L-Asparaginase from E. chrysanthemi expressed in g... - BV FAPESP
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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

L-Asparaginase from E. chrysanthemi expressed in glycoswitch: effect of His-Tag fusion on the extracellular expression

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Author(s):
Effer, Brian [1, 2] ; Lima, Guilherme Meira [1] ; Cabarca, Sindy [3, 4] ; Pessoa, Adalberto [1] ; Farias, Jorge G. [2] ; Monteiro, Gisele [1]
Total Authors: 6
Affiliation:
[1] Univ Sao Paulo, Sch Pharmaceut Sci, Dept Biochem & Pharmaceut Technol, Prof Lineu Prestes 580, BR-05508000 Sao Paulo - Brazil
[2] Univ La Frontera, Fac Engn & Sci, Dept Chem Engn, Francisco Salazar - Chile
[3] Univ Sao Paulo, Lab Appl Struct Biol, Dept Microbiol, Sao Paulo - Brazil
[4] Univ Estadual Campinas, Inst Biol, Campinas - Brazil
Total Affiliations: 4
Document type: Journal article
Source: PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY; v. 49, n. 7 APR 2019.
Web of Science Citations: 0
Abstract

L-Asparaginase (L-ASNase) is an important enzyme used to treat acute lymphoblastic leukemia, recombinantly produced in a prokaryotic expression system. Exploration of alternatives production systems like as extracellular expression in microorganisms generally recognized as safe (such as Pichia pastoris Glycoswitch) could be advantageous, in particular, if this system is able to produce homogeneous glycosylation. Here, we evaluated extracellular expression into Glycoswitch using two different strains constructions containing the asnB gene coding for Erwinia chrysanthemi L-ASNase (with and without His-tag), in order to find the best system for producing the extracellular and biologically active protein. When the His-tag was absent, both cell expression and protein secretion processes were considerably improved. Three-dimensional modeling of the protein suggests that additional structures (His-tag) could adversely affect native conformation and folding from L-ASNase and therefore the expression and cell secretion of this enzyme. (AU)

FAPESP's process: 17/20384-9 - Development of downstream process of humanized L-asparaginase and its characterization
Grantee:Eduardo Krebs Kleingesinds
Support Opportunities: Scholarships in Brazil - Doctorate
FAPESP's process: 16/25896-5 - Biochemical characterization and cytotoxic evaluation of mutant isoforms of L-Asparaginase II from Dickeya chrysanthemi (Erwinia chrysanthemi)
Grantee:Iris Munhoz Costa
Support Opportunities: Scholarships in Brazil - Doctorate
FAPESP's process: 15/07749-2 - Protein engineering and comparison of microbial expression systems of the biopharmaceutical L-asparaginase
Grantee:Gisele Monteiro
Support Opportunities: Regular Research Grants
FAPESP's process: 13/08617-7 - Production of extracellular L-asparaginase: from bioprospecting to the engineering of an antileukemic biopharmaceutical
Grantee:Adalberto Pessoa Junior
Support Opportunities: Research Projects - Thematic Grants
FAPESP's process: 16/15787-4 - Cloning of L-asparaginase from Escherichia coli in a Pichia Pastoris strain with humanized glysosylation
Grantee:Guilherme Meira Lima
Support Opportunities: Scholarships in Brazil - Scientific Initiation