Biochemical characterization of recombinant L-asparaginase (AnsA) from Rhizobium etli, a member of an increasing Rhizobial-type family of L-asparaginases

dc.creatorANGELICA MORENO ENRIQUEZ
dc.creatorZahaed Evangelista Martínez
dc.creatorEDITH GRACIELA GONZALEZ MONDRAGON
dc.creatorARTURO CALDERON FLORES
dc.creatorRoberto Alejandro Arreguín Espinosa de los Monteros
dc.creatorERNESTO PEREZ RUEDA
dc.creatorALEJANDRO HUERTA SAQUERO
dc.date2012-01-05
dc.date.accessioned2023-07-21T19:18:54Z
dc.date.available2023-07-21T19:18:54Z
dc.identifierhttp://cicy.repositorioinstitucional.mx/jspui/handle/1003/1668
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/7737227
dc.descriptionWe report the expression, purification, and characterization of L-asparaginase (AnsA) from Rhizobium etli. The enzyme was purified to homogeneity in a single-step procedure involving affinity chromatography, and the kinetic parameters K(m), V(max), and k(cat) for L-asparagine were determined. The enzymatic activity in the presence of a number of substrates and metal ions was investigated. The molecular mass of the enzyme was 47 kDa by SDS-PAGE. The enzyme showed a maximal activity at 50 degrees C, but the optimal temperature of activity was 37 degrees C. It also showed maximal and optimal activities at pH 9.0. The values of K(m), V(max), k(cat), and k(cat)/K(m) were 8.9 +/- 0.967 × 10⁻³ M, 128 +/- 2.8 U/mg protein, 106 +/- 2 s⁻¹, and 1.2 +/- 0.105 × 10⁴ M⁻¹s⁻¹, respectively. The L-asparaginase activity was reduced in the presence of Mn²⁺, Zn²⁺, Ca²⁺, and Mg²⁺ metal ions for about 52% to 31%. In addition, we found that NH₄⁺, L-Asp, D-Asn, and beta-aspartyl-hydroxamate in the reaction buffer reduced the activity of the enzyme, whereas L-Gln did not modify its enzymatic activity. This is the first report on the expression and characterization of the L-asparaginase (AnsA) from R. etli. Phylogenetic analysis of asparaginases reveals an increasing group of known sequences of the Rhizobialtype asparaginase II family.
dc.formatapplication/pdf
dc.languageeng
dc.relationcitation:Moreno-Enríquez, A., Evangelista-Martínez, Z., González-Mondragón, E. G., Calderón-Flores, A., Arreguín, R., Pérez-Rueda, E., & Huerta-Saquero, A. (2012). Biochemical characterization of recombinant L-asparaginase (AnsA) from Rhizobium etli, a member of an increasing rhizobial-type family of L-asparaginases. J. Microbiol. Biotechnol., 22(3), 292-300.
dc.rightsinfo:eu-repo/semantics/openAccess
dc.rightshttp://creativecommons.org/licenses/by-nc-nd/4.0
dc.sourceJ. Microbiol. Biotechnol., 22(3), 292-300, 2012
dc.subjectinfo:eu-repo/classification/Autores/TYPE II ASPARAGINASE
dc.subjectinfo:eu-repo/classification/Autores/RHIZOBIUM ETLI
dc.subjectinfo:eu-repo/classification/Autores/BIOCHEMICAL CHARACTERIZATION
dc.subjectinfo:eu-repo/classification/cti/2
dc.subjectinfo:eu-repo/classification/cti/24
dc.subjectinfo:eu-repo/classification/cti/2415
dc.subjectinfo:eu-repo/classification/cti/241502
dc.subjectinfo:eu-repo/classification/cti/241502
dc.titleBiochemical characterization of recombinant L-asparaginase (AnsA) from Rhizobium etli, a member of an increasing Rhizobial-type family of L-asparaginases
dc.typeinfo:eu-repo/semantics/article
dc.typeinfo:eu-repo/semantics/publishedVersion


Este ítem pertenece a la siguiente institución