Biochemical characterization and detection of antitumor activity of l-asparaginase from thermophilic Geobacillus kaustophilus DSM 7263
Anti-tumor activity
Geobacillus kaustophilus DSM 7263(T)
L-asparaginase
xCELLigence
Journal
Protein expression and purification
ISSN: 1096-0279
Titre abrégé: Protein Expr Purif
Pays: United States
ID NLM: 9101496
Informations de publication
Date de publication:
11 2022
11 2022
Historique:
received:
21
02
2022
revised:
10
07
2022
accepted:
13
07
2022
pubmed:
22
7
2022
medline:
30
8
2022
entrez:
21
7
2022
Statut:
ppublish
Résumé
L-asparaginases, which are oncolytic enzymes, have been used in clinical applications for many years. These enzymes are also important in food processing industry due to their potential in acrylamide-mitigation. In this study, the gene for l-asparaginase (GkASN) from a thermophilic bacterium, Geobacillus kaustophilus, was cloned and expressed in E. coli Rosetta™2 (DE3) cells utilizing the pET-22b(+) vector. The 6xHis-tag attached enzyme was purified and analyzed both biochemically and structurally. The molecular mass of GkASN was determined as ∼36 kDa by SDS-PAGE, Western Blotting, and MALDI-TOF MS analyses. Optimum temperature and pH for the enzyme was determined as 55 °C and 8.5, respectively. The enzyme retained 89% of its thermal stability at 37 °C and 75% at 55 °C after 6 h of incubation. The enzyme activity was inhibited in the presence of Cu
Identifiants
pubmed: 35863721
pii: S1046-5928(22)00103-6
doi: 10.1016/j.pep.2022.106146
pii:
doi:
Substances chimiques
Asparaginase
EC 3.5.1.1
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
106146Informations de copyright
Copyright © 2022 Elsevier Inc. All rights reserved.