials: AM
authors.lastname: Lopes
authors.affiliation: b Department of Biochemistry and Tissue Biology , Institute of Biology, State University of Campinas - UNICAMP , Campinas , Brazil.
authors.forename: Laura de
authors.initials: L
authors.lastname: Oliveira-Nascimento
authors.affiliation: c Centre of Biological Engineering (CEB), School of Engineering, University of Minho , Braga , Portugal.
authors.forename: Artur
authors.initials: A
authors.lastname: Ribeiro
authors.affiliation: d Biosciences Institute, Coastal Campus, São Paulo State University - UNESP , São Vicente , Brazil.
authors.forename: Carlos Abrunhosa
authors.initials: CA
authors.lastname: Tairum
authors.affiliation: d Biosciences Institute, Coastal Campus, São Paulo State University - UNESP , São Vicente , Brazil.
authors.forename: Carlos Alexandre
authors.initials: CA
authors.lastname: Breyer
authors.affiliation: d Biosciences Institute, Coastal Campus, São Paulo State University - UNESP , São Vicente , Brazil.
authors.forename: Marcos Antonio de
authors.initials: MA
authors.lastname: Oliveira
authors.affiliation: a Department of Biochemical and Pharmaceutical Technology , School of Pharmaceutical Sciences, University of São Paulo , São Paulo , Brazil.
authors.forename: Gisele
authors.initials: G
authors.lastname: Monteiro
authors.affiliation: e Department of Mycology , Federal University of Pernambuco , Recife , Brazil.
authors.forename: Cristina Maria de
authors.initials: CM
authors.lastname: Souza-Motta
authors.affiliation: f Department of Pharmacy , School of Health Sciences, University of Brasilia - UnB , Brasilia , Brazil.
authors.forename: Pérola de Oliveira
authors.initials: PO
authors.lastname: Magalhães
authors.affiliation: g Department of Chemical Engineering , Faculty of Engineering and Science, University of La Frontera , Temuco , Chile.
authors.forename: Jorge Gonzalo Farías
authors.initials: JG
authors.lastname: Avendaño
authors.affiliation: c Centre of Biological Engineering (CEB), School of Engineering, University of Minho , Braga , Portugal.
authors.forename: Artur Manuel
authors.initials: AM
authors.lastname: Cavaco-Paulo
authors.affiliation: h Faculty of Pharmaceutical Sciences , University of Campinas - UNICAMP , Campinas , Brazil.
authors.forename: Priscila Gava
authors.initials: PG
authors.lastname: Mazzola
authors.affiliation: a Department of Biochemical and Pharmaceutical Technology , School of Pharmaceutical Sciences, University of São Paulo , São Paulo , Brazil.
authors.forename: Carlota de Oliveira
authors.initials: CO
authors.lastname: Rangel-Yagui
authors.affiliation: i Department of Biochemistry and Microbiology , Institute of Biosciences, São Paulo State University - UNESP , Rio Claro , Brazil , and.
authors.forename: Lara Durães
authors.initials: LD
authors.lastname: Sette
authors.affiliation: j Department of Civil , Chemical and Environmental Engineering, University of Genoa , Genoa , Italy.
authors.forename: Attilio
authors.initials: A
authors.lastname: Converti
authors.affiliation: a Department of Biochemical and Pharmaceutical Technology , School of Pharmaceutical Sciences, University of São Paulo , São Paulo , Brazil.
authors.forename: Adalberto
authors.initials: A
authors.lastname: Pessoa
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry / Q000378:metabolism / Q000627:therapeutic use; D001215:Asparaginase / Q000737:chemistry / Q000378:metabolism / Q000627:therapeutic use; D001419:Bacteria / Q000378:metabolism; D004339:Drug Compounding; D005658:Fungi / Q000378:metabolism; D015202:Protein Engineering
chemical_list: D000970:Antineoplastic Agents; D001215:Asparaginase
keywords: Acute lymphoblastic leukemia; antineoplastic activity; biopharmaceutical; l-asparaginase; microbial l-asparaginase production