L-asparaginase from Dickeya chrysanthemi : expression, purification and cytotoxicity assessment. . 2022;52(6):668-680. (Impact factor 3.2, Q3), ISSN: 1082-6068. Accession NumberWOS
Microbial L-asparaginases are aminohydrolases that hydrolyze L-asparagine to L-aspartate. They
are used to treat acute lymphoblastic leukemia and Hodgkin’s lymphomas and in food industries.
Increasing demand for L-ASNases is therefore needed. In the current study, the recombinant LASNase
from Dickeya chrysanthemi (DcL-ASNase) was cloned into pET28a (þ) expression vector
and expressed in Escherichia coli as a 6His-tagged fusion protein and purified using Ni2þ chelated
Sepharose chromatography resin, yielding a highly purified enzyme. Kinetics analysis allowed the
determination of its substrate specificity and the physicochemical parameters that affect enzyme
activity. The enzyme showed operational stability at 37 C and 45 C. The immunogenicity of the
purified DcL-ASNase was evaluated by measuring the IgG and IgM levels in rats after injection.
The cytotoxicity DcL-ASNase in selected cancer cell lines and peripheral blood monocytes was
determined. The results showed that the enzyme induces pleiotropic effects, including significant
morphological changes and the formation of apoptotic bodies. No cytotoxic effects were observed
in peripheral blood monocytes at the same concentrations. In addition, gene expression analysis
by RT-PCR of apoptotic biomarkers (Bax, survivin, and Ki-67) allowed the study of the apoptotic
mechanism induced by DcL-ASNase on THP-1 cells.
Glutathione S. transferases (GSTs; EC. 2.5.1.18) are a large family of multifunctional enzymes that play crucial roles in the metabolism and inactivation of a broad range of xenobiotic compounds.…
Background: In the wake of the warning by WHO that the prevalence of dementia may have a rise of 125% in the Middle East by 2050, identification of the genetic risk factors in Arab populations is…