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

Effer, Brian; Lima, Guilherme Meira; Cabarca, Sindy; Pessoa, Adalberto; Farias, Jorge G.; Monteiro, Gisele

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.

Más información

Título según WOS: L-Asparaginase from E. chrysanthemi expressed in glycoswitch: effect of His-Tag fusion on the extracellular expression
Título según SCOPUS: L-Asparaginase from E. chrysanthemi expressed in glycoswitch®: effect of His-Tag fusion on the extracellular expression
Título de la Revista: PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY
Volumen: 49
Número: 7
Editorial: TAYLOR & FRANCIS INC
Fecha de publicación: 2019
Página de inicio: 679
Página final: 685
Idioma: English
DOI:

10.1080/10826068.2019.1599396

Notas: ISI, SCOPUS