Title
l-Asparaginase from Erwinia carotovora: insights about its stability and activity
Author
Marcele Faret, Stephanie Bath de Morais, Nilson Ivo Tonin Zanchin, Tatiana de Arruda Campos Brasil de Souza
Year
2018
Journal
Molecular Biology Reports
Abstract
Enzymatic prospection indicated that l-asparaginase from Erwinia carotovora (ECAR-LANS) posses low glutaminase activity and much effort has been made to produce therapeutic ECAR-LANS. However, its low stability precludes its use in therapy. Herein, biochemical and biophysical assays provided data highlighting the influence of solubilization and storage into ECAR-LANS structure, stability, and activity. Moreover, innovations in recombinant expression and purification guaranteed the purification of functional tetramers. According to solubilization condition, the l-asparaginase activity and temperature of melting ranged up to 25–32%, respectively. CD spectra indicate the tendency of ECAR-LANS to instability and the influence of β-structures in activity. These results provide relevant information to guide formulations with prolonged action in the bloodstream.
Instrument
J-815
Keywords
Circular dichroism, Secondary structure, Chemical stability, Biochemistry