Purification of Dickeya solanil-Asparaginase and Study of the Influence of TiO2 and ZnO Nanoparticles on Its Enzymatic Activity
Autor: | Selma Allouache, Mohamed Azarkan, T Mostefaoui, Farida Bendali |
---|---|
Jazyk: | angličtina |
Rok vydání: | 2019 |
Předmět: |
l-asparaginase
Biomedical Engineering Fast flow Bioengineering Dickeya 02 engineering and technology Dickeya solani L asparaginase 03 medical and health sciences Ammonia chemistry.chemical_compound Hydrolysis Zinc oxide Imagerie médicale radiologie tomographie Titanium(IV) oxide 030304 developmental biology chemistry.chemical_classification 0303 health sciences Chromatography biology Sciences bio-médicales et agricoles 021001 nanoscience & nanotechnology biology.organism_classification Enzyme chemistry Zno nanoparticles Instrumentation médicale Nanoparticles L-asparaginase 0210 nano-technology |
Zdroj: | BioNanoScience |
Popis: | l-asparaginase catalyzes the hydrolysis of l-asparagine to l-aspartate and ammonia. In the present study, we describe the production, purification, and preliminary characterization of Dickeya solanil-asparaginase. The purification of l-asparaginase was done using ion-exchange chromatography on HiTrap Q-Sepharose Fast Flow followed by FPLC-gel filtration chromatography on Superdex 75 pg. The presence of the enzyme was confirmed by enzymatic activity measurements along with SDS-PAGE analysis. The purified l-asparaginase was shown to exhibit specificity towards l-asparagine, while being inactive against l-glutamine. This data allows us to suggest that the l-asparaginase from Dickeya solani might be clinically more relevant than, e.g. the l-asparaginase isolated from E. coli which hydrolyzed also l-glutamine and produces l-glutamate, a neurotoxic agent. We also demonstrated that the enzymatic activity was enhanced in the presence of TiO2 and ZnO nanoparticles, making them good candidates to improve l-asparaginase activity. Indeed, the results obtained show that the TiO2 nanoparticles increased the activity of l-asparaginase by a factor of 6.0 while the ZnO nanoparticles increased it twice. SCOPUS: ar.j info:eu-repo/semantics/published |
Databáze: | OpenAIRE |
Externí odkaz: |